SYMPOSIUM ON SCIENCES PROCEEDINGS ABSTRACTS

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ANKARA UNIVERSITY
FACULTY OF PHARMACY
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INTERNATIONAL
SYMPOSIUM ON
PHARMACEUTICAL
SCIENCES
PROCEEDINGS
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ABSTRACTS
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i lac Uretiminde 83 yillik Deneyim
Hammadde Uretimi
Parenteral ve Liyofilize ilag Uretiminde Uzmanlik
AB-iyi ilag Uretim Kurallarina Uyum

Ankara Universitesi Eczacilik Fakultesi Yayinlari No: 91
ANKARA UNIVERSITY
FACULTY OF PHARMACY
8 INTERNATIONAL SYMPOSIUM
ON PHARMACEUTICAL SCIENCES
ISOPS-8
PROCEEDINGS AND ABSTRACTS
JUNE 13-16, 2006
ANKARA-TURKEY

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Participants and Guests,
It is an honor to have all the participants in 8 th International Symposium of
Pharmaceutical Sciences. This symposium was organized triennially by Ankara
University, Faculty of Pharmacy until 8 th symposium then it was decided to be held
every three years.
It will be a great pleasure to be your host in Ankara.
As our leader Ataturk said "We believe that there will be no future for human without
science". Countries that have science and technology also have economical power.
There are no borders for knowledge so science is the most important factor that
brings people and countries together.
A development of science and technology mostly depends on academia due to the
fact that research and teaching both take considerable components in where science
and technology be alive for further developments and we believe that without
researcher's ideas and knowledge, university concept cannot be realized in high
standards.
In Turkey, we have an enormous increase in scientific publications according to
Science Citation Index. Turkish scientists believe that the increase in scientific
development will not be high enough and one of the most important things is to
produce the scientific research into technological development by the factors of
innovation. Because science brings the meaning of better living conditions along with
technology development and also broadens the aspects of quality of life.
Dear guests,
Turkey is developing rapidly in many areas. After the official acceptance as a
candidate member of EU resulted from Helsinki Meeting in 1999, many regulations
have been taken place in the several areas including politics, economy, and
science nationwide. As you all may know, universities are the initial institutions that
are expected to adapt themselves into mentioned EU regulations, and fortunately, I
would like to say that universities in Turkey are the institutions that can be adapted
themselves most easily to the concepts of EU. Therefore, universities will have a
significant assignment to expand the issues of EU related to economy, politics, law,
and mostly in scientific developments in the adaptation process to these new
guidelines.
Turkey is geographically located in the junction of Europe, Asia and Middle East, and
also has young population that will bring the dynamism and enthusiasm in the
cultural adaptation progress to EU as being a modern and unique Islamic country by
secularism. Besides this, as a member of NATO which is a crucial organization that
assumes responsibility and an important status for the safety of Europe, Turkey
took an imperative role since and during the "cold war" era that construct Turkey as
an indispensable country which plays an imperative role within the Islamic and Turki
countries. So that, due to this commendable perception, the integration of Turkey
to EU will hold several beneficial issues for both sides as Turkey is-being considered in
the bridge role between cultures and religious in terms of economy, politics, humanity
and of course, scientific development.
It is considered the fact by EU that Turkey has some time for fully integration to the
union unless some of the main aspects would be anticipated as the necessity in
which we deem that the incorporation process will take much shorter time than
expected due to our potential of developments and background of historical
experience.

CONTENTS
Page
Proceedings of Plenary Lectures 1
Abstracts of Clinical Pharmacy Workshop 83
Abstracts of Industrial Pharmacy Workshop 89
Abstracts of Oral Presentations 97
Abstracts of Poster Presentations-I (June 14 th , 2006) 115
Abstracts of Poster Presentations-ll (June 15 th ,2006) 243
Participant List 371
Author Index 391

PROCEEDINGS
OF
PLENARY LECTURE

PL-
IMPACT OF CHIRALITY ON ENVIRONMENTAL ANALYSIS
H. Y. Aboul-Enein
Pharmaceutical and Medicinal Chemistry Department, Pharmaceutical, The Pharmaceutical and
Drug Industires Research Division, National Research Center (NRC), Dokki, Cairo 12311,
EGYPT
Analysis of chiral pollutants by an achiral chromatographic assay does not provide the
comprehensive information about their side effects as enantiomers of chiral pollutants may have
different toxicities. Liquid chromatography (LC) including high-performance liquid
chromatography (HPLC) and capillary electrophoresis (CE) has emerged as one of the best
chromatographic methods used for the enantioselective analysis of pesticides and agrochemicals.
In LC, different classes of the CSPs can be used with a variety of mobile phases. Wide choice of
the mobile phases in LC also enhances the range of its applications. Liquid chromatographic
methods can be used for the enantioselective analysis of the organochlorine pollutants by
coupling it with many of the advance detection techniques such as mass spectrometer (MS),
optical detectors etc. Therefore, due to the rapid advancement of the stereoselective LC, it is
becoming a reliable modality. Several examples for the chiral analyses of these chiral
agrochemicals will be presented. A sound and uniform policy should be developed regarding the
regulation of chirality of pesticides, insecticides, fungicides and other agrochemicals at
International level. It is very urgent to develop certain guidelines before exposing the notorious
chiral compounds to our environment. Therefore, the environmental authorities world wide
should come forward on this issue and initiate a guideline and formulate a harmonical policy on
the use of chiral agrochemicals.
Ali I, Aboul-Enein HY (2004) Chiral pollutants: Distribution, toxicity and analysis by
chromatography and capillary electrophoresis, Johan Wily & Sons, Chichester, UK.

PL-
IAM.PC: A USEFUL TOOL FOR PREDICTING MEMBRANE PERMEABILITY
A. Adejare
Department of Pharmaceutical Sciences, Philadelphia College of Pharmacy, University of the
Sciences in Philadelphia, Philadelphia, Pennsylvania 19104, USA
Research efforts in our laboratory include design and syntheses of compounds which can be used
in the treatment of Alzheimer's disease (AD). The mechanisms being investigated are inhibition
of y-secretase, an enzyme believed to be key in the formation of amyloid plaques; and n-methyl-
D-aspartate (NMDA) receptor antagonism as overexcitation is linked to neurodegeneration.
Oral administration is clearly preferred in terms of drugs for treating AD because of the target
population. Questions that have arisen from our studies include how we can predict whether a
novel compound obtained will be absorbed if taken orally and if so whether or not it can cross
the blood brain barrier and therefore reach desired central nervous system (CNS) sites. There
are many biological models for addressing those questions. However, we are more interested in
predictions based on physicochemical properties of the compounds. We therefore began a
project to determine compound membrane permeability based on physicochemical properties.
The Lipinski's rule of 5 already addresses the key parameters for the prediction of oral
bioavailability of a drug. 1 The parameters taken into consideration include molecular weight,
hydrogen bonding abilities, and log P. Molecular weight and hydrogen bonding abilities can be
determined and are less subject to interpretation. We believe that log P is too simplistic a
measure of complex interactions between a compound and biological membrane and that
immobilized artificial membrane (IAM) is a much better measure. This is because
phosphatidylcholine (PC) found on biological membranes can be used as packing on IAM
columns. The resulting columns are therefore better able to simulate interaction of the drug with
biological membrane. 2
Figure 1. Permeation of solute through biological membrane as compared with IAM.PC surface.
As part of our investigation, pharmaceutical profiling studies were conducted on compound 1, a
novel prototype member of a y-secretase inhibitors class. These studies included determination
of solubility, dissociation constant (pKa), octanol/water partition coefficient (log P) and the
capacity factor (k'i A M) on IAM.PC chromatographic columns. The compound was very slightly
soluble in water (0.12 ± 0.05 mg/mL) but the solubility increased considerably in basic medium
(0.27 ± 0.06 mg/mL). The other values obtained were pKa (10.36 ±0.11) and log P (3.36 ±
0.16) that was determined by shake flask method and (3.31 ± 0.01) determined by high
performance liquid chromatography (HPLC). The experimentally determined log P values
correlated well with the calculated one of 3.44. The observed log k'i A M value of (2.79 ± 0.04)
indicates that the compound can reasonably be expected to have high membrane permeability
and therefore good absorption profile if taken orally.

o
H II
M N-S —S
II
O
Compound I
F
In follow-up studies, the partitioning of structurally diverse and functionally unrelated set of 21
compounds was studied using IAM.PC. The aqueous capacity factors (log IC'IAMW) of the
compounds were correlated with their logarithms of octanol/water partition coefficient (log P) to
determine whether molecular binding on IAM columns can predict permeability of drugs
through biological membranes such as intestinal membrane. Good correlation was obtained
between log k'i A Mw and log P of the compounds (r = 0.952). For a subset of 8 compounds, log
k'lAMW showed better correlation with log % of human absorption than did log P. We therefore
now reconstruct the "box" within which a lead should fall to be a true lead for drug discovery
purposes for compounds meant for oral administration.
$ False lead
HB
Optimal properties for orally octive compounds are found within the
logk' 1AM -HB-MW box
Though many in-vitro physicochemical based techniques are available to predict membranes
permeability; IAM.PC is a promising tool for easy and reproducible prediction. Prediction of
blood-brain permeability using this technology is being examined.
Financial support of USA NIH Grant #s 7R15NS036393-04 and 1R15NS050177-01A1 are
gratefully acknowledged.
1. Lipinski, C. A.; Lombardo, F.; Dominy, B. W.; Feeney, P. J. Experimental and Computational
Approaches to Estimate Solubility and Permeability in Drug Discovery and Development
Settings. Adv.Drug Deliv. Rev. 1997, 23, 3-25.
2. Pidgeon, C.; Ong, S.; Liu, H.; Qiu, X.; Pidgeon, M.; Dantzig, A. H.; Munroe, J.; Hornback, W.
J.; Kasher, J. S.; Glunz, L. et al. IAM Chromatography: An in Vitro Screen for Predicting Drug
Membrane Permeability. J. Med. Chem. 1995, 17, 590-594.
3. El-Gendy, A. M.; Adejare, A. Membrane Permeability Related Physicochemical Properties of
a Novel y-Secretase Inhibitor. Int. J. Pharm. 2004, 280, 47-55.

PL-3
ANTIBIOTIC RESISTANCE MECHANISMS IN HELICOBACTER PYLORI
Y. Akyon
Hacettepe University, Faculty of Medicine, Department of M icrobiology and Clinical
Microbiology, 06100, Sihhiye, Ankara, Turkey
Helicobacter pylori is a Gram negative infectious agent that chronically infects the gastric
mucosa of half the human population. Although most of the infected individuals are
asymptomatic H. pylori is the causative of gastritis and peptic ulcer disease and the early risk
factor for gastric cancer (1,2). Infection occurs preferentially in early childhood and despite host
immune responses, once established tends to persist for years or decades. The discovery that
most stomach diseases are a consequence of a Helicobacter pylori infection has completely
changed the management of stomach diseases. Antimicrobials are the treatment of choice in
addition to proton pump inhibitors (PPIs) or ranitidine bismuth. According to European
Maastrich 2-2000 report; Patients who have H. pylori whom are diagnosed duodenal or gastric
ulcer, MALT lymphoma, atrophic gastritis, stomach cancer, patients whom had a newly stomach
resection, patients who have first degree relatives with stomach cancer and if the patient
him/herself wants treatment are the first group patient which will receive H. pylori treatment.
The second group of patients who will receive treatment are; functional dyspeptic patients,
patients who will be treated with anti-acid drugs for a long period (patients with
gastroesophageal reflux disease), patients receiving anti-inflamatory drugs (3). There is no ideal
drug regimen to cure H. pylori infection. Metronidazole, clarithromycin, amoxycillin and
tetracyclin are the antimicrobials for the treatment of infection. Besides combination therapy
with antimicrobials, H2-receptor antagonists, protone pump inhibitors (PPI) or bismuth salts are
also added to the treatment protocole. H. pylori eradication rate is 80-90 % in the patients where
standart drug regimen is applied. 10-20 % of treatment failure is due to; patient in compliance to
the treatment, not completing the treatment due to drug side effects and most importantly the
resistance of H. pylori to the chosen antimicrobial or antimicrobials (4). According to Maastrich
2-2000 concensus a combined therapy with clarithromycin, amoxycillin and a PPI is the first
choice treatment regimen for H. pylori infection. The second choice of regimen is; quadraple
therapy with metronidazole, tetracycline, bismuth salt and PPI(3). The quinolones, furazolidone
and rifabutin are the choice of drugs that can be chosen by the doctor, if both therapy regimens
were a failure. We are now faced with the problem of antimicrobial resistance, which is the main
cause of treatment failure. H. pylori acquires resistance essentially via point mutations, and
today this phenomenon is found with most antibacterials. The most important resistance to
consider is that to clarithromycin, since it is the first-choice antibacterial and clarithromycin
resistance is highly clinically significant. Quadruple therapy or triple therapies with amoxicillinmetronidazole
or tetracycline-metronidazole and a PPI or ranitidine bismuth can then be used
despite a possible resistance to metronidazole if the strain is resistant to clarithromycin.
Resistance to both clarithromycin and metronidazole may lead to the use of other combinations,
i.e. amoxicillin-rifabutin, amoxicillin-levofloxacin or amoxicillin-furazolidone. Resistance to
any of these drugs means their use must be avoided. In some instances, it may also be advisable
to prescribe amoxicillin as the sole antibacterial, or to use a quadruple therapy with furazolidone
instead of metronidazole.

Metronidazole is a drug from the 5-nitroimidazole group. The 5-nitroimidazoles are prodrugs,
they need to be activated within the target cell. H. pylori nitroreductase enzymes activate
metronidazole. rdxA and frxA genes produce the nitrodeductase enzymes. The effect of
metronidazole is on molecules like DNA, RNA, proteins, fatty-acids. Resistance to
metronidazole in H. pylori is by the inactivation of nitroreductase and/or falvadoxin
oxidoreductase. The frame shift mutations in rdxA gene increases the minumum inhibitory
concentration (MIC) levels. The mutations in the frxA gene is responsible of lower level of MIC
resistance (5,6). Clarithromycin, from the macrolide group of antibiotics, is the first choice of
drug in the treatment of H. pylori. Clarithromycin inhibits the protein synthesis by binding to
the 50S subunit of the ribosome. The resistance in clarithromycin is due to point mutations. The
mutation occurs in the 23S rRNA gene 2142 (A2142G) -2143 (A2143G) bases, mostly
adenineguanine and rarely adenine->cytosine (7). In recent years there are new reports of
point mutation sites in the 23S rRNA gene. Amoxicillin is a cell wall synthesis inhibitor.
Resistance to beta-lactam antibiotics is due to their beta-lactamase production, the structural
difference in the penicillin binding proteins (PBP) or differentiation of the other proteins
responsible for cell wall synthesis (10, 11, 12). There are 7 PBP in H. pylori (13, 14, 15). Mittl et
al has defined a beta-lactamease specific to H. pylori (16). However, the resistance to
amoxicillin in H. pylori could not be related to the defined beta-lactamases (17). PBP-D could
not be identified in resistant strains (18). Recent findings show that in PBP1A the change of
serine to arginine causes high level of resistance to amoxicillin (19). Tetracyclines shows its
antibacterial activity by inhibiting the protein synthesis by binding to the 30S subunit of
ribosomes. The resistance to tetracyclines in H. pylori is due to the mutations in 16S rRNA (20).
The resistance in the 16S rRNA is reported to be in the change of 3 base pairs; AGA 92 6-928
TTC and this could be detected by PCR-RFLP (restiction fragment lenght polymorphism) (21).
Although it is theoretically possible to cure a drug-resistant H. pylori infection, a practical
limitation is the availability of the drugs in certain countries. Furthermore, the progressive
increase in drug resistance warrants the need for new antibacterials in the near future.
1. Blaser MJ. Helicobacter pylori, microbiology of a "slow" bacterial infection. Trends
Microbiol 1993; 1: 255-260.
2. Blaser MJ, Parsonnet J. Parasitizm by the "slow" bacterium Helicobacter pylori leads to
altered gastric homeastasis and neoplasia. J Clin Invest 1994; 94: 4-8.
3. Malfertheiner P, Megraud F, O'Morain C, et al. Current concepts in the management of
Helicobacter pylori infection - the Maastricht 2-2000 Consensus report. Alimet
Pharmacol Ther 2002; 16: 167 - 80.
4. Goh KL. Update on the management of Helicobacter pylori infection, including drugresistant
organisms. J Gastroenterol Hepatol 2002; 17: 482-487.
5. Sisson G, Jeong JY, Goodwin A, et al. Metronidazole activation is mutagenic and causes
DNA fragmentation in Helicobacter pylori and in Escherichia coli containing a cloned
H. pylori rcbcA + (nitroreductase) gene. J Bacteriol 2000; 182 (18): 5091-5096.
6. Mukhopadhyay AK, Jeong JY, Dailidiene D, Hoffman PS, Berg DE. The rdxA
ferredoxin gene can down-regulate frxA nitroreductase gene expression and is essential
in many strains of Helicobacter pylori. J Bacteriol 2003; 185(9): 2927-2935.
7. Owen RJ. Molecular testing for antibiotic resistance in Helicobacter pylori. GUT 2002;
50: 285-289.
8. Megraud F. Resistance of Helicobacter pylori to antibiotics and its impact on treatment
options. Drug Res Updates. 2001; 4: 178 - 86.
9. Fujimura S, Kato S, Iinuma K, Watanabe A. In vitro exposure to macrolide antibiotics in
Helicobacter pylori starins isolated from children. J Infect Chemother 2004; 10: 128 —
130.

10. Livermoore DM. Beta-Lactamases in laboratory and clinical resistance. Clin Microbiol
Rev. 1995; 8: 557-584.
11. Spratt BG, Cromie KD. Penicillin-binding proteins of gram-negative bacteria. Rev Infect
Dis. 1988; 10: 699-711.
12. Maki H, Murakami K. Formation of potent hybrid promoters of the mutant 11m gene by
IS256 transposition in methicillin-resistant Staphlococcus aureus. J Bacterid. 1997; 179:
6944 - 6948.
13. DeLoney CR, Schiller NL. Competition of various beta-lactam antibiotics for the major
penicillin-binding proteins of Helicobacter pylori: antibacterial activity and effects on
bacterial morphology. Antimicrob Agents Chemother. 1999; 4: 2702 - 2709.
14. Krishnamurthy P, Parlow MH, Schneider J, et al. Identification of a novel penicillinbinding
protein from Helicobacter pylori. J Bacteriol. 1999; 181: 5107 - 5110.
15. Harris AG, Hazell SL, Netting AG. Use of digoxigenin-labelled ampicillin in the
identification of penicillin-binding proteins in Helicobacter pylori. J Antimicrob
Chemother. 2000; 45: 591 - 598.
16. Mittl PR, Luthy L, Hunziker P, Grutter MG. The cystein-rich protein A from
Helicobacter pylori is a beta-lactamase. J Biol Chem 2000; 275: 17693 - 17699.
17. Dore MP, Osato MS, Realdi G, Mura I, Graham DY, Sepulveda AR. Amoxicillin
tolerance in Helicobacter pylori. J Antimicrob Chemother. 1999; 43: 47 - 54.
18. Dore MP, Graham DY, Sepulveda AR, Realdli G, Osata MS. Sensivity of amoxicillinresistant
Helicobacter pylori to other penicillins. Antimicrob Agents Chemother 1999;
43(7): 1803-1804.
19. Gerrits MM, Schuijffel D, van Zwet A A, Kuipers EJ, Vanderbroucke-Grauls CMJE,
Kusters JG. Alterations in penicillin-binding protein 1A confer resistance to beta-lactam
antibiotics in Helicobacter pylori. Antimicrob Agents Chemother. 2002; 46: 2229 -
2233.
20. Trieber CA, Taylor DE. Mutations in the 16S rRNA genes of Helicobacter pylori
mediate resistance to tetracycline. J Bacteriol 2002; 184(8): 2131-2140.
21.Ribeiro ML, Gerrits MM, Benvengo YHB, et al. Detection of high-level tetracycline
resistance in clinical isolates of Helicobacter pylori using PCR-RFLP. FEMS Immun
Med Microbiol. 2004; 40: 57 -61.
22. Engin D, Ercis S, Ozarslan E, Ozen H, Giine§ D, Has9elik G, Demir H, §imek H
Antibiotic susceptibilities of Helicobacter pylori strains isolated in Turkey. 11 th
International Workshop on Campylobacter, Helicobacter and Related Organisms, 1-5
Eyliil 2001, Freiburg, Germany.

PL-4
QUALITY NEEDS FOR THE DEVELOPMENT OF TRADITIONAL HERBAL
MEDICINAL PRODUCTS
R. Bauer
Institute of Pharmaceutical Sciences, Karl-Franzens-Universitat Graz, Universitatsplatz 4, A-
8010 Graz, Austria
The use of herbal products as medicine has long tradition all over the world [1], In recent years,
safety, efficacy and quality of these products have become important issues for health
authorities. In the European Union, the use of traditional herbal medicinal products has recently
been regulated in Directive 2004/24/EC [2], It requires 30 years of medicinal use with at least 15
years of use related to the European Union. As long as the efficacy of these medicinal products
is plausible on the basis of long-standing use and experience, clinical studies and pre-clinical
tests are not obligatory for such products from the regulatory point of view. However, quality
needs to be demonstrated in any case. Tests for identity, purity and content are obligatory, since
adulterations and substitutions are still common practice. As long as corresponding monographs
are available, these tests have to be performed according to pharmacopoeias. Therefore, for
TCM drugs it may be necessary to consult the English Edition of. However, the preparation of
monographs for Chinese herbal drugs in the European Pharmacopoeia is in progress. In addition
to microscopy, fingerprint analysis by TLC or HPLC can be used to guarantee identity and
consistent quality. Corresponding monographs have already been developed for Chinese plants
[3], In the Pharmacopoeia of the Peoples Republic of China HPLC methods are more and more
used to determine the amount of relevant constituents. Tests for contaminations with heavy
metals, pesticides, fungicides, micro-organisms and mycotoxins are reasonable as well [4]. Some
examples of medicinal plants from Asia will be presented, by which the different quality needs
for the development of traditional herbal medicinal products will be demonstrated.
1. C. Bodeker et al. (2005) WHO Global Atlas of Traditional, Complementary and
Alternative Medicine. World Health Organization, Kobe.
2. M. Silano, M. De Vincenzi, A. De Vincenzi, V. Silano. Fitoterapia 2004, 75: 107-116.
3. H. Wagner und R. Bauer (Eds.) Chinese Drug Monographs and Analysis. Verlag fur
Ganzheitliche Medizin Dr. Erich Wiihr GmbH, Kotzting (1996 - 2006)
4. R. Bauer. Drug Information Journal 1998, 32, 101-110.

PL-
THE EFFECT OF REACTIVE OXYGEN SPECIES (ROS) ON AIRWAYS
V. Bauer 1 . Y. Ito 2 , S. Matyas 1 , V. Pucovsky 1 , F. Bauer 3
1 Inst. Exp. Pharmacol. SASc, Bratislava, Slovakia, 2 Fac. Med. Kyushu Univ. Fukuoka, Japan,
3 Neonat. Dept. Distr. Hosp., Nove Zamky, Slovakia
ROS are key factors playing important role in tissue damage under different pathological
conditions including those of airways, particularly in patients with reduced antioxidant defenses
(ROP, IRDS, ARDS, BPD, COPD, inflammation, etc.) and epithelium damage. Knowledge of
pathophysiological mechanisms of these disorders could be helpful in rational prevention and
therapy. Since ROS might affect smooth muscle, epithelium, endothelium, innervation, membrane
lipids, receptors, transmitter systems, prostanoid production, Ca 2+ homeostasis, etc., the effects of
exogenous H2O2 or O2" (produced by xanthine/xanthine oxidase - X/XO and pyrogallol), 'OH
(produced by FeS0 4 /H 2 0 2 or FeSOVascorbic acid), H2O2 and HOC1 (generated by electrolysis
of the bathing solution - EL) were recorded on the muscle tone, excitatory and inhibitory
neurotransmission of guinea pig and cat tracheal strips or rings, as well as on [Ca 2+ ]j, membrane
potential and membrane currents. In intact preparations, the muscle tension was unaffected by
O2"", while H 2 0 2 , *OH and EL produced a biphasic response, contraction followed by relaxation
accompanied by an increase in TBARS and a decrease in non-protein thiols. The enhanced
trachealis tension was inhibited by antioxidants (e.g. N-acetylcysteine, stobadine, trolox, manitol).
Removal of epithelium or blockade of NO synthase by L-NOARG reduced the relaxation and
unmasked or enhanced the ROS induced trachealis contraction. H2O2 increased free [Ca 2+ ]j (both
in the absence and presence of Ca 2+ e, which effect developed faster and was larger in the presence
of Ca 2+ e) with a subsequent augmentation of muscle tone, the stimulation-evoked e.j.p.s and
contractions, as well as enhanced Ca 2+ and voltage-sensitive potassium conductance. DEDTCA
(inhibitor of endogenous SOD) ameliorated the inhibitory action of pyrogallol and X/XO on the
relaxatory action of an NO generator SNAP and the noncholinergic-nonadrenergic (NANC) i.j.p.s
and relaxations (due to peroxynitrite formed from 0 2 *~ and NO). Blockade of cyclooxygenase
(COX) by indomethacin enhanced the amplitude and duration of contractions on the intact and
denuded preparations. Nordihydroguaiaretic acid (inhibitor of lipoxygenase - LOX) pretreatment
did not alter the responses elicited by ROS in intact preparations and reduced their action on the
denuded ones. Our results suggest that a) various ROS due to mainly the Ca 2+ j release contract
tracheal smooth muscle with simultaneous activation (K + conductance) and release of epithelium
derived relaxing factors (NO and VIP), b) epithelium possesses ROS scavenging capacity which
is under physiological conditions high enough to protect smooth muscle from some of their
actions, c) O2*" have multiple actions, presumably due to its dismutation to H2O2 and presence of
both O2" and H2O2 simultaneously. While H2O2 seems to be responsible for elevation of muscle
tone and augmentation of smooth muscle contraction elicited by nerve stimulation, O2*" inhibits
muscle tone, cholinergic and NANC neurotransmission, and d) COX products participate in
relaxation and LOX products in contraction caused by ROS in airways.

PL-
SUPRAMOLECULAR MATERIALS USED IN THE HIGH-THROUGHPUT
SCREENING TECHNOLOGIES FOR DRUG DESIGN
G. Bazylak
Department of Pharmaco-Bromatology, Faculty of Pharmacy, Collegium Medicum, Nicolaus
Copernicus University, Jagiellonska 13, PL-85-067 Bydgoszcz, Poland
The use of calixarene derivatives as the self-assembled building blocks to form a variety of welldefined
nanonstructures including membrane mimicking materials, vesicles, nanotubes,
nanofibers, and gelators for extension of the label-free high-throughput screening (HTS)
methodologies of combinatorial drug candidate binding to various membrane and membrane
receptors has been reviewed. Especially, recent advances which enable understanding of chiral
behaviour and subsequent ability to control the molecular recognition phenomena of calixarene
derived soft nanostructures and resulting impact of these nanomaterials on the future progress in
the HTS wafer-scale micro-/nanofluidic and high-density nanoarrays technologies has been
addressed. In addition the increasing capability of the cell-mimicking self-assembled calixarene
and/or glycolipid functionalized conjugated polymers for construction of the robust, flexible,
sequencing, massively parallel HTS nanodevices enabling early detection and eradication of
bacterial toxins, viruses and other pathogenic agents present in human environment and foodstuffs
have been discussed.

MODEL STRUCTURES FOR OBTAINING SOME ANALOGUES WITH BIOLOGICAL
ACTIVITIES
I. C. Chirita. A.- V. Missir
Department of Pharmaceutical Chemistry, University of Medicine and Pharmacy "Carol
Davila", Bucharest, Traian Vuia 6, sect. 2, 020956, Romania
Analogue research plays an important role in medicinal chemistry. Having identified a new
target in molecular biology, many similar lead compounds have been synthesized resulting in
closely related products on the market.
Another approach is the further optimising of an existing drug in order to improve the original
one; in many cases these two approaches overlap. In this paper it is analysed the importance of
analogue research. Applying this method of drug design, many new compounds with a great
selectivity of action, with less adverse reactions, and a better phrmacokinetic profile with an
increasing of action duration were synthesized. We specify that the few examples which were
presented in this work are far to be an exhaustive way of exploring this subject. The mentioned
examples are just a few of the latest researches in the classes of drugs which contain therapeutic
compounds widely used by the patients (drugs affecting the cardiovascular and gastrointestinal
function, used in the therapy for hypercholesterolemia and dyslipidemia e.g.). There weren't
brought in attention the researches in the field of antibiotics. In conclusion, we can say that there
are no general rules to determinate the way of research in this field, the resulted compounds
being dependent on the marketing conditions, the company strategy and last, but not least, it
depends on the inventive capacities of the people involved.
PL-

PL-8
THE UNDERGRADUATE AND POSTGRADUATE TRAINING OF CLINICAL
PHARMACY IN ROMANIA
A. N. Cristea
Dept. of Pharmacology and Clinical pharmacy, Faculty of Pharmacy, University of Medicine
and Pharmacy "Carol Davila", Str. Traian Vuia 6, Bucharest, Romania
1. Postgraduate (PG) education of Clinical pharmacy
In Europe, the first Clinical pharmacy lecture for PG has been introduced by the Chief
pharmacist Joaquin BONAL in 1974 in Spain, at St. Paul hospital, Barcelona. In Romania, the
first Clinical pharmacy lecture for PG, for the chief pharmacists in hospitals, has been introduced
in 1985 in Bucharest, at the department of Pharmacology, Faculty of Pharmacy, UMF "Carol
Davila". PG Clinical pharmacy training is organized in modules of daily lectures for 2 weeks or
the equivalent of 1 day/week, on different topics of Clinical pharmacy (e.g. Counseling of the
patient in the community pharmacy). The first Romanian informative book about the clinical
pharmacy was published in 1984 (Simiti I., under redaction, Clinical pharmacy elements, Edit.
Dacia, Cluj- Napoca, 1984). The first Romanian applicative book about the clinical pharmacy is
published in 2006 (Cristea A.N., under redaction, Clinical pharmacy, vol. 1.
2. Undergraduate education (UG) of Clinical pharmacy
Clinical pharmacy started to be taught in universities in: 1986 in France and 1992-1997 in
Romania (introduced successive at Cluj, Tg. Mure§, Ia§i, Bucureti). In Romania in some
universities (Ia§i, Bucureti) Clinical pharmacy teaching started as facultative lectures and has
been than transformed into obligatory lectures in 2001, according with the local adaptation of the
restructuring of the teaching plan. In some universitary centers (Bucure§ti, Ia§i) Clinical
pharmacy education was created and developed at the Pharmacology department by teachers
with pharmacological formation and the departments became departments of Pharmacology and
Clinical pharmacy. In other centers the teachers of Clinical pharmacy were formed independent
of the Pharmacology departments. UG training in Clinical pharmacy for students consists of:
exercises of scientifically analyses of wrong or incoherent medical prescriptions, commented
clinical cases of pharmacotherapy, visits at tne hospital (under the guidance of hospital clinical
pharmacy specialists).
3. Vocational training in Clinical pharmacy
PG education for specialization (vocational training) in Clinical pharmacy started 1987 in France
and 1994 in Romania. In Romania the specializations for pharmacists has been approved by the
Ministry of Health in 1993 and are: clinical pharmacy (3 years), pharmaceutical laboratory (2
years), pharmaceutical industries (2 years) ana general pharmacy (2 years), beginning with 1994.
The organization of the specialization system: admission through national contest, attestation
through exam at the university center, national contest for positions in hospitals. Since 2005
general pharmacy exists no longer as specialization due to the harmonization with the
pharmaceutical law in the European Community, where this specialization does not exist.
Curriculum for vocational training in Clinical pharmacy consist of: 2 years lectures and practical
activities and the third year of practice in a hospital, at the patient's bed. The best two specialists
in Clinical pharmacy, from the first class 1996 completed a training programme for clinical
pharmacy at Salford, in UK, Hope Hospital Pharmacy department, with the support of ESCP
(European Society of Clinical Pharmacy). They are hired by contest at the biggest emergency
hospital in Bucharest and at now they are councilors* in drug field and are coordinating the
training in clinical pharmacy for students and PG.
1. Cristea A.N. Clinical Pharmacy, vol 1: Clinical pharmacy in community pharmacy, Edit.
Medicala, Bucharest, 2006.

PL-
RECENT TRENDS IN ION CHROMATOGRAPHY
P.R. Haddad , G.W. Dicinoski
Australian Centre for Research on Separation Science, School of Chemistry, University of
Tasmania, Private Bag 75 Hobart, 7001, Tasmania, Australia
Ion chromatography (IC) is the premier technique for the separation and analysis of inorganic
anions and is also highly useful for the determination of other ionic species including inorganic
cations, organic acids and bases, carbohydrates, amino acids. Despite the fact that IC is now a
mature technique there have been some very significant developments in the technique over the
last five years [1], These developments will be reviewed, with particular attention to the areas of
high speed separations, miniaturisation of columns (including to the microchip scale), and
powerful software to assist in method development. High speed separations have been a longstanding
goal of IC and the introduction of monolithic column technology has been exploited as
a means of decreasing analysis times. Silica-based and polymeric monolithic materials can be
converted into high-efficiency ion-exchangers either by direct functionalisation with appropriate
ion-exchange functionalities, coating with ionic surfactants, or coating with functionalised
nanoparticles. All of these approaches have been implemented for IC and their use to achieve
separations in less than 60 s will be described. There has been a recent trend towards
miniaturisation of IC columns and a number of capillary-based ion-exchange systems have been
reported [2]. These have used both monolithic and packed column systems. Decreasing the
column diameter has, in turn, necessitated the design of new hardware components, such as
eluent generators and suppressors. Further miniaturisation of IC onto microfluidic platforms has
also been undertaken and progress in this area will be reviewed. Some significant advances have
been made in the design of specialised software for method development in IC. Software for the
simulation and optimisation of isocratic separations has been available for several years [3],
However, many IC separations are now performed using gradient elution and there is a need for
method development software for such separations. New approaches to this problem will be
described, including methodology which enables isocratic retention data to be used to reliably
predict retention times (and to therefore optimise separations) for any elution profile comprising
isocratic steps and linear gradients. This software enables very rapid selection of optimal
gradient elution conditions for a desired group of analytes. IC is now being used for a very broad
range of new applications and two recent application areas of significance will be discussed.
The first is the determination of very low levels of perchlorate ion in foods and other samples.
The second is the use of IC for post-blast identification of improvised explosives of the type
used commonly in terrorist attacks. These two examples illustrate the manner in which IC
continues to develop as an analytical technique.
1. P.R. Haddad. Today's Chemist at Work, Feb (2004) 38.
2. P. Zakaria, J.P. Hutchinson, N. Avdalovic, Y. Liu, P.R. Haddad. Anal. Chem., 11 (2005) 417.
3. J.E. Madden, M.J. Shaw, G.W. Dicinoski and P.R. Haddad. Anal. Chem. 74 (2002) 6023.

PL-1
DRUG DELIVERY ACROSS PULMONARY BIOLOGICAL BARRIERS
C. Ehrhardt', C.-M. Lehr 2
'School of Pharmacy and Pharmaceutical Sciences, University of Dublin, Trinity College,
Dublin 2, Ireland, 2 Saarland University, Department of Biopharmaceutics and Pharmaceutical
Technology, Germany
Inhalation of medicinal aerosols to the lung for the systemic delivery of drugs has developed as
one of the most promising non-oral, non-invasive routes for delivery of drugs to the systemic
circulation. The reasons for the attention that pulmonary delivery is now receiving are three-fold;
i) the increasing numbers of therapeutic peptide and protein pharmaceuticals being developed
with their associated difficulties in conventional oral delivery, ii) recognition that the rate of
absorption from the lung provides improved pharmacokinetic profiles and iii) new developments
in the fields of inhaler devices, allowing superior targeting to specific regions in the lung. In
parallel with the ascension of pulmonary drug delivery, cell-based in vitro models emerged in
the biopharmaceutical scene. Cell lines such as Caco-2 were established as screens for the
permeability characteristics of new drug entities and are meanwhile frequently applied in drug
discovery programs. These cell culture models have been advanced to work at high throughput
capacity, yielding in vitro data that are able to predict in vivo absorption in humans. What has
been successfully shown at the intestinal barrier still remains a formidable task in the respiratory
arena. There are a number of reasons accountable for this delay; first of all, the lack of published
clinical data on absorption rates. Although many studies employing dozens of compounds are
available in public domain, the differences in study design such as choice of inhaler device and
thus area of deposition in the lung are making it impossible to use these data for an IVIVC (in
vitro-in vivo correlation). Accordingly, absorption data from animal experiments (mostly
conducted in rats) is used for correlational purposes. However, it remains elusive if instillation
into rat lungs is a good predictor for aerosolisation into human lungs. A second concern may be
the choice of appropriate cell models. Hitherto, IVIVC studies on pulmonary drug absorption
have been published using bronchial cell lines, Calu-3 and 16HBE14o-, and the intestinal Caco-2
model. Intriguingly, all three models displayed a good predictability for absorption kinetics in rat
lungs. However, absorption into the systemic circulation after inhalation is thought to occur
mainly across the large surface of the alveoli; not across the rather small surface of the
conducting airways. With major phenotypic and morphologic differences between epithelial
cells of the airways (including bronchi) and the alveoli, it remains to be investigated if there are
significant differences in the drug permeation behaviour across different epithelial regions of the
lung. Unfortunately, there is no continuous cell line available that resembles the alveolar type I
epithelial cells (which occupies > 95% of distal airspaces) and has the ability to form confluent
cell monolayers, therefore,, labour-intensive primary culture is the only method to mimic the
alveolar epithelial barrier in vitro. In conclusion, the first step to elucidate drug absorption
mechanisms in the lung is made, but there is still a wide gap to bridge to predict in vivo
absorption from in vitro studies.

In order to optimize seizure control, combination therapy with two or more AED is common and
epileptic patients will often use other medications for the management of associated conditions.
This situation can induce clinically important drug interactions. Anticonvulsivants frequently
interact with other anticonvulsivants given concurrently and if some interactions may be
favourable and require careful control of plasma concentrations to achieve optimal results,
combination therapy may have adverse effects. AED also frequently interact with other drugs.
Among old generation drugs, most of them (carbamazepine, phenytoin, phenobarbital and
primidone) are potent inducers of hepatic enzymes (CYP and glucuronyl transferase (GT)
isoenzymes). By this mechanism, they decrease markedly the plasma levels and the
pharmacological activity of other drugs such as: lamotrigine, tiagabine, several steroidal drugs
(oral contraceptives), cyclosporin A, oral anticoagulants and many cardiovascular, antineoplastic
and psychotropic drugs. Valproic acid is not an enzyme inducer but it may cause interactions by
inhibiting the metabolism of various drugs such as phenobarbital and lamotrigine.
Carbamazepine may reach toxic levels when administered with potent inhibitors of CYP3A4
such as erythromycin and ritonavir. As phenytoin is metabolized by CYP2C9 and CYP2C19,
coadministration with drugs such as fluvoxamin and ticlopidine may lead to phenytoin toxicity.
Recently developed AED are less likely to interfere with the activity of CYP or GT isoenzymes
but other drugs (gabapentin, lamotrigine, levetiracetam, vigabatrine, topiramate) can modify
their metabolism. At high dosage, topiramate may stimulate the metabolism of oral contraceptive
steroids. As patients with epilepsy are more prone than the general population to develop
psychiatric disorders, antiepileptic and antipsychotic drugs are often prescribed together and
interactions may occur, depending mostly on the induction or inhibition of CYP450 but also GT
enzymes and protein binding. Carbamazepine for instance decreases the plasma concentrations
of risperidone, olanzapine, clozapine and haloperidol. These interactions are clinically
significant because the decrease in plasma levels of antipsychotics may lead to re-emergence of
psychopathology. AED enzyme inducers can also decrease the plasma concentrations of
tricyclic antidepressants and most of the benzodiazepine drugs, the clinical significance of this
latter being less significant because of the wide therapeutic index of benzodiazepines. In
conclusion, management of drug interactions is not easy and the clinical significance of them is
often difficult to establish. Each potential interaction should be evaluated using patient-specific
parameters. In many cases the patient only needs to be carefully monitored. If necessary the
dosages will be adjusted to compensate for the predicted effects of the interaction. It will also be
essential to inform patients of potential hazards associated with over the counter medicines and
herbal products. Among AED, the lowest interaction potential is associated with the renally
eliminated drugs gabapentin and levetiracetam.
18

PL-1
DRUG DELIVERY ACROSS PULMONARY BIOLOGICAL BARRIERS
C. Elirhardt', C.-M. Lehr 2
'School of Pharmacy and Pharmaceutical Sciences, University of Dublin, Trinity College,
Dublin 2, Ireland, 2 Saarland University, Department of Biopharmaceutics and Pharmaceutical
Technology, Germany
Inhalation of medicinal aerosols to the lung for the systemic delivery of drugs has developed as
one of the most promising non-oral, non-invasive routes for delivery of drugs to the systemic
circulation. The reasons for the attention that pulmonary delivery is now receiving are three-fold;
i) the increasing numbers of therapeutic peptide and protein pharmaceuticals being developed
with their associated difficulties in conventional oral delivery, ii) recognition that the rate of
absorption from the lung provides improved pharmacokinetic profiles and iii) new developments
in the fields of inhaler devices, allowing superior targeting to specific regions in the lung. In
parallel with the ascension of pulmonary drug delivery, cell-based in vitro models emerged in
the biopharmaceutical scene. Cell lines such as Caco-2 were established as screens for the
permeability characteristics of new drug entities and are meanwhile frequently applied in drug
discovery programs. These cell culture models have been advanced to work at high throughput
capacity, yielding in vitro data that are able to predict in vivo absorption in humans. What has
been successfully shown at the intestinal barrier still remains a formidable task in the respiratory
arena. There are a number of reasons accountable for this delay; first of all, the lack of published
clinical data on absorption rates. Although many studies employing dozens of compounds are
available in public domain, the differences in study design such as choice of inhaler device and
thus area of deposition in the lung are making it impossible to use these data for an IVIVC (in
vitro-in vivo correlation). Accordingly, absorption data from animal experiments (mostly
conducted in rats) is used for correlational purposes. However, it remains elusive if instillation
into rat lungs is a good predictor for aerosolisation into human lungs. A second concern may be
the choice of appropriate cell models. Hitherto, IVIVC studies on pulmonary drug absorption
have been published using bronchial cell lines, Calu-3 and 16HBE14o-, and the intestinal Caco-2
model. Intriguingly, all three models displayed a good predictability for absorption kinetics in rat
lungs. However, absorption into the systemic circulation after inhalation is thought to occur
mainly across the large surface of the alveoli; not across the rather small surface of the
conducting airways. With major phenotypic and morphologic differences between epithelial
cells of the airways (including bronchi) and the alveoli, it remains to be investigated if there are
significant differences in the drug permeation behaviour across different epithelial regions of the
lung. Unfortunately, there is no continuous cell line available that resembles the alveolar type I
epithelial cells (which occupies > 95% of distal airspaces) and has the ability to form confluent
cell monolayers, therefore, f labour-intensive primary culture is the only method to mimic the
alveolar epithelial barrier in vitro. In conclusion, the first step to elucidate drug absorption
mechanisms in the lung is made, but there is still a wide gap to bridge to predict in vivo
absorption from in vitro studies.
15

PL-1
MANGANOPORPHYRINS, A PRECIOUS TOOL FOR THE SYNTHESIS OF MODELS
OF METABOLITES
O. Lafont, F. Estour
Faculte de Medecine et de Pharmacie de Rouen 22 boulevard Gambetta 76183, Rouen cedex 1,
France
Porphyrin catalysts are biomimetic catalysts which, due to their structural analogy with active
site of cytochrome P-450 , constitute precious tools for the synthesis of models of metabolites.
Mn (TDCPP) CI, in particular, is a second generation porphyrin, which is robust enough to be
used in the presence of hydrogen peroxide, as oxigen donor, and imidazole, as co-catalyst, for
preparative purposes. The chemical diversity obtained with this kind of catalysts proved to be
better than what was obtained with simple oxidative agents and the amounts of compounds
which were isolated were higher than what could be obtained via ex-vivo biological pathways.
Examples of stereodelective and/or regioselective xenobiotics oxidations (i.e ; methyloctalones,
chromenes, acronycine) will be described.
R J = R 2 = R 3 =H
R* = OCH 3 ;R 2 = R 3 =H
R 1= H; R 2 =CH 3 CO; R 3 =
(V
R 1 = R 2 = H; R 3 = N(CH 3 ) 2
Methyloctalones
16

PL-1
PHARMACOKINETIC INTERACTION MECHANISMS AND CLINICAL
RELEVANCE FOR DRUGS ACTING ON THE CENTRAL NERVOUS SYSTEM
J. Fontaine
Department of Physiology and Pharmacology, Institute of Pharmacy CP 205-7, University of
Brussels (ULB) - 1050 Brussels - Belgium
The administration of several drugs is frequent and the probability of a drug interaction increases
with the number of drugs received by a patient. A drug-drug interaction may be defined as the
phenomena occurring when the effect or pharmacokinetic of a drug is altered by prior
administration or coadministration of another drug.
Two types of drug interactions can be defined:
(i) Pharmacokinetic interactions are those in which one drug alters the rate or extent of
absorption, distribution or elimination (metabolism or excretion) of another drug.
(ii) Pharmacodynamic interactions are those in which one drug induces a change in a
patient's response to another drug without altering its pharmacokinetics (non
alteration of plasma concentrations).
The clinical relevance of drug interactions are often under- or overestimated, underestimation
being probably more common because most patients who receive potentially interacting drugs
do not develop severe adverse responses. Indeed, the clinical consequence of a drug interaction
can vary largely from one patient to another. The outcome can be harmful if the toxicity of a
drug is increased but a reduction in its efficacy may also be a problem. In rare cases, interactions
can be beneficial for the patient. Anyway, an understanding of the basic principles leading to
drug interactions will enhance the ability to predict possible outcomes resulting from the
coadministration of 2 drugs known or suspected to interact. In this context, the knowledge of the
pharmacological and pharmacokinetic properties of drugs will be essential. We will focus on
pharmacokinetic drug interactions which can occur by altered absorption, distribution,
metabolism or excretion and their main mechanisms will be briefly presented. Most drug
interactions are due to alterations in enzymatic processes of phase I (oxydative reactions of
which six isoenzymes are responsible for about 90% of all the metabolic activity of hepatic
CYP450 enzymes) and of phase II (glucuronidation, sulfation, methylation reactions). Some
drugs and other xenobiotics are capable of increasing the CYP450 isoenzyme activity, a process
called "enzyme induction"; the result is a decrease of plasma concentrations of drugs that are
substrates for that isoenzyme. Conversely drugs that inhibit a CYP450 isoenzyme activity may
decrease the metabolism and increase plasma concentrations of drugs that are substrates for that
isoenzyme. Inhibition of drug metabolism is perhaps the most commonly reported mechanism
responsible for the interaction between two drugs. The time course for the onset and offset of
these interactions can vary considerably and the same drug interaction can have a different time
course depending on the route of administration and the patient himself. It is well known that
some patients are at higher risk for drug interactions either because of the types of drugs they
receive or because of the disease itself. On the other hand, the most dangerous modifications are
those of plasma concentrations of drugs with a narrow therapeutic index while patients with
seizure disorders are probably one of the highest risk groups for drug interactions. In this
context, we have chosen to describe clinically important drug interactions of antiepileptic drugs
(AED).
17

In order to optimize seizure control, combination therapy with two or more AED is common and
epileptic patients will often use other medications for the management of associated conditions.
This situation can induce clinically important drug interactions. Anticonvulsivants frequently
interact with other anticonvulsivants given concurrently and if some interactions may be
favourable and require careful control of plasma concentrations to achieve optimal results,
combination therapy may have adverse effects. AED also frequently interact with other drugs.
Among old generation drugs, most of them (carbamazepine, phenytoin, phenobarbital and
primidone) are potent inducers of hepatic enzymes (CYP and glucuronyl transferase (GT)
isoenzymes). By this mechanism, they decrease markedly the plasma levels and the
pharmacological activity of other drugs such as: lamotrigine, tiagabine, several steroidal drugs
(oral contraceptives), cyclosporin A, oral anticoagulants and many cardiovascular, antineoplastic
and psychotropic drugs. Valproic acid is not an enzyme inducer but it may cause interactions by
inhibiting the metabolism of various drugs such as phenobarbital and lamotrigine.
Carbamazepine may reach toxic levels when administered with potent inhibitors of CYP3A4
such as erythromycin and ritonavir. As phenytoin is metabolized by CYP2C9 and CYP2C19,
coadministration with drugs such as fluvoxamin and ticlopidine may lead to phenytoin toxicity.
Recently developed AED are less likely to interfere with the activity of CYP or GT isoenzymes
but other drugs (gabapentin, lamotrigine, levetiracetam, vigabatrine, topiramate) can modify
their metabolism. At high dosage, topiramate may stimulate the metabolism of oral contraceptive
steroids. As patients with epilepsy are more prone than the general population to develop
psychiatric disorders, antiepileptic and antipsychotic drugs are often prescribed together and
interactions may occur, depending mostly on the induction or inhibition of CYP450 but also GT
enzymes and protein binding. Carbamazepine for instance decreases the plasma concentrations
of risperidone, olanzapine, clozapine and haloperidol. These interactions are clinically
significant because the decrease in plasma levels of antipsychotics may lead to re-emergence of
psychopathology. AED enzyme inducers can also decrease the plasma concentrations of
tricyclic antidepressants and most of the benzodiazepine drugs, the clinical significance of this
latter being less significant because of the wide therapeutic index of benzodiazepines. In
conclusion, management of drug interactions is not easy and the clinical significance of them is
often difficult to establish. Each potential interaction should be evaluated using patient-specific
parameters. In many cases the patient only needs to be carefully monitored. If necessary the
dosages will be adjusted to compensate for the predicted effects of the interaction. It will also be
essential to inform patients of potential hazards associated with over the counter medicines and
herbal products. Among AED, the lowest interaction potential is associated with the renally
eliminated drugs gabapentin and levetiracetam.

PL-1
FUNCTION AND REGULATION OF MULTI-DRUG RESISTANC RELATED
PROTEINS IN HUMAN LUNG CELLS
H. Foth, A. Raemisch, A Torky, F. Glahn, E. Stehfest
Institute of Environmental Toxicology, Martin Luther University, D-069097 Halle-Saale,
Germany
Certain particles, as asbestos particles or respirable cadmium particles or cigarette smoke
particles are considered to have a particular health risk. The mammalian lung is thus exposed to
a multiplicity of chemicals which are chemically highly reactive and may elicit a direct toxic
damage to the contact tissue. Any lung damage induced by noxious environmental chemicals
depends not only on parameters of exposure but also on lung specific metabolism and ability of
lung to adapt to noxious compounds and to repair lesions. Another important point for lung
response to exposure towards factors from the environment is the ability of lung to cope with
stressors. Here transport mechanisms for the elimination of toxic xenobiotics and their
metabolites outside the cell might be crucial. Otherwise accumulation of these compounds, such
as metals, may affect a number of regulatory and other functions, ultimately leading to cell
death. Specific membrane associated proteins, known as efflux pumps, are required to remove
these undesirable molecules from the cellular environment.
Multidrug resistance associated proteins are important factors in resistance of cells against a
variety of substrates. Currently, the MRP-subfamily contains 9 members which are all
transporters but with a different substrate spectrum. The members of the MRP-subfamily, which
are known so far, can be distinguished according to structural features of transmembrane
spanning domains. One group has 5 protein domains inserted into the cell membrane and
comprises the isoforms MRP1, MRP2, MRP3, MRP6 and MRP7. The other group has four
transmembrane protein domains and comprises MRP4, MRP5, MRP8 (ABCC11) and
MRP9(ABCC12). The latter group is characterized by lack of a N-terminal membrane spanning
domain. The isoforms MRP 1-6 are known to be drug transporters. In lung cancer cell lines
MRP1 and 3 protein levels are related to the sensitivity against several chemotherapeutics.
MRP1 is overexpressed in lung cancer cell lines and there was no difference recorded between
non small cell lung cancer, NSCLC, and small cell lung cancer, SCLC. MRP1 expression is
functionally related with resistance to doxorubicin, VCR, VP-16 and cisplatin. MRP1 substrates
are also colchicines, etoposide, rhodamine arsenite and methotrexate.
Many physiological functions of MRP 1 transporter are meanwhile known or are at least very
likely. Often, MRP's are organic anion transporters and mediate an efflux of substrates out of the
cells. The function of MRP1 is the efflux of unconjugated bilirubin and leukotriene transport and
it is involved in the acquired resistance to arsenate. MRPs often transport anionic or neutral
drugs conjugated to glutathione, glucuronate or sulfate. MRP1, MRP2 and MRP3 are
transporters for neutral organics drugs coupled with GSH. MRP3 acts as a transporter for
acetomenophen conjugates with glucuronate and GSH. Under physiological conditions MRP3
substrates are monovalent bile salts like cholate, taurocholate and glycocholate. MRP 4 was
identified as nucleotide analogue pump for phosphonylmethoyethyl-adenine (PMAE) and purine
analogues. In MRP4 transfected cells the efflux of cGMP and cAMP as well as prostaglandin El
and E2 is increased.
19

Some non steroidal antiinflammatory drugs, ibuprofen, indomethacin and diclofenac inhibit
prostaglandin transport by MRP 4. MRP 5 is also an organic anion efflux pump that prefer
nucleotide analogues. It was identified to transport GSH conjugates and GSH itself. Like MRP4,
MRP5 can use cAMP and cGMP as substrate and participates in homeostatsis of intracellular
cAMP- and cGMP- pools. Transfected MRP5 overexpressing human embryonic kidney 293
cells acquire a partial resistance against cadmium chloride and potassium antimonyl tartrate. The
regulation of MRP1 expression is much less clear than its known functionality and putative
important role for cellular defense. MRP transporters mediate a variety of physiological
functions which are closely related to a unidirectional transport of compounds. However for
lung, MRP activity will promote absorption of compounds into the body because at least MRP1
is active at the basolateral site of bronchial epithelium. Such an effect may have enormous
impact on non respiratory functions of the lung and may offer a deeper insight into mechanisms
of harm. The final pathophysiological reaction in lung toward exposure of harmful compounds
may lead to hyperreactivity of the bronchial epithelium, apoptosis or proliferation and finally
lung cancer.
It is unclear whether the functional control of genes and response to factors associated with
regulation of MRP transporters are different between individuals and between normal epithelium
of upper and lower respiratory tract. Taken into account that species variability exists with
respect to distribution and regulation of MRPs and that tumor cells lines often overexpress MRP
isoforms it is important to focus on the putative function of MRP in lung epithelium in normal
human lung cells. Our study aimed to prove cell culture models for their suitability to undertake
long lasting experiments and to compare upper airway epithelial cells and cells from peripheral
sites of human lung. We report on the options to obtain primary human bronchial epithelial
(NHBEC) and peripheral lung cells (PLC) from cases of open chest surgery with lung resections.
The culture duration for a total number of 99 cases was substantial, more than 54 % of bronchial
cell culture could be followed for more than 3 generations (> 10 weeks), about 20% of
specimens were re-cultivated for more that 5 generations (> 20 weeks). The intra-cellular
distribution of the different MRP isoforms in relation to their physiological and non
physiological function is still a point of discussion. For this purpose we used normal human lung
cells (bronchial epithelial cells, NHBEC, and peripheral lung cells, PLC) as well as tumor cell
cultures as test tools to investigate the intracelluar localisation of these proteins under classical
culture conditions and under air-liquid interface by means of indirect fluorescence microscopy.
Characterisation of the cultured cells as lung epithelial cells was performed by means of
immunohistochemical analysis. MRP1 and 3 were localised to the cellular membrane in all
tested lung cell types. In contrast to that MRP2, 4 and 5 could be described as intracellular
proteins in NHBEC and PLC. All MRP1-5 isoforms could be characterized in A549 tumor cell
line as membrane proteins. In order to imitate the physiological in vivo circumstances in the lung
we have established a dry/wet method (air-liquid interface) for cell cultivation so that cultured
cells have the option to polarize between air and basal membrane and this might influence the
distribution pattern of MRP1 and 2 in NHBEC. Using confocal laser scanning techniques we
could show that in cells kept under dry/wet conditions MRP1 was found to be localised to basolateral
cell regions while MRP2 was localised to all cell regions. Under classical culture
conditions MRP1 was not localized to particular membrane regions and MRP2 was found to be
an intracellular protein.
The aim of the current study was to get insight into multidrug resistance associated protein
function and regulation in human lung cells during long term culture. It should be checked
whether prostaglandins are modulators of MRP expression and function because they have been
shown to induced MRPs and mdr proteins in other species and tissues than human lung.
Prostaglandins are highly relevant for the control of lung function and therefore it is important to
clarify whether the expression of MRPs in human lung cells is modulated by them.

A549 lung tumor cells expressed MRP mRNA for isoforms 1, 3, 4, 5 much higher than NHBECs
and PLCs, but for MRP5 mRNA levels were lower. Cyclooxygenase inhibitors indomethacin (10
|aM, 24 h) and celecoxib (10 (iM, 24 h) were effective inhibitors of MRP1 transport in A549
cells. MRP transport was measured by 5,6-carboxy-2'7'-dichorofluorescein (CDF) efflux. In
NHBECs the efflux kinetics of CDF were substantially fastened by prostaglandin E2 (PGE2, 72
h). Also MRP1 mRNA expression was increased in NBHECs but not in PLCs and A549 after
PGE2 treatment. PLCs and A549 did not respond. MRP3 mRNA levels were increased by PGE2
in NHBEC (2.6 fold) and PLC (2.9 fold) compared to paired controls but means were not
significantly different. In NHBEC, PLC and A549 mRNA for MRPs 1, 3, 4 and 5 remained
unchanged after treatment with prostaglandin F2a. Cadmium chloride (5, 10 pM) was an
effective inducer of caspase 3/7 activation in NHBECs with a 5 - 9 fold rise of activity.
Prostaglandin E2 did not change cadmium-induced caspase 3/7 activation in NHBECs and had
no own effect on caspase 3/7. MRPs in bronchial epithelium are candidates for an inflammation
induced activation of MRP transport activity which in fact means absorption of compounds from
the epithelial lining into the body.

PL-1
PLANTS AS A RESOURCE OF NOVEL ANTIBACTERIALS AND RESISTANCE
MODIFYING AGENTS
S. Gibbons
Centre for Pharmacognosy and Phytotherapy, The School of Pharmacy, University of London, 29-39
Brunswick Square, London WC1N 1AX, UK
Multidrug-resistant bacteria continue to provide considerable challenges in terms of their treatment and
containment. Of major concern in terms of morbidity and mortality are methicillin-resistant
Staphylococcus aureus (MRSA) and Mycobacterium tuberculosis. Certain species of Gram-negative
bacteria which are intrinsically resistant to many antibiotics such as Pseudomonas and Acinetobacter
species, are likely to be the true 'superbugs' of the future.
Many of these bacteria express membrane bound multidrug efflux pumps which confer specific and
multidrug-resistance (MDR) to a wide array of antiseptics such as quaternary ammonium compounds
(QACs), which can make eradication from the clinical setting difficult. Some of these mechanisms such
as the NorA system in S. aureus are true multidrug-efflux pumps which extrude many structurally
unrelated antibiotics including certain fluoroquinolones. Tuberculosis (TB) is also on the increase and
MDR variants in Mycobacterium tuberculosis include those that express the IniA system which exports
isoniazid and ethambutol, two of the main front line antibiotics that are used in combination to treat TB.
There is therefore an urgent need to characterize novel natural products and synthetic compounds which
are antibacterial, and also to find compounds that could inhibit these resistance mechanisms and,
potentially, be used in combination with existing antibiotics to restore their efficacy. This would be
valuable as it has been shown that the use of a bacterial resistance modifying agent in combination with
an antibiotic has the ability to reduce the occurrence of antibiotic resistant variants. This would lessen
the burden of antibiotic resistance.
Plants are an unexploited source of antimicrobial substances' 2 and at present there are no examples of
single chemical entity plant natural products that are used clinically as antibacterials. This is highly
unusual given the use of herbal products to treat bacterial infections, examples of which include
cranberry and bearberry. There are several reasons why plants as a source of chemical diversity for new
antibacterials should be evaluated. Firstly, it is likely that plants have evolved a chemical defence to
protect themselves from bacteria and fungi in their environment. Evaluation of extracts from
subterranean plant parts is likely to provide natural products that have activity against soil organisms of
which the genus Mycobacterium is a major representative. This would indicate potential against TB and
fast growing mycobacterial strains. Secondly, it is highly unlikely that clinically relevant species of
bacteria will have been in contact with a class of plant-derived natural product antibacterial, and
therefore intrinsic resistance is less likely. This also implies a final point, that it is possible that the
mode of action of plant antibacterials may be via a novel and unusual mechanism of action, in addition
to the common modes exerted by fluoroquinolones (target DNA gyrase) or tetracyclines (protein
synthesis inhibition). Our group has
been characterizing plant derived
antibacterials with activity against
MRSA and MDR variants of S.
aureus and has recently shown that
members of the acylphloroglucinol
class of natural product e.g. 1 show
promise as leads. 3 We have also been
using fast-growing species of
Mycobacterium such as M. fortuitum,
M. abscessus, M. phlei, M. aurum and M. smegmatis as models to characterize new antimycobacterial
leads.
22

There is a good correlation between the activities of anti-TB drugs against these fast-growing strains
and this alleviates the need for class-Ill facilities. Compound 2 is a member of the rare homoisoflavonoid
class of natural products and displayed MIC values in the region of 8 (ig/ml against some
of these fast-growing species. 4
Our group is also characterizing compounds that inhibit MDR mechanisms in Staphylococcus aureus
and potentiate antibiotic activity against resistant variants. Some of these metabolites from the
flavonoid (3) and alkaloid groups (4), inhibit ethidium bromide efflux from an MDR strain of S. aureus
possessing the NorA MDR transporter, one of
the major characterized antibiotic pumps in
this species. Compound 3 is unusual in that in
addition to being a potent inhibitor of efflux in
multidrug-resistant Staphylococcus aureus,
this compound also displays antibacterial
activity against MRSA and MDR variants of
S. aureus.
This paper will describe our work on
antibacterial and bacterial resistance
modifying natural products and future
challenges with multidrug-resistant
Mycobacterium and Gram-negative bacteria
will be outlined.
3 4
1. Gibbons, S. (2004) Anti-Staphylococcal Plant Natural Products. Natural Product Reports 21:263-
277.
2. Gibbons, S. (2005) Plants as a source of bacterial resistance modulators and anti-infective agents
Phy to chemistry Reviews 4 (1) 63-78.
3. Gibbons, S., Moser, E., Hausmann, S., Stavri, M., Smith, E. and Clennett, C. (2005) An antistaphylococcal
acylphloroglucinol from Hypericum foliosum. Phytochemistry 66 1472-1475.
4. O'Donnell, G., Bucar, F. and Gibbons S. (2006) Phytochemistry and Antimycobacterial activity of
Chlorophytum inornatum. Phytochemistry 67 (2) 178-182.

PL-1
NEW APPROACHES IN THE DISCOVERY OF BIOACTIVE NATURAL PRODUCTS -
METHODS AND EXAMPLES
M. Hamburger
Institute of Pharmaceutical Biology, Department of Pharmaceutical Sciences, University of
Basel, CH-4056 Basel, Switzerland
Natural products remain a major source of inspiration for the development of new drugs.
Medicinal plants and phytomedicines play a major role in health care in developing and
industrialized countries alike. Over the past two decades, a wealth of new technologies has
become available in analytical chemistry and in the life sciences, but crossfertilization into
natural product research has been rather slow. The lecture will address this issue and review
some of the technologies and possible approaches towards discovery of promising bioactive
molecules. We will argue that a consequential implementation and judicial combination of these
technologies and rigorous approaches is needed to move natural product research into a new era.
Even though academia should not attempt to imitate the industrial approach of target-driven
high-throughput screening (HTS), a state-of-the-art technology platform is required which
combines analytical HPLC with various on-line detectors such as PDA, MS, ELSD, and NMR in
on- or offline mode. Such platforms allow an extensive structural characterization of individual
components in complex extracts without need to resort to preparative isolation. The efficient
tracking of bioactivity in an extract remains a major challenge. Various approaches for
interfacing analytical separation with bioassays have been described in the literature. Some of
these will be discussed, including HPLC-based activity profiling, biosensors and other on-line
assays. In many cases, assays with high information content and complex endpoints, such as
phenotypical screens, are to be preferred over biochemical assays, in the primary screening of
extracts as well as for the localization of active compounds. The molecular targets for these leads
can be studied with the tools of molecular and cell biology. The postgenomic era offers a range
of new tools and approaches. Their common feature is that they permit an essentially unbiased
and global investigation which does not need to be hypothesis-driven. For example, effects of
extracts and compounds in a living system can be studied by genomwide expression profiling
with RNA microarrays. HPLC-based profiling will be illustrated with the example of the antiinflammatory
plant Isatis tinctoria, while phenotypical screening and subsequent
characterization of signaling pathways will be discussed with the example of neurotrophic
alkaloids from Paecilomyces militaris. The use of expression profiling in target characterization
will be described with our ongoing studies on Cimicifuga racemosa.
1. Potterat, O., Hamburger, M. Natural Products in Drug Discovery - Concepts and
Approaches for Tracking Bioactivity, Curr. Org. Chem., in press.
24

PL-1
A DIABETES II COST MODEL TO PROJECT SAVINGS ACCRUING FROM
DISEASE MANAGEMENT PROGRAMS
A. G. Hartzema, A. G. Winterstein, S. Snedecor, M. Taylor, R. Segal, R. Franck
University of Florida, Florida.
Diabetes mellitus, a disease continually increasing in prevalence, if left uncontrolled leads to
debilitating complications, including renal failure and blindness. Glycosylated hemoglobin
(HbAlc), an indicator of glycemic control over a 3-month time period, is the preferred measure
for determining whether an individual's diabetes is controlled. The level of glycemic control or
HbAlc determines the progression of the disease and the onset of micro- and macrovascular
complications. Diabetes treatment therefore has a substantial preventive component. It has been
suggested that early prevention can not only reduce the incidence of complications but also
avoid healthcare expenditures that are associated with the treatment of these complications. The
purpose of this research is to develop an economic model that allows for the projection of cost
savings that may accrue from diabetes II disease management programs.
Evaluation of the DiabetikSmart Program
The study considered all adult patients with a medical diagnosis of diabetes II, who are enrolled
in the Medipass program, and who reside in South Florida eligible for participation. Patients
with HIV, hemophilia, or with gestational diabetes were excluded. Even though various
recruitment methods were used, patient accessibility and the dependence on voluntary patient
participation compromises the generalizability of the study population to the universe of Florida
Medicaid patients. This is however, a common problem in disease management programs and
should not detract from the fact that certain populations may participate and benefit from quality
improvement initiatives. HCN enrolled a sample of 502 patients between 9/5/2002 and
6/25/2003. Cross-check with Medipass eligibility data (performed by AHCA) suggested that 443
patients had been eligible at study entry. Of 443 patients, 351 (79%) had a baseline HbAlc value
within the pre-defined acceptable time period surrounding the enrollment date. The remaining
patients were omitted from further analysis. Of the 351 patients, 226 had at least one follow-up
visit with the study nurse. Of 351 patients that constituted the study population, 167 (47.5%)
patients had HbAlc values collected within a pre-defined acceptable follow-up period of
between three and nine months after the initial visit. Patients who dropped out were slightly
younger, had slightly lower HbAlc values, and were more likely Caucasian. Comparison of the
167 patients who completed the study and had valid HbAlc values available suggested a
significant improvement in glycemic control over the course of the study period. Baseline values
of 8.4% with a standard deviation (std.) of 2.14 decreased an average of 0.7% to mean levels of
7.7 (std. 1.70). This difference was found to be statistically significant (p

Table 1. HbAlc frequencies for baseline and follow-up (completers analysis)
Baseline
Follow-up
Frequenc
y Percent Frequency Percent

The first two parameter categories were obtained by retrospective analysis of Medicaid claims
data. Incidence estimates for each defined complication were based on diabetes type 2 patients
with continuous eligibility of at least 24 months or death. First onset of complications was
defined by a preceding year of continuous eligibility without claims for the complication of
interest. As incidences in this model rely on claims they only represent events with respective
charges to Medicaid. While this is appropriate for the economic prediction its validity in an
epidemiologic sense is limited. Annual incidence estimates for selected complication are
displayed in table 3. Cost estimates were obtained by averaging cost for claims associated with
each respective complication over defined time windows. Costing periods varied based on the
average duration of the complication as did the type of claims that were included. All
complications except ESRD, background retinopathy, and prosthesis for LEA were expected to
generate cost for no longer than one year. Initiation of nursing home, home healthcare, and select
drug claims after onset of a complication were included up to one year. Annual cost per selected
complication is listed in table 3. Estimates of risk reduction based on HbAlc levels were
obtained from the literature. Ascertainment followed established methods for systematic reviews
with defined key word searches and standardized review criteria. If multiple risk estimates for a
given complication were available, summative meta-analytic methods (random effect models)
were used to obtain a pooled estimate along with 95% confidence intervals. Incidence estimates
for each complication are displayed in table 3.
Table 3. Model parameters
Incidence
(per 1000 Probability
Direct Cos ts (2003$)
pat-yrs) per year RR (95% CI) Mean Median
AMI 2.85 2.843 xlO" 3 1.16(1.09- 1.27) $26,583.88 $11,980.48
LEA 1.63 1.627 xlO" 3 1.30 (1.10- 1.50) $34,096.58 $16,799.00
Prosthesis N/A N/A N/A $1,810.66 $370.12
Ulcer 54.92 5.344 xlO' 2 1.30(1.20- 1.50) $2,117.68 $51.05
Background Retinopathy 15.32 1.520 xlO" 2 1.25 (1.19 - 1.30) $67.29 $20.12
Proliferative Retinopathy 6.47 6.445 xlO" 3 1.25 (1.19- 1.30) $163.26 $41.75
ESRD year 1 4.30 4.289 xlO" 3 1.22(1.13 - 1.32) $17,078.71 $8,024.26
ESRD subsequent years N/A N/A N/A $13,598.89 $6,569.36
Kidney Transplant 1.91 1.908 xlO" 3 N/A $22,695.90 $23,263.14
Transplant Maintenance N/A N/A N/A $13,680.11 $13,279.43
ESRD Mortality 217.42 1.954 xlO" 1 N/A $0.00 $0.00
All-Cause Mortality 85.68 8.211 xlO" 2 1.17(1.10-1.27) $0.00 $0.00
Patients' transition over five years was simulated in a Markov model. A hypothetical cohort of
1000 patients acquired the various complications based on the above listed incidence estimates.
Incidence estimates varied based on the HbAlc-specific risk estimates when different HbAlc
levels were assumed. Since most complications accrued the largest proportion of cost at onset
(hospitalization) mid-cycle corrections were only used for non-proliferative retinopathy, endstage
renal disease and cost for prostheses after LEA. Mortality was assumed to be independent
of glycemic control for the cost analysis. A separate model to assess the effect of glycemic
control on mortality was developed instead. The discount rate was set at 3%. Table 4 reports
discounted cost savings for the population recruited for the DiabetikSmart program for a 5-year
follow-up period, analyzing only patients who completed the study.

Table 4. Projected 5-year cost savings (discounted) for the DiabetikSmart program (completers)
Model % HbAlc Reduction Mean Diff in Cost Population Savings
AMI 0.7042 $28.85 167 $4,817.28
LEA 0.7042 $39.04 167 $6,519.64
Nephropathy 0.7042 $100.29 167 $16,749.01
Retinopathy 0.7042 $2.51 167 $419.42
Ulcer 0.7042 $52.53 167 $8,771.89
Total Savings $223.22 $37,277.24
Table 5 illustrates the potential cost impact of a 1% reduction in HbAlc for the entire Florida
Medicaid diabetes type 2 population for either a 5-year follow-up period or patients' lifetime. It
is estimated that $39 million could be saved over 5 years if glycemic control was improved by
1%, and $116 million could be saved over patients' lifetime.
Table 5. Projected 5-year and lifetime cost savings (discounted) for the Florida Medicaid
diabetes type 2 population
5-Year Mean Savings Per Patient, Discounted
Standard 1% HbAlc Mean
Complication Care decrease Savings STD Median Total Savings
AMI $278.04 $237.07 $40.96 $65.68 $21.20 $5,050,630.73
LEA $250.80 $195.36 $55.44 $84.93 $29.87 $6,835,448.84
ESRD $784.33 $641.91 $142.42 $156.71 $93.03 $17,560,330.43
Retinopathy $18.34 $14.77 $3.57 $7.26 $1.53 $439,732.05
Ulcer $321.59 $247.00 $74.59 $457.21 $2.54 $9,196,804.46
All Complications $316.98 $543.32 $199.67 $39,082,946.49
Lifetime Mean Savings Per Patient, Discounted
Standard 1% HbAlc Mean
Complication Care decrease Savings STD Median Total Savings
AMI $622.54 $532.33 $90.21 $144.67 $46.65 $11,122,870.42
LEA $854.39 $667.60 $186.79 $365.04 $89.38 $23,031,034.47
ESRD $3,069.23 $2,524.87 $544.36 $675.05 $335.82 $67,118,494.22
Retinopathy $112.49 $92.77 $19.72 $35.23 $8.36 $2,431,236.99
Ulcer $551.05 $448.71 $102.34 $628.54 $3.47 $12,618,053.84
All Complications $943.42 $1,065.16 $650.49 $116,321,689.95
In conclusion;
The literature on the disease progression in Diabetes patients and complication rates is scarce.
There is a need for long-term follow-up studies that examine the relationship between HbAlc
levels and complication rates. It is also not clear if the general assumption that there is a linear
relationship between HbAlc levels and the incidence of complication rates. Then we found that
a 1% lowering of HbAlc in the Diabetes II population considering a 5-year and a life long
impact will result in a significant cost savings to the third party payer.

PL-1
GENOTOXICITY AND DRUG DEVELOPMENT
J. Harvey
GSK R&D, UK
The registration of a new pharmaceutical requires a comprehensive assessment of its genotoxic
potential in accordance with ICH guidelines [1]. Typically, a new pharmaceutical is assessed in a
number of in vitro and in vivo genotoxicity assays before administration to humans. The in vitro
bacterial and mammalian cell based genotoxicity assays, in particular, were designed to be
highly sensitive to detect various mechanisms of genotoxicity. Together, the results of
genotoxicity assays are used to assess a new pharmaceutical's potential carcinogenicity.
Surprisingly, available data indicate that positive genotoxicity findings are a frequent occurrence
in the drug development process [2], Obviously compounds with clear multiple signals in one, or
more, of the standard genotoxicity tests are unlikely to be suitable for clinical trials and will be
non-developable for the majority of clinical indications. However, the relevance of an isolated in
vitro mammalian genotoxicity positive finding for a compound is more difficult to determine,
especially when using healthy volunteers in early clinical trials where the risk-benefit paradigm
is not relevant. Typically, additional mechanistic studies and supplementary in vivo genotoxicity
assays are conducted to determine the relevance of the in vitro genotoxicity findings in humans.
Recently the relevance of some of the standard genotoxicity assays in predicting carcinogenicity
has been questioned by both pharmaceutical companies and regulatory bodies. However, in the
absence of any suitable alternative genotoxicity assays, in the short term more effective
genotoxicity screens are required to eliminate potentially problematic compounds earlier in
development. While there is encouraging progress in the development of various in silico and
bacterial genotoxicity screens, the development of better eukaryotic and/or mammalian cell
genotoxicity screens has been slow. However, various "genomics" based approaches appear
promising and could well bridge this gap in the current available genotoxicity screens. These
recent developments could eliminate the potentially problematic compounds at earlier stages of
pharmaceutical development.
1. ICH Harmonised Tripartite Guideline (S2A) Guidance on specific aspects of regulatory
genotoxicity tests for pharmaceuticals.
2. Snyder and Green (2001) Mutation Research 488 151 -169
29

PL-1
NON-VIRAL GENE DELIVERY SYSTEMS BASED ON BIODEGRADABLE
CATIONIC POLYMERS
W.E. Hennink. H.K. de Wolf, J. Luten, C.J. Snel, C. Oussoren, G. Storm
Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences, Utrecht University,
PO Box 80082, 3508 TB Utrecht, The Netherlands.
Preclinical research in gene therapy has demonstrated the potential for the treatment of both
acquired and inherited diseases. In order to get the therapeutic gene into the target cell, a
delivery vehicle is required. For this purpose both viral and non-viral vectors are under
evaluation. Non-viral gene delivery systems e.g. based on cationic lipids ('lipoplexes') and
cationic polymers ('polyplexes') can overcome some of problems associated with the current
viral based therapies. A variety of cationic polymers have been proposed for polyplex formation,
each with its own transfection efficiencies (1). However, most of these polymers are nonbiodegradable
and rather toxic. Therefore, in recent years there is a growing interest in the
design of transfectants based on non-toxic and biodegradable polymers. Biodegradability
prevents intracellular accumulation of the polymer and, moreover, degradation of the polymer
can be used as a tool for intracellular release of plasmid DNA. Luten et al (2) reported on the
synthesis and gene delivery characteristics of a biodegradable polyphosphazene bearing the
cationic side group 2-dimethylaminoethylamine (DMAEA) (Figure 1).
Figure 1: poly(DMAEA)phosphazene
Polyplexes based on this polymer showed good in vitro transfection (COS-7 cells) even in the
presence of serum. Therefore, further investigation of these polyplexes for in vivo application
was warranted. The gene delivery characteristics of poly(DMAEA)phosphazene polyplexes were
evaluated in tumor bearing mice after intravenous administration. Neuro 2A could be effectively
transfected using poly(DMAEA)phosphazene polyplexes. Optimal transfection was observed at
an N/P ratio of 20. Under these conditions polyplexes with a size of 100 nm and a positive zeta
potential were formed. Polyplexes prepared from this polymer and a plasmid encoding for
luciferase were intravenously administered in mice bearing a subcutaneous tumor consisting of
the same cell line (Neuro 2A). No acute toxicity was observed. A remarkable finding is that
luciferase expression in the tumor tissue was at least 100 fold higher than in all other tissues. A
biodistribution study, using 35S labeled plasmid, showed considerable tumor accumulation of
these non-shielded polyplexes (3). In conclusion, poly(DMAEA)phosphazene is an attractive
gene carrier for in vivo application showing tumor selective gene expression.
1. M. D. Brown, A. G. Schatzlein and I. F. Uchegbu, Int. J. Pharm. 229:1-21 (2001).
2. J. Luten, J. H. Steenis, R. van Someren, J. Kemmink, N. M. E. Schuurmans-
Nieuwenbroek, G. A. Koning, D. J. A. Crommelin, C. F. van Nostrum and W. E.
Hennink, J. Contr. Rel. 89:483-497 (2003).
3. De Wolf, H.K., Luten, J., Snel, C.J., Ousorren C„ Hennink, W.E., and Storm, G, J. Contr.
Rel 709,275-287,2005.
30

PL-1
PRECLINICAL EVALUATION OF PLANT PHENOLICS FOR HEALTH BENEFICIAL
EFFECTS
R. Hiltunen, H. Vuorela, H.J.D. Dorman
Division of Pharmaceutical Biology, Faculty of Pharmacy, University of Helsinki, P.O.Box 56
(Viikinkaari 5 E), FIN-00014 University of Helsinki, Finland.
Phenolic compounds, especially polyphenols comprise a large group of secondary plant
metabolites of varying chemical structures. In the plant kingdom these numerous and nearly
ubiquitous chemicals have an important role in plant physiology e.g. to ensure pollen transport
for survival of plant species and to ensure tissue protection against the damaging effects of UVradiation
and plant diseases.
In plants, phenolics arise from the shikimate and acetate pathways. Up to now more than 8000
polyphenols structures are known and much more are still awaiting discovery. The chemical
structures of plant phenolics vary from simple phenols and phenolic acids, e.g. hydroxycinnamates,
to more complex flavonoids, lignans, lignins and condensed tannins. Among the
plant phenolics, flavonoids comprise the most common and widely distributed group with many
subclasses. More than 5000 flavonoid structures are currently known. In plants phenolics occur
as either hydrophobic free aglycones, mostly water-soluble glycosides bound to the different
types of sugar structures or are esters of aliphatic alcohols.
Numerous plants rich in phenolics have been used in folk medicine over hundreds of years.
Phenolics containing drugs and preparations thereof have been used as cardioactive, diuretic,
antispasmodic, antidiarrhoeal, hypotensive, hypertensive, coagulant, anticoagulant, hypo- and
hyperlipidaemic herbal ingredients and more recently as a source of antioxidants. In addition to
their free radical scavenging activity, polyphenols have been ascribed a number of other
beneficial effects in a variety of disease states, including cancer, cardiovascular diseases and
neurodegenerative disorders amongst others. Flavonoids, hydroxy-cinnamates and other
polyphenols are reported to have multiple biological activities including antibacterial, antiinflammatory,
antimutagenic, antiallergic, antiviral, estrogenic and vasodilatory effects.
Polyphenols and their metabolites have been identified to be inhibitors of numerous enzymes e.g.
phospholipase (A2), cyclooxygenase, lipooxygenase, glutathione reductase, xanthine oxidase and
different types of lipid and protein kinases, e.g. phosphoinositide 3-kinase, Akt/protein kinase B
(Akt/PKB), tyrosine kinases, protein kinase C, and mitogen activated protein kinase (MAP
kinase)'.
During last years, especially after the beginning of the new millennium, the interest in studying
phenolic compounds of plant origin has increased dramatically especially after the largely
accepted opinion and claims that polyphenols are responsible for the health beneficial effects of
fruits, vegetables and berries. Also the knowledge about reactive oxygen and nitrogen species
and their harmful effects on cell and tissue levels and a link to degenerative diseases have
affirmed the interest among researcher groups. Today, on average, every second hour a new
study about the plant phenolics is finished and published and brought to light. Since the year
2000 tens of thousands of publications all over the world approaching the problems of plant
phenolics have annually been published.
31

The chemistry, nutritional properties and nowadays also antioxidative and many other health
beneficial attributes of phenolic compounds have been extensively discussed in the literature. In
general, polyphenols are considered as antioxidants needed to prevent the formation oxygen and
nitrogen reactive species and to prevent free radical from causing damage to critical intracellular
targets such as lipids, proteins and other biomolecules, even to DNA.
There are an increasing number of publications, which call into question the relevance of the
antioxidant protective effect of dietary polyphenols and their metabolites in relation to
cardioprodective, chemopreventive and neuroprotective health benefits. Williams et al. predicted
in 2004 that flavonoids and their in vivo metabolites do not necessarily act as conventional
hydrogen-donating antioxidants but may exert modulatory actions in cells through actions upon
protein kinase and lipid kinase signalling pathways 1 .
For many reasons, all of the proposed beneficial effects of plant phenolics in a number of
diseases and degenerative states cannot be explained only by the antioxidant properties e.g.
reducing capacities or their influence on intracellular redox status. One reason is that once
digested, polyphenolic compounds are decomposed and altered via conjugations, oxidative and
cytochrome-related metabolism. This metabolic modification may result in decreased antioxidant
activity. Another reason is the absorption of phenolics and the distribution of phenolics origin
metabolites into cells and organs. A number of studies have demonstrated that the poor
bioavailability of phenolics does not always offer adequate concentrations of antioxidants in
organs for prevention of harmful oxidative reactions only by reduction. The concentrations of
polyphenols encountered in vivo can, however, be sufficiently high to have pharmacological
activity at receptors, enzymes, and transcription factors.
In 2004, Walle et al. examined the absorption, bioavailability, and metabolism of resveratrol
after oral doses in six human volunteers 2 . The absorption of a dietary relevant oral dose of 25 mg
was at least 70% with a peak plasma level of resveratrol and its metabolites of about 2 pM. That
is considered to be enough to initiate cell transduction. It is known that many of the extracellular
signaling molecules act as very low concentrations, typically at levels less than 10" 8 M. In
general, however, higher concentrations (5 - 200 pM) of resveratrol, its derivatives and the other
polyphenolics are needed to induce apoptosis or to obtain antiproliferative activity in cell
models 3 . Resveratrol has recently been suggested as a potential cancer chemopreventive agent.
The suggestion is based on its inhibitory effects on cellular events associated with cancer
initiation, promotion and progression 4 . Resveratrol has also been detected to inhibit, in a dosedependent
manner, free-radical formation (median effective dose ED 50 = 27 pM) when assayed
in human promyelocytic leukemia (HL-60) cells. Resveratrol was also found to inhibit COX-1
with an ED50 of 15 mM. In the literature 5 , numerous examples of molecular targets have been
identified through which resveratrol and its derivatives act upon the cell. In transcription
regulation e.g. NF-KB, P-catenin and API are down regulated. In cell cycle arrest cyclin A, B1
and D are down regulated and Cdk2, p21 and p27 are up regulated. In induction of apoptosis
and in inhibition of growth p53, casapses, ceramide and Bax are up regulated and Bcl-2, BcI-xL,
TNF, IL-ip, IL-6 and survivin are down regulated.
Quercetin one of the major bioflavonoid in the human diet is considered as potentially
carcinogenic but recently also as an anti-cancer agent. Kuo et al. (2004) investigated the roles of
surviving and p53 in quercetin-treated human lung carcinoma cells. They found out that
quercetin at concentration levels from 20 to 80 pM induced the cytotoxicity and apoptosis in test
cells in a concentration dependent manner, inhibited cell growth, increased the fraction of G2/M
phase and raised cyclin B1 and phospho-cdc2 proteins 6 . In another study 7 quercetin was found to
down regulate the expression of mutant p53 in human breast cancer cell lines and to inhibit the
expression of p53. This inhibition was found to arrest the cells in the G2/M phase of the cell
cycle.

In this study the quercetin concentration at which tumor cell growth was inhibited (IC50) ranged
from 7 nM (melanoma cell lines MNT1, M10, Ml4) to 100 pM in head and neck malignant cell
lines HTB43 and CCL135. Nowadays several promising plant-derived phenolics are in clinical
trials through the auspices of the U. S. National Cancer Institute as potential cancer
chemopreventive agents, including curcumin, genistein, soy isoflavones, green
tea/epigailocatechin gallate and resveratrol.
During the last years in connection with one of our research projects, the search for and
characterization of new natural antioxidants, a number of extracts and sub-extracts of plants
8 9
belonging to Lamiaceae, Apiaceae, Zingiberaceae and Pinaceae families were studied ' Plant
phenolics are said to be multifunctional antioxidants, and they might act at several levels in the
oxidative sequence. These multiple potential mechanisms of antioxidant action make the diverse
group of phenolic compounds an interesting target in the search for health-beneficial
phytochemicals, and also offer a possibility to use phenolic compounds or extracts rich in them
in lipid-rich foods in order to extend their shelf life. Numerous techniques are available to
evaluate the antioxidant activities of compounds and complex mixtures such as plant extracts.
Despite these various methods, just one procedure cannot identify all possible mechanisms
characterizing an antioxidant (see refs. in 8). Thus a well-devised battery of assays should
include a wide and varied selection of all types of techniques. Generally, such assays can by
categorized as non-radical-based systems, e.g. iron (III) reduction and quantification of oxidation
by-products, or free radical-based systems. The latter can further be divided into synthetic radical
systems, e.g. those using DPPH* and ABTS* + , and biologically relevant free radical systems, e.g.
hydroxyl radical-based assays. In a study de-odourised aqueous extracts of four commonly
consumed herbs belonging to the Lamiaceae family, i.e. oregano (Origanum vulgaris L.),
rosemary (Rosmarinus officinalis L.), sage (Salvia officinalis L.) and thyme (Thymus vulgaris
L.), were investigated for their antioxidant properties. Various experimental models were used
for the characterisation of the activity, e.g. iron reduction and chelation capacities, DPPH*,
ABTS' + and *OH radical-scavenging activities and the capacity of the extracts to inhibit copperinduced
oxidation of human low-density lipoproteins (LDL) ex vivo.
Extracts in general show varying degrees of reductive and radical scavenging capacity, and are
capable of extending the lag-time in LDL oxidation. In our study, the antioxidant protection of
LDL was evaluated as the delay in the onset of oxidation (effect on the lag-phase) as well as the
capacity to participate in the on-going peroxidation in the LDL particles (effect on the slope of
the propagation phase). The highest prolongation of the lag-phase was observed with the extracts
highest in the total phenolic content (sage and rosemary). An improved HPLC post-column
methodology for the identification of free radical scavenging phytochemicals in complex
mixtures was developed during the project. Pycnogenol®, a commercial (Horphag Research,
Switzerland) water-soluble maritime pine (Pinus maritime L.) bark extract, demonstrated potent
antioxidant activity throughout a range of model antioxidant assays 10 . A local Scots pine (Pinus
sylvestris L.) bark extract isolated identically produced marginally more potent free radical
scavenging activity than Pycnogenol against DPPH*. Among ca. 100 plant extracts rapeseed,
raspberry, and pine bark demonstrated not only antioxidant activity but also antimicrobial,
antiinflammatory, and antimutagenic properties as well as increased Caco-2 cell permeability".
It was shown that rapeseed and pine bark phenolics and raspberry anthocyanins were good or
excellent antioxidants toward oxidation of phosphatidylcholine membrane (liposomes), rapeseed
oil (crude) phenolics were effective radical scavengers (DPPH test), and both raspberry and pine
bark phenolics inhibited LDL oxidation. None of the tested extracts were mutagenic or toxic to
Caco-2 cells or macrophages.

Ethyl ether extracts of two plant known to contain phenolics, Ligusticum chuarvciong hort (syn.
L. wallichii Franch.) and Angelica sinensis (Oliv.) Diels (Apiaceae) which are largely used in
traditional Chinese medicine to treat atherosclerosis and hypertension were studied for their
protective effects on endothelial cell damage using human umbilical vein endothelial cells
(ECV304) 12 . The cell viability increased from an average 42% (of control) to 90% when the
concentration of the extracts was increased from 0 to 300 pg/ml and time from 0 to 12 hours.
The extracts protected the ECV304 cells both dose- and time-dependently when induced by
hydrogen peroxide. Western blot analysis revealed that the extracts significantly increased the
phosphorylation of extracellular signal-regulated protein kinase (ERK). The relative ERK
activity increased dramatically when the concentration of the extracts increased from 0 to 300
(ig/ml. Extracts also promoted eNOS expression indicating that the extracts protected the cells
from being damaged through the eNOS pathway. The extracts also increased the enzymatic
activity of SOD, CAT and GPX significantly at higher concentration levels (200 and 300 pg/ml)
but not at lower levels (0 and 100 (.ig/ml). These results could be additional evidence that
Ligusticum chuanxiong and Angelica sinensis may prevent both cardiovascular and
cerebrovascular diseases. These two extracts were also studied for their effects on rat vascular
smooth muscle cell (VSMC) proliferation. The extracts significantly inhibited proliferation and
protein synthesis of VSMC in a dose and time-dependent manner. The extracts were found to
inhibit VCMS proliferation by arresting Gi to S progression.
1. Robert J. Williams, Jeremy P. E. Spencer, and Catherine Rice-Evans (2004), Free Radical
Biology & Medicine, Vol. 36, No. 7, pp. 838- 849.
2. Thomas Walle, Faye Hsieh, Mark H. DeLegge, John E. Oatis, Jr., and U. Kristina Walle (2004):
Drug metabolism and disposition drug, Vol. 32, No. 12, pp. 1377—1382.
3. Mariella Roberti, Daniela Pizzirani, Daniele Simoni, Riccardo Rondanin, Riccardo Baruchello,
Caterina Bonora, Filippo Buscemi, Stefania Grimaudo and Manlio Tolomeo (2003), J. Med.
Chem. 46, 3546 - 3554.
4. Meishiang Jang, Lining Cai, George O. Udeani, Karla V. Slowing, Cathy F. Thomas, Christopher
W. W. Beecher, Harry H. S. Fong, Norman R Farnsworth, A. Douglas Kinghorn, Rajendra G.
Mehta, Richard C. Moon, John M Pezzuto (1997), Science (Washington, D.C.) 275(5297), 218-
220.
5. Kathryn A. Roupe, Connie M. Remsberg, Jaime A. Yanez and Neal M. Davies (2006), Current
Clinical Pharmacology 1,81-101
6. Pao-Chen Kuo, Huei-Fang Liu, Jui-I Chao (2004), Journal of Biological Chemistry 279(53),
55875-55885.
7. Davis W. Lamson and Matthew S. Brignall (2000), Alternative Medicine Review 5(3), 196 - 208.
8. H.J.Damien. Dorman, Anna Peltoketo, Raimo Hiltunen, Matti J. Tikkanen (2003) Food
Chemistry 83, 255-262
9. H. J. Damien Dorman, Oliver Bachmayer, Muberra Kosar, and Raimo Hiltunen (2004), J. Agric.
Food Chem. 2004, 52, 762-770
10. Muberra Kosar, Damien Dorman, K. Baser, R. Hiltunen (2004), Chromatographia 2004, 60,
December (No. 11/12), 635 - 638.
11. Satu Vuorela, Kari Kreander, Maarit Karonen, Riina Nieminen, Mari Hamalainen, Anna Galkin,
Leena Laitinen, Juha-Pekka Salminen, Eeva Moilanen, Kalevi Pihlaja, Heikki Vuorela, Pia
Vuorela, and Marina Heinonen (2005) J. Agric. Food Chem., 53 (15), 5922 -5931
12. Yong Zhong Houa, Guang Rong Zhaoa, Jie Yanga, Ying Jin Yuana, Guo Guang Zhua, Raimo
Hiltunen (2004) Life Sciences 75, 1775-1786
13. Yong Zhong Houa, Guang Rong Zhaoa, Ying Jin Yuana, Guo Guang Zhua, Raimo Hiltunen
(2005) Journal of Ethnopharmacology, 100, 140-144.

NOVEL METHOD FOR ASSIGNMENT OF ABSOLUTE CONFIGURATION BASED
ON AN INDUCED AXIAL CHIRALITY AND ITS APPLICATION TO NATURAL
PRODUCTS
PL-20
S. Hosoi
Department of Pharmacognosy and Chemistry of Natural Products, School of Pharmaceutical
Sciences, Kyushu University of Health and Welfare, 1714-1 Yoshino-cho, Nobeoka 882-8508,
Japan
Spectroscopic methods for elucidation of absolute stereostructure, especially using NMR and
CD, have undergone great advancement in recent years. Among them, the exciton-coupled CD
(ECCD) method has been extensively applied to various organic molecules as a microscale
method to establish their absolute configurations in a nonempirical manner [1], Although its
application was initially limited to compounds having two or more functional groups, it has been
extended to compounds with a single functional group. In the course of our stereochemical
studies of chiral monofunctional molecules by the ECCD method, we expected that axial
chirality would be induced if a chiral alcohol was covalently attached to an achiral biaryl
chromophore. Here we present a novel strategy for determining the absolute configuration of
chiral alcohols with a unique chromophoric reagent 1, 3-cyanocarbonyl-3 '-methoxycarbonyl-
2,2 '-binaphthalene.
Induced CD
determination of absolute
configuration
Condensation of chiral alcohols with 1 gave the corresponding esters in good yields. The CD
spectra of the derivatives exhibited bisignate CD curves, indicating that the alcohol's chiral
information was definitely transmitted to the binaphthyl chromophore and induced axial
chirality. According to the analysis of the relationship between exciton chirality and absolute
configuration, the alcohols employed were found to be classified into two groups, A and B,
depending strongly upon the structure on the vicinal carbons. Group A is made up of compounds
lacking an unsaturation or an oxygen atom on the vicinal carbons, whereas group B is made up
of compounds possessing either an unsaturation or an oxygen atom at the vicinal position. In
group A, the exciton chirality of (7)-alcohols was negative and that of (S)-alcohols was positive.
On the contrary, the signs of exciton chirality were opposite in group B.
Next:, molecular mechanics calculations were performed using CONFLEX-MM2 to assess the
most stable conformation of the derivatives. The optimized conformers were enantiomeric in
each antipodal pair. In the most stable conformers, the calculated screw senses between the two
longitudinal 'B b electric transition moments of 2-naphthoate groups were in good accordance
with those expected from their exciton chiralities. That is, in both groups (A and B), the esters
showing positive exciton chirality had a clockwise turn, whereas the turns of those with negative
exciton chirality were counterclockwise.

Inspection of the molecular models of the most stable conformers revealed that, irrespective of
the groups (A or B) and of notation of the absolute configuration of the carbinol carbon (R or S),
the bulkier substituent occupied such a position as to avoid steric repulsion with the methyl ester
group in all derivatives. This suggested that the transmission of the chirality occurred in a
thermodynamically controlled fashion. Thus, taking a combination of the relative bulkiness of
the substituents on the carbinol carbon and the priority of these substituents in the sequence rule
into account, the relationship between the sign of exciton chirality of the binaphthyl derivative
and the absolute configuration of the original alcohol can be summarized as shown below. When
the sign of exciton chirality is positive and the orders of the substituents in bulkiness and in the
sequence rule are the same, the configuration of the alcohol is assigned to be S, whereas when
the sign is positive and the orders are opposite, the configuration is assigned to be R. In the case
that the sign is negative, the relationship between the absolute configuration and the combination
of the orders is the reverse. On the basis of the above results, we proposed a new rule to
determine the absolute configuration of chiral mono-alcohols [2]. We mentioned above a close
relationship between the sign of exciton chirality and the screw sense the two longitudinal *Bb
Sign of exciton (+)
chirality .
OR
opposite
(R) (S) (S)
O : Actual bulkiness Q : Sequence rule L: Large; S: Small
electric transition moments of 2-naphthoate groups in the most stable conformer of the
binaphthyl derivative calculated by molecular mechanics: the esters showing positive exciton
chirality had a clockwise (C) turn, whereas those with negative exciton chirality showed a
counterclockwise (CC) turn. In 2002, Kobayashi and co-workers used this relationship to
determine the absolute configuration of serratezomine A [3], whose secondary alcohol at C8 was
converted to our binaphthyl ester 2. The ester showed a negative exciton chirality [4]: a split CD
curve having a negative Cotton effect at 256 nm and a positive one at 234 nm, indicating the
screw sense of the two naphthyl groups to be a counterclockwise turn. As this result was in
agreement with the more stable conformer predicted by calculations [4], they concluded the
absolute configuration at the C8 position to be S; this conclusion was also supported by the
modified Mosher's method [5]. Apparently, the relative bulkiness of the two substituents of a
secondary alcohol is reflected in the calculation, and the predicted screw sense of the most stable
conformer is expected to the same as that predicted from the observed exciton chirality. Thus,
without consideration of the relative bulkiness of the substituents, which we discussed above, the
absolute configuration of a secondary alcohol can be determined as follows: (1) convert a
secondary alcohol in question to the corresponding binaphthyl ester and measure the CD
spectrum; (2) calculate the most stable conformer of the binapthyl ester derived from either
enantiomer of the secondary alcohol using CONFLEX; (3) compare the sign of the exciton
chirality and the predicted screw sense: for example, if the screw sense obtained from a
calculation based on the i-isomer is clockwise and the observed sign of exciton chirality of the
ester is positive, the absolute configuration of the alcohol can be assigned as R. However, since
the observed CD spectrum is the sum of the contributions from all conformers in the solution,
not only the energetically most stable conformer but also the global population ratio of stable
conformers with clockwise and counterclockwise turns of the derivatives should be considered.

In the above case, further molecular mechanics calculations for the SS-isomer of 2 indicated that
the conformers with a counterclockwise turn, that is, with the same screw sense as that of the
most stable conformer, predominated (C/CC=45/55). The absolute configuration at C8 could be
therefore be safely assigned as S. The practical usefulness of the present technique was
demonstrated by its application to several biologically active natural products [6].
Absolute configuration %
used for calculation
i H I X=H: serratezomine A
Calculated screw C
sense
Observed exciton + +
chirality
Actual absolute R S S
configuration
1. (a) Harada, N.; Nakanishi, K. Circular dichroic Spectroscopy—Exciton Coupling in
Organic Stereochemistry, University Science Books: Mill Valley, CA, 1983. (b)
Nakanishi, K.; Berova, N. In Circular Dichroism—Principles and Applications', Berova,
N., Nakanishi, K„ Woody, R. W., Eds.; Wiley-VCH: New York. 2000; pp 337-382.
2. Hosoi, S.; Kamiya, M.; Ohta, T. Org. Lett. 2001, 3. 3659-3662.
3. Morita, H.; Arisaka, M.; Yoshida, N.; Kobayashi, J. J. Org. Chem. 2000, 65, 6241—
6245.
4. Morita, H.; Kobayashi, J. J. Org. Chem. 2002, 67, 5378-5381.
5. Ohtani, I.; Kusumi, T.; Kashman, Y.; Kakisawa, H. J. Am. Chem. Soc. 1991,113, 4092-
4096.
6. Hosoi, S.; Serata, J.; Kiuchi, F.; Sakushima, A.; Ohta, T. J. Nat. Prod. 2004, 67, 1568-
1570.

EDUCATING THE PHARMACIST AS A MEMBER OF THE HEALTHCARE TEAM
PL-1
S.A. Hudson
Professor of Pharmaceutical Care, Department of Pharmaceutical Sciences, School of Pharmacy,
University of Strathclyde, Glasgow, Scotland UK
The concept of a healthcare team is the premise on which quality in the delivery of health care is
based. The integration of one profession's contribution to overall patient outcomes follows on
from when a health profession has established its own place for its services to patients; its
standards of practice are well enough defined for the profession to feel unthreatened as part of a
larger system of care with other health professions. As care becomes increasingly sophisticated
in terms of the application of technology or in terms of the social context of its delivery there is
increasing attention to the success of teamwork. 'Integrated care' is a term that has arisen from
recognition of the increased complexity of multi-agency contributions to outcomes, in terms not
only of the clinical success but also the social impact, on quality of life and the patient's rights to
autonomy. 'Integrated care' in its restricted application to health care is based on the premise
that "A single service cannot provide the care required". The emphasis on quality in health care
delivery has prompted the need to view the teamwork in terms of'the patient's journey' through
the system and through time as a chronic disease progresses.
The patient's journey
The success of the patients' journey across clinical settings and over time requires attention to
achieving good continuity of care, agreement on standards of practice (national guidelines, local
protocols), interdisciplinary and intra-disciplinary co-operation, referral mechanisms, consistent
delivery of health messages, management of treatment with maximum convenience to the
patient, concordance with the patient, transfer and monitoring of care through documentation
and follow up of the consequences of changes in treatment. The direction of health service
developments is therefore one of increasing co-operation between professions, sharing of skills
and overlapping of responsibilities. This direction is entirely compatible with the development of
(a) pharmaceutical care as the provision of care to develop a quality system of medicines usage
and (b) clinical pharmacy as a patient-centred application of pharmacists' knowledge and skills.
Pharmaceutical care has become relevant to healthcare professionals other than pharmacists, as
medicines use has become increasingly a team responsibility. Prescribing a medicine initiates a
complex series of consequences with the responsibilities for successful outcomes being divided
and transmitted to various members of the health care team and to patients themselves. While
pharmaceutical care is a way of including pharmacists more directly in therapeutic decisionmaking
it also offers prescribes, as initiators of treatments, the means to mobilise a wider team
in the assessment of drug effectiveness, in patient education and in monitoring for unwanted
effects. The response of the healthcare team to more complex treatments is a wider range of
contributions into decision-making and that teamwork potentially offers better monitoring and
evaluation of treatment decisions.
Services providing pharmaceutical care in different European countries are developing rapidly
but variously, according to local opportunities. Published descriptions and evaluations of
services remain scant and are inadequate in providing clear descriptions of the educational needs
of pharmacists who are integrating their services within the healthcare team approach. However
it is clear that the level of maturity of professional services demanded by integrated care
throughout the patient's journey requires initiatives in the educational formation of the
pharmacist - what we may call 'the pharmacist's journey'
38

The pharmacist's journey
The pharmacist's journey requires the acquisition of the attitudes, knowledge and skills
necessary for engagement with patients, teamwork, confidence to achieve change,
documentation, assumption of responsibility and the sharing of knowledge. The building of the
fully integrated professional contributor to the multidisciplinary team requires the appropriate
attitudes to be developed at the earliest opportunity in the pre-graduate education. The patientcentred
attitude must also include the early attainment of effective communication skills.
Interaction with other healthcare team members should ideally occur before graduation but often
relies on the postgraduate practical period to provide the necessary block period of contact in the
patient-care setting - the clinical attachment to a pharmacist role model working inside a health
care team.
Educational initiatives that help to form the pharmacist as an effective health care team member
At Strathclyde, we have developed a scheme with health service colleagues which recognises
(accredits) clinical pharmacists who provide us with a network of 70 accredited clinical
pharmacy supervisors - each having demonstrated that they are working inside multidisciplinary
teams. Exposure of the student to pharmacist role models (teacher-practitioners and practitionerresearchers)
is not only important but essential to the 'cloning' of patterns of effective
professional behaviour (examples of best practice). The postgraduate programmes at Masters,
DPharm and PhD levels require good clinical settings to provide research and development
opportunities. Participation in practice research requires data collection in the patient care areas,
which is dependent on effective co-operation in the clinical setting. Prescribing courses have
been introduced by Schools of Pharmacy to train pharmacists in new prescribing roles that have
become legally recognized in the UK since 2003. The extension of practice into 'supplementary
prescribing' has created a structured, documented system for pharmacist-doctor-nurse
collaboration that can now be taught. Prescribing is also being integrated into the undergraduate
pharmacy degree programme. Full integration within the health care team is demonstrated by
the maintenance of successful professional interactions. In the postgraduate period, the
pharmacist now builds on the attitudes and skills put in place early in the pharmacist's journey,
at the start of the university pharmacy curriculum. The continued professional development
(CPD) of the practising pharmacist is now about the introduction of new services and not just the
traditional topping up of knowledge. This part of the pharmacist's journey is also dependent
upon exposure to pharmacy role models and, increasingly, CPD draws on learning alongside
those from other disciplines.
Project activity is important because research is at the leading edge of the pharmacist's journey.
In the pre-graduate or postgraduate period, practical projects linked to progressive pharmacy
services are an effective way of integrating students within the multidisciplinary team. Project
topics should be carefully chosen so that they are designed to give the student the opportunity to
interact with other team members
'Health services research' is the generic (multidisciplinary) term for research into patients'
needs and patient care. The pharmacy profession has an essential goal to advance health services
research in pharmaceutical care and the use of medicines. The integration of academic and
practitioner skills requires health services research specialists to work closely with a cohort of
collaborating 'practitioner-researchers', at the leading edge of the pharmacist's journey. The
ability of the professional pharmacist to engage with research is important in the development of
the capacity to deliver new patient services.

PL-
CLINICAL PHARMACIST IN CHRONIC DISEASE MANAGEMENT
F. V. izzettin
Clinical Pharmacy Division, Department of Pharmacology, Faculty of Pharmacy, University of
Marmara, 81010, Haydarpa§a, Istanbul, Turkey
Since 1960's pharmacy practice has been improving from a solely product-oriented to a more
patient-oriented practice. In this context pharmacists not only involve in the process of drug
dispensing, but also in the clinical use of drugs; thus, play an important role in rationalizing drug
therapy through many services provided to patients and other members of the health-care team.
The clinical pharmacy services include general functions (e.g. medication history taking, patient
counseling, drug monitoring etc.), specific functions (e.g. detecting and preventing of side
effects, drug interactions etc.) and specialty functions (e.g. pediatric pharmacy, drug information
services etc.). After the implementation of these services in different areas of patient care and in
parallel with the increased knowledge and skills, the term "pharmaceutical care" was introduced
by Hepler and Strand as: "the responsible provision of drug therapy for the purpose of achieving
definite outcomes that improve a patient's quality of life" (1). These outcomes can be the cure of
a disease, elimination or reduction of patient's symptomatology, arresting or slowing of a disease
process, or preventing a disease or symptomatology.
The clinical pharmacist provides pharmaceutical care through the following clinical functions:
• Collection, documentation and interpretation of patient information.
• Identification of patient's drug-related problems (DRP) and establishment of a desired
therapeutic outcome for each DRP.
• Listing the therapeutic alternatives and selection of the best therapeutic plan and
individualization of the therapeutic regimen.
• Designing and implementation of drug monitoring plan and following-up the results.
Major elements for the provision of quality pharmaceutical care are: knowledge, skill, and
function of the staff; systems for data collection and documentation; efficient work flow
processes; references, resources and equipment; communication skills; and quality
assessment/improvement programs. In the literature several studies supply evidence that the
clinical pharmacist, through pharmaceutical care services play an imperial role in drug therapy
management, improve therapeutic outcomes and reduce the health-care costs. Between 1998 and
2002 over the 5 year period, pharmaceutical expenditures rose by 106% in USA (2). Studies
yield that for any dollar spent on pharmaceuticals, another dollar of spending results from "drug
misadventures" (3). The positive impact of pharmacists' improving medication adherence of
patients and prescription behaviors of physicians has been proved by several studies (4, 5).
Better health outcomes result in reduction of drug-induced adverse events and related hospital
emergency room visits and length of hospital stay (2). Patients with chronic disease are often on
multidrug therapy (polypharmacy) which can lead to a decrease in medication adherence and
increase in morbidity, adverse drug events, hospital admissions, deaths and health care costs (2).
As it is estimated that over 50% of patients don't take their medications properly (6), provision
of pharmaceutical care to the patients with chronic disease becomes essential. Pharmacists
working in different settings, including community pharmacies have a positive impact on health
care outcomes especially in chronic diseases by monitoring treatments plans, providing patient
education and promoting cost-effective therapy (7, 8).
40

In order for the pharmacists to take these responsibilities; some changes towards improvement of
patient-oriented services would be necessary in areas such as: regulations, "reimbursement
structure, education, practice, and patient expectations, inter-professional relationships with
other members of the health-care team, responsibilities/initiatives of professional associations,
physical structure of the pharmacies and the attitude of pharmacists (9, 10, 11). Depending on
the literature and our researches on role of pharmacist in chronic disease management we can
conclude that pharmacists as drug-specialists can improve health outcomes in chronic diseases
and clinical pharmacy interventions have positive input on patient behaviors, leading to a
reduction in health-related expenses and improvement in patient outcomes. In this lecture, the
principles of patient education and drug monitoring which are the key elements of
pharmaceutical care will be discussed and examples from our researches and others in the
literature (12) on the impact of pharmaceutical care provision to patients with chronic diseases
such as diabetes mellitus, hypertension, tuberculosis, hyperlipidemia will be presented.
1. Hepler CD, Strand LM. Opportunities and responsibilities in pharmaceutical care. American
Journal of Hospital Pharmacy 1990; 47:533-42.
2. DaVanzo J, Dobson A, Koeing L, Book R. Medication therapy management services: a
critical review. J Am Pharm Assoc 2005; 45(5):580-7.
3. Manasse HR. Medication use in an imperfect world: drug misadventuring as an issue of
public policy. Part 1. Am J Hosp Pharm 1989; 46:929-44.
4. Lipton HL, Byrns PJ, Soumerai SB et al. Pharmacists as agents of change for rational drug
therapy. Int J Technol Assess Health Care 1995; 11:485-508.
5. Indritz ME, Artz MB. Value added to health by pharmacists. Soc Sci Med 1999; 48:647-60.
6. www.accp.com/position/paperl l.pdf. Accessed March 24, 2006.
7. Yanchick JK. Implementation of a drug therapy monitoring clinic in a primary care setting.
Am J Health-Sys Pharm 2000; 57(4):S30-S34.
8. Wagner EH. The role of patient care teams in chronic disease management. BMJ 2000;
320:569-72.
9. Desselle S, Rappaport HM. The identification of pharmaceutical care practice standards in
the community pharmacy setting. Journal of Phannaceutical Care 1997; 1.
10. Tanna N. Progress made towards implementing pharmaceutical care. The Pharmaceutical
Journal 2002; 269:166-9.
11. Simpson D. Pharmaceutical care: The Minnesota model. The Pharm J 1997; 258:899-904.
12. Doucette WR, McDonough RP, Klepser D, McCarthy R. Comprehensive medication therapy
management: identifying and resolving drug related issues in a community pharmacy. Clin
Ther 2005; 27(7): 1104-11.

PL-3
EXISTENCE OF DOSE RESPONSE THRESHOLDS FOR DNA DAMAGE INDUCTION
BY ALKYLATING AGENTS
G. J.S. Jenkins
Swansea School of Medicine, Swansea University, Singleton Park Swansea SA28PP, UK.
During the safety evaluation of chemical compounds, the detection and quantitation of toxic
effects are critical for the setting up of exposure standards. When toxic effects are detected, the
exposure concentrations at which they occur and the types of dose response relationships
observed will represent important components in the regulation process. However, regulatory
action will be influenced by the type of endpoint showing a response and whether there are
concentrations below which a toxic response may not be observed, i.e. whether the toxic
response is thresholded. The main stance of the regulatory authorities over the past 3 decades has
been that genotoxins (DNA damaging agents) exhibit a linear relationship between exposure
dose and mutagenic response. This has been interpreted such that it can be assumed that there are
no safe levels of DNA damage, however low their abundance within the DNA. It appears
obvious to many people that thresholds (or NOEL's) do exist and that organisms can tolerate low
levels of DNA damage, just as they exist in an environment inducing low levels of endogenously
formed DNA damage. Although accepted on a theoretical basis, there has been little
experimental evidence to date, demonstrating thresholds for genotoxins. In our research group,
we are concerned with establishing if genotoxic thresholds can be induced by exposure of cells
to exogenous chemical agents (particularly alkylating agents). The demonstration and acceptance
(by the scientific and regulatory community) of the concept of a threshold of genotoxic activity
can have important economic implications for the use of individual compounds and allow the
setting of safe exposure levels. The clear unambiguous demonstration of a threshold of
genotoxic activity indicates that a compound will not produce mutations (or chromosomal
effects) below a critical exposure level thus reducing the potential for the induction of cancer or
congenital abnormalities at these exposures. Obviously, the biological basis for the potential
threshold is extremely important in understanding the mechanisms involved and for accepting
the plausibility that a threshold exists. In our research group we are particularly interested in the
mechanisms responsible for thresholded effects of alkylating agents in particular. We have
shown that the N7G inducing alkylating agents EMS and MMS have non-linear (thresholded)
dose responses for mutation induction (using the hprt mutation assay) and for chromosome
abnormalities (using the micronucleus assay). However, the related 0 6 G inducing alkylators
ENU and MMU do not show such non-linear responses and have linear dose responses. This
data indicates that the N7G adduct is repaired in a manner which leads to a thresholded dose
response, i.e. the repair appears to be saturated at higher doses. Therefore low dose exposures to
N7G inducing alkylators appear to be safe, whereas low dose exposures to 0 6 G alkylators may
not be safe, as they show linear responses. Importantly, it appears that DNA repair contributes to
the process of linear v non-linear dose responses. The exact repair pathway responsible for the
non-linear dose response of the N7G alkylators is not known as yet, but experiments with repair
deficient cell lines should answer this question. Once the particular DNA repair pathway is
identified, it will be possible to monitor the repair status (e.g. polymorphisms of key repair
genes) of any exposed populations (e.g industrial exposures) to ensure that the workers exposed
to these alkylators have adequate repair responses.
42

PL-
PARAOXONASE; FROM TOXICOLOGICAL FINDINGS TO CLINICAL
IMPORTANCE
A. Karakaya
Department of Toxicology, Faculty of Pharmacy, Ankara University, Ankara, Turkey
Since the discovery of human serum paraoxonase (PON1), the enzyme has been the subject of
various field of research. In the past, PON1 was identified as an enzyme which catalyze the
hydrolysis of esters such as organophosphate (OP) compounds and also arylesters. This enzyme
is one of the best-characterized enzyme or gene products that toxicologist think that it plays an
important role in toxic response to the effects of environmental chemicals and pharmaceuticals.
Although the enzyme PON1 and its gene have been named because of its ability to catalyze the
hydrolysis of paraoxon and other organophosphate compounds, in recent years, there is a
growing evidence that this enzyme and the other related family members participate in several
other important catalytic functions as well. Indeed, PON1 has been recognized as an antioxidant
enzyme, which plays an important role in lipid metabolism preventing the accumulation of the
lipid peroxides. Flere, we will briefly review developments in the field of PON1 research and
discuss the role of this organophosphate-hydrolizing enzyme in cardiovascular medicine.
Definition of the enzyme and the protective effect against OP's
The Paraoxonase (PON1) is an enzyme including in the group of "A-esterases (Arylesterases)"
which catalyzes the hydrolysis of esters such as organophosphate compounds. The enzyme
PON1 and its gene have been named because of its ability to catalyze the hydrolysis of
paraoxon, which is the metabolite of paration. This enzyme has protective role by hydrolyzing
and detoxifying several organophosphate insecticides, warfare agents such as sarin and tabun
and several carbamates and aromatic carboxylic acid esters which are very important
environment pollutants.
The paraoxonase phenotype polymorphisms and historical considerations
Mazur (1946) is the first investigator to report that organophosphorus compounds could be
hydrolyzed enzymatically by several animal tissues (1). Aldridge (1953) described the general
characteristics of the serum paraoxonases of several mammalian species and he proposed the
definition of "A-esterases" for the enzymes hydrolyzing organophosphates and arylesters (2).
The relationship between arylesterase activity and paraoxonase activity remained a rather
complex problem for many years.
It was thought that hydrolysis of phenyl acetate (arylesterase activity) was catalyzed by a
different human serum protein than that catalyzing the hydrolysis of paraoxon (paraoxonase
activity). Because, the frequency distribution histograms showed arylesterase activities to be a
unimodal distribution, whereas the paraoxonase activities followed a bimodal distribution.
Eckerson et al. (1983) showed that the ratio of paraoxonase activity in the presence of 1 M NaCl
divided by the arylesterase activity gave trimodal distributions (3). Finally it was possible to
prove by expression of human PON1 that one protein was responsible for both esterase activity
(4). There are variations in the frequencies reported for several populations. The gene frequency
for the B allele in our population was found as 0.368 (5). It was suggested that individuals with
low serum PON1 activity are more susceptible to poisoning by toxic OP oxons than individuals
with higher serum enzyme levels (6).
43

The PON1 gene family and polymorphism
Paraoxonase or PON family of genes are located on the long arm human chromosome 7 in the
order PON1, PON3 and PON2. All three PON genes are next to one another. PON1 and PON3
are mainly expressed in the liver and excreted in the blood where they are associated with the
cholesterol-carrying particles high-density lipoprotein (HDL) (the "good cholesterol"). PON2 is
not present in blood, but is expressed widely in a number of tissues, including the liver, lung,
brain and heart. Of the paraoxonase family, PON1 is the most investigated and best understood
member. PON1 was one of the early genes identified as an environmentally relevant gene, in that
it is important in determining an induvidual's sensitivity or resistance to exposure from specific
organophosphorus (OP) insecticides. The coding region of human PON1 gene contains two
polymorphic sites: one at position 192 [glutamine (Q) to arginine (R) substitution] and a second
at position 55 [leucine (L) to methionine (M) substitution]. The polymorphisms affect the
hydrolytic activity of the PON1 isoenzymes with respect to certain substrates, such as paraoxon
and lipid peroxides (6). Q192R polymorphism is responsible for a striking substrate specific
difference in the hydrolytic activity of the enzyme (7). The L55M polymorphism is located in
the N-terminal side of PON1, which plays a role in the binding of PON1 to HDL and may thus
alters the ability of PON1 to form a complex with HDL (8). The L55M polymorphism has not
been found to affect catalytic efficiency but to affect the PON1 enzyme concentration in plasma
(9). It has been shown that genotyping analysis of these two polymorphism is potentially
important in respect of definition of population who has risk of poisoning with the
organophosphate compounds such as agriculture workers and warfare agents exposed
individuals (e.g. the effect of the PON1 polymorphisms on activity may explain why some Gulf
War Veterans have developed Gulf War syndrome and some have not, despite similar exposure.
In the Veterans, paraoxon hydrolysis was less than 50% of that found in the controls) (10).
The protective role of PON 1 in LDL oxidation
Recent progress in paraoxonase research for clarifying its physiologic role has shown that this
enzyme has a very important physiologic role other than the protective role against chronic OP
poisoning. Much of these enzymes' activity is associated with HDL and PON1 may play a role
in lipid metabolism preventing the accumulation of the lipid peroxides. Although the physiologic
substrate of PON1 is unknown, a protective role against the oxidative degradation of serum
lipoproteins has been attributed to this enzyme. Indeed, in vivo and in vitro biochemical studies
of this enzyme have shown that human serum PON1 is located on HDL and seems to protect
low-density lipoprotein (LDL) oxidative stress by hydrolyzing lipid peroxides (11).
PON1 and its relationship to diseases
Atherosclerosis and coronary heart diseases (CHD):
Biochemical studies of this enzyme have shown that serum paraoxonase is located on highdensity
lipoprotein (HDL) and although its natural substrates are unknown, it is proposed to have
a responsibility for the antioxidative activity of HDL. In vitro studies indicate that PON1 can
significantly lower lipid peroxide generation during LDL oxidation and therefore, may provide
HDL-associated protection against atherosclerotic process. In humans, serum PON1 activity and
serum HDL susceptibility to oxidation or atherosclerosis are inversely related. Some studies also
suggest that a relationship exist between PON1 polymorphism and CHD, whereas other studies,
in different populations, have not found such an association (12-14).
Diabetes Mellitus (DM): Low serum paraoxonase activity has been thought as a typical feature
of type I and type II diabetes implicating the enzyme in the etiology of the disease and its
complications. On the other hand, it was suggested that PON1 and PON2 polymorphisms have
been associated with the presence of several diabetic complications (15,16).
Disorders in lipid metabolism (Familial hypercholesterolaemia (FH), Tangier disease, HDL
deficiency)
Cerebrovascular disease and neurological disease (Alzheimer's disease)
Gulf War syndrome (Neurological symptoms in veterans of the Persian Gulf War)

The importance of paraoxonase as an antioxidant enzyme
Studies in PON 1-knockout mice and in PONl-overexpressing mice, as well as in vitro studies
demonstrated that PON1 is associated with reduced oxidative stress. PON1 can hydrolyze
specific oxidized lipids in lipoproteins, in macrophages, and in atherosclerotic lesions. Human
and rabbit PON3 are also HDL-associated and they protect LDL against oxidation. PON2
overexpression in cells was shown to lower intracellular oxidative states and to reduce the cells
ability to oxidize LDL. In patients with hypercholesterolemia, statins therapy (may act as
antioxidants) was also demonstrated to be associated with increased serum PON1 activity (17).
The role of PON1 in drug metabolism
PON1 may be involved in the metabolism of pharmaceutical drugs; it was found that the diuretic
spironolactone and the hypocholesterolemic drugs mevastatin, lovastatin and simvastatin are
hydrolyzed by PON1. Glucocorticoid g-lactones and the antibacterial pro-drug prulifloxacin can
be bioactivated in vivo by PON1 (18-20).
The last and future considerations in paraoxonase research
1. The investigations of novel PON1 substrates such as lactones and cyclic carbonate esters, (e.g.
more recently PON1 has been suggested to protect against homocysteinylation by hydrolyzing
homocysteine thiolactone)
2. The role of PON1 in the metabolism of drugs and other different xenobiotics and its
pharmacogenomic and toxicogenomic aspects
3. The consequences of its polymorphisms with respect to susceptibility to disease development
or toxic injury
4. The other PONs (PON2 ve PON3)
5. PON1 as a possible therapeutic and prophylactic agent
(Recombinant "native" PON1 may be useful in reducing the risk of OP poisoning and the risk of
cardiovascular disease in individuals that have very low PON1 activity)
1. Mazur A (1946) J Biol Chem 164:271-289
2. Aldridge WN (1953) Biochem J 53:110-117
3. Eckerson HW et al. (1983) Am J Hum Genet. 35:1126-1138
4. Sorenson RC et al. (1995) Proc Natl Acad Sci USA 92:7187-7191
5. Karakaya A et al (1991) Pharmacogenetics 1:58-61
6. Costa and Furlong (2002) Paraoxonase (PON1) in Health and Disease: Basic and
Clinical Aspect, Kluwer Academic Publishers
7. Davies HG et al (1996) Nat Genet 14:334-336
8. Furlong CE et al (1991) Biochem 30:10133-10140
9. Brophy VH et al (2001) Am J Hum Genet 68:1428-1436
10. Mackness B et al (2000) Biochem Biophy Res Comm 276:729-733
11. Aviram M (1999) Mol Med Tod 5:381-386
12. Imai Y et al (2000) Atherosclerosis 149:435-442
13. Mackness MI et al (2002) Atheroscler Suppl 3:49-55
14. Karakaya A et al. (1999) Chem Bio Inter 118:193-200
15. Mackness B et al (2000) Clin Sci 98:355-363
16. Mackness B et al (2002) Eur J Clin Invest 32:259-264
17. Mira R et al (2004) Lett Drug Des & Disc 1:269-274
18. Billecke S et al (2000) Drug Metab Dispos 28:1335-1342
19. Biggadike K et al (2000) J Med Chem 43:19-21
20. Tougou K et al (1998) Drug Metab Dispos 26:355-359

PL-
HYPHENATED ELECTRO ANALYTICAL METHODS AS «IN VITRO » TOOLS FOR
PREDICTIVE DRUG METABOLISATION STUDIES
J-M Kauffmann. B.Blankert
Universite Libre de Bruxelles, Institut de Pharmacie, Campus Plaine CP 205/6, 1050 Bruxelles,
Belgium
In the field of drug discovery it is of critical importance to reduce as early as possible the risks of
attrition rates in later stages of drug development. Phase I drug metabolism frequently involves
oxidative processes. Drug metabolism and toxicity are very commonly linked to redox
biochemical processes including phase I oxidative biotransformation and oxidative stress that is
secondary to metabolic activation. The on-line combination of electrochemical cells with mass
spectrometry (EC-MS) or electrochemical cells with liquid chromatography coupled to mass
spectrometry (EC-LC-MS) are powerful instrumental approaches to study both quantitatively
and qualitatively the redox pattern of drug candidates. By studying model compounds
(promethazine, promazine, clozapine) it was possible to point out that electrochemical oxidation
mimics (several but not all) biological phase I redox reactions (e.g., N- dealkylation, N-, S-, C-
oxidations). Compound oxidation was studied across a range of fixed potentials (vs. Pd) and
product formation was studied by examining potential-dependent mass shifts. Of particular
interest is the fact that glutathione (GSH) is not electroactive at the studied graphite electrode
allowing its co-injection in the experimental set up and the identification of GSH conjugated
species (related to phase II metabolites) [1,2]. The coupling of metalloenzymes to amperometric
electrodes is another attractive approach for studying drug metabolisation and identifying
reaction inhibitors or activators. The enzyme horseradish peroxidase is often used as a model
system mimicking enzymatic biotransformations. The enzyme has been immobilised onto solid
carbon electrodes for the oxidation of selected drug compounds in the presence of hydrogen
peroxide. Such a biosensor allows the quantitative assay of the studied compound but it permits
also the screening of thiol species which react with the oxidised product generated at the
biosensor-solution interface.
The present communication is intended to highlight the interest in implementing in
pharmaceutical research such hybrid technologies for the early study of drug oxidation pattern
and the reactivity of the oxidized products towards endogenous thiol compounds [3].
1. Prediction of clozapine metabolism by on-line electrochemistry/liquid chromatography/
mass spectrometry, S. M. van Leeuwen, B. Blankert, J-M Kauffmann, U. Karst
Anal.Bioanal.Chem., 382(2005) 742-750.
2. Electrochemical, chemical and enzymatic oxidations of phenothiazine B.Blankert, H.
Hayen, S.M. van Leeuwen,U. Karst, E. Bodoki, S. Lotrean, R. Sandulescu, N. Mora
Diez, O. Dominguez, J. Arcos J.-M. Kauffmann. Electroanalysis, 17 (2005) 1501-1510.
3. Biosensors in Drug Discovery and Drug Analysis D.Yu, B.Blankert, J-C Vire, J-M
Kauffmann Anal.Letters, 38 (2005) 1687-1701.
46

PL-
COMBINATORIAL BIOSYNTHESIS OF PODOPHYLLOTOXIN
M.K. Julsing 1 , N.P. Vasilev 1 , A. Koulman 2 , W.J.Quax 1 , O. Kayser'
'Department of Pharmaceutical Biology, GUIDE, University of Groningen, the Netherlands,
2 AgResearch, Palmerston North, New Zealand
Podophyllotoxin (POD) is a synthetic precursor derived from Podophyllum hexandrum for the
semi synthesis of the antineoplastic drugs Etoposide and Tenoposide. The drug POD is obtained
by isolation from plant source that has already endangered status in parts of Asia. The synthesis
is too expensive, why an alternative route of production must be found. As an alternative the
conversion of the biosynthetic precursor deoxypodophyllotoxin (DOP) to POD with recombinant
human liver Cytochrome P450 enzymes was investigated. Main metabolic step was
hydroxylation of DOP in the position C7. E. coli DH5a was engineered to express Cytochrome
P450 3A4 and seven other enzymes with a NADPH Reductase for bioconversion. The
expression system was validated and optimised for high yield of protein expression: Expressed
proteins were characterised to determine activity, identity and purity. Metabolic conversion was
determined with whole cells expressing the enzyme and membrane fractions of homogenised E.
coli. Reference compounds and DOP were incubated for 2 h for bioconversion and the amount
of metabolised POD was determined by GC-MS, LC-MS and LC-NMR. From all tested
enzymes it turned out that only Cytochrome P450 3A4 was active to metabolise DOP to POD.
Besides POD other peaks were recorded and structure elucidation is still under investigation.
The rate of bioconversion to POD was 5-10% standing in line with published data for
Cytochrome P 450 3A4 (1). According to (2) low metabolisation rates are caused by mechanism
based inhibition of the active centre by POD. To overcome these problem kinetic studies on the
mechanism of inhibition were carried out.
1. Guengrich, FP (2001) in: Principles and Methods of Toxicology, (Hayes W, ed.) Taylor,
Francis, Philadelphia, p. 1625
2. Usia et al. (2005) Life Sciences 76, 2381
47

PL-
DESIGNING NEW THERAPIES FOR DIABETIC VASCULAR DISEASES FROM
CELL BASE ASSAYS
G.L. King
Harvard University, Medical School, Joslin Diabetes Center, Boston, USA.
The idea of protein kinase C (PKC) activation causing diabetic vascular complications was
initiated in my laboratory in 1988-1989 when we found that the exposure of retinal capillary
endothelial cells to pathophysiological levels of glucose in the media activated a cellular
signaling pathway called protein kinase C (PKC). Since PKC activation is known to regulate
many vascular functions, we postulated that hyperglycemia can activate PKC pathway to cause a
variety of vascular dysfunctions in the eye, kidney, and cardiovascular system. PKC is a
serine/threonine kinase that is activated by lipids such as diacylglycerol (DAG) and can also be
activated by calcium. Multiple hormones can activate PKC in a very rapid manner. These
hormones include angiotensin, platelet-derived growth factor (PDGF), histamine, endothelin
(ET), vascular endothelial growth factor (VEGF), and others. Essentially, any cytokine that can
increase diacylglycerol, or calcium influx can activate PKC intracellularly. In addition any
enzyme system or cytokine that can activate phospholipose C, which will increase DAG and
calcium intracellularly, will also activate PKC. Thus, PKC activation is common to multiple
cytokines and cellular actions. One mechanism by which cells are able to bring specificity to the
activation of the PKC is by having the twelve different PKC isoforms located in different sites in
the cell. Elevations of glucose and diabetes have been shown to activate several PKC isoforms
including cc, P'/ 2 , and 5. Since our report sixteen years ago that elevation of glucose levels in the
media of cultured retinal endothelial cells and vascular smooth muscle cells activate PKC
activities, there have been 700 reports confirming these findings in all vascular tissues of many
species of animal and diabetic patients. Various vascular pathologies have been shown to be due
to PKC activation. These activities include increases in capillary permeability, basement
membrane thickening, fibrosis, abnormality of blood flow, increase vascular contractility,
activation of monocytes, and other vascular disorders associated with diabetic micro-vascular
and cardiovascular complications. In an effort to design therapy based on normalizing PKC
activation on the vasculature induced by diabetes, we have focused on designing and testing an
isoform selective inhibitor of PKC P'/2. In 1996, Jirousek et al reported that fourteen membered
macrocycles containing bis indolylmaleimide or Ruboxistaurin (RBX) can inhibit PKC (3
isoform specifically. We reported that the inhibitory effects of this compound RBX (IC 50 = 5.5
nm) appears to be fifty times more potent for PKC (3 Vi isoforms than any other PKC isoforms
(IC 50 250-500nm) levels. The half-life of RBX in the plasma is relatively short at one to two
hours in human subjects. The major metabolite of RBX is desmethyl RBX, which has a much
longer high life and has a similar inhibitory pattern on PKC isoforms as the original compound.
These findings suggest that RBX maybe be useful for treatment of diabetic vascular
complications. Animal studies involving mice and rats have recently demonstrated that RBX is
able to normalize capillary permeability, blood flow, and decrease angiogenesis in the retina of
diabetic animals. In the kidney, the use of RBX can decrease mesangial expansion, expression
of TGF beta, CTGF, and renal function abnormalities such as hyper-filtration and proteinuria in
diabetic animals. Clinically, multiple Phases one, two, and now three studies have shown that 32
mg/d of RBX, is able to normalize blood flow, retinal capillary permeability, and preserve visual
acuity in patients with diabetic macular edema.
48

In the kidney diabetic patients who continue to have proteinuria in spite of optimal treatment by
angiotensin inhibitors were able to significantly reduce proteinuria and decrease the loss of
estimated GFR after one year of treatment with RBX. This is compared to diabetic patients
treated with angiotensin converting enzyme inhibitors or angiotensin receptor blockers. In Phase
three trials, RBX at 32 mg/d was able to preserve visual acuity in diabetic patients with macular
edema. Significant side effects have not been observed in 3000 patients who so far have taken
RBX in more than ten clinical trials for up to three years. Unlike diabetic retinopathy and
nephropathy, Phase two and Phase three clinical trials for diabetic neuropathy did not show any
positive effect. These data suggest that PKC beta isoform activation is an important mechanism
by which hyperglycemia and diabetes are causing diabetic retinopathy and nephropathy, unlike
diabetic neuropathy. Further clinical studies will be needed to determine PKC (3 isoform's role
in diabetic cardiovascular diseases.

STRATEGY IN THE ISOLATION, STRUCTURE ELUCIDATION AND BIOLOGICAL
ACTIVITIES OF SAPONINS AS POTENT ANTITUMOR, IMMUNOMODULATING
AND ANTIMICROBIAL AGENTS
M.-A. Lacaille-Dubois
Laboratoire de Pharmacognosie, Unite de Molecules d'Interet Biologique, UMIB, UPRES-EA
3660, Faculte de Pharmacie, Universite de Bourgogne, 7, Bd Jeanne d'Arc, 21079 Dijon Cedex,
France
Saponins are an important class of natural products which are structurally described as
glycosylated triterpenoids or steroids. They are widely spread throughout the plant or marine
animal kingdom. The principal uses of saponin drugs in therapy are related primarily to the
antiedematose, antiphlogistic, hepatoprotective, veinotonic, expectorant and CNS stimulating
activities. The scientific progress of chromatographic and spectroscopic techniques mainly 2D-
NMR and many new in vitro and in vivo test systems resulted in the characterization of a large
number of bioactive saponins including cytotoxic, antitumor, immunoadjuvant, antifungal,
antibacterial, antiviral, analgesic, antiinflammatory, hypocholesterolemic, antihepatotoxic and
hypoglycemic compounds. Our laboratory has focussed in recent years on the isolation, structure
elucidation and biological studies of new saponins as potent antitumor, immunomodulating and
antifungal agents mainly from the African plant biodiversity. This lecture will highlight some
relevant examples of our recent research on new bioactive saponins from Polygalaceae,
Mimosaceae, Caryophyllaceae, Apiaceae and Dioscoreaceae.'" 3 Our strategy combines isolation
procedures by successive chromatographic steps (flash chromatography, medium-pressure liquid
chromatography or MPLC, semi-preparative HPLC), structural elucidation by FAB/MS,
ESI/MS, and 2D NMR spectroscopy (COSY, TOCSY, NOESY, HSQC, HMBC) and in vitro
biological assessment. The bioassays included hemolysis on sheep erythrocytes, proliferation of
Jurkat cells and splenocytes, apoptosis induction on Jurkat cells, cytotoxicity on human HT-29
and HCT 116 colon cancer cells and antifungal activity against three human pathogenic yeasts of
Candida species by using the broth dilution test. The discussion will also focus on the
understanding of their mechanism of action and structure/activity relationships.
1. Mitaine-Offer A.-C., Miyamoto T., Jolly C., Delaude C., Lacaille-Dubois M.A. Helv. Chim.
Acta 2005, 88, 2986-2995.
2. Sautour M„ Miyamoto T., Lacaille-Dubois M.A. J. Nat. Prod. 2005, 68, 1489-1493.
3. Haddad M., Laurens V., Lacaille-Dubois M.A. Bioorg. Med. Chem. 2004, 12, 4725-4734.
PL-
50

PL-
BIOCHEMISTRY/MOLECULAR BIOLOGY OF FORMATION OF HEALTH-
PROTECTING MEDICINAL LIGNANS AND RELATED PHENOLICS
M. Jourdes, L. B. Davin, N.G. Lewis
Institute of Biological Chemistry, Washington State University, Pullman, WA 99164-6340, USA
The remarkable diversity and properties of plant phenolics encountered in nature has ultimately
led to their growing application in health-protection and disease treatment. Such applications
can range from antioxidant properties (e.g. chlorogenic acid), to the widespread treatment of
various viruses/cancers (e.g. podophyllotoxin and nor-dihydroguaiaretic acid derivatives), to
antibacterial and flavor agents (e.g. eugenol, chavicol), etc. These compounds, in turn, are either
products of the phenylpropanoid pathway leading to monolignols, or are derived as offshoots
from same. Our laboratory has as a central goal defining the biochemical steps in many of these
pathways, with some of the more applied reasons for this being: to identify biotechnological
means to increase their naturally occurring levels; to introduce their biochemical pathways into
normal dietary foodstuffs, or to identify improved cell culture conditions for their optimal
formation in bioreactors. In this contribution, we describe our progress in defining the identify
and mechanistic basis of various enzymatic steps in their pathways. This includes how arogenic
acid is formed in planta, and how this compound differs from bacterial systems, as well as in
mechanistically defining the enzymology associated with chlorogenic acid (e.g.p-coumaryl CoA
shikimate quinic acid transferase) and monolignol (e.g. cinnamyl alcohol dehydrogenase)
formation. Beyond the monolignol forming pathway, carbon can be specifically diverted into
various natural products, such as to the bioactive dihydromonolignols, allyl- and
propenylphenols, and lignans, depending on the species and tissues involved. We also describe
the progress made in understanding the mechanistic enzymology associated with the recently
discovered chavicol synthase, allylic double bond reductases, dirigent proteins, pinoresinollariciresinol
reductases, secoisolariciresinol dehydrogenase, enantiospecific polyphenol oxidases.
From a more practical perspective, the prospects for the beneficial utilization of these
biochemical processes are evaluated.
51

THE POWER OF LC-MS IN PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
H. Lingeman, H. Irth
Vrije Universiteit Amsterdam, Amsterdam, The Netherlands
PL-3
In bio- and pharmaceutical analysis sample preparation, separation and detection play an
important role. In order to optimize sensitivity, selectivity and throughput a number of
approaches can be used. The present lecture will focus on novel analytical methodologies that
allow the selective detection of drugs, peptides and proteins in complex biological matrices.
Mainly two different approaches will be described: (i) on-line immunoaffinity preconcentration
or receptor-based concentration and LC-MS(MS) and (ii) post-column ligand-exchange MS.
With respect to sample preparation it will be clear that SPE is the dominant approach in the
future. Developments with respect to new sorbent materials, new formats, new mixed mode
phases, and approaches to minimize ion suppression in MS will be highlighted, for both
pharmaceutical and bio-analytical applications. With respect to protein analysis, one of the
important features is digestion of the sample before a chromatographic separation can be
performed. In addition to the available time-consuming off-line procedures, nowadays, on-line
extraction procedures for digested proteins in a gel spot and fully on-line procedures for
denaturated proteins and biotoxins by on-line SEC - digestion - LC-MS are described.
Regarding the separation, the use of high temperature LC will significantly improve the
throughput in analytical systems. The potential of using this approach in an on-line combination
with an enzymatic screening system for bioactive compounds will enhance the selectivity, the
sensitivity as well as the throughput. In order to improve the selectivity during an analytical
separation an example will be described using the quantitative analysis of proteins in biological
fluids by on-line immunoaffinity chromatography - protein digestion - LC-MS. With respect to
detection the selectivity of the MS can be improved by using special reactions. In additional to
the biochemical-screening assays mentioned above, the application of ESI-MS for the selective
detection of metal ligands after a post-column continuous-flow ligand-exchange reaction is one
of the possibilities. Finally, the potential of hyphenated systems will be shown by introducing a
prototype of a comprehensive chromatography - MS system for metabolomics. A combined
hybrid comprehensive system based on LC x (LC-MS/MS / GC-MS/MS) will be described.
52

PL-31
BETA-BLOCKER EFFECTS ON DIABETES-INDUCED CARDIOMYOPATHY
J.H. McNeill, V. Sharma
Division of Pharmacology and Toxicology, Faculty of Pharmaceutical Sciences, 2146 East Mall,
The University of British Columbia, Vancouver, Canada, V6T 1Z3
Patients with diabetes develop heart failure even in the absence of valvular or ischemic heart
disease. In the 1970's, it was proposed that there was a diabetic cardiomyopathy, a disease of
heart muscle which occurs as a direct result of the diabetic state. Diabetic cardiomyopathy is
now recognized as a clinical entity whose prevalence in the diabetic population is alarmingly
high. Diabetic cardiomyopathy manifests clinically as a diastolic dysfunction which progresses
to systolic dysfunction. The compliance of the heart is decreased by increased collagen crosslinking,
subendocardial glycogen deposition and interstitial fibrosis. The contractility of the heart
is decreased by a number of factors including abnormal calcium handling, alterations in a wide
range of signaling pathways (Protein kinase C, protein kinase B, RhoA/ Rho kinase), widespread
changes in gene expression (target genes of PPAR and FOXO, fetal gene program induction),
oxidative stress, mitochondrial dysfunction and cell death. There are also widespread structural
abnormalities. P-blockers such as metoprolol improve cardiac function and reduce mortality in
patients with heart failure. The mechanism of this effect involves improvement in cardiac
hypertrophy, decreased apoptosis and necrosis, anti-arrhythmic effects and improvement in
cardiac substrate utilization. The normal heart derives 20-40% of its energy from pyruvate
oxidation and 60-80% from fatty acid oxidation. Increases in fatty acid oxidation feed back and
inhibit glucose oxidation; this is known as the Randle Cycle. The diabetic heart relies almost
exclusively on fatty acid oxidation as an energy source. This 'switch' in energy metabolism is
believed to be harmful for two reasons. Firstly, fatty acid oxidation requires more oxygen per
molecule of ATP generated and is therefore less efficient. Secondly, as the influx of fatty acids
into the cytoplasm overwhelms the capacity of the cardiomyocyte to metabolize them,
lipotoxicity occurs. Lipotoxicity is a major mechanism by which diabetic cardiomyopathy
occurs. It has been suggested that inhibition of fatty acid oxidation, with a subsequent
improvement in glucose oxidation may be a potential therapeutic strategy for heart failure. P-
blockers such as metoprolol and carvedilol have been shown to inhibit fatty acid oxidation and
stimulate glucose oxidation in patients with dilated cardiomyopathy. In microembolism-induced
heart failure in dogs, chronic treatment with metoprolol decreased the activity of carnitine
palmitoyltransferase 1 (CPT-1), the rate-limiting step of fatty acid oxidation. CPT-1 converts
long-chain acyl-coenzyme-A's into acyl-carnitines. These can then enter the mitochondria to be
reconverted to acyl-coenzyme A's and oxidized. CPT-1 is subject to inhibition by malonyl
coenzyme A (malonyl CoA). Malonyl CoA is synthesized from acetyl CoA by acetyl CoA
carboxylase (ACC). ACC is regulated by protein kinase A (PKA) and AMP-activated protein
kinase, which phosphorylate and inhibit ACC. The streptozotocin (STZ) diabetic rat is a model
of poorly controlled type 1 diabetes. This model develops a diabetic cardiomyopathy similar to
that seen clinically 6 weeks after induction of diabetes. We hypothesized that chronic treatment
with metoprolol would improve cardiac function, inhibit fatty acid oxidation and stimulate
glucose oxidation in the diabetic heart.
53

Rats were divided into 4 groups: control (C), control treated (CT), diabetic (D) and diabetic
treated (DT). The CT and DT groups were treated with 75mg/kg/day metoprolol (this is
equivalent to a human dose of lOOmg per day). Six weeks after the induction of diabetes, heart
function and metabolism was measured using the isolated perfused working heart. Hearts were
perfused with Krebs buffer containing 5.5mmol glucose, 0.8mmol palmitate and lOOpU insulin.
Chronic treatment with metoprolol improved cardiac function in the diabetic hearts. This was
associated with a decrease in fatty acid oxidation and an increase in glucose oxidation.
Surprisingly, in the control treated hearts, fatty acid oxidation was increased and glucose
oxidation, decreased. Glycolysis was reduced by 50% in the diabetic hearts. Chronic treatment
with metoprolol had no effect on glycolysis. However, the coupling of glycolysis to glucose
oxidation was improved. When perfusions were carried out in the absence of insulin, glucose
oxidation was negligible but the fatty acid oxidation pattern was preserved. Myocardial
energetics were not significantly altered by either diabetes or chronic metoprolol treatment.
Metoprolol, when added to the perfusate at a concentration of 2000ng/ml, inhibited fatty acid
oxidation and stimulated glucose oxidation. When these experiments were repeated in the
absence of insulin, glucose oxidation was negligible but the inhibition of fatty acid oxidation was
preserved. The total activity of CPT-1 was increased in the diabetic heart, and decreased by
chronic metoprolol treatment in both control and diabetic hearts. The sensitivity of CPT-1 to
malonyl CoA inhibition was increased in the diabetic heart and decreased by metoprolol
treatment. The effect of metoprolol on CPT-1 activity would be expected to decrease fatty acid
oxidation whilst the effect of metoprolol on CPT-1 sensitivity would be expected to increase
fatty acid oxidation. Results of malonyl CoA level measurements are awaited. AMPK activity
was not altered either by diabetes or by metoprolol treatment, although AMPK phosphorylation
was increased in the diabetic hearts. ACC phosphorylation was increased by metoprolol
treatment in both control and diabetic hearts. Increased ACC phosphorylation would be expected
to increase fatty acid oxidation. These results indicate that metoprolol is a direct acute inhibitor
of fatty acid oxidation. Chronic treatment with metoprolol improves heart function in diabetic
cardiomyopathy. This is associated with an inhibition of fatty acid oxidation and stimulation of
glucose oxidation in the diabetic hearts, but the opposite pattern in controls. Metoprolol
decreases total CPT-1 activity but also decreases CPT-1 sensitivity to malonyl CoA inhibition.
This partly explains the complexity of the observed pattern of metoprolol's effects. Future
studies will elucidate the mechanism of this effect, and compare the acute metabolic effects of
metoprolol with the chronic effects.
Supported by a grant in aid and a program grant from the Heart and Stroke Foundation of BC
and Yukon.VS is the recipient of a Graduate Student Scholarship from the CIHR/Rx&D
Foundations.

PL-32
NATURAL RESOURCES FOR PREVENTION AGAINST DIABETIC
COMPLICATIONS
Y. Okada
Department of Natural Medicine & Phytochemistry, Meiji Pharmaceutical University, 2-522-1
Noshio, Kiyose-shi, Tokyo 204-8588, Japan
Diabetes is a chronic and systemic disease that triggers life-changing complications in virtually
every system of the body. Several candidate mechanisms contributing to diabetic complications
have been proposed. These mechanisms accepted widely include the poliol pathway, glycation
and so on. Furthermore, the hyperaggregability of platelets, as well as their hypercoagulability
in diabetic patients have also been reported. With a view of developing new bioactive
components, herbal medicines and foodstuffs were examined for the activity of prevention of
diabetic complications. We examined a lot of materials to find out naturally occurring substances
for prevention against the complications of diabetes by using the screening tests mainly on
aldose reductase inhibitory activity 1 "" 3 ' and glycation inhibitory activity, and found some
materials are useful. I would like to introduce a part of results from our studies obtained so far,
mainly on the style of Zea mays L. (Corn Silk) and Turkish plant, Cistus laurifolius L.
Style of Zea mays L. (Corn Silk)
From the result of screening test, the style of Z. mays was elucidated for active components, and
the effectivity of water extract on diabetic nephropathy was investigated by using streptozotocin
(STZ) induced diabetic rats.Isolation of Compounds from Z. mays
The style of Zea mays L. was purchased from Mikuni & (Osaka, Japan), and extracted with
MeOH and water to the corresponding extract and then separated by various chromatography to
yield 20 compounds including new flavone C-glycosides, chrysoeriol 6-C-P-boivinopyranosyl-7-
O-p-glucopyranoside (1) 4) and chrysoeriol 6-C-P-fucopyranoside (2) 5) , having Advanced
Glycation End products (AGE) formation inhibitory activity and a new sesquiterpene. Their
structures were elucidated by means of chemical and spectral analysis.
Inhibition Test on CML Formation in vitro
Bovine serum albumin (BSA) was incubated with lOOmM glucose in the presence or absence of
test compound for 7 d in 0.1 M phosphate buffer (pH 7.4) at 37°C. After incubation, the level of
CML was measured by CML-specific ELISA.
5

Effect of Streptozotocin (STZ) Induced Diabetic Nephropathy
Extraction of Plant Material
The style of Z. mays was extracted with water twice at 80 °C for 2 h and then lyophilized to
give a water extract.
Experimental Animals
Male Wistar rats (7 weeks old) were used and carried out in accordance with the guidelines of
the Institutional Ethics Committee for Animal Research, Meiji Pharmaceutical University.
Experimental Procedure
Rats were housed for 7 d in an environmentally controlled facility with 12 h light-12 h dark
cycle and free access to general food and water before use. They were randomized into control
group (non-diabetic, NC), non-administered group (diabetic, DM-NT) and administered group
(diabetic, DM-T). Diabetes was evoked by intravenous injection of STZ (40 mg/kg, dissolved in
50 mM sodium citrate buffer). Administered groups were fed tap water containing water extract
of style of Z mays at the concentration of 0.15 % (w/v). At 12 weeks, urine was collected for 24
h using a metabolic cage, followed by collecting a blood sample and kidney.
Plasma glucose (PG), hemoglobin Ale (HbAlc), fructosamine (FRA), Creatinine clearance
(Ccr), urinary albumin excretion (UAE) and other clinical parameters were also examined.
Results and Discussion
UAE (mg/d) Ccr (ml/min /lOOg body weight)
DM-T 1.70±1.23 0.70±0.11**
DM-NT 2.38±1.35* 0.89±0.15*
NC 0.63±0.60 0.38±0.20
*p

Effects of Turkish Plant Materials on AR.
The inhibitory effects on the h-AR were determined by the method described in the previous
papers. The human muscle AR recombinant activities were assayed spectrophotometrically by
measurind the decrease in absorption of NADPH at 340 nm for 3 min with DL-glyceraldehyde
as substrate.
Results and Discussion
The results of the inhibitory actions of 40 kinds of MeOH extracts obtained from Turkish plants
on h-AR. Of the 12 kinds of extracts with an inhibitory activity of exceeding 60 % on h-AR, the
MeOH extracts of Potentilla recta, Tripleurospermum callosum, and Cistus genus, C. lauriforius
(leaf and fruit), C. creticus (leaf and fruit), C. monspeliensis (leaf), C. salvifolius (leaf) and C.
parviflorus (leaf) exhibited a potent inhibition in duplicate tests.
The extract derived from the leaf of C. laurifolius that exhibited an AR inhibitory effect was
fractionated by column chromatography and HPLC to yield three known compounds with
monitoring by assay of the inhibitory effect on h-AR. Inhibitory effect of one of isolates,
quercetin 3-O-methyl ether was almost as potent as that of the positive control eparlestat, which
is a well known remedy for treating the complications of diabetes in Japan.
As presented above, this compound is thought to be a promising aldose reductase inhibitor which
warrants further investigation.
1. Y.Okada, N.Miyauchi, K.Suzuki, T.Kobayashi, C.Tsutsui, K.Mayuzumi, T.Okuyama,
Natural Medicines, 48, 324-329 (1994).
2. Y. Iwahori, S.Enomoto, Y.Okada, J. Tanaka, T.Okuyama, Natural Medicines, 53, 138-
140(1999).
3. Y.Okada, K.Tachibana, N.Miyauchi, T.Okuyama,Proceedings of the International
Symposium on Natural Medicines, 0ct.28-30, 1997, Kyoto, Japan. 295-303, 1998,
Elsevier Science B.V.
4. R.Suzuki, Y.Okada, T.Okuyama, J.Nat. Prod., 66(4), 564-565 (2003).
5. R.Suzuki, Y.Okada, T.Okuyama, Chem. Pharm. Bull., 51(10) 1186-1188 (2003).
6. R.Suzuki, Y.Okada. T.Okuyama, Biol. Pharm. Bull., 28(5) 919-920 (2005).
7. Y.Okada. A.Guvenf, C.S. Erdurak, M.Cokun, T.Okuyama, Biol. Pharm. Bull., 27(7)
1140-1143 (2004).
5

PL-33
THE WILD MEDICINAL AND AROMATIC PLANT TRADE IN TURKEY
"THE MOST TRADED SPECIES"
N. Ozhatay
Department of Pharmaceutical Botany, Faculty of Pharmacy, University of Istanbul, 34116,
Beyazit, Istanbul, Turkey
Wild collected medicinal and aromatic plants fall into two basic categories: taxa collected for
local use / traditional use and taxa commercially traded both internally and externally (through
export).
INTERNAL TRADE:
The principle markets for medicinal and aromatic plants within Turkey are bazaars and market
stalls, herbalists and to the pharmaceutical industry. Their principle uses are thought to be as (I)
herbal teas" mainly Labiatae species such as Salvia, Sideritis, Stachys\ (II) raw material in the
production of Helva" Coven - Gypsophila & Ankyropetalum roots, as a whitener in the
production of helva and raw material in the production of salep "Tuber, one of nearly 40 species
of orchids is used in the production of ice-cream and hot winter drink."
EXTERNAL TRADE (EXPORT):
Turkey exports approximately 30.000 tones of medicinal and aromatic plants per annum,
generating nearly 50 million US dollars from the trade per annum. These figures indicate that
Turkey is the third largest exporter of medicinal and aromatic plants of wild origin of any
country on earth after China and India. The five principle species in export trade:
1. Ceratonia siliqua (Carob), fruit, 34% (percentage of overall quantity)
2. Origanum sp., Satureja sp., Coridothymus sp., and Thymbra sp. (Oregano); herba, 20%
3. Capparis sp.(Caper), flower buds, 15%
4. Laurus nobilis (bay Laurel) 10%
5. Miscellaneous 9%
(The fifth largest export category, it is within this category that materials of all non-categorized
species are exported.)
5

PL-34
ELECTROCHEMICAL DNA BIOSENSORS
M. Ozsoz, A. Erdem, D. Ozkan Ariksoysal, P. Kara Kadayifcilar, H. Karadeniz, G. Yalcin, S.
Cavdar, B. Meric
Department_of Analytical Chemistry, Faculty of Pharmacy, Ege University, 35100 Bornova-
Izm ir, Turkey
The analysis of nucleic acids is of great importance and it has found wide acceptance not only in
operate analytical applications but also in diagnostic testing. A increasing interest has appeared
in the development of a simple rapid and user-friendly method which is containing of DNA
sequence analysis and mutant gene analysis to allow early and precise diagnoses of infectious
agents for routine clinical tests such as detection of inherited disease.
The objective of our work to develop a unique biosensor that detected hybridisation directly
rather than direct sequencing of DNA. Electrochemical genosensors are attractive devices for
converting the hybridization event into an analytical signal for obtaining sequence-specific
information in connection with clinical, environmental or forensic investigations.
Electrochemical genosensors rely on the immobilization of a single-stranded (ss) DNA sequence
(the "probe") for hybridizing with its complementary ("target") strand to give rise to
voltammetric signals (in connection to a suitable electrochemical label). The aim of our projects
were therefore to prove that a DNA recognition interface could be developed that exploited the
difference in persistence amount of ssDNA and dsDNA to detect specific target sequences of
DNA based on guanine oxidation signal, and that were also simple to prepare, simple to use,
reliable, selective and microfabrication-compatible so that they could be included in a lab-onchip
device.
In our laboratory, we described different type of an electrochemical DNA detection procedure
based on oxidation signals of guanine, mediator signal or nanoparticle signal to detect DNA
damage or an inherited disease such as Factor V Leiden Mutation, Hepatitis B,... etc.
All of these promising studies have shown that DNA genosensors will play an important role for
the design of DNA microaarays.
1. Ozsoz M., Erdem A., Kerman K., Ozkan D., Tugrul B., Topcuoglu N., Ekren H.,Taylan
M., Anal. Chem., 2003, 75, 2181-2187
2. Palecek E., Kizek R., Havran L„ Billova S., Fojta M., 2002a. Anal. Chim. Acta 469, 73-
83.
3. A. Erdem, K. Kerman, B. Meric, U.S. Akarca, M. Ozsoz, Anal.Chim. Acta., 422
(2000)139.
4. Dilsat Ozkan Ariksoysal, Hakan Karadeniz, Arzum Erdem, Aylin Sengonul, A.Arzu
Sayiner, Mehmet Ozsoz, Analytical Chemistry, 77 : 4908-4917, 2005
5. S.R.Mikkelsen, Electroanalysis, 8 (1), 15-19 (1996)
6. G. Marrazza, I. Chianella, M. Mascini, Anal Chim Acta, 387, 297-307 (1999).
7. A. Erdem, M. Ozsoz, Electroanal., 14(14), 965-974 (2002).
8. Boon E.M., Ceres D.M., Drummond T.G., Hill M.G., Barton J.K., 2000. Nat. Biotechnol.
18, 1096-1100.
9. Carpini G., Lucarelli F., Marrazza G., Mascini M., 2004. Biosens. Bioelectron., 20, 167-
175.
5

PL-35
TARGET-DRIVEN DRUG DISCOVERY IN CANCER
M. Ozturk
Bilkent University, Faculty of Science, Department of Molecular Biology and Genetics, 06800
Ankara
Cancer is a disease of cells having the capacity for uncontrolled growth and tissue invasion, due
to genetic and epigenetic changes. Today, specific changes that occur in many different cancer
types are known, as result of intense research activities in molecular biology and genetics.
Changes that are known in cancer cells can be grouped into five main groups: (1) self sufficiency
for proliferation; (2) insensitivity to growth inhibitory signals; (3) resistance to apoptosis or
programmed cell death; (4) immortality or limitless proliferation potential; (5) ability for tissue
invasion and metastasis. Molecular details of the first four characteristics just stated are well
known. Cancer cell acquire these features as a result of aberrant function of genes that are
broadly classified as "oncogenes" and "tumour suppressor genes". Oncogenes usually allow
cancer cells to proliferate, to resist to apoptosis, and to gain immortality. On the other hand,
tumour suppressor genes are those that inhibit such cellular processes in normal cells, but fail to
do so in cancer cells due to mutations or other problems. Proteins that are encoded by oncogenes
and tumour suppressor genes play key roles in pathways that regulate different cellular processes
that I have just mentioned. These pathways are usually initiated by an extracellular signal that
activates a cellular receptor. Then, receptors allow the amplification of the signal in the
cytoplasm by enzymatic activity or protein-protein interactions. Cell nucleus is the terminal
place that the signaling reaches and results in upregulation and/or donwregulation of a set of
genes. Consequently, cell will respond to an external stimulus by a change in its comportment.
In cancer cells, due to mutations or epigenetic changes, aberrations occur in different signaling
pathways. Signals that stimulate cell proliferation and survival are amplified, whereas the
antagonistic signals are blocked. During recent years, screening of molecules that interfere with
cancer signaling pathways has led to the discovery of many drug candidates. Drug candidates
that act externally are usually monoclonal antibodies or recombinant proteins, whereas those
acting in cancer cells are small chemicals either natural or obtained synthetically. Both
approaches have been proven to be effective by the development of new drugs that are now
widely used in as cancer therapeutics.

PL-36
MODELING OF INTERACTIONS WITH THE MULTIDRUG RESISTANCE
TRANSPORTER P-GLYCOPROTEIN
I. Paieva', M. Wiese 2
1 Center of Biomedical Engineering, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria
2 Institute of Pharmacy, University of Bonn, 53121 Bonn, Germany
Introduction
P-glycoprotein (P-gp) belongs to the highly conserved ATP-binding cassette (ABC) protein
superfamily. In humans it is coded by the ABCB1 gene. P-gp is naturally expressed in several
organs where it functions as a natural detoxification system. The protein is actively involved in a
variety of processes such as lowering drug oral bioavailability, preventing CNS drugs from
penetrating across the blood-brain barrier (BBB), decreasing intracellular accumulation of
anticancer and other cytotoxic agents, HIV-protease inhibitors, antibiotics and many others.
Thus, P-gp is responsible for multidrug resistance (MDR) of the tumor cells to a broad spectrum
of agents. This ranks it as one of the major MDR proteins with significant physiological and
pharmacological relevance.
Aim
The studies of P-gp interactions with drugs address two main purposes from a pharmacological
point of view: (i) better understanding of the processes involved in the MDR phenomenon and
(ii) identification of compounds that could potentially interact with P-gp at earlier stages of the
drug development process.
Methods
P-gp and its interactions with drugs are intensively investigated by a number of experimental and
modeling techniques. Depending on the available experimental data and the level of knowledge
about the structure and function of the protein, modeling studies undergo several stages. The
absence of 3D structural data of the protein obtained at a reasonable resolution predetermines the
relatively high portion of applications of the so-called ligand-based drug design approaches [1]:
the classical QSAR analyses of Hansch and Free-Wilson; the 3D-QSAR approaches CoMFA
(Comparative Molecular Field Analysis) and CoMSIA (Comparative Molecular Similarity
Indices Analysis) (Sybyl, Tripos Inc., USA); HINT (Hydrophatic INTeractions) force field
(HINT, EduSoft L.C., USA); pharmacophore modeling by GASP (Genetic Algorithm Similarity
Program) (Sybyl, Tripos Ass., USA); homology and protein modeling using the relevant
program modules in Sybyl and MOE (Chemical Computing Group Inc., Canada).
Results and Discussion
A number of QSAR and 3D-QSAR models have been developed based on experimental MDR
reversing activities in vitro of numerous classes of compounds representing different generation
of MDR modulators (phenothiazines and related compounds, propafenones and benzofurans,
triazines, anthranilamides) proven to interact with P-gp and to modulate MDR in resistant tumor
cells. The QSAR models mostly employed structural fragments as descriptors and correlated
them with activity values in the Free-Wilson analysis to estimate the most significant structural
features in homologues series of compounds [2,3], The first 3D-QSAR models applied CoMFA
and HINT and outlined the essential role of hydrophobic fields alone and in combination with
steric and electrostatic fields for the MDR-reversal effect of the compounds investigated [4-11],

It has been demonstrated that hydrophobicity considered as a space directed molecular property
and not as an integral lipophilicity (logP) is the appropriate structural descriptor for modeling the
P-gp-drug interactions. The same has also been shown for some BBB penetrating drugs [12].
Using CoMSIA the role of the hydrogen-bond (HB) acceptor molecular fields has additionally
been revealed [3,10]. These results are in agreement with the putative location of the drug
binding sites on the P-gp transmembrane domains (TMs) that are involved predominantly in
hydrophobic and HB interactions.
Recently data on interactions of drugs with particular binding sites of P-gp have become
available. This initiated pharmacophore modeling of P-gp ligands. In an attempt to explain the
broad structural variety of the P-gp drugs that interact with the verapamil binding site a flexible
alignment by GASP has been performed using structurally diverse MDR substrates and
inhibitors, including the P-gp substrate Rhodamine 123 [8,11,13], Several pharmacophore points
in a particular space arrangement have been identified: two hydrophobic planes, three HBacceptors
and one HB-donor. Pharmacophore patterns of various drugs have been obtained and
different binding modes were suggested for some of them. It has been shown that the binding
affinity of the drugs correlates with the number of the pharmacophore points simultaneously
involved in the interaction with P-gp, thus underlying the relevance of the developed
pharmacophore model. Using the same program a pharmacophore model of drugs that interact
with the Hoechst 33342 binding site of P-gp has also been developed [8,11,14]. The
pharmacophore models obtained for Rhodamine 123 and Hoechst 33342 have been suggested to
relate to the putative R- and H-binding sites of P-gp and the corresponding parts of TMs have
been proposed [14].
The high resolution X-ray structures of the nucleotide-free forms of the bacterial lipid ABC
transporter MsbA from Escherichia coli and Vibrio cholerae obtained recently by Chang [15,16]
have given a start to the homology modeling of P-gp. Aiming to better understand the structurefunction
relationships of P-gp two protein models have been developed: (i) a cross-linking model
based on the distance constraints obtained from the experimental cross-linking studies of Loo
and Clarke [17,18] and (ii) a homology model based on the X-ray structure of the nucleotide-free
Escherichia coli MsbA (4.5 A resolution) transporter [14]. Analyzing the location of the amino
acids involved in the putative R- and H-sites on TMs it has been shown that the orientation of
the residues differ in both models: in the homology model the binding sites face the membrane,
in the cross-linking model they are directed to the pore. This result presumes that the TMs
undergo rotation and translation during the transition of the protein from the state presented by
the homology model to the state presented by the cross-linking model. Thus, the models are
suggested to correspond to two different functional states of the protein: the ATP-free (the
homology model) and the ATP-bound (the cross-linking model). The results qualitatively agree
with the P-gp electron density maps at 20 A resolution reported by Rosenberg et al. [19].
The modeled TMs arrangement in the nucleotide-bound state of P-gp (based on the 8 A resolved
electron density map) published recently [20] allows the assignment of TMs in correspondence
with the cross-linking model (Fig. 1). As seen from the cross-linking model (Fig. IB) seven TMs
(4, 5, 6, 9, 10, 11, and 12) face the pore - the same numbers are assigned to the TMs forming the
pore in Fig. 1A. The assignment of the other TMs is done according to the homology model of
the nucleotide-free functional state [14] and the recent results of Loo et al. [21,22] showing that
TM2 and TM11 and TM5 and TM8 should remain close to each other after the repacking of the
protein. The largest gaps in the arrangement are between TM5 and TM6, and TM4 and TM12 in
agreement with the electron density map of P-gp resolved at 20A [19].
6

A
i r ^
B
11
12 J & 10
#
5 6
Figure I. Top views of: (A) the arrangement of TMs according to Rosenberg et al. [20]; the helical spiral ribbons in
the original picture are overlaid by cylinders; the corresponding TMs in both halves are 1-7, 2-8, 3-9, 4-10, 5-11,
and 6-12; (B) the cross-linking model of P-gp proposed in [14]; the amino acids presumed to be involved in binding
of Hoechst 33342 and Rhodamine 123 on TM5 and TM6 respectively face the pore.
The very recent results on the automatic identification of the protein binding pockets generated
by the program SitelD (Sybyl, Tripos Inc., USA) confirm the possibility for existence of
multiple binding sites on the protein. The mostly involved amino acids correspond to those
identified in the experimental studies. The binding pockets have been identified in a number of
low energy conformations of the whole P-gp, each of the halves and the pairs TM5-TM6 and
TM11-TM12. The conformers have been obtained by molecular dynamics using the P-gp
homology model based on the MsbA from Vibrio cholerae (3.8 A resolution).
We thank our colleagues from University of Bonn and Bulgarian Academy of Sciences and especially Christoph
Globisch, Ivanka Tsakovska, Romy Fleischer, and Iglika Lessigiarska whose efforts contributed to the above
results.
The work has been supported by grants from Deutsche Forschungsgemeinschaft, Alexander von Humboldt
foundation, Bulgarian National Science Fund (grant L-1416), Graduiertenkolleg 804.
1. Wiese, M„ Pajeva, I. Curr. Med. Chem. 2001, 8, 685-713.
2. Pajeva, I., Wiese, M. Quant. Struct.-Act. Relat. 1997, 16, 1-10.
3. Globisch, C. et al., Bioorg. Med. Chem. 2006, 14, 1588-1598.
4. Pajeva, I., Wiese, M. J. Med. Chem. 1998, 41, 1815-1826.
5. Pajeva, I., Wiese, M. Quant. Struct.-Act. Relat. 1998,17, 301-312.
6. Tsakovska, I., Pajeva, I. SAR QSAR Environ. Res. 2002, 13, 473-484.
7. Fleischer, R„ Wiese, M. J. Med. Chem. 2003, 46,4988-5004.
8. Pajeva, I. et al. Actual. Chim. Therapeut. 2005, 31, 167-180.
9. Pajeva, I., Wiese, M. In: Molecular Modeling and Prediction of Bioactivity, Gundertofte, K.,
Jorgensen, F. S. (Eds.), Kluwer Acad./Plenum Publishers, NY, 2000, 414-416.
10. Tsakovska, I. et al., Biotechnol. Biotechn. Eq. 2003, 17, 163-169.
11. Pajeva I. et al. Med. Chem. Res. 2005, 14, 106-117.
12. Lessigiarska, I. et al. SAR QSAR Environ. Res. 2005,16, 79-91.
13. Pajeva I., Wiese, M. J. Med. Chem. 2002,45, 5671-86.
14. Pajeva, I. et al. J. Med. Chem. 2004, 47, 2523-2533.
15. Chang, G„ Roth, C. B. Science 2001, 293, 1793-1800.
16. Chang, G. J. Mol. Biol. 2003, 330, 419-430
17. Loo, T.W., Clarke, D. M. J. Biol. Chem. 2001,276, 36877-36880.
18. Loo, T. W„ Clarke, D. M. Proc. Natl. Acad. Sci. USA 2002, 99, 3511-3516.
19. Rosenberg, M. F. et al. J. Biol. Chem. 1997, 272, 10685-10694
20. Rosenberg, M. F. et al. J. Biol. Chem. 2005, 280, 2857-2862
21. Loo, T. W. et al. J. Biol. Chem. 2004, 279, 7692-7697.
22. Loo, T. W. et al. J. Biol. Chem. 2004, 279, 18232-18238.
63

PL-37
ELECTROCHEMISTRY OF NUCLEIC ACIDS AND PROTEINS:
TOWARDS SENSORS FOR GENOMICS AND PROTEOMICS
E. Palecek, V. Ostatna, V. Dorcak, L. Havran, M. Masarik, M. Trefulka, K. Cahova, D.Krstic, Z.
Pechan, F. Jelen, M. Fojta
Institute of Biophysics, Academy of Sciences of the CR, 61265 Brno, Czech Republic
Progress in genomics and particularly in the Human Genome Project in the 1990's greatly
stimulated interest in new methods capable to unravel the genetic information stored in the
nucleotide sequence of DNA. After the construction of the first DNA arrays (chips) with optical
detection attempts to develop DNA chips with electrochemical detection (which can be simpler
and should be less expensive) have become popular among electrochemists. Since the middle of
the 1990's the electrochemistry of nucleic acids and the development of DNA sensors have been
a booming field involving large number of laboratories all over the world. Similar increase in the
interest in electrochemistry of proteins can be expected if the electrochemical analysis meets the
requirements of proteomics. In this talk we wish to show that such expectations are not
unrealistic.
Electrochemistry of nucleic acids
Electroactivity of nucleic acids was discovered about 45 years ago. It was shown that at mercury
electrodes adenine (A) and cytosine were reduced in DNA while guanine (G) produced an
anodic signal due to oxidation of G reduction product 1 2 . Later oxidation of G and A at carbon
electrodes was demonstrated. Signals at mercury electrodes were highly sensitive to structural
changes in double-stranded DNA 3 ' 4 '
5 . Already in the beginning of the 1980's electroactive
labels were covalently bound to DNA and used as probes of the DNA structure 2 4; \ In the
middle of 1980's DNA-modified electrodes were introduced, which then have become popular,
particularly in relation to the development the DNA sensors.
Covalent labels of DNA. DNA and RNA are naturally electroactive but both their oxidation and
reduction are electrochemically irreversible, occurring at highly positive or highly negative
potentials. Electroactive labels were introduced into DNA to obtain electrochemical signals at
potentials closer to the potential of zero charge and/or to increase the sensitivity of the analysis.
Ferrocene, daunomycin, viologen, thionine etc. were used as DNA labels. These labels require
solid state organic chemistry and could hardly be used for labeling of longer NAs, such as
plasmid and chromosomal DNAs, viral RNAs, etc. Already in the beginning of the 1980's we
introduced electroactive osmium labels which can be applied for end-labeling of DNA x 6 .
Osmium tetroxide complexes with nitrogen ligands (Os vm ,L) can be used for DNA labeling in
an average biochemical or molecular-biological laboratory without any special equipment. They
produce redox signals at mercury, amalgam and carbon electrodes; in addition electrocatalytic
signals can be observed at mercury and amalgam electrodes 5 ' 1 . Recently we have found that
sixvalent osmium complexes (Os VI ,L) can be used for labeling of oligoribonucleotides and of
some polysaccharides (M. Trefulka and E. Palecek, unpublished).
Sensors for nucleotide sequencing by DNA hybridization.
In the 1990's advances in nucleotide sequencing of the human genome stimulated interest in
DNA hybridization sensors. In addition to optical detection, electrochemical detection showed
great promise because of simple design, low energy requirements and low cost. By the end of the
20th century, a number of electrochemical sensors were available, in which both the DNA
hybridization and its detection were performed at the same surface 8 .

In the beginning of the 2000's a new conception was introduced, in which the hybridization was
performed at one surface (optimized for DNA hybridization) and the DNA detection at another
surface 5 '
9 . At present various electrochemical sensors for DNA hybridization are available
enabling determinations of specific nucleotide sequences 5 l0,
' DNA point mutations
and lengths of long repetitive sequences 5 lj
' in amplified DNA fragments with remarkable
sensitivities. Determination of a specific DNA sequence in eukaryotic genomes without
amplification of DNA remains, however, a great challenge.
3 10 12
Thiol end-labeled oligodeoxynucleotides (HS-ODN). Single-stranded (ss) ODN probes
immobilized on solid surfaces are commonly used in DNA hybridization sensors 5 ' 10, l4 ' 15 16 ' 17 .
Direct linkage of thiol-end-labeled ODNs HS-ODNs at gold surfaces, facilitating formation of
DNA self-assembled monolayer (SAM), is one of the most frequently used ways of DNA
immobilization in these sensors 12 ' 6 . Such monolayers resemble the well-known self-assembly
of alkanethiols on gold surfaces x ,6 . Recently we have shown that HS-ODN form selfassembled
monolayers (SAMs) at mercury electrodes providing signals due to (a) reduction of
bases and (b) reduction of the Hg-S bond. In addition HS-ODNs produce specific signals in
cobalt solutions suitable for protein analysis; these signals appear at less negative potentials than
those of cysteine-containing peptides and proteins offering a chance for investigation of DNA-
• • 18
protein interactions .
Electrochemistry of proteins
Small conjugated proteins containing redox centers produce fast reversible electrode processes
,9 . Present trends in proteomics require new sensitive methods for the analysis of all proteins not
limited by their size and reversibility of their electrochemical processes.
Electrocatalysis at mercury electrodes. All proteins and peptides so far tested have been able to
catalyze hydrogen evolution at mercury electrodes providing analytically useful signals 20, 21 .
Nanomolar and subnanomolar concentrations of peptides and proteins can be determined by
constant current chronopotentiometric stripping analysis (CPSA) producing peak H at highly
negative potentials. Peptides and proteins produce peak H regardless of presence or absence of
cysteine in their molecules. However, presence of cysteine strongly influences peak H and its
20; 22
dependence on pH. This peak sensitively reflects changes due to protein aggregation and
denaturation 20 2j
' and appears potentially useful in studies of mutant proteins 20 . It is also
sensitive to the peptide and protein redox states (V. Dorcak and E. Palecek, unpublished).
Oxidation at carbon electrodes. Tryptophan and tyrosine residues in peptides and proteins are
oxidizable at carbon electrodes. Using CPSA or square wave stripping voltammetry well
developed peaks can be obtained. Oxidation and reduction signals were applied to study avid in
and streptavidin 24 bovine serum albumin 23 and other proteins. These signals are potentially
useful in protein sensors.
Aggregation of a-synuclein is involved in Parkinson's and Alzheimer's diseases. Such
aggregation was induced in vitro and studied by circular dichroism (CD), fluorescence, atomic
force microscopy and by electrochemical methods at mercury and carbon electrodes. Using peak
H a-synuclein can be determined at subnanomolar concentrations and its aggregation traced at
mercury and carbon electrodes 20; 22 . Early changes in a-synuclein properties, preceding its
aggregation were detected by means of peak H and other electrochemical methods. These preaggregation
changes, which are supposed to be critical for the development of the Parkinson
disease, were not detectable by tyrosine oxidation signals and by CD or fluorescence
measurements. Mutants of a-synuclein showed differences in their pre-aggregation behavior.
6

DNA-protein interactions. DNA binding proteins play a central role in many aspects of genetic
activity in an organism, such as transcription, replication, DNA repair, rearrangement,
packaging, etc. It is therefore extremely important to investigate the processes of DNA-protein
binding and the nature of complexes formed between DNA and proteins. In comparison to DNA
and RNA, proteins are much less regular and therefore understanding them is more difficult. In
the past two decades we witnessed a great expansion in high-quality structures of DNA-binding
proteins 25 . These structures and particularly those of their complexes with DNA have provided
valuable insights into the principles of binding, including how specific nucleotide sequence is
recognized by the protein and how DNA structure is usually modified on protein binding. In
addition to the high resolution X-ray crystal analysis a number of methods have been used in
studies of DNA protein interactions. On the other hand, application of electrochemical analysis
to studies of DNA-protein interactions was until very recently almost completely missing.
Considering that both DNA and proteins are electroactive and can be analyzed with high
sensitivity, application of electrochemical analysis in DNA-protein analysis appears natural and
very promising. Examples of electrochemical methods for investigation of DNA-protein analysis
4 ' 20 will be presented.
This work was supported by grants of the Grant Agency of the Czech Republic 203/06/1685, of the the
Academy of Sciences of the Czech Republic IAA50040513, A100040602 and AVOZ50040507 and
Ministry of Education, Youth and Sports, CR, LC06035.
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Nature 188, 656-657.
2. Palecek, E. (2005). Polarography of DNA. Retrospective view. In Electrochemistry of nucleic
acids and proteins. Towards electrochemical sensors for genomics and proteomics. (Palecek, E.,
Scheller, F. & Wang, J., eds.), pp. 1-17. Elsevier, Amsterdam.
3. Palecek, E. (1976). Premelting changes in DNA conformation. Progress in Nucleic Acid
Research and Molecidar Biology 18, 151-213.
4. Palecek, E. & Fojta, M. (2005). Electrochemical DNA sensors. In Bioelectronics (Willner, I. &
Katz, E, eds.), pp. 127-192. Wiley-VCH, Weinheim.
5. Palecek, E. & Jelen, F. (2005). Electrochemistry of nucleic acids. In Electrochemistry of nucleic
acids and proteins. Towards electrochemical sensors for genomics and proteomics. (Palecek, E.,
Scheller, F. & Wang, J., eds.), pp. 74-174. Elsevier, Amsterdam.
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7. Yosypchuk, B., Fojta, M., Havran, L., Heyrovsky, M. & Palecek, E. (2006). Voltammetric
behavior of osmium-labeled DNA at mercury meniscus-modified solid amalgam electrodes.
Detecting DNA hybridization. Electroanal. 18, 186-194.
8. Palecek, E. & Fojta, M. (2001). Detecting DNA Hybridization and Damage. Anal. Chem. 73,
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proteins. Towards electrochemical sensors for genomics and proteomics (Palecek, E., Scheller,
F. & Wang, J., eds.). Elsevier, Amsterdam.
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DNA hybridization on membrane-modified carbon electrodes. Anal. Lett. 38, 2493-2507.
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and cobalt-phenanthroline as redox indicators of DNA hybridization at gold electrodes. Front.
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13. Fojta, M., Havran, L., Vojtiskova, M. & Palecek, E. (2004). Electrochemical detection of DNA
triplet repeat expansion. J. Am. Chem. Soc. 126, 6532-6533.
6

14. Drummond, T. G., Hill, M. G. & Barton, J. K. (2003). DNA-based electrochemical sensors. Nat.
Biotechnol. 21, 1192-1199.
15. Finklea, H. O. (2000). Self-assembled monolayers on electrodes. In Encyclopedia of Analytical
Chemistry (Meyers, R. A., ed.), Vol. 11, pp. 10090-10115. Wiley, New York.
16. Tarlov, M. J. & Steel, A. B. (2003). DNA-Based Sensors. In Biomolecular Films. Design,
Function, and Applications. (Rusling, J. F., ed.), pp. 545-608. Marcel DeKker, New York.
17. Thorp, H. H. (2004). Electrocatalytic DNA oxidation. In Long-Range Charge Transfer in DNA
II, Vol. 237, pp. 159-181.
18. Ostatna, V., Jelen, F., Hianik, T. & Palecek, E. (2005). Electrochemical responses of thiolated
oligodeoxynucleotides in cobalt-containing solution. Electroanal. 17, 1413-1420.
19. Armstrong, F. A. (2002). Voltammetry of proteins. In Bioelectrochemistry (Wilson, G. S., ed.),
Vol. 9, pp. 11-29. Wiley-VCH, Weinheim.
20. Palecek, E. (2005). Electroactivity of proteins and its possibilities in biomedicine and proteomics.
In Electrochemistry of nucleic acids and proteins. Towards electrochemical sensors for genomics
and proteomics. (Palecek, E., Scheller, F. & Wang, J., eds.), pp. 690-750. Elsevier, Amsterdam.
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(Palecek, E., Scheller, F. & Wang, J., eds.).
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aggregated a-synuclein involved in Parkinson's disease. Electroanal. 16, 1172-1181.
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serum albumin. D.c. polarography, stripping voltammetry and constant current
chronopotentiometry. J. Electroanal. Chem. in press.
24. Havran, L., Billova, S. & Palecek, E. (2004). Electroactivity of avidin and streptavidin. Avidin
signals at mercury and carbon electrodes respond to biotin binding. Electroanal. 16, 1139-1148.
25. Luscombe, N. M., Austin, S. E., Berman, H. M. & Thornton, J. M. (2000). An overview of the
structures of protein-DNA complexes. Gen. Biol. 1.
6

THE FARNESOID X RECEPTOR AND DRUG DISCOVERY: FROM FUNCTIONS
UNRAVELING TO THERAPEUTIC EXPLOITATION
R. Pellicciari
Dipartimento di Chimica e Tecnologia del Farmaco, Universita di Perugia, via del Liceo, 1,
06123 Perugia, Italy.
PL-38
During the last years great progresses have been made in the elucidation of properties, functions
and therapeutic applications of nuclear receptors with a particular attention reserved to a group
referred to as 'adopted orphans' since natural ligands have recently been identified. Among
them, the Famesoid X Receptor (FXR) has been shown to be highly expressed in liver, intestine,
kidneys and identified as a bile acids 'sensor' with chenodeoxycholic acid (CDCA) as the most
potent natural ligand. Upon activation, FXR heterodimerises with the RXR receptor and
regulates a cohort of genes that function to repress bile acid synthesis and import in hepatocytes,
to stimulate bile acids export from cells and to protect hepatocytes from bile acids toxicity.
Recently, it has been discovered that FXRactivation also modifies the transcriptional activity of
a variety of transcription factors controlling gluconeogenesis and lipogenesis thus affecting in
concert bile acid, lipid and carbohydrate metabolism.
We have previously reported the synthesis of 6-ECDCA (INT-747) a potent and selective FXR
agonist able to protect against cholestasis as well as liver fibrosis when administered in vivo.
More recently we have been engaged in the identification of FXR receptor partial agonists and
antagonists endowed with gene selective patterns correlated with desired physiological effects.
Their availability can be instrumental in expanding our insight into the FXR physiological roles
and in opening the way to new therapeutic opportunities. Thus, molecular mechanisms
underlying lipid homeostasis and energy balance can be unravelled and intriguing possibilities
for the use of FXR modulators in the metabolic syndrome revealed.
1. Pellicciari, R, et al. J Med Chem. 2002; 45(17): 3569-72.
2. Costantino, G, etal. Bioorg Med Chem Lett. 2003; 13 (11): 1865-8.
3. Pellicciari, R, et al. J Med Chem. 2004; 47(18):4559-69.
4. Costantino, G, et al. J Med Chem. 2005; 48 (9):3251-9.
5. Mi, LZ, etal. Mol Cell. 2003; 11 (4): 1093-100.
6. Fiorucci, S, et al. Gastroenterology. 2004; 127(5):1497- 1512.
7. Fiorucci, S, et al. J Pharmacol Exp Ther. 2005; 313(2):604-12
8. Fiorucci, S, et al. JPharmacol Exp Ther. 2005; 314(2):584-595.
9. Rizzo, G, et al. Mol Pharmacol. 2005; 68(2):551-8.
10. Pellicciari, R, et al. J MedUhem. 2005; 48(17): 5383-403.

PL-39
THE IMPLEMENTATION OF THE EUROPEAN BACHELOR-MASTER SYSTEM IN
PHARMACY EDUCATION IN FLANDERS (BELGIUM)
B. Rombaut
Dean, School of Pharmacy, Vrije Universiteit Brussel, Laarbeeklaan 103, B-1090 Brussels,
Belgium
After she signed the Bologna Declaration in 1999, the Flemish Minister of Education (Belgium is
a Federal State, in which the communities are responsible for education) started the process to
reform the Flemish higher education system according to the the Declaration principles, of which
the most important are : (i) the adoption of a system readable and comparable degrees; (ii) the
introduction of an education system essentially based on two cycles;(iii) the establishment of a
credit system (ECTS) and, (iv) the promotion of student mobility.
As a consequence, already on the 2 nd of April 2003, the act of the Flemish Community on the
implementation of the Bachelor-Master (BaMa) system became Law. In this Act, it is also stated
that universities are responsible for the content of the Pharmacy programme and that educational
goals should be defined.
A Working Group on the implementation of BaMa was installed by the four Schools of Pharmacy
of the Flemish Universities (KUL; UG; VUB and UA). The objectives of this Working Group
were: (i) developing a strategy to increase the intake of students; (ii) implementation of the BaMa
model in pharmacy education; (iii) adaptation of the programme to the new role and tasks of the
pharmacist in the contemporary society; (iv) to increase collaboration between universities.
The Working Group finally proposed the following model, which is now implemented in the four
Schools of Pharmacy: (i) Bachelor in Pharmaceutical Sciences: a (three years) programme of 180
ECTS. This Bachelor programme is a so-called Academic Bachelor with no civil effect (no
professional implication). Students having obtained this degree can enter two comparable
Masters: (ii) a Master in Pharmaceutical Care of 120 ECTS (two years) or (iii) a Master in Drug
Development, again of 120 ECTS (two years). In both Masters, a masterthesis of 24 ECTS and a
6-month training in a community pharmacy are mandatory. Holders of both Masters also receive
the title of Pharmacist.
Masters in Pharmaceutical Care possess the knowledge, skills, competences, and attitudes of a
community pharmacist in the contemporary society. This means less attention to manufacturing
drugs and analysis of drugs, but more pharmaceutical care, pharmacotherapy and communication
skills.
The Master in Drug Development has again the knowledge, skills, competences and the attitudes
to act as a community pharmacist (but at a slightly lower level). But, as consequently more
training in drug development.

PL-40
PHARMACOEPIDEMIOLOGY AND ITS IMPLICATION ON PHARMACY AND
PHARMACEUTICAL CARE
M. Schaefer
Charite-Universitatsmedizin Berlin, Germany
Since 1983, pharmacoepidemiology has been seen as "the study of medicines as determinants of
health and disease in populations and defined to be the science of practice of drug safety and a
bridge science drawn from the disciplines of pharmacology, therapeutics, epidemiology and
statistics" (Spitzer 1991).
Its basic question is to evaluate the relationship between beneficial effects and risks of a drug
therapy in a defined population in order to improve the therapeutic outcome and reduce adverse
drug reactions.
It seems reasonable that pharmacoepidemiological methods are not only used in medicine to
provide a data basis for decision making but also within pharmacy. This goes in line with the
future development of the profession which is to adapt its role in a continuously changing
political and social environment.
Strategies of pharmacists to define their future role include
- more patient-orientated attitude and activities like patient counselling
- more professional responsibility for the safe and efficient use of medicines and
- pharmacists' care for the therapeutic outcome of drug treatment.
Usually these strategies are described by the concept of Pharmaceutical Care. They also
constitute the requirements for an effective use of pharmacoepidemiological methods and
outcomes within the process of drug use. This concept also allows the pharmacists to function as
a safeguard in order to enhance a rational drug use by patients and to contribute to a measurable
therapeutic outcome of drug treatment, usually in close cooperation with the physician.
Although the evolution of Pharmaceutical Care as a concept follows basic principles worldwide
it has to be adapted to the national framework and system.
For the acceptance of Pharmaceutical Care by the sickness funds and society in general it is
crucial that pharmacists can give evidence for the direct as well as indirect beneficial effects of
Pharmaceutical Care. There is also a need for educating and training pharmacists sufficiently in
order to enable them to provide this service. The successful implementation of Pharmaceutical
Care as a practice requires an effective management of data generated in the care process which
also can provide a powerful database for pharmacoepidemiological research. New information
technologies will lead to a closer and more effective cooperation between health care
professionals. Properly used they will have the potential to support Pharmaceutical Care as an
indispensable function within health care.

One prerequisite of Pharmaceutical care is to keep a medication history for those patients who
join the Pharmaceutical Care programme of a pharmacy. As a next step a medication profde can
be generated form each indiviual medication history by a suitable software programme. A
medication profile is a list of medicines prescribed for (Rx) and purchased by (OTC) an
individual patient usually displayed over a period of 6 months and structured according to the
ATC-Classification.
The point of supply to the patient is marked and hence the expected duration of use calculated
considering number of tablets, capsules etc. per package, their strength and the recommended
dose per day for the individual patient.
Based on these data pharmacists and other health care professional can check the medication
according to the appropriateness of the drug, dose and frequency through a drug review process
and obtain the following information:
Duplication of prescriptions
Interactions between medicines used at the same time
Compliance with chronic medication
- Adverse drug reactions (if there is within a plausible time frame a consecutive use of a
medicine indicating that an adverse reaction is being treated)
- Contra indications (given that a disease state or a previous allergic reaction have been
documented for this individual)
By using additional information from the direct communication with patients pharmacists will be
able to reveal so-called drug-related problems.
Drug-related problems comprise all problems coming along with the drug use by a specific
patient and usually require an intervention by the dispensing pharmacist often in cooperation
with the prescribing physician. The identification of drug-related problems in the community
pharmacy thus contributes to drug safety and is focussed on an efficient drug use with a definite
outcome.
Although the detection and solution of drug-related problems is an important step of the
Pharmaceutical Care process it can also be used as a professional activity to introduce elements
of Pharmaceutical Care where a permanent practice is not established yet.
The process of Pharmaceutical Care requires a standardized way of data documentation. This
includes
- individual patient data (age, gender, contraindications, allergies etc.)
- medicines supplied to the patient and dosis prescribed or used
any drug-related problem occurring during treatment
The main groups of drug-related problems comprise the following:
- unsuitable drug choice
- unsuitable use by the patient, incl. compliance
- unsuitable dosage

- drug-drug interactions
- adverse drug reactions
- other drug-related problems
. patient-related
. communication-related
. technical and logistic problems
The problems documented in the Bavarian study in 1998 were categorised using the coding
system described resulting in the following problem categories:
- inappropriate choice of drug Rx 42,2% OTC 49,6%
- inappropriate drug use by the patient
(including compliance) Rx 18,8% OTC 23,7%
- inappropriate dosage Rx 16,7% OTC 4,1%
- drug interactions Rx 3,8% OTC 4,1%
- adverse drug reactions Rx 4,6% OTC 4,3%
other logistic problems Rx 13,8% OTC 13,9%
These results can be specified for different groups of drugs as well as specific drugs. They also
indicate priorities in identifying and avoiding drug-related problems in community pharmacies.
The degree of intervention is different for various problem categories dependent on the kind of
problem reported. However, there is probably a selection bias due to the fact that pharmacists
were more likely to report problems they could solve. The study also allowed an estimation to
which degree problems could be solved (86.5% completely, 11.1% partly and 2.5% unsolved)
and which time it took to detect and solve the problem with or without contact with a physician.
The detection and solution of drug-related problems has significant financial implications as can
be seen from the following model calculation which starts from the number of prescription in a
given country and considers assumptions from literature with regard to the prevalence of such
problems:
Estimate of avoided costs for drug-related problems
detectable by a systematic drug review (Germany)
Number of prescriptions per year
470,000,000
from that 2% with drug-related problems
9,400,000
from that 30% potentially dangerous to health 2,820,000
from that 30% leading to stay in hospital
from that 30% avoidable by drug review
7 days in hospital per case €
846,000
253,800
908,375
cost per hospital day €
291
cost reduction (hospital stay) € 517,777,380
per year by avoiding
drug-related problems
Source:Kommunikationsplattform im Gesundheitswesen. Kosten-Nutzen-Analyse: Neue Versichertenkarte unci
Elektronisches Rezept, May 2001

Apart from these findings, documented data of patient characteristics, medicines prescribed for
and problems experienced by them also generate a data base which can be used for
pharmacoepidemiological studies. Whereas activities on the individual level aim at an
improvement of the drug use by a patient data on aggregated level provide evidence about the
prevalence of certain events like ADRs, interactions and medication stops:
Data from medication profiles
on individual level
(Pharmaceutical Care)
Detection of ADRs
Check for interactions
individual adaption of doses
Check for compliance
Documentation of
medication stops
Check for individual characteristics
(e.g. Cyp 450 variations)
on aggregated level
(Pharmacoepidemiology)
Prevalence of ADRs
Prevalence of Interactions
Dose range
Comparison of compliance rates
Prevalence of
medication stops
pharmacoepidemiological
evaluation
However, to use information from medication profiles for pharmacoepidemiological purposes
requires that data from as many profiles as possible and from different pharmacies can be linked
and aggregated. To achieve this, the documentation has to be stardardized and interfaces must be
defined clearly.
Conclusion
The documentation of drug use by individual patients as well as their drug-related problems is an
effective mean to improve practice of care to patients using medicines. On the other hand these
data generate a powerful data base which can be used for pharmacoepidemiological studies
given that this is not prohibited by considerations concerning the individual right of data
protection. Therefore pharmacists have to communicate clearly the benefical effects of the
documentation of drug use for patients.
In the future, the introduction of electronic Smart Cards and electronic patient records will
increase the amount of health-related data which is in principle available for the risk-benefit
evaluation of medicines. However, this will only be possible if general methodological issues of
the documentation and the quality of data are taken into consideration at a very early stage of
building the e-health environment.

aligned, further portions of both molecules
form a flat hydrophobic surface of very
similar shape (region D in Fig. 2) and the
hydroxy groups at C-6 and C-10,
respectively are in positions (E in Fig. 2)
where they might donate hydrogen bonds
to a common acceptor site. When
structural analogues from both groups of
terpenoids were compared, this common
pattern was found to be present in all
active convulsants, while inactive
compounds showed deviations which
should explain their inactivity [5].
Fig. 2. Molecular models of PIC (left) and ANI (right)
aligned with respect to the oxygen atoms labelled A, B and C.
Further regions contributing to the biophore model described
in the text are labelled D and E. Modified, according to [5].
In a cooperative project with Y. Ozoe et al. (Shimane,
Japan), it was shown that some of the seco-prezizaane-type
compounds isolated by us from Illicium species possess
conspicuous selectivity towards insect GABA-ligated
chloride channel (Fig. 3). Compounds in the upper left
corner of the diagram show negligible affinity to the
mammalian ion channel while they bind with high affinity to I
the insect receptor. These compounds may thus be
considered very promising lead structures towards new
selective insecticides [6],
These findings prompted us to conduct a 3D-QSAR study
with the aim of identifying the structural requirements for
this selective activity.
To this end, we used the Quasar approach [7] which is based
on the construction of a receptor surface model and
calculation of each of the compounds' interaction energy
with this "pseudo-receptor".
The structures of 30 terpenoids (13 seco-prezizaanes, 17
picrotoxanes/picrodendranes) were aligned according to our
biophore hypothesis and submitted to Quasar. For each set
of binding data (Musca domestica and Rattus norvegicus), an
independent binding site model was constructed. Fig. 4
shows the excellent correlation of the experimental data with
those predicted by the Quasar models.
>
A
A
A
o
A
O
° O „
0 20 40 60 80 I OO
Rattus norvegicus
Fig. 3. Displacement of [ 3 H]-EBOB
from the binding site at rat brain and
housefly head GABA receptor-coupled
CI" channels by seco-prezizaane
sesquiterpenes from Illicium species (10
HM each). Data were taken from [6],
Open circles: Low affinity towards both
receptors, open triangles: High affinity
towards both receptors; filled triangles:
Medium to high affinity towards the
insect, negligible activity towards the
mammalian receptor.
A
/
/ . /
/
• yy-'

Figure 5 shows the two different pseudo-receptors with two seco-prezizaanes (insect-selective and nonselective),
viewed from an angle analogous to that in Figure 2. The overall characteristics of the binding
sites in insects and mammals are predicted to be quite similar with the major exception that the rat
receptor model possesses an additional site of polar interactions on the right side which interacts
specifically with the oxirane/oxetane oxygens of picrotoxinin- and anisatin-type convulsants, respectively.
The insect-selective compounds mentioned above do not possess such structure elements and form
favourable interactions with the insect receptor via other structure elements which explains their high
selectivity.
Housefly
Fig. 5. Pseudoanisatin (insect-selective;
A) and veranisatin A (non-selective, B)
shown inside the fly (top) and rat
(bottom) receptor surface models (from
IB]).
Receptor particles: Blue: Salt bridge
negative, SB-; Red: Salt bridge positive,
SB+, Yellow: Hydrogen bond acceptor,
Acc; Green: Hydrogen bond donor
(Don); Grey (small spheres):
Hydrophobic, neutral; Yellow-brown
(small spheres): Hydrophobic positive
(Hy-); Red-brown (small spheres):
Hydrophobic, negative (Hy-).
Note the interaction of the P-lactone ring
oxygen of A with the SB+ and Don
particles on the right side of the rat
receptor (arrows). Especially the SB+
particle is moved towards the oxetane
ring oxygen as compared to B.
On the background of these results, it appeared an interesting task to obtain further information on the
structure of the binding site in an atomistic context. Based on the reported structure of a closely related
ion channel, the nicotinic acetylcholine receptor (nAChR) [9], a model of the insect GABAA-receptor
coupled ion channel was generated by homology modelling. Taking into account the known information
on Ala302 being involved in picrotoxinin binding, and the information on the likely properties of the
binding site from our Quasar study, it was possible to identify futher amino acid residues in the vicinity
of Ala302 which should contribute to the binding site.
Figure 6 shows a picture of the channel model in which only these residues are shown along with the
channel proteins' backbones. According to our results, the binding site should be located at the
cytoplasmic end of the channel pore, at the interface between the M2-helices of two protein subunits.
Assuming that the opening mechanism of the GABA-gated chloride channel is similar to that
reported for the nAChR-gated sodium channel [9], binding in this area would inevitably hinder
the movement of the M2 helix required to open the channel pore. This would be in agreement with
electrophysiological data which have clearly shown that picrotoxinin does not block open channels but
binds to and stabilizes a closed state of the channel [10],

•
1. Lin, J.H., Chen, I.W., Deluna, F.A. J. Pharm. Sci, 1994, 83 (12), 1741-1746.
2. Samdancioglu, S., Calis, S., Sumnu, M., Hincal, A.A. Drug Dev. and Ind Pharm., 2006 (In
press).
3. Samdancioglu, S., Calis, S., Sumnu, M., Hincal, A.A. Eur. J. Pharm. Sci. (Suppl), 2004,
23(1), S52, PO-55.
4. Samdancioglu, S., Calis, S., Sumnu, M., Hincal, A.A. 15^ International Symposium on
Microencapsulation, 18-21 September, 2005, Parma, Italy. Abstract.
5. Samdancioglu, S., Calis, S., Sumnu, M., Hincal, A.A. Pharmaceutical Sciences Fair &
Exhibition, 12-17 June, 2005, Nice, France. Abstract.
6. Samdancioglu, S., Calis, S., Kir, S., Sumnu, M. FABAD J. Pharm. Sci. 2003, 28 (4), 183-
192.

Figure 5 shows the two different pseudo-receptors with two seco-prezizaanes (insect-selective and nonselective),
viewed from an angle analogous to that in Figure 2. The overall characteristics of the binding
sites in insects and mammals are predicted to be quite similar with the major exception that the rat
receptor model possesses an additional site of polar interactions on the right side which interacts
specifically with the oxirane/oxetane oxygens of picrotoxinin- and anisatin-type convulsants, respectively.
The insect-selective compounds mentioned above do not possess such structure elements and form
favourable interactions with the insect receptor via other structure elements which explains their high
selectivity.
Housefly
Fig. 5. Pseudoanisatin (insect-selective;
A) and veranisatin A (non-selective, B)
shown inside the fly (top) and rat
(bottom) receptor surface models (from
[8]).
Receptor particles: Blue: Salt bridge
negative, SB-; Red: Salt bridge positive,
SB+, Yellow: Hydrogen bond acceptor,
Acc; Green: Hydrogen bond donor
(Don); Grey (small spheres):
Hydrophobic, neutral; Yellow-brown
(small spheres): Hydrophobic positive
(Hy-); Red-brown (small spheres):
Hydrophobic, negative (Hy-).
Note the interaction of the P-lactone ring
oxygen of A with the SB+ and Don
particles on the right side of the rat
receptor (arrows). Especially the SB+
particle is moved towards the oxetane
ring oxygen as compared to B.
On the background of these results, it appeared an interesting task to obtain further information on the
structure of the binding site in an atomistic context. Based on the reported structure of a closely related
ion channel, the nicotinic acetylcholine receptor (nAChR) [9], a model of the insect GABAA-receptor
coupled ion channel was generated by homology modelling. Taking into account the known information
on Ala302 being involved in picrotoxinin binding, and the information on the likely properties of the
binding site from our Quasar study, it was possible to identify futher amino acid residues in the vicinity
of Ala302 which should contribute to the binding site.
Figure 6 shows a picture of the channel model in which only these residues are shown along with the
channel proteins' backbones. According to our results, the binding site should be located at the
cytoplasmic end of the channel pore, at the interface between the M2-helices of two protein subunits.
Assuming that the opening mechanism of the GABA-gated chloride channel is similar to that
reported for the nAChR-gated sodium channel [9], binding in this area would inevitably hinder
the movement of the M2 helix required to open the channel pore. This would be in agreement with
electrophysiological data which have clearly shown that picrotoxinin does not block open channels but
binds to and stabilizes a closed state of the channel [10],

Fig. 6. Homology model of the insect GABAA-gated chloride channel showing the overall topology and a
molecular surface representation. The channel is shown as viewed from the inside of the cell membrane. The
surface is coloured by residue properties: Green: hydrophobic, blue: polar, positive charge; light blue: polar,
neutral. In the left picture, only residues in the vicinity of A302 (arrows) are shown. Likely binding cavities for
sesquiterpenoids are marked by asterisks in the right picture.
Similar studies into the structure of chloride channels of mammals and other organisms are in
progress. If should be possible to elucidate in detail the reasons for the differential binding
preferences of certain sesquiterpenoids which will be important with respect to the design of new
insecticides and antiparasitic compounds.
References:
1. Schmidt, T.J., Miiller, E„ Fronczek, F.R. J. Nat. Prod. 64, 411-414 (2001).
2. Schmidt, T.J. Current Org. Chem. 3, 577-605 (1999).
3. Hosie, A.M.. Ozoe, Y., Koike, K., Ohmoto, T„ Nikaido, T., Sattelle, D.B. Br. J. Pharmac. , 119,
1569-1576(1996).
4. Ozoe, Y., Akamatsu, M., Higata, T., Ikeda, I., Mochida, K„ Koike, K., Ohmoto, T., Nikaido, T.
Bioorg. Med. Chem., 6,481-492 (1998).
5. Schmidt, T.J., Okuyama, E., Fronczek, F.R. Bioorg. Med. Chem., 7, 2857-2865 (1999).
6. Kuriyama, T„ Schmidt, T.J., Okuyama. E., Ozoe, Y. Bioorg. Med. Chem., 10, 1873-1881 (2002).
7. Vedani, A., Dobler, M. J. Med. Chem. 45, 2139-2149 (2002).
8. Schmidt, T.J., Gurrath, M„ Ozoe, Y. Bioorg. Med. Chem., 12, 4149-4167 (2004).
9. Miyazawa. A., Fujiyoshi, Y„ Unwin, N. Nature, 423, 949-955 (2003).
10. Newland, C. F„ Cull-Candy, S.G. J. Physiol., 447, 191-213 (1992).

PL-43
BISPHOSPHONATE LOADED MICROPARTICULATE SYSTEMS FOR
IMPLANTATION
S. §amdancioglu, S. M. Sumnu
Hacettepe University, Faculty of Pharmacy, Department of Pharmaceutical Technology, 06100
Ankara, Turkey
Alendronate sodium (AS), an aminobisphosphonate, is a potent inhibitor of osteoclast-mediated
bone resorption and is used for the treatment of bone disorders, osteoporosis and Paget's disease of
the bone (1). The aim of this study was to formulate and evaluate the AS loaded microparticulate
systems for implantation in the treatment of osteolysis in orthopedics (by providing high local drug
levels while avoiding the systemic therapy) and also in the treatment of bone loss in the area of
dental prostheses. Therefore, chitosan microspheres were prepared by an emulsion polymerization
technique (2,3) and the PLGA microspheres were prepared using o/w single emulsion-solvent
evaporation (2,4) and w/o/w double emulsion-solvent evaporation method. The natural polymers,
sodium alginate (SA) and chitosan, were used for the preparation of SA and chitosan treated sodium
alginate (CTSA) beads (5). The beads were prepared by physical crosslinking of calcium ions to SA
polymer Particle size, loading efficacy, surface characteristics and in vitro release characteristics
were examined. AS amount in microparticles was determined by pre-column HPLC method using
9-fluorenylmethyl chloroformate derivatization at 266 nm (6). After the examination of the SEM
photographs of microspheres, chitosan microspheres were observed to have spherical structure and
smooth surface characteristics while PLGA microspheres were observed to have spherical porous
surface structure. Particle sizes of PLGA micropsheres are in the range of 33.98-74.36 pm and
chitosan microspheres are in the range of 109.28-111.28-pm. Loading efficacy was found to be
3.30% for chitosan microspheres and 7.70% for PLGA microspheres. It was observed that the 85%
of AS had been released at the end of the third day from chitosan microspheres, whereas 58% was
released at the end of the fifth day from PLGA microspheres. The surface morphology of SA beads
examined by SEM photographs indicated a porous, rough and also a rigid surface for SA
formulations. By increasing the amount of SA, the beads had a more spherical surface
characteristic. CTSA formulations had no spherical surface characteristic. However, CTSA beads
had some cracks and less porous structure on their surface than SA beads. Particle sizes of SA beads
are in the range of 919.06 ± 12.82-1867.64 ±15. 69 pm and CTSA beads are in the range of 1018.60
± 16.73- 2060.34 ±17. 23 pm. Loading efficacy was found to be 0.70-2.71 % for SA beads and
4.01-6.26 for CTSA beads. It was observed that the 95-100% of AS had been released at the end of
sixth hour from SA beads, whereas 100% was released at the end of the twentieth hour from CTSA
beads. At a higher SA concentration (6%), AS loading efficacy was found to have increased and
also the addition of chitosan increased the drug loading efficacy and particle size of the beads. The
microparticles in this study can provide the local delivery of AS to specific regions in dental and
orthopedic application. Local application of microparticles as implants may prevent bone resorption
during dental and orthopedic procedures.
Therefore, AS loaded microparticulate systems can be used for the treatment of osteolysis in
orthopedics. This study has shown that chitosan microspheres, PLGA microspheres and SA beads
can be employed as delivery systems for AS and it was considered that AS containing
microparticulate systems seem to be promising for local application in osteolysis.

1. Lin, J.H., Chen, I.W., Deluna, F.A. J. Phann. Sci, 1994, 83 (12), 1741-1746.
2. Samdancioglu, S., Calis, S., Sumnu, M., Hincal, A.A. Drug Dev. and Ind Pharm., 2006 (In
press).
3. Samdancioglu, S., Calis, S., Sumnu, M., Hincal, A.A. Eur. J. Pharm. Sci. (Suppl), 2004,
23(1), S52, PO-55.
4. Samdancioglu, S., Calis, S., Sumnu, M., Hincal, A.A. 15^ International Symposium on
Microencapsulation, 18-21 September, 2005, Parma, Italy. Abstract.
5. Samdancioglu, S., Calis, S., Sumnu, M., Hincal, A.A. Pharmaceutical Sciences Fair &
Exhibition, 12-17 June, 2005, Nice, France. Abstract.
6. Samdancioglu, S., Calis, S., Kir, S., Sumnu, M. FABAD J. Pharm. Sci. 2003, 28 (4), 183-
192.

PL-44
THE PHARMACY PROFESSION - A STRATEGIC APPROACH
I. Ustel
Freelance Consultant, Ankara, Turkey
Pharmacy is the profession that optimizes the drug utilization cycle with special emphasis on
continuous health promotion. "Reprofessionalization" of pharmacy should be achieved through
outcome-focused and evidence-based programmes built on the pillars of the cultural infrastructure
within the framework of "managed pharmaceutical care". Pharmacy profession should be tailored to
add more value to the quality-of-life of the public in a cost-effective manner. That is, the rational
for the pharmacy profession must be defined and demonstrated in the light of the factual
contribution to health as well as economic and psycho-social dynamics both at macro and micro
levels.
The competency-mix of the pharmacists should be geared towards future public demand to ensure
service delivery above the determined effectiveness and efficiency threshold. A paradigm shift is
needed to redefine the pharmacy profession for assuring the agility needed to realize the new roles.
Shared value framing, participative visioning, interactive mission development, integrated strategy
formulation, creative action plans, and focused benchmarking are the means to this end. Change
management methodology targeted at reforming pharmacy practice could only be accomplished by
approaching the process of reprofessionalization from the strategic perspective taking into account
both the strengths and weaknesses of the profession as well as the opportunities and threats
stemming from the interacting external factors. A spectrum of scenarios has to be developed to
prepare feasible contingency plans.
Leaders of pharmacy should built a competency base to frame realistic strategic models. Tools for
such modelling are Hoshin Kanri, quality function deployment, the servqual model, and the
balanced scorecard.

CLINICAL PHARMACY
WORKSHOP

CP-1
IDENTIFYING TARGET GROUPS FOR PHARMACEUTICAL CARE IN PRACTICE
R. P. Dessing
Ligusterweg 1, NL-2202-AD NOORDWIJK The Netherlands
Introduction:
A general approach to pharmaceutical care.
Keypoints in the pharmaceutical care process.
Is it possible to practise according to pharmaceutical care standards in community pharmacy
A close analysis of the pharmacy population: The needs and wants
Personal involvement in the PC process: Role and attitude of the pharmacist
Discussion points during the workshop:
- - Which tools are available in your pharmacy to execute a specific PC programme
- - Suggest a structure or organisational form that favours a possible success
- - Identify factors which obstruct such a process.
- - Is a drug just a commodity and have patients their 'own resposibility'
Reflection;
A specific medicine is not just a material product, but it is a composition of intellectual, material,
cultural, social and ethical elements. The pharmaceutical product represents a way of thinking and
today's public opinion about the good life. The medicine as a combination of technology, politics
and culture is the result of a process where many other responsible persons are involved. By
establishing a pharmaceutical practice, purchasing the medicine and dispensing it to clients, the
pharmacist proceeds on this track and becomes part of the original concept, as designed by
researchers, manufacturers, politicians and business managers. The responsibility which extends
from the acting person (i.e. the pharmacist) covers both the benefits and possible risks that are
connected with the actual use of medicines. This responsibility will not change by promoting your
practice as 'commercial' or as 'caring' because responsibility is measured according to the
compliance to certain basic principles.
85

CP-3
SYSTEMATIC APPROACH TO DRUG-THERAPY
F.V. Izzettin
Clinical Pharmacy Division, Department of Pharmacology, Faculty of Pharmacy, University of
Marmara, 81010, Haydarpa§a, istanbul, Turkey
In this short lecture, "the systematic approach to drug-therapy" will be presented. Systematic
approach to drug-therapy consists of the following steps:
1. Data collection: The pharmacist takes the medical as well as the medication history of the
patient.
2. Identification of the problem(s) (Is the problem drug-related or not).
3. Identification of the therapeutic end-points.
4. Construction of alternative therapeutic plans (including pharmacological and nonpharmacological
therapy options).
5. Choosing the most appropriate therapeutic plan for the individual patient.
6. Implementation of this therapeutic plan in the total medical care of the patient.
7. Monitoring the efficacy and safety of the implemented therapeutic plan.
8. If the plan fails in efficacy and/or safety, the pharmacist considers the next best alternative
for the individual case.
8

INDUSTRIAL PHARMACY
WORKSHOP

IP-2
BiR YERLi FIRMANIN IHRACAT GiRi§IMLERiNE RUHSATLANDIRMA
ACISINDAN BAKI§
S. Bingol
Mustafa Nevzat Ila San. A.§., istanbul, Turkey
Yerli ila firmalarinin ihracat giri§imleri gefmi yillarda daha ziyade Tiirki Cumhuriyetleri, Kuzey
Afrika iilkeleri gibi konvansiyonel pazarlar olarak tanimlanan ulkelerle kisith kahrken gunumiizde
AB ulkeleri, ABD, Kanada gibi regiile pazarlara dogru geni§lemeye ba§lamitir. Bu geni§lemede
Turk Ila9 Endiistrisi'nin siirekli yatirimlarla ula§mi§ oldugu ytiksek teknolojik diizey , Giincel Iyi
ila Uretimi kurallarina uygunluk ifinde Uretim, kaliteye verilen onem ve bilgi birikimi biiyiik rol
oynamaktadir.
Bu sunumda yerli ilaf firmalari ifin yeni bir pencere olan ABD pazari uzerinde odaklanilacak;
bu iilkeye ihracat yapmayi hedefleyecek bir ila firmasinin ruhsatlandirma afisindan saglamasi
gereken hususlar ozetlenecek; FDA'in jenerik bir uriine pazarlama yetkisi verebilmek i9in
kar§ilanmasini bekleyecegi kriterler uzerinde durulacaktir.
92

IP-3
BiR YERLI FIRMANIN IHRACAT GIRl§iMLERiNE RUHSATLANDIRMA
A^ISINDAN BAKI§
A. Angin
Abdi Ibrahim Ila San. A.§., Istanbul, Turkey
Yerli ila firmalarimn diinyaya a9ilmadaki en onemli yolu iiretim yapilan tesislerin, ulkelerin GMP
gerekliliklerini kar§ilayacak standartlari saglamasindan ve devaminda da ruhsatlandirma prosediirii
iin istenilen formatta ba§vurunun yapdmasindan ge9mektedir. Yerli ila firmalari fogunlukla
iilkelere kendi irketlerini kurarak, bir distributor aracihgi ile veya fason uretici olarak girmeyi
tercih etmektedirler ve buna gore de iilkede ruhsatlandirma afisindan izlenen yol degi§ebilmektedir.
Yerli ila9 firmalarimn ihracat giri§imleri artik sadece CIS ulkeleri veya Tiirki Cumhuriyetlerle
sinirli kalmamakta, Avrupa ve Amerika gibi regiilasyonlan geli§tni§ iilkelere dogru hizla
geni§lemektedir. Bu da Turk ila Endiistrisi'nin 1984 yilinda GMP Yonetmeligi'nin yiiriirlUge
girmesinden gunumiize kadar surdiirdugii yatirimlarla AB ulkeleri ile kiyaslanabilir teknolojik
diizeye ula§masinin bir sonucudur. Bunu takiben ruhsatlandirma yonetmeliginin Avrupa Birligi'ne
uyumu, Turkiye'de geli§tirilen ve ruhsat dosyasi oluturulan Uruniin standartlanni da Avrupa
standard arina taimi§tir.
93

IP-3
BiR YERLI FIRMANIN iHRACAT GIRi§iMLERiNE RUHSATLANDIRMA
A^ISINDAN BAKI§
A. Angin
Abdi Ibrahim ila San. A.$., istanbul, Turkey
Yerli ila firmalarinin dUnyaya a9ilmadaki en onemli yolu iiretim yapilan tesislerin, iilkelerin GMP
gerekliliklerini kar§ilayacak standartlari saglamasindan ve devaminda da ruhsatlandirma prosedurii
i

IP-
AVRUPA RUHSATLANDIRMA KRITERLERINi SAGLAMADA TURK iLA£
FiRMALARININ KAR§ILA§ABiLECEKLERI ZORLUKLARLA BA§ETME
STRATEJlLERl
Z. Ulusoy
Nobel Ila San. ve Tic. A.§., istanbul, Turkey
2005 yilinin Ekim ayinda balayan Avrupa Birligi (AB) ile miizakere slireci dogrultusunda, AB'ye
uyum 9ali$malarinin hizlandigi Turkiye'de, ila ruhsatlandirma prosediirleri de 2006 yili itiban ile
bir o kadar AB'ye yakinlatirilmi!jtir. Bu sunumda ama5lanan, Turkiye'de AB'ye uyum
9ei\e\esinde ger9ekle§tirilen bu degi^ikliklerin, Turkiye Ila Sanayi'nin AB pazarina giri§ siiratini
belirleyen ruhsatlandirma stratejilerinde neleri degi§tirdigini, AB'nin kendi i9inde yaadigi
degi§ime de iik tutarak mercek altina almaktir.
9

IPiDARI
KURUMLARDA §EFFAFLIK: ILAC SANAYi BOYUTU
C. Buharali
Istanbul Ekonomi Dani§manhk Ltd. $ti., istanbul, Turkey
Ila sanayi ulusal ekonomilerde onemli bir yere sahiptir. Onemi sadece yarattigi ekonomik
hareketlilikten kaynaklanmamaktadir. ila sanayi insanlarin sagliklarini dogrudan etkileme
olanagina sahip oldugu ifin onemlidir. Bir ilacin varligi veya yoklugu bazen yaamla olum
arasindaki ince

ABSTRACTS OF
ORAL
PRESENTATION

0-2
PATIENT-COUNSELLING PRACTICES AND ATTITUDES OF PHARMACY STAFF
AND PHARMACY STUDENTS ON EMERGENCY CONTRACEPTION PILLS
1 2 2 2 2 2 . 1
S. Apikoglu Rabus , A. Dinfer , F. Donmez , A. Ekiz , O. Giiler , B. Reiazi , F.V. Izzettin
1 2
Clinical Pharmacy Department, Clinical Pharmacy Student Research Group, Marmara University
Faculty of Pharmacy, Istanbul, Turkey
Background and Objective: Availability of emergency contraception pills (ECPs) has brought the
responsibility of proper and complete patient counselling in order to prevent misuse. This survey
was conducted to assess the present status of ECP-counselling practices and the attitudes of
community pharmacists, pharmacy technicians and final-grade pharmacy students.
Design: For this cross-sectional study, a survey was designed based on the work by Aneblom et al
(1). It was consisting of four domains: "reproductive health", "information", availability" and "risk
behaviour". 5-point Likert scales were used as respond scales. A total of 56 pharmacies representing
most of the geographic regions of Istanbul were visited; information was gathered from 40
pharmacists and 16 pharmacy technicians. Besides, 70 final-grade pharmacy students were also
invited to fill in the questionnaire. Setting: Marmara University, Faculty of Pharmacy, Clinical
Pharmacy Department Main Outcome Measures: Scores of counselling on various ECP-related
issues. Scores of attitudes on ECPs. Results: As the ECP-counselling practices, the pharmacists
more often counselled on side-effects, pregnancy tests and others measures of contraception than
the pharmacy technicians; while the pharmacy technicians reported on the timeframes more often
than the pharmacists. The "reproductive health" domain of the questionnaire did not differ much
between the three groups. All the groups agreed that ECPs were positive and ethical; while, they
were all uncertain (neither agree nor disagree) that the young ones could take the responsibility for
the ECP use. Similarly, the pharmacists and the pharmacy-technicians were uncertain that ECP
would increase women's control on reproductive health. All groups agreed that information on ECP
should be supplied to everyone regardless of gender and age. The "availability" domain revealed
different results. Pharmacists and pharmacy students were uncertain about the positivity of nonprescription
availability of ECPs. The pharmacists agreed that ECPs should only be dispensed with
prescription. All the groups agreed that ECPs should only be dispensed to those over 18.
Conclusions: Pharmacists, pharmacy technicians and pharmacy students agreed that ECPs were
positive and everyone should be informed about them; while they agreed to restrict their dispensing.
1. Aneblom G, Lundborg CS, Carlsten A, Eurenius K, Tyden T., Patient Educ Couns 2004;
55:129-35.
100

0-3
QUALITY CONTROL IN ANALYTICAL RESULTS
N. E. Basci
Department of Analytical Chemistry, Faculty of Pharmacy, Hacettepe University, 06100, Ankara,
Turkey
The value of chemical measurements depends upon the level of confidence that can be placed in the
results. Quality control is one of the most important steps to ensure the reliability of the analytical
results. The chemical testing community is adopting quality principles which, whilst not actually
guaranteeing the quality of the data produced, increases the likelihood of it being based and fit for
its intended purpose. Those most widely recognised and used in chemical testing fall into three
groups and are applied according to a laboratory's individual needs. The first step is the use of
methods validated to guarantee that the requirements for a specific intended use are fulfilled. The
second level of quality assurance is the implementation of an internal quality control program (IQC)
to monitor performance of the procedures when they are used during routine work. Finally, the
highest level of quality assurance is the participation in external quality assessment programs. IQC
is one of the most important elements contributing to quality assurance in the laboratory. Since in
general quantitative methods are the most used, guidelines and recommendations provided by the
literature are usually related to IQC in this type of analytical procedures. Different IQC parameters
have been defined according to the aim of the method (qualitative or quantitative). IQC parameters
for chromatographic methods and the acceptance criteria used to check the IQC data obtained are
described and discussed. Thus, IQC parameters and acceptance criteria to assess the quality control
data obtained in chromatographic procedures have been presented. These criteria can be also
applied to other fields of analytical and clinical chemistry and toxicology. The use of control charts
is the most extended procedure to establish an internal quality system for quantitative methods. The
most important control rules used for the interpretation of these control charts are based on the use
of control limits. Strategies for the implementation of an IQC in a laboratory have been presented,
taking into account recommendations and requirements described by official organizations and
quality standards.
11

0-6
THE PHENOLIC COMPOUNDS OF POSIDONIA OCEANICA (L.) DELILE BY
DIFFERENT PARAMETERS
M.Z. Haznedaroglu, U. Zeybek
Ege University, Faculty of Pharmacy, Dept. of Pharmaceutical Botany 35100 Bornova -
Izmir/Turkey
Posidonia oceanica (L.) Delile (Posidoniaceae) is a phanerogam widely distributed in the Aegean
and Mediterranean seas. The phenolic compounds of the leaves were investigated previously
(1,2,3,4, 5). Major components of the plant obtained from other sources had been tested for various
activities such as antiHIV, anticarcinogenic, immunostimulant, antiulcerogenic, antioxidant and
antidiabetic with promising results (3). In this study, twenty six phenolic compounds were analysed.
Concentrations of these compounds during different seasons and at different times of the day were
evaluated in samples from different regions growing in two depths. The leaves, the roots and the
rhizomes were examined separately. The quantification of phenolic acids including chicoric acid
was determined by HPLC. As a result, phloroglucinol (highest quantity in root, summer, Southwest
Aegean Sea, 10 m depth, evening parameters); ferulic, caffeic and coumaric acids (highest
concentration in older leaves, autumn, West Aegean Sea, 10 m depth, evening parameters); chicoric
acid (highest quantity in young leaves, autumn, Southwest Aegean Sea, 15 m depth, morning
parameters) were found to be the most important phenolics in the plant.
Authors are thankfull to TUBITAK and EBILTEM due to their support by a research project
TBAG-2205(102T103) and 2002/BIL/039; and to E.U ARGEFAR for laboratory conditions.
1. Agostini, S., Desjobert, J. M., Pergent, G. (1998). Phytochemistry 48:611-17.
2. Cuny, P., Serve, L., Jupin, H., Boudouresque, C.F. (1995). Aquat. Bot. 52:237 242.
3. Dumay, O., Costa, J., Desjobert, J. M., Pergent, G. (2004). Phytochemistry 65: 3211-3220
4. Haznedaroglu, M. Z. (1999). Ege Denizi'nde yayilis gosteren Posidonia oceanica (L.)
Delile bitkisi uzerinde arastirmalar. Master Thesis Ege U. Saglik Bilimleri Enst. Bornova -
Izmir.
5. Serve, L. Piovetti, L., Combout, G. (1984). GIS Posidonia International Workshop 137-144.
10

0-7
NOVEL ELECTROCHEMICAL REDOX INDICATOR, ECHINOMYCIN FOR
DETECTION OF DNA HYBRIDIZATION ON GOLD SURFACES
H. Karadeniz 1 , A. Erdem 1 , F. Jelen 2 , M. Ozsoz', E. Palecek 2,
'Analytical Chemistry Department, Faculty of Pharmacy, Ege University, 35100 Bornova, Izmir,
Turkey, institute of Biophysics, Academy of Sciences of the Czech Republic, Kralovopolska 135,
612 65 Brno, Czech Republic
Many electrochemical studies are dealing with the interactions of DNA with redox indicators
characterizing the structure of immobilized DNA (1-5). These indicators are usually represented by
small electroactive substances such as organic dyes, (1,3), anthracycline antibiotics (6), cationic
metal complex compounds with aromatic ligands, e.g. [Co(phen) 3 ] 3+ (7), or bis-intercalators (8,9).
The DNA indicators interact in a different way with single-stranded (ss) and double-stranded (ds)
DNA providing different electrochemical responses for dsDNA and ssDNA. The performance of
such redox indicators frequently depends on the nature of the electrode used as a transducer in the
sensor. In this report, a bis-intercalator echinomycin (ECHI) and a simple intercalator [Co(phen)3] J+
were used as a novel electrochemical redox indicators to detect DNA hybridization at gold
electrodes (AuE). In order to minimize the nonspecific adsorption of oligonucleotides (ODN), the
thiol-derivatized oligonucleotides were immobilized onto AuE in the first step, and the exposition
of 6-mercapto-l-hexanol followed in the second step of this procedure (10). In this arrangement
good reproducibility and discrimination between single stranded probe and double stranded hybrid
DNA were obtained. In addition, DNA single-base mismatch (DNA point mutation) was detected
by means of ECHI. The detection of DNA hybridization using ECHI and [Co(phen) 3 ] 3+ as redox
indicators were compared. Our results show that both ECHI and [Co(phen)3] 3+ enable selective
monitoring of DNA hybridization at AuE surface. Moreover, using ECHI as a redox indicator we
were able to sensitively detect single-base mismatch in the DNA duplex.
1 - Erdem, A., K. Kerman, B. Meric, U. S. Akarca & M. Ozsoz. Electroanalysis 11, 586-587 (1999).
2. Erdem, A., K. Kerman, B. Meric & M. Ozsoz, Electroanalysis 13, 219-223 (2001).
3. Erdem, A., K. Kerman, B. Meric, U. S. Akarca & M. Ozsoz, Anal. Chim. Acta 422,139-149 (2000)
4. Palecek, E. & M. Fojta, Anal. Chem. 73, 74A-83A (2001)
5. Palecek, E. & F. Jelen, in (Palecek, E., Scheller, F., Wang, J., Eds.) Perspectives in Bioanalysysis. Vol. 1
Electrochemistry of nucleic acids and proteins. Towards electrochemical sensors for genomics and proteomics,
Elesevier, New York, 2005, pp. 74-173
6. Wang, J., M. Ozsoz, X.H. Cai, G. Rivas, H. Shiraishi, D.H. Grant, M. Chicharro, J. Fernandes & E. Palecek,
Bioelectrochem. Bioenerg. 45, 33-40 (1998)
7. Millan, K. M. & S. R. Mikkelsen, Anal. Chem. 65, 2317-2323 (1993)
8. Jelen, F., A. Erdem & E. Palecek, J. Biomol. Struct. Dyn. 17, 1176-1177 (2000)
9. Jelen, F„ A. Erdem & E. Palecek, Bioelectrochem. 55, 165-167 (2002)
10. Heme, T. M. & M. J. Tarlov, J. Am. Chem. Soc. 119, 8916-8920 (1997)
This work has been supported by the Turkish Academy of Sciences in the framework of the Young Scientist Award
Program (KAE/TUBA-GEBIP/2001-2-8) and the grant A100040602 from the Grant Agency of the Academy of
Sciences of the Czech Republic.
1

0-8
PREVENTION AND TREATMENT OF SEVERE FUNGAL INFECTIONS IN HAEMATO-
ONCOLOGY: SINGLE CENTRE EXPERIENCE
1 2 1 1 3
H. Koblihova , E. Faber , V. Popelkova , J. Vlcek , V. Scurti for the SFIM working group
1
Department of Social and Clinical Pharmacy, Faculty of Pharmacy, Heyrovskeho 1203, 500 05
2
Hradec Kralove, Czech Republic, Department of Hemato-Oncology, University Hospital, I.P.
3
Pavlova 6, 775 20 Olomouc, Czech Republic, Centro Studi SIFO, Consorzio Mario Negri Sud, Via
Nazionale 8/A, 660 30 Santa Maria Imbaro (CH), Italy
Invasive fungal diseases are typically difficult to prevent, diagnose, and treat, particularly among
patients with prolonged neutropenia and severe graft-versus-host disease. This project is an ESCP
Hospital Pharmacy Project "Severe Fungal Infections Management and Outcome": it is an observational
study to evaluate the management of haemato-oncology patients with or at risk of severe fungal
infections and to compare the different therapeutic approaches throughout Europe. National data from
one centre in Czech Republic are presented. The study was designed as 6-months observational
prospective study and was settled at the department of Haemato-Oncology (42 beds), University
Hospital, Olomouc, Czech Republic. Epidemiological, clinical and therapeutic information using
patient's medical record was collected at recruitment i.e. 1 st day of using antifungal drug, after 7 and 15
days and at discharge. Invasive fungal infections (IFI) were defined according to an international
classification [1]. An electronic archive built using File MakerPro6 was available to input and analyse
the data. From March to September 2005, 49 (25 male) patients (pts) have been included in the survey.
Patients' median age was 49 years (range 22-75), median hospitalisation period was 22 days (2-55).
They had all haematologic malignant disease: mainly leukaemia (20pts), lymphoma (19), myeloma (6)
and others (4). 14 pts received previously stem cell transplantation (12 allogeneic). All patients received
antifungal drugs as prophylaxis. No IFI was observed during the study. The most administered
antimycotic was fluconazole (59%), followed by itraconazole, voriconazole and ketoconazole (31%,
8%, and 2% respectively). The antimycotic therapy was changed to 22 pts (45%) during hospitalization
(for some patient more than once) and suspended to 21 pts. None of the pts received combination of
antimycotics. Overall 82 various changes (substance, dosage or route of administration) in antifungal
therapy were noticed. Reasons for changing the therapy were: improvement of clinical condition
(21.9%), non-toleration or occurrence of adverse reactions (19.5%), non-response to the administration
(18.3%), laboratoiy results obtaining (15.9%), shift due to the discharge (13.4%) and worsening of
clinical condition (11.0%).The data in our hospital documented the use of antimycotics as prophylaxis
in patients with haemato-oncology malignancies. Despite no observed IFI, several reasons for changing
the antifungal therapy during hospitalization were found. Management of patients with or at risk of
severe fungal infections is a complex issue; it has to be targeted and reflect the evolution of patient's
clinical condition.
1. Ascioglu S., Rex H., de Pauw B., et al. Defining opportunistic invasive fungal infections in
immunocompromised patients with cancer and haematopoietic stern cells transplants: an international
consensus. Clinical Infectious Diseases 2002 (34): 7-14.
10

0-9
OUTLIERS AND PHARMACOKINETIC VALIDATION OF DATA IN
BIOEQUIVALENCE STUDIES
C. Mircioiu, D. Miron, I. Mircioiu, F. Radulescu
University of Medicine & Pharmacy "Carol Davila", Bucharest, Romania
Validation of bioanalytical and pharmacokinetic data and implicit elimination of outliers
results from combined biological, pharmaceutical and statistical points of view. All
statistical methods are based more or less on comparison of deviations from mean with
standard deviation. The specific aspect of the problem appears from the fact that main
component of deviations issues in almost all cases from biological variability. At this
chapter we have to take into consideration intravariability and intervariability of in vivo
dissolution, of absorption, of metabolism and elimination. Since biological variability
component is greater then all other factors, apparently would be not possible to separate
pharmaceutical technology factors bias and variability from biological factors. In a crossover
experiment, following an intelligent combination of variables is possible to undertake
this separation. The method is less applicable in the analysis of outliers. The paper
proposes some algorithms applicable in such an analysis, starting from a fundamental
characteristic of biological variability: though high, this variability has a continuous time
course. Sudden changes are not coming form biological factors. As concrete cases are
presented the analysis deviations inside pharmacokinetic curves, deviations between curves
of the same subject in different periods of the experiment, deviations between pairs of
corresponding points and deviations between global pharmacockinetic parameters (areas
under curves), deviations between subjects etc. A special presented case refers to drugs
with active metabolites, to comparisons between pharmacockinetics of parent drug and
metabolites. All examples are taken from experiments undertaken by authors, finalization
of paper being a guide for deciding on the elimination of some data as outliers, based both
on statistical and pharmacokinetic arguments.
1

O-IO
ANTIOXIDANT AND ANTICHOLINESTERASE EVALUATION OF SELECTED
TURKISH SALVIA SPECIES
L Orhan 1 . M. Kartal 2 , Q. Naz 3 , A. Ejaz 3 , G. Yilmaz 4 , Y. Kan 5 , B. §ener\ M. I. Choudhary 3
'Department of Pharmacognosy, Faculty of Pharmacy, Gazi University, 06330 Ankara, Turkey,
^Department of Pharmacognosy, Faculty of Pharmacy, Ankara University, 06330 Ankara, Turkey,
3 H.E.J. Research Institute of Chemistry, University of Karachi, 75270 Karachi, Pakistan,
department of Pharmaceutical Botany, Faculty of Pharmacy, Ankara University, 06330 Ankara,
Turkey, ^Department of Field Crops, Faculty of Agriculture, Selcuk University, 42070 Konya,
Turkey
Since Salvia species (Lamiaceae) have been recorded to be used against memory loss in European
folk medicine, we herein examined in vitro anticholinesterase and antioxidant activities of 56
extracts prepared with petroleum ether, chloroform, ethyl acetate and methanol extracts obtained
from fourteen Salvia species (S. albimaculata Hedge & Hub, S. aucheri Bentham var. canescens
Boiss & Heldr, S. candidissima Vahl. ssp. occidentalism S. ceratophylla L., S. cryptantha Montbret
& Bentham, S. cyanescens Boiss & Bal., S. frigida Boiss, S. forskahlei L., S. halophila Hedge, S.
migrostegia Boiss & Bal., S. multicaulis Vahl., S. sclarea L., S. syriaca L., S. verticillata L. ssp.
amasiaca) growing in Turkey. The antioxidant activity were assessed by both chemical and
enzymatic methods against l,l-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging method and
xanthine/xanthine oxidase (XO) system generated superoxide anion radical inhibition using
microplate-reader assay. Anticholinesterase effect of the extracts was tested against both
acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) at concentrations of 0.2 and 1
mg/ml using microplate-reader assay based on Ellman method. Our data indicates that nonpolar
extracts of Salvia species for anticholinesterase activity and the polar extracts for antioxidant
activity are worth further phytochemical evaluation for identifying their active components.
108

0-11
IMPROVEMENT OF THE DISSOLUTION RATE OF NITRENDIPINE USING A PULSE
COMBUSTION DRYER SYSTEM
L. Wang 1 ' 2 , F.-De.Cui 2 , H. Sunada 1
1 Faculty of Pharmacy, Meijo University, 150 Yagotoyama, Tempaku-ku, Nagoya 468-8503, Japan,
2 Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang 110016, P.R. China
Nitrendipine (NTD), a dihydropyridine calcium channel blocking agent, is used to treat a variety of
cardiovascular disorders. Due to its low aqueous solubility (about 2pg/mL), leading to the poor
absorption of nitrendipine after oral administration, we prepared solid dispersions of nitrendipine to
improve its dissolution rate and solubility. Solid dispersions (SD) of nitrendipine (NTD) were
prepared by the pulse combustion dryer system, HYPULCON using AEROSIL 200 and Tween 80
as carriers. The physicochemical properties were investigated and the mechanism of SD formation
and dissolution rate improvement was discussed. Powder X-ray diffraction and DSC evaluation
showed that NTD in SDs was dispersed in an amorphous state when the concentration of Tween 80
solution was 5%. FT-IR Spectroscopy obtained with the SDs indicated the presence of hydrogen
bonding between the drug and carriers. The dissolution rate of NTD in SDs with Tween 80 was
remarkably improved compared with original drug and a high supersaturation level of the drug was
observed. At the end of dissolution test (60min) for SD prepared with 5% Tween 80 solution, the
concentration of NTD were 23.73pg/ml, about 73 times that of original NTD crystals. This may due
to the amorphization of the drug and the solubilization of Tween 80. It was proved that
HYPULCON can be used to prepare solid dispersion particles with improved properties.
10

0-12
DEVELOPMENT AND CHARACTERIZATION OF NOVEL SERS SUBSTRATES
U. Tamer 1 , C. Shannon 2
'Department of Analytical Chemistry, Faculty of Pharmacy, Gazi University, 06330, Etiler, Ankara,
Turkey,
2 Department of Chemistry and Biochemistry, Auburn University, Auburn, Alabama,
36849, USA
Noble metallic colloid nanoparticles (e.g. gold, silver) have been successfully applied to the label
technologies because of their easily controllable size distribution, long-term stability, and
biocompatibility. This work presents an effective and simple way to produce desired size and shape
nanoparticles for Surface Enhanced Raman Scattering. Highly sensitive nanoparticles can be used
for SERS immediately after preparation. Nanoparticles have been characterized UV-vis, SEM,
AFM. The SERS activity of these nanoparticles was tested using various analytes. In addition,
interaction between biomaterials and nanoparticles was also investigated. Treatment of biomaterials
with nanoparticles not only enhances the normal Raman signal by several orders of magnitude but
also further reduces the fluorescence backgrounds via interaction between the analyte and SERS
substrate. Some of the challenges facing this detection method will be also discussed.
10

0-13
NOVEL TARGETS IN ANTIMALARIAL DRUG DISCOVERY: NATURAL PRODUCTS
INHIBITING THE ENZYMES OF PLASMODIAL TYPE II FATTY ACID BIOSYNTHESIS
(FAS-II)
D. Ta§demir
Centre for Pharmacognosy and Phytotherapy, School of Pharmacy, University of London, London
WC1N 1AX, UK
Malaria is the most prevalent human parasitic disease, which claims the lives of at least 3000
children each day. Treatment of Plasmodium falciparum malaria, the most widespread and the
deadliest form of malaria, has depended for decades on the use of chloroquine or the antifolate
combination pyrimethamine-sulfadoxine. However, the spread of drug-resistance to these and all
other widely available and affordable antimalarial drugs underlines the need for new lead compounds
active against novel targets. The completion of the P. falciparum genome sequencing project (1), at
the same time with that of human and Anopheles mosquito, has revealed a number of biochemical
processes previously unknown to Plasmodium. This includes the shikimate-, non-mevalonate
isoprene- and type II fatty acid (FAS-II) biosyntheses, which take place in a recently discovered
plastid-like organelle (apicoplast) in Plasmodium (2). The plasmodial FAS-II is similar to that found
in plants and bacteria, and characterized by the presence of a number of monofunctional enzymes. In
contrast, humans use a type-I FAS system where each enzymatic reaction is achieved by a certain
domain of a large, single protein. Because of this difference and the vital role of fatty acids, the
enzymes involved in fatty acid biosynthesis of P. falciparum emerge as valuable targets for malaria
drug discovery. ZyFabG (ketoacyl-ACP reductase), PyFabI (/ra«-enoyl-ACP reductase), and P/FabZ
(beta-hvdroxyacyl-ACP dehydratase) are three key enzymes of plasmodial FAS-II system. Our
earlier studies have focused on the identification of both plant- and marine-derived small-molecule
inhibitors of recombinant P^Fabl enzyme (3,4) that has been heterologously expressed (in E. coli)
and purified. After the discovery of a flavonoid as the first natural inhibitor of this enzyme (3), we
have evaluated a large flavonoid library for their inhibitory activity against /yFabl, as well as
/^FabCi and PfFabZ. Several polyhydroxylated flavonoids and tea catechins bearing a galloyl
function at C-3 position have appeared as very potent inhibitors of all three enzymes. This is a very
unique aspect, as it is quite unlikely for the malaria parasite to develop resistance to the drug by
introducing mutations at all three enzymes simultaneously. This lecture will summarize our efforts to
discover new-age antimalarial agents from Turkish plants and marine organisms that specifically
target the individual enzymes of plasmodial FAS-II system.
1. Gardner, M.J. et al. (2002) Nature 419: 498-511.
2. Fadden, G.I. et al. (1996) Nature 381: 482.
3. Kirmizibekmez, H. et al. (2004) Planta Med. 70: 711-717.
4. Tasdemir D. et al. (2005) Phytochemistry 66: 355-362.

0-14
ANTIPROLIFERATIVE EFFECT OF SOME DITHIOCARBONIC ACID DERIVATIVES
(XANTHATES) ON DIFFERENT TUMOR CELL LINES
M. Topalov 1 , G. Momekov 1 , S. Yanev 2
'Department Pharmacology and Toxicology, Faculty of Pharmacy, Medical University - Sofia and
2 Department of Drug Toxicology, Institute of Physiology, BAS, Sofia, Bulgaria
Xanthates (alkyl and aryl derivatives of dithiocarbonic acid, ROCS2K) are well known metal ions
chelating agents with variety of biological properties as antitumor, antiviral and antioxidant effects.
The cytotoxic effect of different xanthates (R= 2, 8, 12, cyclohexyl and tricyclodecan-9-il (D609)
on malignant human cell lines alone or in combination with fatty acid and epirubicine were studied.
The following cell lines were used: SKW-3, HL-60, HL-6O/DOX, K-562, LAMA-84; human
leucocytes. Cells were seeded at 3x10 5 cells/ml in growth RPMI medium supplemented with 10%
FCS and L-glutamine (2.5 mg/ml), incubated in humidified atmosphere containing 5%C02 and
95% air at 37°C. Cell viability was checked by MTT test after 72h incubation without or with
different concentrations of xanthates; the oligonucleosomal DNA fragmentation (a key hallmark
feature of apoptosis) was determined by horizontal agarose gel electrophoresis of DNA, ethidium
bromide staining and UV-transillumination. Xanthates tested in this study exhibited pronounced
cytotoxic effect against different human tumor cell lines. Xanthate cytotoxicity depends on: the cell
line - most pronounced against SKW-3 > LAMA-84 > HL-60; less effective against K-562 and
HL6O/DOX. Xanthates structure (IC 50 ): most active are D609 (23 - 200 pM); C12 (50 - 160 pM)
and C8 (32 - 120 pM); CYCLO (65 - 175 pM) and C2 (70 - 200 pM) are less active. K-562 and
HL-6O/DOX are less responsive to xanthate cytotoxicity (only D609 and C12 are active in higher
concentrations). HL-60 lauric acid cytotoxicity (IC 50 = 150 pM) was potentiated by non toxic D609
concentration (25 pM). D609 and C12 applied in not toxic concentrations (6.25 to 25 pM) to HL-
6O/DOX cell line could not potentiated the cytotoxic effect of epirubicine. C8 (in 10 and 20 pM
concentrations for 24h) induced apoptosis in SKW-3 cells. Xanthates were not toxic to human
leucocytes in concentrations up to 200 pM. In order to elucidate the mechanistic peculiarities,
underlying the differential susceptibility of the distinct human cell lines tested, we evaluated their
effects on: NO production with or without LPS stimulation; ROS (H 2 0 2 ) production before or after
PMA stimulation; phosphatidylcholine-phospholipase C and sphingomyelinase activity; xanthates
metabolism.

0-15
ANTIAGING SKIN CARE PRODUCTS- AN UPDATE
Y. Yazan
Anadolu University, Faculty of Pharmacy, Department of Pharmaceutical and Cosmetic Technology,
Eski$ehir, Tiirkiye
During the past few years, many novel and effective skin care therapies have become available to
correct and protect against the effects of skin aging. Purposes of the antiaging skin care products are:
- To maintain the long endurance of healthy and good-looking skin,
- To reduce or slow down the symptoms of aging,
- To solve some skin disorders.
Unwelcome changes in aging skin are brought about by the relentless pull of gravity and cumulative
damage to DNA, collagen and cell membranes by free radicals produced from normal cellular
metabolism, environmental elements and exposure to solar radiation. Almost all of the currently
marketed skin care products incorporate one or more of the following agents: Sunscreens, Moisturizers,
Alpha-Hydroxy Acids (AHAs), Beta-Hydroxy Acid (BHA), Retinoids, Retinol, Furfuryladenine,
Antioxidants, Human Growth Factors, Bleaching Agents, Copper and Botanical Agents. Although it is
almost impossible to eliminate exposure to UV radiation entirely, it can be minimized by proper use of
an effective sunscreen in all cosmetic products. Moisturizers usually incorporate emollients to smooth
the skin surface by working their way into the non-living outer layers of the skin, filling spaces between
the layers and lubricating; and humectants to help skin cells absorb and retain moisture in these layers.
AHAs are commonly used at low concentrations in cleansers, moisturizers and toners, and at higher
concentrations as light peel solutions. There is only one clinically important BHA, salicylic acid. Retin-
A (retinoic acid or tretinoin), originally approved for the treatment of acne, has been shown to be
effective for the prevention and treatment of sun-induced skin aging. Vitamin A, or retinol, is converted
to retinoic acid (tretinoin, Retin-A) in living cells. When applied to skin, retinol penetrates better than
retinoic acid and does not produce the same irritating effects. Long-term treatment with furfuryladenine
has been shown to improve hydration and texture of the skin, as well as reduce fine wrinkles and
improve blotchiness. Antioxidants are often incorporated into skin care products to protect the skin from
free radical damage produced by normal aging, pollution, and UV radiation from sun exposure.
Harvested as a by-product of tissue-cultured human skin, various human growth factors have been
shown to reduce the number and depth of wrinkles and fine lines, as well as improve skin texture and
elasticity when used over time. Among the most commonly used agents for "bleaching" brown marks,
liver spots, melasma, etc are hydroquinone and kojic acid. Copper which is found in most biological
systems, acts as a cofactor in collagen and elastin production, production of new blood vessels and
deposition of glycosaminoglycans in the skin. A variety of botanical agents may be incorporated in skin
care preparations. These agents are usually labeled as "extracts" and although there may be a rationale
for their use based on folklore or in some cases in vitro studies, in most cases there is little if any
scientific evidence that they are any more efficacious than placebo. Guidance and maintenance of
accurate delivery systems are needed besides the active agents developed. As a conclusion, large variety
for the selection of raw materials and the production method, size of the delivery system and trigger for
release leads to the need for optimum design for the optimum cosmetic efficiency and safety.
113

ABSTRACTS OF
POSTER PRESENTATIONS
- I -

P-2
EFFECT OF RALOXIFENE TREATMENT ON THE LEVEL OF APE/REF-l mRNA
EXPRESSION IN BRAIN CORTEX OF OVARIECTOMIZED FEMALE RATS
G. Durmaz. S.E. Turunc, A. Yalcin
Department of Biochemistry, Faculty of Pharmacy, Ege University Bornova 35100 Izmir, Turkey
Raloxifene is a selective estrogen receptor modulator (SERM) and currently used to prevent and to
treat osteoporosis in post-menapausal women. Raloxifene can pass blood-brain barrier and act as an
estrogen agonist/antagonist in the brain. SERMs regulate neuronal differentiation and exert
neuroprotective actions via modulating synaptic formation and synaptic plasticity. Similar to
estrogen treatment, raloxifene is suggested to be neuroprotective with its regulator effects in the
brain. Ref-1 is a multifunctional enzyme involved in the base excision DNA repair of
apurinic/apyrimidinic sites, which is common form of DNA damage after oxidative stress. Ref-1
functions primarily as an endonuclease that removes apurinic /apyrimidinic lesions due to oxidative
stress. Increasing evidence suggests that increased expression of Ref-1 may be neuroprotective. It
has been reported that ovariectomy promotes oxidative stress in the rat brain. It has been shown that
the expression level of Ref-1 seems to correlate with cellular sensitivity to ROS and oxidative DNA
damage. In this present study, we investigated the effect of raloxifene treatment (LY139481) on the
expression levels of Ref-lmRNA in the cortex of ovariectomized female rats. By using reverse
transcription polymerase chain reaction (RT-PCR), we found that Ref-1 mRNA expression is upregulated
by raloxifene treatment in the brain cortex. The results of the present study may support
the possible neuroprotective effect of raloxifene in relation to the up-regulated level of Ref-1 in
neurodegenerative pathologies linked to oxidative stress in postmenopausal women.
1. Zhao, L., O'Neill, K., Brinton, R.D. (2005), Selective estrogen receptor modulators (SERMs)
for the brain: Current status and remaining challenges for developing NeuroSERMs, Brain
Research Reviews, 49(3): 472-493.
2. Sperk, G. (1994), Kainic acid seizures in the rat, Progress in Neurobiology, 42(1): 1-32.
3. Yalcin, A., Kanit, L., Durmaz, G., Sargin, S., Terek, C.H., Tanyolac, B. (2005), Altered level of
apurinic/apyrimidinic endonuclease/redox factor-1 (APE/Ref-1) mRNA in the hippocampus of
ovariectomized rats treated by raloxifene against kainic acid, Clinical and Experimental
Pharmacology and Physiology, 32(8):611-614.
118

P-
RADICAL SCAVENGING ACTIVITY OF FERULIC ACID (TVMAS^-HYDROXY-S-
METHOXYCINNAMIC ACID)
M. Elmastas, I. Demirtas
Department of Chemistry, Faculty of Arts and Science, University of Gaziosmanpasa, 60240,
Tokat, Turkey
Polyphenols recently have received increasing attention because of some interesting new findings
regarding of their biological activities. From pharmacological and therapeutic points of view, the
antioxidant properties of polyphenols, such as free radical scavenging and inhibition of lipid
peroxidation, are the most crucial (1). Even though a variety of herbs are known to be sources of
phenolic compounds, studies isolating polyphenols and evaluating their antioxidative effects have
rarely been carried out. Phenolic compounds are secondary plant metabolites and naturally present
in almost all plant materials, including food products of plant origin. These compounds are thought
to be an integral part of both human and animal diets (2). Hydroxycinnamic acid is the major
subgroup of phenolic compounds. Hydroxycinnamates are phenylpropanoid metabolites and occur
widely available in plants. Ferulic acid is a ubiquitous phenolic acid of low toxicity in plant
kingdom, and it can be easily absorbed and metabolized in human body. Ferulic acid shows a lot of
biological activities, including antioxidant and anti-inflammatory activities. The free radical
scavenging activity of ferulic acid was measured by the l,l-diphenyl-2-picryl-hydrazil (DPPH»).
This activity was measured by the spectrophotometric analysis (3). In this study, we evaluated the
possible free radical scavenging activity of ferulic acid; a-tocopherol, butylated hydroxyanisole
(BHA) and butylated hydroxytoluene (BHT) were used as the reference compounds. In different
concentration of DPPH radical scavenging activity of ferulic acid and standards were analyzed.
BHA, BHT and a -tocopherol were used as references for radical scavengers. The scavenging effect
of ferulic acid and standards on the DPPH radical decreased in the order of Ferulic acid > a-
tocopherol >BHA> BHT, which were 95.2, 94.5, 93.7, 85.6 % at the concentration of 75 pg/mL in
methanol, respectively. Free radical scavenging activity of these samples also increased with an
increase in their concentration. Present study has clearly showed that ferulic acid was an effective
free radical scavenging activity in vitro when it is compared to standard references compounds such
as BUT, BHA, a-tocopherol, a natural antioxidant. Therefore, ferulic acid is used as radical
scavenging agent in food and pharmaceutical industries.
1. Jing, H., Yao, L.Y., Li, J.S., Song, Y.Q, Chao, W., Research progress of pharmacology of
sodium ferulate. Northwest J. Pharmacy. 2002, 17, 236-238.
2. Bravo, L., Polyphenols: Chemistry, dietary sources, metabolism and nutritional significance.
Nutrition Reviews, 1998, 56, 317-333.
3. Sanchez-Moreno, C., Larrauri, J.A., Saura-Calixto, F., A procedure to measure the antiradical
efficiency of polyphenols. J. Sci. Food Agric. 1998, 76, 270-276.
119

EVALUATION OF TOTAL ANTIOXIDANT CAPACITY AND OXIDATIVE STRESS IN
BREAST CANCER
1 1 2 1
D. Erten-Sener , A. Gonen , M. Akinci , M. Torun
l
Department of Biochemistry, Faculty of Pharmacy, Gazi University, 06330, Etiler, Ankara,
Turkey, Department of Surgery, Ankara Educational and Research Hospital, Ankara, Turkey.
As a concequence of aerobic metabolism small amounts of reactive oxygen species (ROS) are
constantly generated in organisms. Low levels of ROS are indispensable in many biochemical
processes; however, over production or inadequate removal of ROS results in oxidative stress.
Oxidative stress is considered to be involved in the pathophysiology of all cancers. The aim of this
study is to examine oxidative stress and antioxidant status in patients with breast cancer by
evaluation of the serum levels of total antioxidant capacity (TAC) and lipid peroxidation pruducts
as malondialdehyde (MDA) and lipid hydroperoxide and to investigate the relationship between
these parameters, oxidative stress and serum lipids and lipoproteins. Serum TAC, MDA, lipid
hydroperoxide, HDL-cholesterol, VLDL-cholesteroI, LDL-cholesterol, total cholesterol,
triacylglycerol (TAG), albumin and uric acid levels of 56 breast cancer patients in different clinical
stages and 18 healthy women were determined in this study. Serum TAC levels were determined
according to the ABTS radical cation (ABTS- + ) decolorization assay described by Re et al.
Significantly lower levels of TAC were detected in patients with breast cancer in comparison to
controls (2.01 ±0.01 mmol/1 and 2.07±0.03 mmol/1 respectively; p

P-
OXIDANT/ANTIOXIDANT STATUS IN PATIENTS WITH BENIGN AND MALIGN
TUMOR
1 1 2 1 1
A. Gonerup , D. Erten §ener , S. Asian , B. §im$ek , M. Torun
'Department of Biochemistry, Faculty of Pharmacy, Gazi University, Hipodrom Ankara,
2
Turkey, Department of Surgery, Ankara Oncology Educational and Research Hospital,
Demetevler Ankara, Turkey
Breast cancer is one of the most common cancers in women of developed and developing countries.
Oxidative stress caused by increased free radical generation and/or a decreased antioxidant level in
the target cells and tissues has been suggested to play an important role in carcinogenesis. In this
study, we aimed to determine the extent of oxidative stress by measuring malondialdehyde, nitrate
and nitrite, lipid hydroperoxide and total antioxidant capacity in blood and tissue of patients with
malignant breast tumor (n=15) and benign breast disease (n-15). Lower serum and tissue
malondialdehyde levels were found decreased in breast cancer patients as compared to benign
group (p

A COMPARATIVE STUDY OF LIPID PEROXIDATION AND ANTIOXIDANT ENZYME
ACTIVITIES DURING CORONARY ARTERY SURGERY BYPASS GRAFTING
A. Gonenc , A. Hacievki', A. Soyagir 2 , M. Torun', M.N. Orman 3 , B. §im§ek'
1
Department of Biochemistry, Faculty of Pharmacy, Gazi University, Hipodrom Ankara,
2
Turkey, Department of Cardiac Surgery, Giiven Hospital, Kavaklidere Ankara, Turkey,
3
Department of Biometry, Faculty of Veterinary, Ankara University, Di§kapi Ankara, Turkey
Reperfusion after ischemia induces a sequence of events ultimately leading to cellular damage and
organ dysfunction. Several studies have proposed the essential role of reactive oxygen species in the
pathogenesis of myocardial ischemia-reperfusion injury. The aim of this study was to investigate
the difference in oxidative stress in off-pump versus on-pump coronary artery bypass surgery. In the
present study, in serial blood samples, plasma malondialdehyde as index of lipid peroxidation, red
blood cells glutathione peroxidase and superoxide dismutase were measured to compare the extent
of oxidative stress in 30 patients undergoing off-pump coronary artery bypass grafting, 12 patients
undergoing on-pump coronary artery bypass grafting and 18 healthy controls. In off-pump coronary
artery bypass grafting group, MDA levels increased significantly from 2.87±0.62 nmol/mL before
anesthesia and 2.87±0.65 nmol/mL after anesthesia to 3.05±0.66 nmol/mL after ischemia (p

EFFECTS OF EMBRYONIC NEURAL STEM CELL THERAPY ON BLOOD TOTAL
ANTIOXIDANT CAPACITY, CATALASE ACTIVITY AND MDA LEVELS OF CHRONIC
SPINAL CORD INJURY RAT
S. GUleli 1 , T. Dagci 2 , S. Konyahoglu 2
'Ege University, Faculty of Pharmacy, Department of Biochemistry, Bornova-35100 Izmir, Turkey,
2 Ege University, Faculty of Medicine, Department of Physiology, Bornova-35100 Izmir, Turkey
Neural stem cells (NSCs) have great potential as a therapeutic tool for the repair of a number of
central nervous system (CNS) disorders. NSCs can either be isolated from embryonic and adult
brain tissues or be induced from both rat and human embryonic stem cells. These cells proliferate in
vitro through many passages without losing their multipotentiality. Primary trauma to the spinal
cord triggers a cascade of cellular and molecular events that promote continued tissue damage and
expansion of the lesion for many days to months following the initial injury. Therefore, the final
outcome of spinal cord injury (SCI) is related to the extent of the initial physical damage and
ensuing secondery events that lead to the death of neurons. These secondary events are edema,
hemorrage, inflammation reaction, lipid peroxidation and oxidative stress damage. In fact,
considerable evidents has emerged pointing to the critical contribution of all of these factors in
secondary SCI. In this study, we examined the sequence of oxidative stress events following spinal
cord injury and effect of NCSs on chronic SCI rat by measuring total antioxidant capacity (TAC),
catalase (Cat) activity and malondialdehyde (MDA) levels in rat plasma. Cat activity was evaluated
using by Aebi method. MDA level was measured by thiobarbituric acid (TBA) method and TAC
was determined by trolox equivalent antioxidant capacity method. Our results demostrated that,
plasma TAC, Cat activity and MDA levels of chronic SCI and stem cell applied chronic SCI rats are
not significantly different between each other. Stem cell therapy in chronic SCI rat does not change
oxidative stress biomarkers, we studied.

IN VIVO ANTIOXIDANT EFFECTS OF TRYPTOPHAN IN LIVER AND KIDNEY OF
EXPERIMENTAL DIABETIC RAT
A.Z. Karabay, Z. Biiyukbingol
Ankara University, Faculty of Pharmacy, Department of Biochemistry, 06100 Tandogan-Ankara-
Turkey
Oxidative stress, defined as an imbalance between oxidants and antioxidants plays major role in
development and progression of diabetes and its complications. It is known that various indolic
compounds can act as free radical scavengers. In the present study, possible antioxidant effects of
tryptophan (Trp) against diabetes induced oxidative stress in liver and kidney tissues were
examined in an experimental diabetic model. Streptozotocin (STZ) was used as diabetogenic agent;
diabetes was induced 72 hours after intavenous (i.v) injection of a single 40 mg/kg dose of STZ.
After 8 weeks of diabetes induction Trp treatment was started and continued for 4 weeks. Rats were
divided into four groups: control, diabetic, Trp treated control and Trp treated diabetic. Trp treated
rats were subdivided into 3 groups and received different doses of Trp orally for 4 weeks (25, 50
and 100 mg/ kg body weight/ per day). At the end of 12 weeks tissue samples of kidney and liver
were taken from each group and lipid peroxide and nitrite levels were measured. Increased nitrite
and lipid peroxide levels were found in liver and kidney tissues of diabetic rats when compared with
the control group. Trp administration decreased nitrite and lipid peroxide levels significantly in both
kidney and liver tisues of diabetic rats in a dose dependent manner. Trp treatment could not restore
the elevated nitrite and lipid peroxide levels to control values. These findings suggest that Trp can
partially affect the impaired antioxidant system but can not restore it completely in diabetic kidney
and liver.
P-
124

STUDY REGARDING THE INFLAMMATORY AND ENDOTHELIAL STATUS IN
OBESE PATIENTS
N. Mitrea, D. Margina
University of Medicine and Pharmacy, Faculty of Pharmacy, Department of Biochemistry,
Bucharest, Romania
The aim of the study was to asses the modifications in inflammatory markers of the endothelial
dysfunction associated with the weight gain.
We analyzed 60 non-insulin-dependent type 2 diabetic patients, 37 (58.73%) females and 26
(41.26%) males. For each patient, the age, sex, body mass index (BMI), fasting plasma glucose
(FPG), plasma total cholesterol, HDL cholesterol (HDL), interleukin 6 (IL6) and C-reactive protein
(CRP) levels were recorded. The patients were divided into three groups, according to their BMI:
control group (n=ll), BMI

P-10
SCREENING AND EVALUATION OF RAT KIDNEY ALDOSE REDUCTASE
INHIBITORY ACTIVITY OF SOME PYRIDAZINE-3-ONE DERIVATIVES
M. Sarikava', M. Siikiiroglu 2 , B. ^alikan-Ergun 2 , E. Banoglu 2 , N. Da§-Evcimen', S. Siizen J
'Ankara University, Faculty of Pharmacy, Department of Biochemistry, 06100, Tandogan, Ankara,
Turkey, 2 Gazi University, Faculty of Phannacy, Department of Pharmaceutical Chemistry, 06330,
Etiler, Ankara, Turkey, Ankara University, Faculty of Pharmacy, Department of Pharmaceutical
Chemistry, 06100, Tandogan, Ankara, Turkey
Aldose Reductase (AR) is an enzyme that catalyzes the conversion of glucose to sorbitol, which is
in turn converted to fructose by sorbitol dehydrogenase. The increased glucose flux through this
metabolic pathway has been linked to the development of diabetic complications such as
neuropathy, nephropathy, retinopathy and cataract. Inhibitors of AR thus seem to have the potential
to prevent or treat diabetic complications (1). AR inhibitors belong to different chemical classes
mainly (i) hydantoins, (ii) carboxylic acid derivatives and (iii) pyridazinone analogues. At present
however, side effects related to toxicity or inadequate pharmacokinetic profiles have rendered most
of the drug candidates undesirable (2). In this study, the pyridazine-3-one analogues were evaluated
for their ability to inhibit rat kidney AR by an in-vitro spectrophotometric assay. The introduction
of a pyrazole ring on pyridazinone; a compound investigated in this study, led to a marked decrease
in inhibitory potency. This finding is paradoxical to many effective AR inhibitory compounds poses
similar ring system to pyrazole such as thiazole (3), oxadiazole (4), pyrolidine (2), hydantoin (1).
Moreover introduction of acetic acid side chain on pyridazinone did not improve the AR inhibitory
activity that was an unexpected result. According to our AR inhibitory screening study results on
pyridazine-3-one derivatives, we embarked on the synthesis of more derivatives in order to discover
more active molecules.
1. Suzen S, BUyukbingol E. Curr. Med. Chem., 10, 1329, 2003.
2. Pau A, Asproni B, Boatto G, Grella G.E, Caprariis P.De, Costantino L, Pinna G.A. Pharm. Sci.,
21, 545, 2004.
3. Maccari R, Ottana R, Curing C et al. Bioorg. Med. Chem. 13, 2809, 2005.
4. Klebe G, Kramer O, Sotriffer C. Cell. Mol. Life Sci., 67,783, 2004
126

p-11
THE ANTIOXIDANT / PROOXIDANT PARADOX IN BLACK TEA INFUSIONS
D. Siir-Altiner, B.Yenice
Department of Biochemistry, Faculty of Pharmacy, University of Marmara, Haydarpasa, Istanbul,
Turkey
Our aim in this study was to elucidate the in vitro antioxidant and prooxidant actions of black tea
(Camelia sinensis L.) infusions. For this purpose we examined the effect of different tea
concentrations on lipid oxidation (1), protein oxidation (2), carbohydrate damage (3) and DNA
damage (4). It was determined that tea at low concentrations showed antioxidant activity on all
parameters. At high concentrations this activity has stopped to increase on lipid oxidation
parameter; has decreased on protein oxidation and DNA damage parameters; has turned into a
prooxidant activity on carbohydrate damage.
Table 1. In vitro effect of tea concentrations on antioxidant potential
Parameters Control Tea Concentrations
% 0.13 % 0.25 % 0.50 %1 % 2 %4 %8
Lipid oxidation
(% inhibition) * 13.59±0.14a 22.36±0.31b 44.65 ±0.27c 48.08±0.30d 52.61±0.31e 56.64±0.42f 56.62±0.51f
Protein oxidation
(% inhibition) • 74.00±0.95a 76.88±O.IObc 77.51 ±0.15c 77.95±0.I0c 75.76±0.14ab 74.89±0.14a 73.58±0.90d
Carbohydrate
damage (A; ]2 ) 0.383±0.001a 0.379±0.140b 0.384±0.001ac 0.385 ±0.001cd 0.388±0.001d 0.397±00le 0.398±0.00le 0.401±0.001f
DNA
damage (A532) 0.406 ± 0.700a 0.035±0.001b 0.041±0.001c 0.050±0.001d 0.089 ±0.001e 0.109±0.010f
A
A
* . Values contain the control values
Values are expressed as arithmetic average ± standard deviation (n- 3)
Letters on the same line show statistical differences (p

P-14
DOES RISEDRONATE CAUSE OXIDATIVE STRESS IN THE OVARIECTOMIZED
RATS
S. Yalin'. U. Comelekoglu 2 , S. Bagi 3 , A. Nayci 4 , V. G. Giiler', M. Berkoz', P. Eroglu', R.
5
Hatungil
1 2
Mersin University, Pharmacy Faculty, Department of Biochemistry, Mersin, Turkey, Mersin
3
University, Medical Faculty, Department of Biophysics, Mersin, Turkey, Baskent University,
4
Medical Faculty, Department of Physical Treatment and Rehabilitation, Adana, Turkey, Mersin
5
University, Medical Faculty, Department of Pediatric Surgery, Mersin, Turkey, Mersin University,
Medical Faculty, Department of Physiology, Mersin, Turkey
Osteoporosis is a systemic skeletal disease characterized by low bone mass and micro architectural
deterioration of bone tissue, with a consequent increase in fragility and susceptibility to fractures.
Risedronate is a bisphosphonate medication used in the prevention and treatment of osteoporosis.
Twelve-week-old female rats were assigned randomly to a control rats (CON, n^lO) and
ovariectomized rats receiving risedronate (OVX-RIS, n=10). OVX-RIS rats were anaesthetized
with ketamine and underwent bilateral ovariectomy via ventral incision. Ten weeks after
ovariectomy, risedronate sodium (0.5 mg/kg body weight) was given via gavage two times a week
for 12 weeks to the OVX-RIS group. At the end of treatment period, rats were decapitated and then
liver tissues were removed. The determination of malondialdehyde (MDA) levels was performed by
the method of Yagi Tissue catalase (CAT) activity was measured by the method of Aebi. Although
catalase activity did not show any significant difference between the groups. MDA concentration
was significantly decreased in the OVX-RIS group (p

CHANGES IN THE LEVEL OF GLUTATHIONE THAT ACTS AS AN INTERMEDIARY
TO THE PROTECTIVE EFFECTS OF MELATONIN PRETREATMENT IN CEREBRAL
INJURY INDUCED BY RADIATION IN RATS
S Yard unci 1 , O. Yildirim 2 . F. Akbiyik 3 , S. gomoglu 4 , M. Akmansu 5 , G, Bozkurt 6 , S. Siirucu 6
P-15
'Ankara University, Faculty of Medicine, Department of Physiology, 2 Ankara University, Faculty
of Science, Department of Biology, 3 Hacettepe University, Faculty of Medicine, Department of
Biochemistry, SSK Di§kapi Aducation and Research Hospital, 2nd Neurological Clinic,
4 Gazi
University, Faculty of Medicine, Department of Radiation Oncology,
5 Hacettepe University,
Faculty of Medicine, Department of Brain Surgery, 6 Hacettepe University, Faculty of Medicine,
Department of Anatomy, Turkey.
In this study we investigated whether pretreatment with melatonin was protective against the injury
of the central nervous system in rats receiving LD50 radiation. The rats were randomized into 4
groups for this purpose. First group was control rats, second group was melatonin received animals,
third group was only irradiated rats and forth group was melatonin received before irradiated
animals. The rats in the second group exposed to the similar procedure V2 hour after receiving
melatonin. The rats in the third group received 675 cGy of whole body irradiation (LD 50 ) following
the injection of saline. The animals in the forth group received whole body irradiation following the
preadininistration of melatonin. Cerebral cortexes of rats in the fourth group were taken out and
cortical MDA and GSH levels were measured. The significant increases in the levels of cortical
MDA with the application of radiation was indicating that oxidative damage had an important role
in this injury. Following the irradiation, the decreases observed in the level of cortical GSH were
considered to be related to the increases in the consumption and/or utilization of glutathione in
CNS. On the contrary, significant increases were detected in the levels of plasma total glutathione
following irradiation in melatonin pretreated rats.. The results obtained from this study
demonstrated that pretreatment with melatonin was beneficial in decreasing the degree of damage
that develops in CNS following irradiation. The beneficial effect of melatonin can be related with
its protecting the CNS from oxidative injury and preventing the decrease in the level of cortical
glutathione.
1

P-16
PREVALENCE OF PATIENT REPORTED ADVERSE EFFECTS OF STATINS -
PARTICULAR EMPHASIS ON MYALGIA
1 2 2 2 3 3
S. Apikoglu Rabu , E. Bozkurt , C. Demircioglu , S. Tandogan , D. Aydin , G. Uzel , F.V.
izzettin
1 2
Clinical Pharmacy Department, Clinical Pharmacy Student Research Group, Marmara University
Faculty of Pharmacy, Cardiovascular Surgery Department, Kosuyolu Heart and Research Hospital,
Istanbul, Turkey
Background and Objective: Statins are one of the most commonly prescribed drugs due to their
established cardiovascular benefits both in primary and secondary prevention. Despite their good
safety profiles, in clinical practice higher number of patients, than those reported in the clinical
trials seem to complain from side effects, particularly of myalgia. The aim of this study is to
examine the patient-reported real-world prevalence of statin side effects with particular emphasis on
myalgia. Design: This cross-sectional study was conducted on 176 patients from cardiovascular
outpatient clinics of two different hospitals. Demographic as well as clinical data were collected and
the patients were asked to report the undesirable effects (if any) that occurred after the initiation of
statin administration. The probability that the myalgia was due to a statin adverse effect was
estimated using 'Naranjo Probability Scale" (1). Setting: Marmara University, Faculty of Pharmacy,
Clinical Pharmacy Department Main Outcome Measures: The percentage of statin adverse effects
and the Naranjo scores of the particular adverse effect of myalgia. Results: The mean age was 58,19
± 0,87 (SEM) years and 60,2% of the patients were male. The mean duration of statin
administration was 22 ± 3,03 (SEM) months. Most of the patients were on atorvastatin (80,1%) and
this was followed by simvastatin (11,4%). 44,9% of the patients reported to suffer from an adverse
event with the highest percentage (40,3%) reported to have muscle aches and/or joint pain
with/without muscle weakness. This is followed by dizziness (3,4%) and gastrointestinal problems,
sleep disturbances and hypersensitivity reactions, all reported in 2,8% of the patients. Transaminase
elevation of >3 times upper limit of normal was recorded in 3 (out of 176) patients. After
employing the Naranjo Scale, myalgia was estimated to be "possibly" due to statin therapy in
90,1% of those suffering from myalgia, while "probably" statin-induced in 9,9%. As objective
measurements such as creatine kinase levels were not performed, myalgia was failed to be
estimated as a "definite adverse reaction to drug" in none of the patients. Conclusions: Welldocumented
statin side effects particularly myalgia can be encountered in a high percentage of
patients in clinical practice. The pharmacist should direct the patient counseling towards the
monitoring of myopathy symptoms and educate the patient on tolerance techniques.
1. Naranjo CA, Busto U, Sellers EM, Sandor P, Ruiz I, Roberts EA, Janecek E, Domecq C,
Greenblatt DJ. A method for estimating the probability of adverse drug reactions. Clin Pharmacol
Ther 1981;30:239-45.
12

P-17
MID-TERM RESULTS OF SECONDARY PREVENTION IN CORONARY ARTERY
DISEASE: DRUG UTILIZATION PROFILES
1 . 1 1 2 2 3
S. Apikoglu Rabus , F.V. Izzettin , M. Sancar , O. Karakaya , R. Kargin , C. Yakut
1 2
Clinical Pharmacy Department, Marmara University Faculty of Pharmacy, Cardiology
Department, Cardiovascular Surgery Department, Kosuyolu Heart and Research Hospital, Istanbul,
Turkey
Background and Objective: Coronary artery disease (CAD) is an established cause of mortality and
morbidity and effects a considerable portion of the society. Guidelines were developed to ensure the
coverage of all eligible patients of secondary prevention. However, the secondary prevention
practice by aspirin, beta-blockers, angiotensin converting enzyme (ACE) inhibitors and statins still
can be sub-optimal. Also, there exists the patient-induced problem of non-compliance. In this study
we aimed to examine these issues in a small cohort of patients angiographically diagnosed to have
coronary artery disease, some of whom also had experienced a myocardial infarction. Design: In
this prospective study conducted in a cardiovascular specialty hospital, from a subsequent cohort of
patients who admitted to the hospital for coronary angiography performance, 73 patients who were
diagnosed to have CAD were followed up for 5 years. The baseline demographic and clinical data
were collected just before angiography. The baseline drug data were collected at the day of
discharge. The fifth year data were taken from the patients via face-to-face consultations or phone
interviews. Setting: Marmara University. Faculty of Pharmacy, Clinical Pharmacy Department
Main Outcome Measures: The percentage of patients prescribed aspirin, statins, beta-blockers and
ACE inhibitors in accordance to various guidelines and the percentage of patients compliant to their
prescribed therapies at the fifth year. Results: When the Sheffield Tables, National Cholesterol
Education Program-Adult Treatment Panel Guidelines and guideline proposed by Bersot et al for
low-HDL populations were performed, 27, 24 and 45 patients (out of 73) respectively were found to
be eligible for statin therapy. Taking the Bersot guideline, out of the eligible patients 40% were not
initially prescribed a statin, 28,9% were never prescribed a statin while 26,7% were no more taking
a statin and 33,3% kept taking their drugs for five years. Among those who had a myocardial
infarction (n=20) 55% kept taking aspirin, while 45% quitted the drug without any documented
contraindication. Beta-blockers were never prescribed in 30% and ACE inhibitors were never
prescribed in 20% of patients where 35% and 30% of patients discontinued beta-blockers and ACEinhibitors
respectively. Conclusions: Besides the sub-optimal prescription of secondary prevention
drugs, non-compliance seems to be a challenging issue in pharmaceutical care of coronary artery
disease patients. Clinical pharmacist should be the person to optimise the drug adherence.
1

P-18
THE PILOT STUDY OF EFFICACY OF THE CONSERVATIVE AND SURGICAL
GLAUCOMA THERAPY
B. Cadikova'. J. Novak 2 , J. Vlcek'
1
Department of Social and Clinical Pharmacy, Faculty of Pharmacy, Charles University; Hradec
2
Kralove, Czech Republic, Department of Ophthalmology, The Regional Hospital Pardubice, Czech
Republic
Background and Objective: Glaucoma is the third largest cause of worldwide blindness. The
treatment of glaucoma focuses mainly on the intraocular pressure (IOP) reduction and on the
pharmacologic prevention of the development vision's damage. Currently there are two methods of
IOP correction: conservative (pharmacotherapy) and invasive (surgical or laser intervention).The
aim of this study is to evaluate the IOP lowering efficacy of different therapies in the cohort of
glaucoma patients. Design: A five years retrospective cross-section study (2001-2005). Setting:
Department of Ophthalmology, Regional Hospital Pardubice, Czech Republic. Main Outcomes
Measures: Consumption of the ocular hypotensive drugs, the progress of examination's outcomes,
efficiency of glaucoma surgeries. Results: 151 patients with diagnoses H400 (14; 9.03%), H401
(125; 80.65%), H402 (9; 5.81%), H403 (1; 0.65%), H405 (1; 0.65%), H408 (3; 1.94%), H409 (2;
1.29%) treated at Department of Ophthalmology The regional Hospital Pardubice CR were included
in the study of which 38.41% men and 61.59% women (average age: 59 year, median 61 year).
Overall IOP value is 18.58 torr, mean IOP left eyel8.62 torr, mean IOP right eye 18.54 torr (3039
measured eyes). The visual field restriction or damage could be occurred in 624 of 735 eyes
(84.48%). Administrated therapy: beta-blockers, analogs of prostaglandins, parasympathomimetics,
carbonic anhydrase inhibitors and adrenergic agents (41.98%; 26.78%; 15.76%; 14.83% and 0.56%
respectively). The count of drugs used one year before glaucoma surgery is 3.47 per patient and one
year after glaucoma surgery 0.88 per patient. Conclusions: This pilot study suggests that the most
frequent treatment strategy is pharmacotherapy (118 patients, 79.47%), the most frequently
prescribed drugs used for glaucoma treatment are beta-blockers, the most of patients were
administrated combined pharmacotherapy, the most frequently surgery is filtration surgery (86.00%
of all glaucoma surgeries).
1. Rikkert et al.; Intraocular pressure-lowering effects of all commonly used glaucoma drugs: a
meta-analysis of randomized clinical trials; Ophthalmology.; 2005; Jul; 112(7): 1177-85.
1

P-19
EFFECTS OF BUDDY PROGRAM ON ANTIRETROVIRAL DRUG ADHERENCE
1 2
S. Urairat, S. Lerkiatbundit
1 2
Department of Pharmacy, Sathingpra hospital, Songkla, 90110, Thailand, Department of
Pharmacy Administration, Faculty of Pharmaceutical Sciences, Songkla, 90112, Thailand
The purpose of this study was to determine the effects of buddy program on antiretroviral drug
adherence. The study was randomized-controlled study. Thirty-eight non-adherent AIDS patients
were recruited from 4 hospitals through an interview. The subjects were randomly assigned to a
control group (N= 19) and an experimental group (N=19). Both groups received the same
education on AIDS, antiretroviral drugs and self-care. In the experimental group, the subjects chose
their own buddy who received the same information from the researcher, and also the information
on patients' outcome report and a letter asking him/her to take care of the patients' antiretroviral
drug use. The researcher assessed non-adherence by three methods: interview, self report and pill
count. The subjects were followed up for four consecutive visits with one-month interval. The rate
of non-adherence in the experimental group after one and two months after the intervention was
lower then that in the control group. The same pattern was found over three methods of adherence
assessment. The non-adherence rates in the experimental group and the control group were less than
5 % at the first and the second months after the intervention. Two months after the intervention,
non-adherence rates in the control group as assessed by interview, self report and pill count were
1.02 ± 1.27, 1.77 ± 2.16, and 0.70 ± 1.08, respectively. CD4 levels and body weight between
groups were not statistically significant. The patients in the experimental group and buddy were
very satisfied to the buddy program. In conclusion, buddy program significantly reduced nonadherence
but there was no impact on CD4 level and body weight at two months of the intervention.
1

P-20
PREVALENCE OF THE POTENTIAL INTERACTION BETWEEN ORALLY
ADNINISTERED FLUOROQUINOLONES AND DI- OR TRI-VALENT CATION-
CONTAINING COMPOUNDS
1 2 2 1 1 1
S. Pattharachavakul , P. Chayakul , P. Suksanan , T. Boonlon , P. Buttajeen , P. Tanvejsilp
1
^ Faculty of Pharmaceutical Sciences, Prince of Songkla University, Songkhla, Thailand, 90112,
Faculty of Medicine, Prince of Songkla University, Songkhla, Thailand
Co-administration of oral fluoroquinolone and di- or tri-valent cation-containing compounds (e.g.
calcium or aluminum products) could significantly decreased the absorption of fluoroquinolones
and subsequently compromised the therapeutic effect of these medications^. The objectives of
this study are to determine the prevalence of concomitant prescribing of oral fluoroquinolones with
the di- or tri-valent cation-containing compounds and the effect of this interaction on the therapeutic
outcome. A retrospective medical records review of out-patients who were prescribed oral
ciprofloxacin, norfloxacin, levofloxacin and ofloxacin at Songklanagarind Hospital from January to
June 2005 was performed. Data collection included demographics, regimen of fluoroquinolones,
indication of fluoroquinolones, therapeutic outcome of fluoroquinolones, and regimens of
concomitant medications. Patients who were prescribed di- or tri-valent cation-containing
compounds at the same time as oral fluoroquinolones were identified as patients who were
potentially to experience the drug interactions and were defined as cases. Controls were patients
who received oral fluoroquinolones without prescriptions of any di- or tri-valent cation-containing
compounds. Cases and controls were matched for gender, type of fluoroquinolones received,
fluoroquinolones indication. A thousand and three hundred eighty oral fluoroquinolones
prescriptions were reviewed. Of these 1,380 prescriptions, 52 (3.76%) prescriptions were also
concomitantly prescribed di- or tri-valent cation-containing compounds. Successful therapeutic
outcome were shown in 24 (46.2%) of 52 patients in case group and 20(38.5%) of 52 patients in
control group. However; about half of patients in case and control group had inadequate
information for therapeutic efficacy evaluation. In conclusion, prevalence of potentially interaction
between oral fluoroquinolones and di- or tri-valent cation-containing compounds at
Songklanagarind Hospital was quite low. Future studies with more number of subjectes to
investigate the impact of this interaction on therapeutic efficacy of fluoroquinolones are warranted.
1. Nix DE, Wilton JH, Schentag JJ, Parpia SH, Norman A and Goldstein HR. Inhibition of
Norfloxacin Absoption by Antacid and Sucralfate. Reviews Infectious Diseases 1989;
11(5): 1096.
2. Allen A, Bygate E, Faessel H, Isaac L and Lewis A. The effect of ferrous sulphate and sucralfate
on the bioavailability of oral gemifloxacin in healthy volunteers. International Journal of
Antimicrobial Agents; 15(4):283-9
16

COMPARISON OF CLINICAL SIGNIFICANCE RATING OF DRUG INTERACTIONS
FROM THREE REFERENCE SOURCES
J. Pongwecharak, S. Limsuwannachot, P. Thongroichang, A. Awang
Department of Clinical Pharmacy, Faculty of Pharmaceutical Science, Prince of Songkla
University, Hatyai, Songkhla, Thailand 90112
P-21
There are several resources, either hard copies, online databases or websites, for checking drug
interactions for their clinical significance and management. The present study compared three
reference sources providing information on drug interactions. The sources were two textbooks, i.e.,
Drug Interaction Facts (2003 edition) and Managing Clinically Important Drug Interactions (2002
edition), and an online subscribed database, namely, Micromedex Healthcare Series (2003
subscription). Only pairs of drug interaction of which clinical significance was rated as major (level
1) or moderate (level 2) were included. The results of the same pair from each source were
compared. Frequency of concordance was presented. References on which the drug interactions
were based were further checked and classified into a case report, study in healthy volunteers, study
in patients, or others. From the review, there was a total of 1,756 pairs of drug interactions with
clinical significance rating of major or moderate in the three sources. Among the three sources, the
Managing Clinically Important Drug Interactions provided fewest pairs of drug interactions, i.e, 28
pairs of major clinical significance and 127 pairs of moderate level. The corresponding number of
the Drug Interaction Facts and the Micromedex Healthcare Series were comparable. Of all, drug
interactions with major clinical significance constituted 31.5% while 68.5% were graded as
moderate. Only 1.6% of the pairs were rated as major clinical significance in all three sources,
while slightly over a quarter (27.3%) were in agreement in two sources. The corresponding figures
(1.6% and 30.2%, respectively) were similar for those rated as moderately clinical significant. It
was noted that agreement between two sources was more common between the Drug Information
Facts and the Micromedex Healthcare series, i.e., 27.8% for major clinical significance and 31.8%
for moderate level. Among the pairs which had same rating across three sources, types of reference
on which the drug interactions were based in each source differed. The results implied that relying
upon one reference source of drug interaction may be inadequate due to low agreement in clinical
significance rating in various sources. This could affect decision on management of drug interaction
encountered in practice.
1. Abarca J., Malone, DC, Armstrong EP, Grizzle AJ, Hansten PD, van Bergen, RC, et al. J Am
Pharm Assoc 2004; 44 (2): 135-41.
2. Fulda TR, Valuck RK, Zanden JV, Parker S, Byrns PJ. Curr Ther Res 2000; 61 (8): 540-7.
1

P-22
COMPARISON OF THE EFFECT OF GLYCYRRHIZA GLABRA WITH DIFFERENT
ANTIULCER DRUGS ON ASPIRIN INDUCED ULCER IN RATS
1 1 1 2 3 4 . 1
T. Hantash , M. Sancar , B. Okuyan , Z. Cirakli , U. Cevikba ; p. jjras , F. V. Izzettin
I
l Department of Clinical Pharmacy, Faculty of Pharmacy, University of Marmara, Tibbiye Cad.
2
No:49 Haydarpasa, 34817, Istanbul, Turkey, Department of Biochemistry, Bakirkoy State
3
Hospital, Istanbul, Turkey, Department of Pathology, Istanbul Faculty of Medicine, Istanbul
4
University, Istanbul, Turkey, Department of Biochemistry, Faculty of Pharmacy, University of
Marmara, Tibbiye Cad. No:49 Haydarpasa, 34817, Istanbul, Turkey
The aim of the study was to evaluate the possibility of finding a more effective, safer effect and
cheaper agent to prevent and treat nonsteroidal antiinflammatory drugs (NSAID)-induced ulcers,
the effect of Glycyrrhiza glabra (liquorice, family: Leguminosa) decoction was compared with the
effects obtained by omeprazole and misoprostol in rats. Rats were divided into prophylactic and
treatment groups. Liquorice decoction 20-30 ml/kg orally, omeprazole 2.3 mg/kg i.p. and
misoprostol 50 pg/kg orally were administered for 3 consecutive days just a half hour before aspirin
200 mg/kg administration in the case of prophylactic group. In the case of the treatment group,
aspirin 200 mg/kg was administered orally for 3 consecutive days, and then other drugs were
administered at the same doses as the prophylactic group daily for 4 weeks. The animals in
prophylactic study (at 4^ day) and treatment study (at 30*' 1 day) were killed by ketamine overdose
and the stomach including part of the duodenum was removed for histopathological examination.
According to histopathologic evaluation, misoprostol showed statistically significant protection, but
liquorice decoction and omeprazole did not. In the treatment group histopathological examination
showed no significant difference between the three agents used to treat the induced ulcer. Based on
our results we believe that Glycyrrhiza glabra can be used as an effective agent in NSAID-induced
ulcer treatment.
1

DIFFERENT MODELS IN NONSTEROIDAL ANTI-INFLAMMATORY DRUG INDUCED
ULCERS PREVENTION IN RATS
1 . 1 2 3
M. Sancar , F. V. Izzettin , M. Huritoglu , U. Qevikba
i
Department of Clinical Pharmacy, Faculty of Pharmacy, University of Marmara, Tibbiye Cad.
2
No:49 Haydarpasa, 34817, Istanbul, Turkey, Department of Internal Medicine, Vakif Gureba
3
Hospital, Istanbul, Turkey, Department of Pathology, Istanbul Faculty of Medicine, Istanbul
University, Istanbul, Turkey
P-23
Gastrointestinal symptoms are the most common adverse events associated with nonsteroidal
antiinflammatory (NSAID) therapy. There are different strategies for prevention of GI tract
problems caused by NSAIDs. Currently the accepted method for decreasing the risk of serious
NSAID related GI toxicity is to limit NSAID use. However, in fact this is not a realistic option
especially for rheumatic patients. In this study we aimed to determine the efficacy of different
antiulcer drugs for the prevention of NSAID induced ulcers. In this animal study we determined the
efficacy of misoprostol (100 pg/kg/day and 10 pg/kg/day p.o.), omeprazole (5 mg/kg/day and 1.5
mg/kg/day i.p.), ranitidine (40 mg/kg/day and 10 mg/kg/day i.p.), bismuth (70 mg/kg/day and 15
mg/kg/day p.o.), combinations of misoprostol (10 pg/kg/day) plus omeprazole (1.5 mg/kg/day) and
misoprostol (10 pg/kg/day) plus ranitidine (10 mg/kg/day) for the prevention of 50 mg/kg/day s.c.
indomethacin induced ulcer. Each group was concomitantly treated for 5 days with the above
treatment regimens plus indomethacin. After 5 days treatment histopathological and hematological
examination was performed. The following regimens were found to be effective in the prevention of
indomethacin induced gastric lesions: 100 pg/kg misoprostol, 10 pg/kg misoprostol, 5 mg/kg
omeprazole, combinations of 10 pg/kg misoprostol plus 1.5 mg/kg omeprazole and 10 pg/kg
misoprostol plus 10 mg/kg ranitidine. The ulcer scores in groups treated with ranitidine, bismuth
and low dose omeprazole were not found to be different from the indomethacin group score, so that;
the efficacy of these groups was not enough to prevent indomethacin induced ulcers. Our
experimental animal study may be helpful in planning potential prevention therapies for NSAID
induced ulcer.
1

A STUDY ON POINT OF VIEW OF TURKISH DOCTORS ON BIO-EQUIVALENCY AND
BIO-EQUIVALENCY IN TURKEY
E. Bilgener, G. Oz9elikay
Department of Pharmacy Management, Faculty of Pharmacy, University of Ankara, 06100,
Tandogan, Ankara,Turkey
P-24
World medicine industry is the most multi-national industry area. Although the world trade in
medicine was not very high-scaled, it has been improved faster since the 60's. The sub-structure
investments made for the research and development studies by the manufacturer companies and as a
result of the developments in technology, day by day, finding new medicine molecules has been
very effective during the last couple of decades. Years ago, some data protection has arranged in
order to protect the big companies who made research and development investments. The originator
companies have been protected by the help of these arrangements and by the help of avoiding other
firms to produce the newly developed molecules during a specific period of time. At the end,
against the monopolisation and with the idea that the medicine prices might have been highly
increased the other firms were allowed to produce these new molecules. Medicine is an important
product for human health. Although, in the past, there was an idea that the medicine can't have
quality, thanks to the developing technology and the result of many qualitative and quantitative
evaluations, it has been proved that the medicine can have a quality. The companies which have
been developed in producing generic molecules have started to look for some standards through
these experiences and information. The last point that these standards to arrive have been called bioequivalency.
This study has been prepared with the aim of giving information about the point view
of the Turkish doctors about these arrangements and the molecules that have been claimed to be
equivalent or about the arrangements have been done for bio-equivalency, even in order to point out
bio-equivalency and its importance. Fifteen-questioned questionnaire forms which were applied to
127 expert doctors and 209 practitioners in Gorum form the material of this research. According to
the data which are gained at the end of this study, most of the doctors don't believe in the medicine
claimed to be bio-equivalent are really bio-equivalent and most of them stated that they don't use
the generic medicine on themselves or their relatives. After forming the code keys of the
information in questionnaires, their statistical evaluations and the comments has been done by the
help of SPSS pocket programme using computer.
1

P-25
COMMUNITY PHARMACISTS' KNOWLEDGE, ATTITUDE AND HABITS
REGARDING TOBACCO PRODUCTS IN ANKARA: A PILOT STUDY
1 1 2 3
Z.Qalgan , SYegenoglu , E.Tahir , N.Bilir
'Department of Pharmacy Management, Faculty of Pharmacy, Hacettepe University, 06100
2
Sihhiye, Ankara, Turkey, School of Medicine, Hacettepe University, 06100 Sihhiye, Ankara,
Turkey, Department of Public Health, School of Medicine, Hacettepe University, 06100 Sihhiye,
Ankara, Turkey
Tobacco products are cigarette, narghile tobacco, pipe and cigar in general. Cigarette is the most
known and used tobacco product. In Turkey, smoking is a common habit and a significant public
health problem. On the other hand, community pharmacies offer an opportunity for intervening in
this problem because, people ask the pharmacists' advice on their diseases and keep in touch with
them. In addition, since the pharmacists are role models for community members, their attitude
towards smoking is of great importance. With this cross-sectional survey that comprises 83
community pharmacies in Balxjelievler and Balgat districts in the 1. region of Ankara, we aimed to
determine pharmacists' knowledge, attitude and habits concerning tobacco products especially
cigarette, and at the end to make a proposal on possible pregraduate/postgraduate educational
interventions. Written permission for survey was taken from Ankara Chamber of Pharmacists. We
designed a questionnaire with 41 questions under four sections. In the first section, we asked the
pharmacists some of their sociodemographic characteristics. In the second section, questions about
their attitude towards smoking were asked. In the following two sections, questions related to their
smoking habits and their knowledge with respect to smoking were searched. The questionnaire was
pretested on 10 community pharmacies in Ankara. In the light of the results and pharmacists'
comments, the questionnaire was redesigned. Some questions in the questionnaire are:
>Do you smoke
> To which patients do you ask whether they smoke or not
> How do you consult people who come to you to quit smoking
> How soon after you wake up do you smoke your first cigarette (from Fagerstrom test for
nicotine dependence)
> If you do not smoke, what is the reason for this
> In your opinion, which one(s) is/are harmful (passive smoking, narghile tobacco, narghile
tobacco with aroma, pipe, cigar)
> Is smoking in a pharmacy forbidden by law
> What do you want to learn about smoking
1. Heatherton T. F., Kozlowski L. T., Frecker R. C., Fagerstrom K. O. (1991). The Fagerstrom
Test for Nicotine Dependence: A revision of the Fagerstrom Tolerance Questionnaire. British
Journal of Addictions, 86, 1119-1127.
2. Yegenoglu S., Asian D., Evrener §.E., Acar A., Bilir N. (2005). What is Behind Smoking
Among Pharmacy Students: A Quantitative and Qualitative Study from Turkey. Substance
Use & Misuse,41, 405-414.
1

P-2 8
SAFETY RESEARCH ON COMMUNITY PHARMACIES IN KOCAELi
O.N. Erdogan. S. Kaya
Kocaeli University, College of Hereke Omer Ismet Uzunyol, Marshall Campus, Korfez, 41800-
Kocaeli, Turkey
The aim of this research was to understand the precautionary measures of pharmacies in terms of
safety issues of service. There are 322 community pharmacies in Kocaeli. Group sampling was used
to select the community pharmacies which would be included into the study. Four regions among
eight regions were selected to this end. First a pilot study was done in 10 pharmacies to improve the
questionnaire. Some addings and removals were made to questions as to pilot study. Throughout
December 2005, face-to-face questionnaire was applied to 156 of 247 community pharmacists in
four regions; 58 of Gebze, 8 of Korfez, 24 of Derince, and 66 of Izmit. According to the study
findings; 97 pharmacists are woman, 59 are men. One thirds of community pharmacists (33,3 %)
have described that region where service provided was unsafe. There are no any measurements of
16.5 percent of community pharmacies for drugs which need to be protected in cold in case of
electricity cut. Thus in-service trainings need to be provided for pharmacists about safety and
related issues.
144

P-2 9
DRUG DATA EXCLUSIVITY IN TURKEY FROM THE PERSPECTIVE OF
PHARMACEUTICAL COMPANIES
O. Giirson, G. Oz^elikay
Department of Pharmacy Management, Faculty of Pharmacy, University of Ankara, 06100,
Tandogan, Ankara, Turkey
The purpose of this study is to determine conceptions and/or disagreements of local and foreign
drug companies in Turkey about data exclusivity - the most controversial issue between generic and
original pharmaceutical companies all over the world - during the implementation period of the
new drug licensing regulation. The material of this study is a questionnaire replied by members of
Turkish Pharmaceutical Manufacturers' Association (by 2002, this association had 53 local and
foreign members). The questionnaire includes 17 questions and questionnaires were fdled out by
regulatory affairs managers, business development coordinators and corporate affairs directors of
the companies by face-to-face interviews, mails and e-mails.
With evaluating this study, the results are:
1. According to the shared opinion of local and foreign companies (%59.4), generic drug
companies will attach importance to research and development activities in the long term.
Data exclusivity will be able to encourage the generic companies to produce original drugs.
It is impossible to invent an original drug by Turkish local companies today. However, %60
of local companies that replied the questionnaire survey think that data exclusivity has an
encouraging role on research and development.
2. %66.7 of local companies that replied the questionnaire think that equivalent drug policies
will not go on with data exclusivity. However, %94.1 of foreign drug companies think that
equivalent drug policies will go on with data exclusivity.
3. %93.8 of all the companies that joined the study state that informing relevant persons and
institutions about data protection is not sufficient and more objective studies must be done
about data protection.
Although data exclusivity can cause negative effects like decreasing in the market size of local
pharmaceutical companies, in majority both of local and foreign companies perceive the data
exclusivity as a necessary and beneficial change. Because replies of questionnaire state that data
exclusivity has positive effects in the long term like increasing on investments of foreign
pharmaceutical companies in Turkey, encouraging the local companies for research and
development activities.
145

P-
COMPARISON OF BACITRACIN AND SULFAMETHOXAZOLE-TRIMETHOPRIM
DISC METHOD WITH REMEL STREPTEX FOR IDENTIFICATION OF GROUP A
BETA HEMOLYTIC STREPTOCOCCI
M. Ervilmaz. O. Atli, A. Akin
Departmant of Pharmaceutical Microbiology, Faculty of Pharmacy, University of Ankara, 06100
Ankara-Turkey
Group A beta hemolytic streptococcus (GABHS) is the most frequent cause of bacterial
pharyngotonsillitis. The prevalence of Streptococcus pyogenes (GABHS) infections has increased
drastically in the last decade and correlates with increasing resistance to macrolide and tetracycline
groups of antibiotics. The bacitracin and sulfamethoxazole-trimethoprim (SXT) disc method is
preferred for defining GABHS in daily routine tests since it is inexpensive, easy-to-use and has a
high rate of sensitivity although subculturing individual beta-hemolytic colonies among the
crowded bacterial colonies is sometimes very difficult. Remel Streptex is a latex agglutination test
for identification of streptococcal group antigens from cultured organisms. The latex agglutination
test, which is based on specific antigen-antibody reaction, is more sensitive than bacitracin-SXT
method. However its high cost prevents routine laboratory application. We compared Remel
Streptex with the bacitracin and SXT disc method for sensitivity in the presumptive identification of
Streptococcus pyogenes. A total of 110 GABHS isolated from throat cultures were examined. We
used the Remel Streptex as the gold standart method for identifying GABHS. Principle of Remel
Streptex is; extracting group specific antigens from streptococci by nitrous acid in a simple and
short incubation step. Than extract is neutralised and antigens are detected and identified with
suspensions of latex particles coated with group specific antibodies. A positive result is seen as an
aggregation of latex particles. Compared with the Remel Streptex, the sensitivity of bacitracin and
SXT disc method was 93.6%. As a result bacitracin and SXT disc method has a high sensitivity to
identify Streptococcus pyogenes from subcultured plates.
1

P-29
DRUG DATA EXCLUSIVITY IN TURKEY FROM THE PERSPECTIVE OF
PHARMACEUTICAL COMPANIES
O. Giirson. G. Ozselikay
Department of Pharmacy Management, Faculty of Pharmacy, University of Ankara, 06100,
Tandogan, Ankara, Turkey
The purpose of this study is to determine conceptions and/or disagreements of local and foreign
drug companies in Turkey about data exclusivity - the most controversial issue between generic and
original pharmaceutical companies all over the world - during the implementation period of the
new drug licensing regulation. The material of this study is a questionnaire replied by members of
Turkish Pharmaceutical Manufacturers' Association (by 2002, this association had 53 local and
foreign members). The questionnaire includes 17 questions and questionnaires were filled out by
regulatory affairs managers, business development coordinators and corporate affairs directors of
the companies by face-to-face interviews, mails and e-mails.
With evaluating this study, the results are:
1. According to the shared opinion of local and foreign companies (%59.4), generic drug
companies will attach importance to research and development activities in the long term.
Data exclusivity will be able to encourage the generic companies to produce original drugs.
It is impossible to invent an original drug by Turkish local companies today. However, %60
of local companies that replied the questionnaire survey think that data exclusivity has an
encouraging role on research and development.
2. %66.7 of local companies that replied the questionnaire think that equivalent drug policies
will not go on with data exclusivity. However, %94.1 of foreign drug companies think that
equivalent drug policies will go on with data exclusivity.
3. %93.8 of all the companies that joined the study state that informing relevant persons and
institutions about data protection is not sufficient and more objective studies must be done
about data protection.
Although data exclusivity can cause negative effects like decreasing in the market size of local
pharmaceutical companies, in majority both of local and foreign companies perceive the data
exclusivity as a necessary and beneficial change. Because replies of questionnaire state that data
exclusivity has positive effects in the long term like increasing on investments of foreign
pharmaceutical companies in Turkey, encouraging the local companies for research and
development activities.
145
147

P-32
COMPARISON OF BACITRACIN AND SULFAMETHOXAZOLE-TRIMETHOPRIM
DISC METHOD WITH REMEL STREPTEX FOR IDENTIFICATION OF GROUP A
BETA HEMOLYTIC STREPTOCOCCI
M. Eryilmaz. O. Atli, A. Akin
Departmant of Pharmaceutical Microbiology, Faculty of Pharmacy, University of Ankara, 06100
Ankara-Turkey
Group A beta hemolytic streptococcus (GABHS) is the most frequent cause of bacterial
pharyngotonsillitis. The prevalence of Streptococcus pyogenes (GABHS) infections has increased
drastically in the last decade and correlates with increasing resistance to macrolide and tetracycline
groups of antibiotics. The bacitracin and sulfamethoxazole-trimethoprim (SXT) disc method is
preferred for defining GABHS in daily routine tests since it is inexpensive, easy-to-use and has a
high rate of sensitivity although subculturing individual beta-hemolytic colonies among the
crowded bacterial colonies is sometimes very difficult. Remel Streptex is a latex agglutination test
for identification of streptococcal group antigens from cultured organisms. The latex agglutination
test, which is based on specific antigen-antibody reaction, is more sensitive than bacitracin-SXT
method. However its high cost prevents routine laboratory application. We compared Remel
Streptex with the bacitracin and SXT disc method for sensitivity in the presumptive identification of
Streptococcus pyogenes. A total of 110 GABHS isolated from throat cultures were examined. We
used the Remel Streptex as the gold standart method for identifying GABHS. Principle of Remel
Streptex is; extracting group specific antigens from streptococci by nitrous acid in a simple and
short incubation step. Than extract is neutralised and antigens are detected and identified with
suspensions of latex particles coated with group specific antibodies. A positive result is seen as an
aggregation of latex particles. Compared with the Remel Streptex, the sensitivity of bacitracin and
SXT disc method was 93.6%. As a result bacitracin and SXT disc method has a high sensitivity to
identify Streptococcus pyogenes from subcultured plates.
148

P-29
DRUG DATA EXCLUSIVITY IN TURKEY FROM THE PERSPECTIVE OF
PHARMACEUTICAL COMPANIES
O. Giirson. G. Ozfelikay
Department of Pharmacy Management, Faculty of Pharmacy, University of Ankara, 06100,
Tandogan, Ankara, Turkey
The purpose of this study is to determine conceptions and/or disagreements of local and foreign
drug companies in Turkey about data exclusivity - the most controversial issue between generic and
original pharmaceutical companies all over the world - during the implementation period of the
new drug licensing regulation. The material of this study is a questionnaire replied by members of
Turkish Pharmaceutical Manufacturers' Association (by 2002, this association had 53 local and
foreign members). The questionnaire includes 17 questions and questionnaires were fdled out by
regulatory affairs managers, business development coordinators and corporate affairs directors of
the companies by face-to-face interviews, mails and e-mails.
With evaluating this study, the results are:
1. According to the shared opinion of local and foreign companies (%59.4), generic drug
companies will attach importance to research and development activities in the long term.
Data exclusivity will be able to encourage the generic companies to produce original drugs.
It is impossible to invent an original drug by Turkish local companies today. However, %60
of local companies that replied the questionnaire survey think that data exclusivity has an
encouraging role on research and development.
2. %66.7 of local companies that replied the questionnaire think that equivalent drug policies
will not go on with data exclusivity. However, %94.1 of foreign drug companies think that
equivalent drug policies will go on with data exclusivity.
3. %93.8 of all the companies that joined the study state that informing relevant persons and
institutions about data protection is not sufficient and more objective studies must be done
about data protection.
Although data exclusivity can cause negative effects like decreasing in the market size of local
pharmaceutical companies, in majority both of local and foreign companies perceive the data
exclusivity as a necessary and beneficial change. Because replies of questionnaire state that data
exclusivity has positive effects in the long term like increasing on investments of foreign
pharmaceutical companies in Turkey, encouraging the local companies for research and
development activities.
145

P-30
A STUDY ON THE UTILIZATION OF PHARMACOECONOMY
BY DRUG COMPANIES IN TURKEY
H. ilbars, G. Ozelikay
Department of Pharmacy Management, Faculty of Pharmacy, University of Ankara, 06100,
Tandogan, Ankara,Turkey
Pharmacoeconomy is defined as "the health care systems and the diagnosis and analyses of drug
treatment for the public". Pharmacoeconomic study defines measures and compares the cost of
products and services in relation to pharmacy (in other words consumed resources) and the results
thereof (with respect to clinical, economical and human). In this context, study methods concerned
with the minimization of costs, cost effectiveness, cost of illness, cost utilization, cost benefit,
decision analyses, and also life quality and other human evaluations are included.
Pharmacoeconomy is a branch of economy in which; various pharmaceutical products are compared
with recourse to cost-benefit, cost- effectiveness, cost- minimization and cost utilization analyses,
or a treatment method is compared with alternative drug treatments. Pharmacoeconomy plays a
progressive role in the determination of health policies. This study aims at finding out the
perspective of companies operating in the drug industry of Turkey about Pharmacoeconomic study
and what they carry out in line with their perspective. The material of the study consists of the
survey sheets conducted with the 63 members of Drug Industry Employers Trade Union. The
survey sheet includes questions to find out whether the company carries out pharmacoeconomic
studies or not, if the answer is positive then, what kind of analyses are used, and the point of view
of the company on these kinds of studies. On the basis of the results of the study, 85 % of the drug
companies operating in Turkey declared that a department related to Pharmacoeconomy does not
exist in the company. While 76 % of the companies surveyed declare that pharmacoeconomic
studies are of utmost importance and there is necessity to conduct these studies in Turkey, the
survey results revealed out that 82 % of the companies did not conduct any pharmacoeconomic
analyses to date. There is no obligation to conduct pharmaeconomic analyses by drug companies in
our country also remain among the reasons for nonperformance of any phannacoeconomic study.
146

P-31
SATISFACTION OF YEDITEPE UNIVERSITY PRACTICE PHARMACY PATIENTS
C- Uslu, N. Sencan
Yeditepe University, Faculty of Pharmacy, 26 Agustos Yerle§imi, Kayi§dag-Istanbul.
Satisfaction is an individual's judgment about the extent to which a product or service provides a
pleasurable level of consumption-related fulfillment. Patient satisfaction with pharmacy services
has undergone several revisions, but the most recent version includes seven dimensions of
satisfaction: accesssibility , consideration , explanation, Finance , general satisfaction , product
availability and technical competence.While there is not complete agreement on these dimensions
in the previous surveys, there are some common themes that pharmacist should address, if they
wish to maximize patient satisfaction; Access the pharmacist in the pharmacy, the pharmacy itself
(location, hours), product availability, courtesy of the pharmacist, technical competency( no
medication error) and clinical competency. A survey was conducted to measure the satisfaction
Yeditepe University Faculty of Pharmacy Practice Pharmacy patients, to examine what are the
dimensions, students expect of Practice Pharmacy. 15 questions are asked to understand the
different situations and crietirias to assess patient satisfaction/dissatisfaction. Some questions were
designed to compare pairs of symmetrical situations.The survey was conducted with nearly 30 girl
and 30 boy students who stay in the dormitory of Yeditepe University, 26 Agustos Campus, in
March 2006. The results are given in tables and percentages. SPSS 11.5 program is used to avalute
the results. The final report of the survey will be crosschecked and discussed with the real services
and datas of the practice pharmacy.
1

P-32
COMPARISON OF BACITRACIN AND SULFAMETHOXAZOLE-TRIMETHOPRIM
DISC METHOD WITH REMEL STREPTEX FOR IDENTIFICATION OF GROUP A
BETA HEMOLYTIC STREPTOCOCCI
M. Eryilmaz, O. Atli, A. Akin
Departmant of Pharmaceutical Microbiology, Faculty of Pharmacy, University of Ankara, 06100
Ankara-Turkey
Group A beta hemolytic streptococcus (GABHS) is the most frequent cause of bacterial
pharyngotonsillitis. The prevalence of Streptococcus pyogenes (GABHS) infections has increased
drastically in the last decade and correlates with increasing resistance to macrolide and tetracycline
groups of antibiotics. The bacitracin and sulfamethoxazole-trimethoprim (SXT) disc method is
preferred for defining GABHS in daily routine tests since it is inexpensive, easy-to-use and has a
high rate of sensitivity although subculturing individual beta-hemolytic colonies among the
crowded bacterial colonies is sometimes very difficult. Remel Streptex is a latex agglutination test
for identification of streptococcal group antigens from cultured organisms. The latex agglutination
test, which is based on specific antigen-antibody reaction, is more sensitive than bacitracin-SXT
method. However its high cost prevents routine laboratory application. We compared Remel
Streptex with the bacitracin and SXT disc method for sensitivity in the presumptive identification of
Streptococcus pyogenes. A total of 110 GABHS isolated from throat cultures were examined. We
used the Remel Streptex as the gold standart method for identifying GABHS. Principle of Remel
Streptex is; extracting group specific antigens from streptococci by nitrous acid in a simple and
short incubation step. Than extract is neutralised and antigens are detected and identified with
suspensions of latex particles coated with group specific antibodies. A positive result is seen as an
aggregation of latex particles. Compared with the Remel Streptex, the sensitivity of bacitracin and
SXT disc method was 93.6%. As a result bacitracin and SXT disc method has a high sensitivity to
identify Streptococcus pyogenes from subcultured plates.
148

P-33
HELICOBACTER PYLORI DETECTION BY CULTURE AND REAL TIME PCR AND
DETECTION OF CLARITHROMYCIN RESISTANCE BY REAL TIME PCR
M.T. Ozdemir 1 . S. Yildiz 1 , A. Yiice 2 , H. Demir 2 , Y. Akyon 3
'Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Ankara University, 06100,
Ankara, Turkey, Departments of Hacettepe University, Medical Faculty, Pediatric
Gastroenterology, Hepatology and Nutrition Unit, 3 Microbiology and Clinical Microbiology,
06100, Ankara, Turkey
The main cause of failure of Helicobacter pylori eradication therapy is resistance to one of the most
commonly used antimicrobial agent for the H. pylori treatment clarithromycin. Rapid identification
of patients infected with clarithromycin resistant Helicobacter pylori without the need of culture,
plays an important role in the treatment. Our aim was to detect both H. pylori and the
clarithromycin resistance from the biopsy sample by a rapid method. The presence of Helicobacter
pylori and clarithromycin resistance were investigated in 201 gastric biopsy specimens obtained
from children who referred for endoscopy at Hacettepe University ihsan Dogramaci Child Hospital.
Presence of Helicobacter pylori was detected by both culture and Real Time PCR (Light Cycler,
Roche Diagnostics, Germany) methods and the sensitivity of the two methods was compared. A
Real Time probe hybridization melting point analysis assay was used to detect 2143 or 2144 (A-C
or A-G) point mutations in the 23S rRNA gene associated with clarithromycin resistance from the
gastric biopsy samples. In 201 samples 37.31 % was positive by culture and 88.06 % was positive
by Real Time PCR assay. 75 specimens were classified as H. pylori by both Real Time PCR and
culture methods. Real Time PCR results were also in consistant with a total of 24 samples which
provide negative result in the standart culture method. 2143 or 2144 (A-C or A-G) point mutation
was detected in 21.47 % of the samples and classified as clarithromycin resistant. In 7 cases (%
3,95), simultaneous presence of two different genotypes (mutant and wild type) in the same biopsy
material was detected. Although the PCR method is more expensive than standart culture methods,
the reduced time of the experiment, labor effectiveness and sensitivity of the test are major profits
of this technique. Moreover, rapid detection of resistance along with bacteria identification in the
same experiment provides an additional advantage. Real time PCR technique significantly increased
the success rate of H. pylori detection and also provided an opportunity to test clarithromycin
resistance in the same experiment of Helicobacter pylori detection.
1

p-
THE FOLK MEDICINAL PLANTS OF LALAPA§A (EDiRNE -TURKEY)
D.F. Alparslan, G.E. Bulut, E. Tuzlaci
Department of Pharmaceutical Botany, Faculty of Pharmacy, Marmara University,34668,
Haydarpasa, Istanbul, Turkey.
An ethnobotanical investigation was made between 2004-2006 in Lalapa§a (Edirne) located near the
Turkish-Bulgarian border in the Europaean part of Turkey. This presentation includes only the folk
medicinal plants pertaining 51 taxa. These are given in a table (incl. botanical name, local name,
part used, ailment treated, therapeutic effect, preparation, administration and duration of the
treatment). According to this study, the plants are mostly used for stomach ailments (Cerasus
mahaleb var. mahaleb, Cydonia oblonga, Ficus carica subsp. carica, Mentha spicata subsp.
spicata, Origanum vulgare subsp. vulgare, Plantago major subsp. major, Rumex crispus, Sambucus
nigra, Satureja hortensis, Thymus longicaulis subsp. longicaulis var. subisophyllus, Urtica dioica,
Verbascum macrurum, V. ovalifolium subsp. thracicum), diabetes (Cydonia oblonga, Hypericum
perforatum, Matricaria chamomilla var. recutita, Paliurus spina-christi, Plantago major subsp.
major, Prunus spinosa subsp. dasyphylla , Sambucus nigra, Ulmus minor subsp. canescens),
hemorrhoids (.Achillea crithmifolia, Cotinus coggyria, Fraxinus ornus subsp. ornus, Juglans regia,
Malva sylvestris, Matricaria chamomilla var. recutita, Teucrium polium), cold (Cydonia oblonga,
Lavatera thuringiaca, Morns alba, Origanum vulgare subsp. vulgare, Rosa canina), cancer
(Cerasus mahaleb var. mahaleb, Rubus canescens, R. sanctus), eczema (Cotinus coggyria, Ouercus
virgiliana, Urtica urens), heart diseases (Achillea crithmifolia, Paliurus spina-christi, Rosa canina),
prostate ailments (Carduus nutans, Corylus avellana var. avellana, Cotinus coggyria) and wart
(Cichorium intybus, Crataegus monogyna subsp. azarella, Ficus carica subsp. carica).
152

P-33
HELICOBACTER PYLORI DETECTION BY CULTURE AND REAL TIME PCR AND
DETECTION OF CLARITHROMYCIN RESISTANCE BY REAL TIME PCR
M.T. Ozdemir 1 . S. Yildiz', A. Yiice 2 , H. Demir 2 , Y. Akyon 3
'Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Ankara University, 06100,
Ankara, Turkey, Departments of Hacettepe University, Medical Faculty, 2 Pediatric
Gastroenterology, Hepatology and Nutrition Unit, 3 Microbiology and Clinical Microbiology,
06100, Ankara, Turkey
The main cause of failure of Helicobacter pylori eradication therapy is resistance to one of the most
commonly used antimicrobial agent for the H. pylori treatment clarithromycin. Rapid identification
of patients infected with clarithromycin resistant Helicobacter pylori without the need of culture,
plays an important role in the treatment. Our aim was to detect both H. pylori and the
clarithromycin resistance from the biopsy sample by a rapid method. The presence of Helicobacter
pylori and clarithromycin resistance were investigated in 201 gastric biopsy specimens obtained
from children who referred for endoscopy at Hacettepe University ihsan Dogramaci Child Hospital.
Presence of Helicobacter pylori was detected by both culture and Real Time PCR (Light Cycler,
Roche Diagnostics, Germany) methods and the sensitivity of the two methods was compared. A
Real Time probe hybridization melting point analysis assay was used to detect 2143 or 2144 (A-C
or A-G) point mutations in the 23S rRNA gene associated with clarithromycin resistance from the
gastric biopsy samples. In 201 samples 37.31 % was positive by culture and 88.06 % was positive
by Real Time PCR assay. 75 specimens were classified as H. pylori by both Real Time PCR and
culture methods. Real Time PCR results were also in consistant with a total of 24 samples which
provide negative result in the standart culture method. 2143 or 2144 (A-C or A-G) point mutation
was detected in 21.47 % of the samples and classified as clarithromycin resistant. In 7 cases (%
3,95), simultaneous presence of two different genotypes (mutant and wild type) in the same biopsy
material was detected. Although the PCR method is more expensive than standart culture methods,
the reduced time of the experiment, labor effectiveness and sensitivity of the test are major profits
of this technique. Moreover, rapid detection of resistance along with bacteria identification in the
same experiment provides an additional advantage. Real time PCR technique significantly increased
the success rate of H. pylori detection and also provided an opportunity to test clarithromycin
resistance in the same experiment of Helicobacter pylori detection.
149

P-34
MINERAL COMPONENTS OF ROSEHIPS (ROSA SP.) SALING IN HERBALIST
MARKETS IN ANKARA
J. Ahmed 1 , A. GUven 1 , A. Eken 2 , A. Aydin 2
'Department of Pharmaceutical Botany, Faculty of Pharmacy, University of Ankara, 06100,
Tandogan, Ankara, Turkey, 2 Gtilhane Military Medical Academy, Department of Pharmaceutical
Sciences, Department of Toxicology, 06500, Ankara Turkey
The genus Rosa L. (Rosaceae) which naturally growing in Turkey has a wide distribution and
represented by 30 taxon 1 . The false fruit of Rosa L. consists of the achenes enclosed in fleshy cup
shaped hepanthia. The medicinal drug according to DAB 10 consists of the ripened and dried
hypanthia of the false fruit largely freed of the fruit and the hairs attached to the receptacle J and it
contains not less than 0.3 % of ascorbic acid, calculated with reference to the dried drug 2 . This drug
is known in Turkey as "ku§burnu" and used as tonic and against constipation. Also it is used in folk
medicine as antidiabetic 4 . Because of the high amount of ascorbic acid present in the drug, it is used
as anti-cold in the folk medicine. In Anatolia the drug is collected from nature and consumed as
marmelate, fruit juice and tea 4 . In big cities this drug is sold in the herbalist markets. It is important
to have a good quality control for medicinal herbs in order to protect consumers from
contamination. For the analysis of the trace elements of the Rosa L. drug saling in herbalists, we
collected seven sample from different herbalist in five district of Ankara. Two sample has been
collected from nature also analysed. Plant materials and thier water extracts were digested in
Microwave Acid Digestion System. The important minerals of these samples (Zn, Cu, Fe, Ca, Mg,
Pb, Cd, Mo, Mn, As) were determined by Atomic Absorption Spectroscopy (AAS) with Graphite
Furnace System.
1. O. Nilsson, Rosa L., p. 106-126, In: .Davis, P.H. (ed.), "Flora of Turkey and East Aegean
Islands", Vol. 4, Edinburgh University Pres, Edinburgh (1972)
2. European Pharmacopoeia, Fourth Edition, p. 1078, Quality of Medicines of the Council of
Europe, Strasburg (2002)
3. N.G. Bisset, Herbal Drugs and Phytopharmaceuticals, p. 424-426, Medpharm Scientific
Publishers, Stuttgart (1994)
4. T. Baytop, Turkiye'de Bitkiler ile Tedavi (Gefmi^te ve Bugiin), 2. Baski, s. 214-215, Nobel Tip
Kitapevleri (1999)
1

P-35
THE ANTIOXIDANT ACTIVITIES OF EUPHORBIA ACANTHOTHAMNOS,
E. MACROCLADA, E. RIGIDA FROM TURKEY
A. Barla', M. Ozturk 3 , M. Boga \ S. Kiiltiir 2 , S. Oksuz'
Department of General Chemistry, Faculty of Pharmacy, University of Istanbul 34116, Istanbul, Turkey,
2
Department of Pharmaceutical Botanic, Faculty of Pharmacy, University of Istanbul 34116, Istanbul,
Turkey, Department of Chemistry, Faculty of Science and Arts, Mugla University Mugla 48000, Turkey
The family Euphorbiaceae comprises of 331 genus and about 7500 species. The genus Euphorbia
consists about 2000 species [1, 2], 96 species of Euphorbia grown widely in Turkey, 13 of them were
endemic [3], Species of the family Euphorbiaceae are well known to be generally toxic and to show skin
irritant [4, 5], antitumor [6] and tumor promoting effects [7, 8]. Plants of the genus Euphorbia have been
the source of a large number of biologically active compounds. Biological activities including skin
irritant, tumor promoting, and proinflammatory [7, 9]. Some species of the genus Euphorbia have been
used as medicinal plants for the treatment of skin diseases, gonorrhea, migraine and intestinal parasites,
and as wart cures [10], Euphorbia acanthothamnos, Euphorbia rigida and Euphorbia macroclada were
analyzed for their potent antioxidant activity. The antioxidant potential of extracts of E. acanthothamnos,
E. rigida and E. macroclada were evaluated using different complementary antioxidant tests, such as,
l,l-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity test, total antioxidant activity by using
B-carotene bleaching method, metal-chelating activity. In addition, total phenolic and flavanoid contents
were also measured. Acetone and ethanol extracts of E. acanthothamnos and E. macroclada were
exhibited strong total antioxidant activity higher than using known positive standards (BHT, a-
tocopherol, quercetin). Hexane extract of E. acanthothamnos and E. macroclada, and ethanol extract of
E. acanthothamnos were found to be good metal chelating activity as well as known positive standard
quercetin. In addition, total phenolic and flavonoid contents in all extracts of three Euphorbia species
were determined as pyrocatechol and quercetin equivalents, respectively. The correlation were found in
all tests as a result, between l,l-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity test and
B-carotene-linoleic acid assay.
1. Cullen J. (eds.) The Europian Garden Flora. Cambridge University Pres, vol 5. 1997. p. 73.
2. Bruirimitt RK (Compilied by). Vascular Plant Families and Genera. Whitstable Litho Ltd.,Whitstable, Kent.
1992
3. Davis PH, Mill RR, Tan K. (eds) Flora of Turkey and the East Aegean Islands, vol 10. University Pres,
Edinburg, 1988. p. 513. 539.
4. Schmidt RJ, Evans FJ. Contact dermatitis 1980; 6: 204.
5. Gundidza M, Sorg B, Hecker E. J. Ethnopharmacol. 1993; 39: 209.
6. Betancur- Galvis LA, Morales GE, Forero JE, Roldan J. Mem. Inst. Oswaldo Cruz 2002; 97: 541.
7. Evans FJ, Taylor SE. Prog. Chem. Org. Nat. Prod. 1983; 44: 1.
8. Vogg G, Mattes E, Rothenburger J, Hertkorn N, Achatz S, Sandermann Jr. H. Phytochemistry, 1999; 51: 289.
9. Macro JA, Sanz-cervera JF, Yuste A. Phytochemistry 1997; 45: 563.
10. Singla AK, Pathak K. Fitoterapia 1990; 61: 483.
1

p-
THE FOLK MEDICINAL PLANTS OF LALAPA§A (EDIRNE -TURKEY)
D.F. Alparslan, G.E. Bulut, E. Tuzlaci
Department of Pharmaceutical Botany, Faculty of Pharmacy, Marmara University,34668,
Haydarpasa, Istanbul, Turkey.
An ethnobotanical investigation was made between 2004-2006 in Lalapa$a (Edirne) located near the
Turkish-Bulgarian border in the Europaean part of Turkey. This presentation includes only the folk
medicinal plants pertaining 51 taxa. These are given in a table (incl. botanical name, local name,
part used, ailment treated, therapeutic effect, preparation, administration and duration of the
treatment). According to this study, the plants are mostly used for stomach ailments (Cerasus
mahaleb var. mahaleb, Cydonia oblonga, Ficus carica subsp. carica, Mentha spicata subsp.
spicata, Origanum vulgare subsp. vulgare, Plantago major subsp. major, Rumex crispus, Sambucus
nigra, Satureja hortensis, Thymus longicaulis subsp. longicaulis var. subisophyllus, Urtica dioica,
Verbascum macrurum, V. ovalifolium subsp. thracicum), diabetes (Cydonia oblonga, Hypericum
perforatum, Matricaria chamomilla var. recutita, Paliurus spina-christi, Plantago major subsp.
major, Prunus spinosa subsp. dasyphylla , Sambucus nigra, Ulmus minor subsp. canescens),
hemorrhoids (Achillea crithmifolia, Cotinus coggyria, Fraxinus ornus subsp. ornus, Juglans regia,
Malva sylvestris, Matricaria chamomilla var. recutita, Teucrium polium), cold (Cydonia oblonga,
Lavatera thuringiaca, Morus alba, Origanum vulgare subsp. vulgare, Rosa canina), cancer
(Cerasus mahaleb var. mahaleb, Rubus canescens, R. sanctus), eczema (Cotinus coggyria, Quercus
virgiliana, Urtica urens), heart diseases (Achillea crithmifolia, Paliurus spina-christi, Rosa canina),
prostate ailments (Carduus nutans, Corylus avellana var. avellana, Cotinus coggyria) and wart
(Cichorium intybus, Crataegus monogyna subsp. azarellci, Ficus carica subsp. carica).
152

EFFECTS OF POSIDONIA OCEANICA EXTRACT ON GLUCOSE TOLERANCE AND
ALLOXAN- INDUCED DIABETES IN RATS
1 2
M.Z. Haznedaroglu , G. Gokce
1 2
Ege University, Faculty of Pharmacy, Dept. of Pharmaceutical Botany, Dept. of Pharmacology
35100 Bornova - Izmir / Turkey
P-37
Posidonia oceanica (L.) Delile is a phanerogram allocated widely in Aegean an Mediterranean Sea
with a foremost role in ecological system. Phenolic compounds detected in the plant such as
chicoric acid, phloroglucinol, ferulic acid, caffeic acid and coumaric acid were known to have
antioxidant and antiviral properties. AntiHIV and anticarcinogenic activities of these compounds
are also areas of growing interest and a number of studies focusing on the subject are currently
being carried out. In this study, we investigated the hypoglycemic effects of Posidonia oceanica
extract (POE) in means of glucose tolerance and alloxan - induced diabetes. Administration of POE
(100 and 500 mg/kg b.wt.) to normal rats increased glucose tolerance significantly (p

DETERMINATION OF SELECTED TRACE METALS IN CYMODOCEA NODOSA BY
ATOMIC ABSORPTION SPECTROMETRY (AAS)
1 2 2 2
M.Z. Haznedaroglu , C. Tas , C. Akay , S. Cevheroglu
1
Ege University, Faculty of Pharmacy, Dept. of Pharmaceutical Botany 35100 Bornova - Izmir /
2
Tiirkiye, Ministry of National Defence, Army Drug Factory, Diskapi, Ankara, Tiirkiye
P-38
Cymodocea nodosa (Ucria) Aschers (Cymodoceacea) is a submerged marine perennial herb
distributed in shallow sandy parts of the Aegean and Mediterranean seas. This work concerns on the
quantitative analysis of trace elements and metals such as Zn, Cd, Co, Mn, Fe, Cr, Cu, Ca, Ni, Mg,
Pb, Al in Cymodocea nodosa. These elements were investigated in the rhizomes and leaves of the
plant collected from two regions of the Turkish coastline. Plant materials were digested using
microwave heating. Analyses were performed by atomic absorption spectrometry (AAS)
techniques. The concentrations of Zn, Fe, Mg, Ca and Al were higher than other analysed elements
in both tissues of the plant. Rhizomes had lower concentrations of Cd and Pb compared to the
leaves both in Kas and Marmaris.
1

P-39
DETERMINATION OF SELECTED TRACE METALS IN TARAXACUM OFFICINALE BY
ATOMIC ABSORPTION SPECTROMETRY (AAS)
1 2 2 2
M. Z. Haznedaroglu , C. Tas , C. Akay , S. Cevheroglu
Ege University, Faculty of Pharmacy, Dept. of Pharmaceutical Botany 35100 Bornova - Izmir /
2
Turkiye, Ministry of National Defence, Army Drug Factory, Diskapi, Ankara, Tiirkiye
Taraxacum officinale G. H. Weber ex Wiggers is a small perennial herb from the Asteraceae family
that has been investigated for their metal constituents previously (1,2,3,4). The plant is not native to
Turkish flora but it is distributed and identified in different areas especially in gardens. This work
concerns the quantitative analyses of trace elements and metals (Zn, Cd, Co, Mn, Fe, Cr, Cu, Ca,
Ni, Mg, Pb, Al) in the leaves and flowers of Taraxacum officinale collected from the garden of
Army Drug Factory, Ankara. The plant material was digested using microwave heating. The
analyses were performed by atomic absorption spectrometry (AAS) techniques. Leaves and the
flowers were found to have higher concentrations of Zn, Fe, Mn, Mg, Ca and Al in comparison to
the other analysed elements. Cd quantities in leaves and flowers were similar while Pb quantities
analysed from flowers were higher compared to the leaves. As the collection place was close to
traffic we can suggest that the flowers reveal the Pb pollution higher than the leaves.
1. Keane, B., Collier, M.H., Shann, J.R., Rogstad, S.H. (2001). Metal content of dandelion
Taraxacum officinale leaves in relation to soil contamination and airborne particulate matter.
The Science of the Total Environment 281: 63-78
2. Normandin, L., Kennedy, G., Zayeda, J. (1999). Potential of Dandelion Taraxacum officinale as
a bioindicator of manganese arising from the use of methylcyclopentadienyl manganese
tricarbonyl in unleaded gasoline. The Science of the Total Environment 239: 165-171.
3. Pichtel, J., Kuroiwa, K., Sawyerr H.T. (2000) Distribution of Pb, Cd and Ba in soils and plants
of two contaminated sites. Environmental Pollutionl 10: 171-178
4. Tolgyessy, J., Harangozo, M., Dillinger, P. 1993. Determination of Cu, Ni, Zn and Pb contents
in Taraxacum officinale near the highway D-61 Bratislava-Trnava (SR) by radionuclide X-ray
fluorescence analysis. Journal of Radioanalytical and Nuclear Chemistry 176: 451-455
1

P-40
LEAF ANATOMY OF TURKISH SATUREJA L. (LAMIACEAE)
F. Satil'. A. Kava 2
i
Balikesir University, Faculty of Science & Arts, Department of Biology, 10100 Balikesir, Turkey,
2
Anadolu University, Faculty of Pharmacy, Pharmaceutical Botany, 26470 Eskisehir Turkey
There are some taxonomic uncertainties within the Turkish members of Satureja. It is extremely
difficult to distinguish some of Turkey Satureja species because of the great morphological
similarity . A comparative study was undertaken on the leaves of fifteen Satureja L. species in
order to assess anatomical variations which may be a useful feature to identify each one and to
evaluate their significance for the taxonomy of the genus. The specimens are collected from
different regions of Turkey. The anatomical study has been done on fully flowered fresh plants and
herbarium material. The investigated species can be primarily divided into two main groups as
bifacial and equifacial leaf according to mesophyll structure. They can be secondary divided into
two types based on the midrib region in cross section as projecting part towards and without
protruding. Thirdly, transverse sections show that two main vascular bundle types can be identified
according to the presence or absence of scleranchyma. When transverse sections are compared,
some important differences in the protruding midrib, mesophyll and vascular bundle structures are
the most useful distinctive features for Turkish Satureja species.
1. PH. Davis, Satureja L. In: Davis PH, Mill RR, Tan K, eds. Flora of Turkey and the Aegean
Islands. Vol. 7. Edinburgh: Edinburgh University Press, 314-323 (1982).
156

P-41
THE LEAF ANATOMY OF ERICA L. SPECIES NATIVE TO TURKEY
A. Guven, G. Kendir
Department of Pharmaceutical Botany, Faculty of Pharmacy, Ankara University, 06100, Tandogan
Ankara, Turkey
The genus Erica L. (Ericaceae) is represented by more than 700 species in the world, and mainly
found in the South Africa, furthermore Mediterranean and West Europea. This genus is represented
only four species of which one taxon is endemic in Turkey. E. arborea L. and E. manipuliflora
Salisb. are very common all side regions of Turkey; E. bocquetti (Pe§men) P. F. Stevens is endemic
species and E. sicula Guss. subsp. libanotica (C.& W. Barbey) P. F. Stevens has rare distribution in
South West Anatolia. These species are called as a "funda", "puren" or "supiirge fahsi" in Turkey .
Herbal teas prepared from aerials parts of E. arborea and E. manipuliflora have been used as
diuretic, astrigent and treatment of urinary infections in Turkey. 5 % infusion of E. arborea is taken
one glassful after meals for slimming. E. arborea has been exported from Turkey. Erica species
contain many active compounds such as flavonoids, anthocyanoins, coumarins and triterpenic
compounds. These species have been reported to posses cytotoxic activity, anticarcinogenic,
antiulcer and antimicrobial activity. Although a lot of work has been done on Erica species
especially chemical and biological activity, anatomical studies on Erica species are very rare. The
aim of this study is to provide an improved anatomical properties of four Erica species based on
anatomical observations. In this research, microscopical characterstics of the leaves of E. arborea
exported and other Erica species native to Turkey are reported. Microscopical views of transverse
and surface sections from the leaves of each species were illustrated, photographed and described in
detail. Leaves are whorled, channelled beneth or strongly revolute. The simple eglandular trichomes
are located along the chanel part of abaxial surface of leaves. There are unicellular simple
eglandular trichomes on surface of upper epidermis.
1. Chamberlain, D.F., Erica L., Flora of Turkey and East Aegean Islands (Ed.: P. H. Davis) Vol. 6,
p.: 89, Edinburgh University Press, Edinburgh (1972).
2. T. Baytop, Turkiye'de Bitkiler ile Tedavi (Gefmiste ve Bugun), 2. Baski, s: 208, Nobel Tip
Kitapevleri (1999).
1

DPPH RADICAL SCAVENGING EFFECTS OF HIBISCUS TRIONUM (MALVACEAE)
EXTRACTS
P-42
C.S. Kilic 1 . T. Coban 2
'Ankara University, Faculty of Pharmacy, Department of Pharmaceutical Botany, 06100,
Tandogan, Ankara, 2 Ankara University, Faculty of Pharmacy, Department of Pharmaceutical
Toxicology, 06100, Tandogan, Ankara.
Reactive oxygen species (ROS) are implicated in many pathogenic processes. Detoxification of
ROS by antioxidants (AO) therefore affords protection against such diseases. Since a Hibiscus
species (Hibiscus sabdariffa L.), that does not grow in Turkey naturally is proved to possess
antioxidant activity, 1 ' 2 we wanted to investigate whether the only Hibiscus species growing in
Turkey, Hibiscus trionum L. 3 (Venice Mallow) also possessed antioxidant activity or not. The
comparative antioxidant potential of different extracts of H. trionum L. were studied by the 1,1'-
diphenyl-2-picryhydrazyl (DPPH ) scavenging assay. Crude water-lyophilized, methanol, and
aqueous dried extracts from aerial parts were prepared separately. All extracts were assayed in the
concentration range 0,5-0.0625 mg /ml and all extract possessed the strong inhibition on DPPH
radical. Methanol extracts exhibited the highest scavenging activity on DPPH radical with IC50
0.092 mg/mL. Water and aqueous dried extracts showed significant DPPH radical scavenging
activity with IC50 0.22 and IC50 0.36 mg/mL, respectively. This result is rather interesting since this
species is considered to be a noxious weed and farmers are advised to take action against it
immediately. 4 ' 5 By proving with this study that we may benefit from this plant, we may be able to
alter the reputation of this plant.
1. Liu, J-Y, Chen, C-C, Wang, W-H, Hsu, J-D, Yang, M-Y, Wang, C-H, The protective effects of
Hibiscus sabdariffa extract on CCU-induced liver fibrosis in rats, Food and Chemical Toxicology,
44, 336-343 (2006)
2. Duh, P-D., Yen, G-C., Antioxidative eactivity of three herbal water extracts, Food Chemistry, 4,
639-645, 1997
3. Davis, PH, Flora of Turkey and the East Aegean Islands, Edinburgh University Press, Edinburgh,
UK, Volume 2, 402(1967)
4. http://www.agriculture.com/ag/story,jhtml;jsessionid=PZ2ZJJlNSLJNJQFIBQSCCAQAQstory
id=/templatedata/ag/story/data/agWorld_8138.xml
5. http://www.co.weld.co.us/departments/weed_pest/pdf/factSheets/WeldCountyWeedWatch4.Watc
h4.pdf
1

P-43
COMPARATIVE LEAF ANATOMICAL CHARACTERISTICS OF THE TURKISH
ALLIUM SPECIES (SECT. BREVISPATHA, SECT. SCORODON AND SECT.
CODONOPRASUM)
M. Kocvigit, N. Ozhatay
Istanbul University, Faculty of Pharmacy, Department of Pharmaceutical Botany, 34452, Beyazit-
Istanbul
The genus Allium L. represented by 197 taxa in Turkey of which 72 are endemic (endemizm rate
36,5%).They are grouped into 14 sections in this study closely related sections are examined by leaf
anatomical characteristics: Sect. Scorodon C. Koch, Sect. Codonoprasum Reichb., Sect.
Brevispatha Valsecchi.
Sect. Brevispatha (7 wild taxa in Turkey) :
Examined specimens:
A. callidictyon; B6 Sivas, Ula, Tecer village, Tecer Mont. 1500 m, 21.7.1978, ISTE 40989.
A.callimischon subsp. haemostictum; C2 Mugla, Koycegiz, Sandras Mont., 500 m, 28.10.1977,
ISTE 43972.
Sect. Scorodon (13 wild taxa in Turkey)
Examined specimens:
A. sivasicum; A7 Giresun, §ebinkarahisar-Alucra, 18 km from Alucra, 1710 m, 9.7.1982, ISTE
49341.
A.moschatum; Al (A) Balikesir, Marmara adasi, VirankOy hills, 400 m, 24.8.1979, ISTE 43503.
Sect. Codonoprasum (43 wild taxa in Turkey)
Examined specimens:
A.flavum subsp. flavum; Al (E) Kirklareli-Derekoy, 5 km from Kirklareli, 1350 m, 9.9.1976, ISTE
35923.
A. paniculatum subsp. paniculatum; B1 Balikesir, Kazdagi, Kapikule 1350 m 9.8.2004, ISTE
81860.
Leaf; unifacial. Cuticle; bearing a central longitudinal striation over most cells or occasionally a
row of micropapillae. Epidermis; cells in regular files, longitudinally elongated. Palisade and
spongy parenchyma are not fully distinguishable, however at some places single layered palisade
cells are in the sections. Laticifers situated at spongy parenchyma. Stomata; numerous, anomocytic.
Sect. Brevispatha certainly are distinguishable with anatomical characters from the other two
sections, however Sect. Scorodon and Sect. Codonoprasum have similar anatomical characteristics.
1

INVESTIGATIONS OF ANTINOCICEPTIVE AND ANTI-INFLAMMATORY
ACTIVITIES OF PHENOLIC COMPOUNDS OF SIDERITIS BRE VIBRA CTEA TA P.H.
DAVIS
A. Guvenf 1 , E. Kiipeli 2 . i. Cali§ 3
P-44
'Ankara University, Faculty of Pharmacy, Department of Pharmaceutical Botany, Tandogan
06100, Ankara, Turkey, 2 Gazi University, Faculty of Pharmacy, Department of Pharmacognosy,
Etiler 06330, Ankara, Turkey,
3 Hacettepe University, Faculty of Pharmacy, Department of
Pharmacognosy, 06100 Ankara- Turkey
Sideritis L. (Lamiaceae, Labiatae) species are widely used as medicinal plants and as herbal teas in
Turkey in which 45 species of the genus are naturally found. Sideritis brevibracteata P. H. Davis is
an endemic species. Around Alanya (Antalya) the aerials parts of S. brevibracteata are served as
tea. In a previous screening study, it was reported that S. brevibracteata has shown high antioxidant
activity. The methanol, chloroform, n-butanol and remain aqueous extracts from aerial parts of S.
brevibracteata P.H. Davis, were investigated for their in vivo anti-inflammatory and antinociceptive
activity. For the anti-inflammatory activity, carrageenan-induced hind paw edema, PGEi- induced
hind paw edema and 12-0-tetradecanoyl-13-acetate (TPA)-induced mouse ear edema models and
for the antinociceptive activity, p-benzoquinone-induced abdominal constriction test were used, n-
Butanol extract of the plant was shown to possess significant inhibitory activity against
carrageenan-induced hind paw edema model and of/>-benzoquinone-induced writhings in mice. In
the present study, five 8-hydroxyflavone glycosides and one phenylethanoid glycoside isoleted from
«-butanol extract of the aerial parts of S. brevibracteata have been investigated for their activities.
These compounds are hypolaetin 7-0-[6'"-0-acetyl-p-D-allopyranosyl-(l—>2)-P-Dglucopyranoside]
(1), hypolaetin 7-0-[6"'-0-acetyl-|3-D-allopyranosyl-(l—>2)-6"-C-acetyl-P-Dglucopyranoside]
(2), isoscutellarein 7-0-[6"'-

P-45
MINERAL COMPONENTS OF ELDER FLOWERS (SAMBUCUS L.) SALING IN
HERBALIST MARKETS IN ANKARA
O. Mumcu Arisan 1 , A. Giiven 2 , A. Eken 3 , A. Aydm 3
'Hacettepe University, Faculy of Pharmacy, Department of Pharmaceutical Botany, 06100, Sihhiye,
Ankara, Turkey, 2 Ankara University, Faculy of Pharmacy, Department of Pharmaceutical Botany,
06100, Tandogan, Ankara, Turkey, 3 Gulhane Military Academy, Department of Pharmaceutical
Sciences, Department of Toxicology, 06500, Ankara, Turkey
The genus Sambucus L. (Caprifoliaceae) represented by two species in Turkey, mainly found in
East Anatolia and Black Sea Regions'. The medicinal drug of Elder flowers consists of the dried
flowers of Sambucus nigra L. It contains not less than 0,8% of flavonoids, calculated as
isoquercitroside with reference to the dried drug 2 . The drug used as anti-cold, antipyretic and eye
diseases 3 . In Turkey, this dug is known as "miirver" and used as diuretic, diaforetic and anticonstipation
in the folk medicine 4 . In Anatolia the drug is collected from nature and consumed as
tea. In big cities this drug is sold in the herbalist markets. It is important to have quality control for
medicinal herbs in order to protect consumers from contamination. For the analysis of the trace
elements of the Sambucus L. drug saling in herbalists, we collected five sample from different
herbalist in five district of Ankara. Plant material and their water extract were digested in
Microwave Acid Digestion System. The important minerals of these samples (Zn, Cu, Ca, Mg, Pb,
Cd, Mo, Mn, As) were determinated by Atomic Absorption Spectroscopy (AAS) with Graphite
Furnace System.
1. D.F. Chamberlain, Sambucus L., In: Davis, P.H. (ed.), "Flora of Turkey and East Aegean
Islands", Vol. 4, p. 542-543, Edinburgh University Press, Edinburgh (1972).
2. European Pharmacopoeia, Fourth Edition, p. 1098-1099, Quality of Medicines of the Council of
Europe, Strasburg (2002).
3. N.G. Bisset, Herbal Drugs and Phytopharmaceuticals, p. 446-448, Medpharm Scientific
Publishers, Stuttgart (1994).
4. T. Baytop, Turkiye'de Bitkiler ile Tedavi (Ge9mite ve BugUn), 2. Baski, s. 214-215, Nobel Tip
Kitapevleri (1999).
16

THE ANTIOXIDANT ACTIVITY OF STEMS AND ROOTS OF RHUBARB: RHEUMRIBES L.
(I§KIN): AN EDIBLE PLANT
P-46
1,2 . . 1 2 3
M. Oztiirk ' , F. Aydogmu Oztiirk , M. E. Duru , G. Topu
i
Department of Analytical Chemistry, Faculty of Pharmacy, University of Istanbul 34116, Istanbul,
2
Turkiye, Department of Chemistry, Faculty of Science and Arts, Mugla University Mugla 48000,
Turkiye, Department of Chemistry, Faculty of Science, Istanbul Technical University Istanbul 34469,
Turkiye
Rheum ribes L. (Polygonaceae) is locally known as "lgin, usgun or u^gun" and grown mostly in Eastern
Turkiye, Lebanon and Iran. R. ribes is the source of one of the most important crude drugs in Asiatic
regions [1]. Rheum species are medicinally important plants due to anthracene derivatives occurring in
the subterranean parts of the plant [2] and their roots are used as laxative and antipsoriatic drug in Iran
[3]. The roots of the plants are used for the treatment of hemorrhoids and diabetes [4] while the stems
and petioles of the plant have digestive and appetizer usage in Eastern Turkiye. R. ribes, the only native
Rheum species growing in Turkiye which has been studied by chemically and some anthraquinones,
quercetin flavonoids and a stilben were obtained [4, 5], Although the roots, stalk and leaves of extracts
of R. ribes were investigated for their antimicrobial [6] and hypoglycemic activity [7] there is no study
on the antioxidant activity of R. ribes. In this study, the antioxidant activity of chloroform and methanol
extract of roots and stem of R. ribes was studied by using six different antioxidant tests, namely total
antioxidant activity by B-carotene bleaching method, DPPH radical scavenging activity, ferric reducing
power, cupric reducing antioxidant capacity (CUPRAC), superoxide anion radical scavenging and metal
chelating activities comparing with well known standards a-tocopherol, quercetin, BHT and L-ascorbic
acid. All tests are carried out in triplicate. Total antioxidant activity of the chloroform and methanol
extracts of the roots (93.1%, 84.1%) and stems (82.2%, 82.0%) respectively, exhibited strong activity
being higher than known standards BHT (66.2%) and a-tocopherol (65.2%) at 100 pg/mL concentration.
DPPH radical scavenging potential of stems was found to be higher than roots. Except chloroform
extracts of the roots, all extracts were exhibited good metal chelating activity than quercetin. Both
extracts of the roots showed more potent superoxide anion radical scavenging activity than BHT, and
comparable with well known radical scavenger L-ascorbic acid. Also, total phenolic and flavonoid
contents in both extracts of the roots and stems of R. ribes were determined as pyrocatechol and
quercetin equivalents.
1. Kashiwada, Y., Nonaka, G., Nishioka, I. & Yamagishi, T. (1988). Phytochemistry, 27, 1473-1477.
2. Baytop, T. (1999). Therapy with Medicinal Plants in Turkiye (2nd edn). Nobel Tip Kit.: Istanbul,
319-320.
3. Shokravi, A. & Agha Nasiri, K. (1997). Iran J. Chem. & Chem. Eng., 16, 10-15.
4. Meri9li, A.H., Tuzlaci, E. (1990). Fitoterapia, 61, 375.
5. Tosun, F. and Akyiiz-Kizilay, (2003). Ankara. Ecz. Fac. Derg., 32(1), 31-35.
6. Bonjar, G.H.S. (2004). Asian J. of Plant Sci., 3, 82-86.
7. Ozbek, H., Ceylan, E., Kara, M., Ozgokce, F., Koyuncu, M. (2004). Scand. J. Lab. Anim. Sci., 31(2),
113-115.
16

P-47
SCREENING OF ANTIOXIDANT ACTIVITY OF THREE EDIBLE COMMERCIAL
MUSHROOMS (MORCHELLA CONIC A VAR. CONIC A, LEPISTA NUBA AND LACTARIUS
DELICIOUS)
I. Kivrak M. Oztiirk U2 , M.E. Dura A. Tlirkoglu 3
1 Department of Chemistry, Faculty of Science and Arts, Mugla University, Mugla 48000, Tiirkiye, 2
Department of Chemistry, Faculty of Pharmacy, Istanbul University 34116, Istanbul, Tiirkiye, 3 Faculty of
Education, Department of Science Education, Pamukkale University, 20020, Denizli, Tiirkiye
Oxidation is essential to many living organisms for the production of energy to fuel biological processes.
Oxidative damage caused by free radicals may be related to aging and diseases, such as atherosclerosis,
diabetes, cancer and cirrhosis [1]. Almost all organisms are well protected against free radical damage by
enzymes such as superoxide dismutase and catalase, or compounds such as ascorbic acid, tocopherol and
glutathione [2], Synthetic antioxidants have been used in stabilization of foods. The most commonly used
synthetic antioxidants are butylatedhydroxyanisole (BHA), butylatedhydroxytoluene (BHT), and tertbutylated
hydroxyquinone (TBHQ), that are applied in fat and oily foods to prevent oxidative deterioration.
However, BHA and BHT were found to be anticarcinogenic as well as carcinogenic in experimental
animals [3]. The medicinal mushrooms have an established history of use in traditional oriental therapies in
Mugla and Denizli provinces. Morchella conica var. conica (Pers.) Bound., Lepista nuda (Bull.) Cooke
and Lactarius delicious Fr. macro fungi are valuable economically, and this mushroom species is widely
consumed. Although M. conica, L. nuda and L. delicious which are commercial mushrooms were found to
be medicinally active in several therapies such as antitumour, antiviral, and immunomodulating activities
[4], there is no study on the antioxidant activity of these three mushrooms. In this study, the antioxidant
activity of ethanol extracts of M. conica, L. nuda, and L. delicious, which are economically valuable, were
studied by using two complementary antioxidant tests, namely total antioxidant activity by (3-carotene
bleaching method, DPPH radical scavenging activities comparing with well known standards a-tocopherol
and BHT. In addition, total phenolic and flavonoid contents in extracts of the three mushrooms were
determined as pyrocatechol and quercetin equivalents. All tests are carried out in triplicate. Total
antioxidant activities of M. conica, L. nuda, L. delicious, BHA and a-tocopherol have been found to be
86.8%, 81.9%, 82.2%, 96.4% and 98.6%, respectively. Total flavonoid amount of mushrooms has been
found to be 9.17±0.12, 8.21±0.02 and 7.17±0.16 \ig mg" 1 quercetin equivalent, respectively. And phenolic
compound amount has been found to be 63.80±0.25, 48.01±0.09 and 41.93±0.03 (.ig mg" 1 pyrocatechol
equivalent, respectively. Positive correlations were found between total phenolic and flavonoid content in
the mushroom extracts and their antioxidant activities. Edible mushrooms may have potential as natural
antioxidants.
1. Halliwell, B., & Gutteridge, J. M. C. (1984). Biochemical Journal, 219, 1-4.
2. Mau, J.-L., Lin, H.-C., & Song, S.-F. (2002). Food Research International, 35, 519-526.
3. Botterweck, A. A. M., Verhagen, H., Goldbohm, R. A., Kleinjans, J., & Brandt, P. A. (2000). Food
and Chemical Toxicology, 38, 599-605.
4. Wasser, S. P., & Weis, A. L. (1999). International Journal of Medicinal Mushrooms, 1, 31-62.
16

P-48
ANTIOXIDANT ACTIVITY OF TWO EDIBLE OPUNTIA FRUITS
(iOPUNTIA FICUS-INDICA AND O. MACROCENTRA)
I. Kivrak 1 , M. Oztiirk 1 ' 2 . M.E. Duru 1 , M. Harmandar 1
1 Department of Chemistry, Faculty of Science and Arts, Mugla University, Mugla 48000, Turkiye,
2 Department of Chemistry, Faculty of Pharmacy, Istanbul University 34116, Istanbul, Tiirkiye
Increasing world population and urbanization, industrial pollution, increasing interest for fast-food,
synthetic additives used to keep food longer result in an increase in diseases such as cancer. Widely
used in foods, BHT and BHA are known to be toxic. Evidence obtained from the studies and
showing that the main cause behind the increase seen in cancer cases is the use of synthetic matters
obliges people to turn into natural products [1]. The cactus family - Cactaceae - includes about 2000
species of plants distributed in places of desertic or very dry climate, mainly in Central America and
South America, although they have been introduced and adapted to other places of dry and warm
climate, such as Australia, the Mediterranean and East Africa. More rarely, we encounter epifitic
species evolved to live in forests of humid climates, though. Opuntia has been the main genus in the
opuntioid subfamily of Cactaceae [2], Besides being consumed as fruits, Opuntia species spreading
endemically on the coasts of Mediterranean in our country are also used for oriental therapies of
orthopedic problems [3], Although these fruits are widely consumed by the people, no study has
been carried out to determine and analyze their phenolic and flavonoid compounds; hence, we have
decided to conduct this study. For this purpose, O. ficus-indica (L.) Miller and O. macrocentra (L.)
Miller, which are widely consumed in our country, were investigated for antioxidant potential. The
antioxidant activities were determined by using three complementary in-vitro assays inhibition of
DPPH radical, total antioxidant activity, reducing power. Total phenolic contend and total flavonoid
contend also measured. Total antioxidant activity and DPPH free radical scavenging activity of the
both methanol extracts of Opuntia have been found to have almost similar inhibition value to that of
commercially available standard antioxidants (BHT, BHA). So, it is hoped to detect new natural
antioxidant compounds with no cytotoxic effect as alternative to synthetic antioxidant compounds.
1. Botterweck, A. A. M., Verhagen, H., Goldbohm, R. A., Kleinjans, J.,Brandt, P.A. (2000).
Food and Chemical Toxicology, 38, 599-605.
2. Davis, P. H. (1982). Flora of Turkey and the East Eagan Islands, Edinburgh, University
Press, Edinburgh.
3. Honda G, Yesilada E, Tabata M, Sezik E, Fujita T, Takeda Y, Takaishi Y, Tanaka T (1996)
Journal of Ethnopharmacology, 53, 75-87.
16

BOTANICAL INVESTIGATIONS ON SIDERITIS ALBIFLORA HUB.-MOR. & SIDERITIS
LEPTOCLADA O. SCHWARZ & P.H. DAVIS
1 2 1
F.P. Sahin , H. Duman , N. Ezer
i
Department of Pharmaceutical Botany, Faculty of Pharmacy, Hacettepe University, 06100,
2
Sihhiye, Ankara, Turkey, Department of Biology, Faculty of Science and Art, Gazi University,
06500, Teknikokullar, Ankara, Turkey.
P-49
The genus Sideritis L.(Lamiaceae) is distributed in an area stretching from the Mediterranean region
to Europe, Bahama's, Western China and Morocco. In the Flora of Turkey and the East Eagean
Islands, 38 Sideritis species were reported by Huber-Morath in 1982. Since then, 6 species and 2
new records have been described in the flora of Turkey and the number of Sideritis species reached
to 46. The genus Sideritis is devided into two sections in Turkey. Section Hesiodia Bentham is
known with reliable taxonomic characters. Section Empedoclia (Rafin) Bentham which shows a
high level of endemism was reported with a few clear-cut species. As a part of our ongoing studies
on Sideritis species growing in Turkey, in this study we have explained morphological
characteristics of two endemic Sideritis species, Sideritis albiflora Flub.-Mor. and Sideritis
leptoclada O. Schwarz & P.H. Davis, which belong to Empedoclia section and are close to each
other by means of taxanomic features.
This work supported by TUBITAK (TBAG 1853)
16

BOTANICAL INVESTIGATIONS ON TWO PURPLE-VIOLET FLOWERED SIDERITIS
SPECIES: SIDERITISRUBRIFLORA HUB.-MOR. & SIDERITIS OZTURKIIZ. AYTAC &
A. AKSOY
1 2 1
F.P. Sahin , H. Duman , N. Ezer
i
Department of Pharmaceutical Botany, Faculty of Pharmacy, Hacettepe University, 06100,
2
Sihhiye, Ankara, Turkey, Department of Biology, Faculty of Science and Art, Gazi University,
06500, Teknikokullar, Ankara, Turkey.
P-50
The genus Sideritis L. (Labiatae) including more than 150 species worldwide is distributed mainly
in Spain and Turkey. Many species of the genus have been known since the time of Dioscorides and
used in the traditional folk medicine, especially in the Middle East, as herbal tea to treat different
illness. In Turkish flora, 53 Sideritis taxa consist of 39 species, 12 subspecies and 2 varietes are
known. The yellow flowered species is most common. Four taxa have purple-violet flower while
one has white. In this study, botanical investigations were carried out on two purple-flowered
Sideritis species, Sideritis rubriflora Hub.-Mor. and Sideritis ozturkii Z. Ayta & A. Aksoy which
are endemic and used as herbal tea. The characteristics detected during morphological studies were
explained by drawings and original pictures.
This work supported by TUBITAK (TBAG 1853)
16

P-51
A TAXONOMIC DISCUSSION ON SIDERITIS SYRIACA SUBSP. NUSAIRIENSIS (POST)
HUB.-MOR. & SIDERITIS HUBER-MORA THIIGREUTER & BURDET
1 2 1
F.P. Sahin , H. Duman , N. Ezer
i
Department of Pharmaceutical Botany, Faculty of Pharmacy, Hacettepe University, 06100,
2
Sihhiye, Ankara, Turkey, Department of Biology, Faculty of Science and Art, Gazi University,
06500, Teknikokullar, Ankara, Turkey.
The genus Sideritis L. (Lamiaceae) is represented by more than 150 species which are distributed
from Bahama's to Western China and from Germany to Morocco, and mainly found in the
Mediterranean region. The genus Sideritis comprises two sections in Turkey. While section
Hesiodia Bentham includes 4 species, 42 species belong to section Empedoclia (Rafin) Bentham, a
taxanomically difficult section, which usually has no particular element since growing in areas
transitional between two phytogeographical regions in Turkey. Sideritis huber-morathii Greuter &
Burdet is reported to be closely related to Sideritis syriaca subsp. nusairiensis (Post) Hub.-Mor.
This study was carried out in order to provide insight into the morphology of these two close
Sideritis species which belong to section Empedoclia and discuss their taxonomic status.
This work supported by TUBITAK (TBAG 1853)
16

DETERMINATION OF NAPHTHALENE IN HONEY SPECIMENS PROVIDED FROM
NORTH, SOUTH, EAST, SOUTH -EAST AND CENTRAL ANATOLIA BY HPLC
D. Beyoglu. G.Z. Omurtag
Department of Pharmaceutical Toxicology, Faculty of Pharmacy, Marmara University, Haydarpasa
34668, Istanbul, Turkey.
P-52
Naphthalene (NAF) is a chemical compound sometimes found in honey due to an indirect
contamination. The occurrence of NAF depends on factors such as pesticide treatment. NAF causes
a number of illnesses in man and laboratory animals such as haematological disorders and cataract.
In our study, NAF was investigated in honey samples which are provided from markets and street
bazaars. NAF was detected using high performance liquid chromatography (HPLC) with diode
array detector (DAD). HPLC was used because of their facility in detection. In this study, NAF
analysis was carried out in 45 honey samples. Out of 45 samples, 8 of them were purchased from
markets and 37 of them were from street bazaars existing in North, South, East, South -East and
Central Anatolia. NAF was not detected in honey samples. Honey has an important role in our food
chain and in our country's economy. Therefore, foodstuffs are needed to be controlled and analyzed
during food process. All residue analyses for the entire food chain have a lot of importance for
healthy people and healthy generations.
(The results presented in this study were obtained during an investigation supported by the
Scientific Research Projects Commission of Marmara University; project number SAG-
040/230804.)
16

THE ANTIOXIDANT EFFECT OF TAURINE ON HEXAVALENT CHROMIUM INDUCED
OXIDATIVE STRESS IN MICE HEART
I 2 1
1.1. Boftgelmez , T. Soylemezoglu , G. Giivendik
1 2
Department of Toxicology, Faculty of Pharmacy, Ankara University, 06100, Ankara-Turkey,
Institute of Forensic Medicine, Ankara University, 06100, Dikimevi, Ankara-Turkey
P-53
Hexavalent chromium [Cr(VI)] compounds which have extensive industrial uses, exert serious
toxic, carcinogenic and mutagenic effects in human and animals (1,2). The exact mechanism of
Cr(VI) toxicity has not been elucidated yet; however, it has been suggested that intracellular
reduction of Cr(VI) and related free radical reactions play important role in cytotoxicity (3,4). The
aim of this study was to investigate the effect of taurine treatment on Cr(VI)-induced oxidative
stress in mice heart tissue. Experimental groups consisted of four subgroups: (l)control, (2)Cr(VI)-
injected, (3)taurine pre-treatment, (4)taurine post-treatment. As a biomarker of lipid peroxidation,
thiobarbituric acid reactive substances (TBARS) and non-protein thiols (NPSH) levels were
determined. Swiss albino mice were exposed to K.2Cr207 as Cr(VI), intraperitoneally, at a single
dose of 20 mg Cr/kg. Taurine (lg/kg) was administered intraperitoneally either an hour (lh) before
or after Cr(VI) exposure. In the exposure group, it was obvious that TBARS level was significantly
elevated, and NPSH level was slightly reduced as compared with the control group. When taurine
treatment groups were evaluated in this respect, taurine pre- and post- treatments effectively
reduced the Cr(VI)-induced lipid peroxidation. In line with this finding, taurine treatments also
ameliorated the NPSH level supression. Therefore, taurine seems to exert both protective and
antidotal effects on Cr(VI)-induced oxidative stress in mice heart tissue. Treatment with
antioxidants such as taurine could be suggested as a potential strategy in order to prevent or attenute
Cr(VI)-induced damage.
1. Barceloux, D.G. (1999) Clin. Toxicol., 37(2):173-194.
2. De Flora, S. (2000) 21(4):533-541.
3. Liu, K.J., Shi, X. (2001) Mol. Cell. Biochem., 222:41-47.
4. Shi, X., Chiu, A., Chen, CT, Halliwell, B., Castranova, V., Vallyathan, V. (1999) J. Toxicol.
Environ. Health, Part B, 2:87-104.
16

P-54
NEOPTERIN AS A MARKER FOR IMMUNE SYSTEM ACTIVATION IN COAL
MINERS' PNEUMOCONIOSIS
O. Cemiloglu Ulker^, B. Yucesoy^, I. O. Tekin^, A. Karakaya'
1 Ankara University, Faculty of Pharmacy, Department of Toxicology, Ankara, Turkey,
2 Toxicology and Molecular Biology Branch, Health Effects Laboratory Division, National Institute
for Occupational Safety and Health, Morgantown, WV, USA, ^Karaelmas University, Faculty of
Medicine, Department of Immunology, Zonguldak, Turkey
Coal workers' pneumoconiosis (CWP) is an important occupational, fibrotic pulmonary disease that
occurs by chronic inhalation of coal dust. Development of CWP is related with the immune system
activation. CWP is divided into two stages according to the severity of disease: Simple
pneumoconiosis (SP) and progressive massive fibrosis (PMF). Neopterin is a marker associated
with the activation cell-mediated immunity so the levels of neopterin in body fluids are elevated in
cell-mediated immune system diseases. This study was undertaken to find out if the urinary and
serum neopterin levels are useful markers for CWP disease and the disease severity. The mean
serum and urinary neopterin levels were measured in both CWP and control groups. As the control
group, active and retired miners were selected. In the CWP group mean serum neopterin level was
significantly higher than that of the control group ( 9.97 ± 1.73 nmol/1 vs 5.81 ± 0.46 nmol/1,
p

ASSESSMENT OF THE OPTIMUM DOSE FOR CHROMOSOMAL RADIOSENSITIVITY
ASSAY AND INDUCED MICRONUCLEUS FREQUENCIES IN HEAD AND NECK
CANCER PATIENTS
E. Coskun 1 , G. Cakmak Demircigil 1 , F. Cetindag 2 , O. Sunter 2 , H. Edinsel 2 , N.A. Kocabas 1 , S.
Burgaz 1
P-55
^Gazi University, Faculty of Pharmacy, Dept. of Toxicology, Hipodrom, Ankara-Turkey
2 Abdurrahman Yurtaslan Oncology Hospital, Dept.of Radiation Oncology, Head and Neck Group,
Ankara-Turkey
The micronucleus (MN) assay has been used to evaluate the radiation sensitivity and to predict
cancer risk of human subjects. The main aims of this study are to decide the optimum dose for
irradiation and to evaluate the chromosomal radiosensitivity of head and neck cancer (FTNC)
patients (n=41) and healthy controls (C) (n=27) using the MN assay. For the assessment of dose
response curves, whole blood samples of a healthy subject were irradiated with a dose range of 0.5 -
4 Gy g rays ( 60 Co) in vitro and a dose rate of 0.62 Gy/min. 2 Gy has been selected as the optimum
dose which shows an evident MN induction without cytotoxicity by trypan blue test. The mean ±
SD frequencies (%o) of radiation-induced MN were 165.29±46.86 and 151.00±43.38 in HNC
patients and C subjects which is not significantly different. Baseline MN frequencies of HNC
patients (27.85±10.42) were significantly higher than C subjects (9.22±7.26) (p

P-56
MICRONUCLEUS FREQUENCIES IN SURROGATE AND TARGET TISSUES OF
WORKERS EXPOSED TO CRYSTALLINE SILICA CONTAINING DUST
G. Cakmak Demircigil 1 , E. Coskun 1 , N. Vidinli 2 , M. Yilmaz 3 , Y. Erbas 2 , A. Cimrin 4 , R.PF.
Schins 5 , P. J. A. Borm 5 , S. Burgaz 1
'Department of Toxicology, Gazi University, Turkey, 2 National Institute of Occupational Safety
and Health, Ankara, Turkey, 3 Department of Otolaryngology, Medicine Faculty, Gazi University,
Turkey, Department of Chest Disease, Faculty of Medicine, Dokuz Eylul University, Izmir,
Turkey, 5 Particle Research, Environmental Health Research (IUF), Heinrich-Heine University,
Germany.
The International Agency for Research on Cancer (IARC,1997) reclassified respirable crystalline
silica (quartz and cristobalite) inhaled from occupational sources as a human carcinogen, based on
epidemiological and animal studies. Biological activity of crystalline silica has been found to be
variable among different industries where exposure occured. Mining, crushing, grinding,
sandblasting, construction are among high risk activities regarding to the crystalline silica exposure
in developing countries such as Turkey. We have conducted a workplace study which was carried
out among 50 male workers from 7 different workplaces, mainly involved in grinding, mixing,
bagging and sandblasting and 29 healthy male officials matched for age and smoking status as non
exposed control group. Workers were exposed to crystalline silica containing dust for between 4
months and 28 years with an average duration of 7 years.In our study, it has been evaluated the
genotoxicity in crystalline silica exposed workers by micronucleus assay both in peripheral
lymphocytes (PBL) as a surrogate tissue and nasal epithelial cells (NEC) as a target tissue.
Additionaly, the respirable dust and the respirable crystalline silica concentrations of the
workplaces, chest X ray film evaluations of the workers and control group have been made in order
to find out the exposure levels and silicosis risk in such workplaces. Micronucleus frequencies
(%mean±SD) in PBL and NEC cells of workers vs control group respectively were 1,25±0,42 vs
0,56±0,29 and 0,83± 0,23 vs 0,28±0,16. These MN frequencies of workers were significantly higher
than the control group (p

ORGANOCHLORINE PESTICIDE (OCP) AND POLYCHLORINATED BIPHENYL (PCB)
LEVELS IN ADIPOSE TISSUE OF INFERTILE MEN
T. Calik Durmaz M.H. $atiroglu^, B. Aydinuraz^, C. KabukoP, I. Cok 1
P-57
^Gazi University, Faculty of Pharmacy, Department of Toxicology, 06330, Hipodrom,Ankara,
Turkey, 2 Department of Obstetrics and Gynecology, School of Medicine, Ankara University,
06590 Cebeci, Ankara, Turkey, ^Gen-Art Women Health, IVF & Reproductive Biotechnology,
Cinnah Caddesi No: 47, 06680, Qankaya, Ankara, Turkey
Considerable attention has been given in the past few years to the possibility that man-made
chemicals (xenobiotics) in the environment may pose a hazard to human reproductive health.
Several studies have pointed out the fact that chemical compounds, drugs, solvents and
environmental pollutants could mimic or antagonize the effects of steroid hormones, like estrogens
and androgens. These chemicals are called as xenoestrogens. They are widespread used chemicals
like pesticides (i.e. p.p'DDE, o.p'DDT, chlordan, endosulfan,...) dyes and paintings (i.e phenol red)
or the degradation of plastic materials (i.e. bisphenol A). Although direct evidence is lacking,
theorical considerations and epidemiological evidence implicate these compounds as potential
hazards to human and wildlife reproductive health.The present study aims to determine the levels of
some Organochlorine pesticides (OCPs) and Polychlorinated biphenyls (PCBs) in infertile men. For
this purpose adipose tissue samples provided from males who have been living in Ankara at least
for 5 years and who have been diagnosed as infertile. Residual levels of OCPs (a-BHC, (3-BHC, y-
BHC, HCB , Endosulphan I, II, p,p'-DDE , and p,p'-DDT) and seven major persistent PCB
congeners (IUPAC Numbers: 28,52, 101, 118, 138, 153, 180) were measured in 25 infertile men
and 20 healthy men's adipose tissue samples by gas chromatography with electron capture detection
(GC-ECD). In the study the levels of OCPs and PCBs in adipose tissue of infertile men compared
with those provided from controls. No statistically significant differences were found between
control and infertile group (p>0.05) for OCP's. However significant differences (p

DETERMINATION OF FUMONISIN BI AND B 2 IN SOME MEDICINAL PLANTS BY
HPLC
M.U. Dumlu 1 . G.Z. Omurtag 2
1 2
Marmara University, Faculty of Pharmacy, Department of Pharmacognosy, "Department of
Pharmaceutical Toxicology, 34668 Haydarpasa, Istanbul, Turkey
P-58
Fumonisin Bi (FBi) and B 2 (FB 2 ) being mycotoxins, are secondary metabolites produced by several
species of the fungus Fusarium, such as Fusarium verticillioides (Sacc.) Nirenberg (ex F.
moniliforme Sheldon) and Fusarium proliferatum. FBi is a common fungal contaminant of cereals
and other agricultural products (medicinal plants etc.). Also, FBi is the most prevalent of
fumonisins. It is known to be toxic to animals and potential carcinogen to humans. FB| is known to
be the cause of equine leukoencephalomalacia in horses and porcine pulmonary syndrome. FB]
inhibits cell growth, is hepatocarcinogenic in rats and it has been statistically associated with high
incidence of esophageal cancer in humans. The objective of this study was to measure the potential
level of FBi and FB 2 contamination in some medicinal plants that are especially consumed in
Turkey. FBi and FB 2 were detected using the high performance liquid chromatography (HPLC)
with fluorescence detection after derivatization with o-phthaldialdehide (OPA). The total number of
home-made available medicinal plant samples such as lime, coriander and rosehip analyzed in this
research was 41. The origin of the whole samples was the Black Sea region of Turkey. The
minimum detectable amount for the OPA derivatives of FBi and FB 2 were 1 ng per injection and
2.5 ng per injection, respectively. Medicinal plant specimens were FBi and FB 2 free.
1

P-59
CURRENT LEVELS AND TRENDS OF ORGANOCHLORINE POLLUTANTS IN
HUMANS AND ENVIRONMENT IN TURKEY
E. Durmaz, T. Qalik Durmaz, i. £ok
Gazi University, Faculty of Pharmacy, Department of Toxicology, 06330, Hipodrom, Ankara,
Turkey.
Chemical production and trade which had been started in early 20 th century have been increased for
last three decades that resulted public and official concern regarding the potential risks by chemicals
and pesticides. Persistent Organochlorine pesticides (OCPs) and poly chlorinated biphenyls (PCBs)
are the more important groups of Persistent Organic Pollutants (POPs) and many of these
compounds have been or continue to be used in large quantities and, due to their environmental
persistence, have the ability to bioaccumulate and biomagnify. Humans can be exposed to
organochlorine compounds through diet, occupational accidents and the environment. Exposure to
these chemicals, either acute or chronic, can be associated with a wide range of adverse health
effects such as body weight loss, thymic atropy, chloracne, impairment of immune responses,
carcinogenesis and adverse reproductive effects to wildlife as well as laboratory animals. OCPs
were produced in large number in the 1940-1950s and global production increased year by year. In
Turkey, OCPs have been started to be used against pests in 1945, large quantities of these chemicals
were used during 1960s and 1970s, and since 1983 usage of these chemicals, excluding Endosulfan,
have been severely restricted. OCP residues have been monitored in the Turkish population by
carrying out regional surveys at given time intervals since 1976 in Turkey. PCBs were first
manufactured commercially in 1929 and concern about the presence of PCBs in environment began
in the 1960s. Because of the bioaccumulation and toxicity, usage of these chemicals for different
purposes has been restricted or banned in most countries, since the beginning of the 1970's. PCBs
are restricted for use in closed systems and banned after 1996 in Turkey. There are very limited data
of PCB contamination levels both in humans and environment in Turkey The presenting study aims
to determine the changes of OCP levels from 1970s to present and current levels of PCBs in
humans and environment in Turkey. The results have been discussed in terms of regions and OCPs
and PCBs in which analyses had been made.
1

P-60
VALIDATION OF A REVERSED-PHASE HPLC METHOD FOR THE ANALYSIS OF
SODIUM BENZOATE IN SOFT DRINKS AND JAMS
G. Altiokka 1 , B.Ergun 2 . K. Kircali 1
'Department of Analytical Chemistry, Faculty of Pharmacy, Anadolu University, 26470, Eski§ehir,
Turkey and 2 Department of Pharmaceutical Toxicology, Faculty of Pharmacy, Anadolu University,
26470, Eskiehir, Turkey
A rapid, simple and sensitive method is described for the determination of the preservatives sodium
benzoate in soft drinks, various jams and ketchup. The method utilizes high performance liquid
chromatography (HPLC) followed by diode array detection. Chromatographic separation was
achieved using a CI8 reversed phase column and methanol : water (70:30) adjusted to pH 3.45 with
glacial acetic acid as mobile phase, 0.45 ml.min"' flow rate and UV detection at 245 nm.
Amoxicillin (IS) was used as the internal standard. The retention time for sodium benzoate and IS
were 5.01 and 12.07 min, respectively. The method is selective, reliable reproducible with relative
standard deviation (RSD) of 0.66 and linear in the range of 50-450 ng/mL sodium benzoate. The
limit of detection (LOD) and limit of quantification (LOQ) were 61 pg/mL and 203 pg/mL,
respectively. The proposed method can be used for the routine analysis of sodium benzoate in soft
drinks and jams.
16

P-61
FENVALERATE EXPOSURE ALTERS THYROID HORMONE STATUS
IN SELENIUM AND/OR IODINE DEFICIENT RATS
B. Giray, A. Caglayan, p Erkekoglu, F. Hincal,
Hacettepe University, Faculty of Pharmacy, Department of Toxicology, Ankara TURKEY
Considering the potential adverse effects of selenium and iodine deficiencies, and taking into
account the heavily but improperly controlled usage of insecticides and their endocrine disrupting
effects, this study was undertaken to investigate the effects of fenvalerate (FV), a pyrethroid
insecticide, on thyroid hormone parameters in both healthy and selenium and/or iodine deficient
rats. Three week-old Wistar rats were fed either with selenium deficient diet, or iodine deficiency
was introduced by 1% sodium perchlorate- containing drinking water for 7 weeks. During the last
week of feeding period test groups received 100 mg/kg/d, i.p., FV. In all groups, plasma total T 4 ,
total Ti and TSH levels were determined. Plasma T4 was found to be lowered by iodine deficiency
(ID) and by selenium plus iodine deficiency (ISeD), while a significant increase was observed in
selenium deficient (SeD) rats. FV exposure had no effect on the T 4 levels of healthy controls, but
caused significant increases both in ID and ISeD, and a significant decrease in (SeD). T 3 levels did
not change in SeD and ISeD, while a significant decrease was obvious in ID. FV exposure,
however, caused dramatic increases (75%- 200%) in T3 of all groups including control rats.
Significant elevations in TSH levels were observed in ID and ISeD, but no alteration was observed
in SeD. FV exposure caused insignificant decrease of TSH in healthy controls, no effect in SeD and
significant elevation in ID and ISeD. These results, thus, showed that the widely used pyrethroid
insecticide FV has the potential to change significantly thyroid hormone parameters both in normal
and deficiency states and consequences of its thyroid status modifying effect may be of critical
importance particularly in sensitive individuals and patients with thyroid dysfunction.
1

P-62
GENETIC POLYMORPHISM OF XPD GENE AND HEAD AND NECK CANCER
1 2 2 3 4
G. Gtivenc. . M. Turanli , E. Corner! , P. Demirbas , H.S. Siizen
1 2
Institute of Health Sciences, Ankara University, 06500, Besevler, Ankara, Turkey, Department of
Otolaryngology/ Head and Neck Surgery, Ankara Oncology Hospital, Demetevler, Ankara, Turkey,
3
4
Institute of Biotechnology, Ankara University, 06500, Besevler, Ankara, Turkey, Department of
Toxicology, Faculty of Pharmacy, Ankara University, 06100, Tandogan, Ankara, Turkey.
Tobacco smoke contains an array of potent chemical carcinogens of reactive oxygen species they
may produce DNA bulky adducts, crosslinks, oxidative or base DNA damage and DNA strand
breaks. The xeroderma pigmentosum group D (XPD) protein is well-characterized DNA helicase
necessary for the nucleotide excision repair of bulky DNA lesions, such as those induced by
cigarette smoking. Inter- individual differences in DNA repair capacity have been demonstrated
using a variety of phenotypic assays including reduced repair among patients with squamous cell
carcinoma of the head and neck cancer (SCCHN). Polymorphisms of the nucleotide excision repair
gene XPD gene candidates for influencing cancer susceptibility. We conducted a case-control study
of 50 SCCHN patients and 50 control subjects matched for age and sex to investigate the role of
Lys751 Gin polymorphism in SCCHN. Genomic DNA was isolated from the white blood cells of
whole blood samples of patients with head and neck cancer and controls without any cancer.
Genotypes were determined by polymerase chain reaction coupled restriction fragment length
polymorphism methods in cases and volunteers. (7). Forty-two percent of the patients were
homozygous for Lys/Lys genotype. Forty percent were heterozygous Lys/Gln, and 18% were
mutant genotype. The distribution of genotypes (Lys/Lys, Lys/Gln, and Gln/Gln) among controls
was 36%, 54%, and 10%, respectively. Although there was not a significant association in variant
alleles of XPD (OR= 1.543, 95% CI 0.437-5.448, p= 0,299), frequency of Gln/Gln genotype was
higher in the cases (18%) than controls (10%). This study evaluates the influence of genetic
polymorphism at XPD gene loci on head and neck cancers among Turkish cancer patients. Our
preliminary results suggest that the Lys751Gln genotype may not be a risk factor for
development of head and neck cancer. These findings are limited due to the relatively small
numbers in the subgroups and need to be verified in larger samples.
Supported by Ankara University, Institute of Biotechnology, Project No: 2001-K-120-240.
1

P-63
EFFECTS OF CO- SOLVENT ON MINOXIDIL SKIN ABSORPTION
M. Adrangi 1 , M. Kazemipour 2 . M. Ansari 3 , S. Ahmadi 3
1 Department of Pharmaceutics, School of Pharmacy, Tabriz Medical Sciences University, Tabriz,
Iran, 2 Department of Chemistry, School of Science, Azad University, Kerman Branch, Iran, J
Department of Pharmaceutics, School of Pharmacy, Kerman Medical Sciences University, Kerman,
Transdermal delivery system for the local therapeutics would be expected to avoid the systemic
absorption and to make the substantial penetration into deeper tissues, such as muscle, of topically
applied drugs as much as possible. As a main barrier against transdermal absorption, the stratum
corneum could be overcome by a lot of promising physical and/or chemical approaches. Minoxidil
usually as a 2% topical solution is applied for treatment of alopecia areata. Because of its low
solubility, some co-solvents may be used that they can influence permeation of minoxidil through
the skin. Ethanol is known as an efficient permeation enhancer. Tata et al (1) found that the relative
concentrations of propylene glycol and ethanols as a binary solvent system had significant effects
on skin penetration of 2% solutions of minoxidil. Coldman et al. (2) found that up to relatively high
2-propanol composition, penetration of fluocinolone increased dramatically with an increase in the
fraction of the volatile solvent when administered in the open application. The aim of this work was
to investigate effect of combination of ethanol, propylene glycol and isopropyl alcohol on
penetration of minoxidil through mouse skin in vitro. Some 2% solutions of minoxidil in a mixture
of 10, 15, and 20% of propylene glycol and or isopropanol in 70% ethanol were prepared. The
release study was performed on the isolated skin of mice called "Surie mice"using a static Franz
diffusion cell. Minoxidill released was estimated spectrophotometrically. The study indicated that
the skin penetration of minoxidil increases directly on relation to propylene glycol concentration,
while we are urged to consider a strict percentage of excipient using isopropanol if we regard the
maximal skin penetration, the optimum concentration is 15%. by using more isoprpanol
concentrations minoxidil absorption will decrease, so we can use more concentration of isoprpanol
for more restricted pentration to deep layers of the skin. Thus, rearding the physicochemical
properties and the extent of released drug in proportion of time, some suggestions can be made for
the modification of formulation so that they can be helpful toward much more safety for the patients
suffering from hair loss.
1. TataS., Weiner N., Flynn G. J. Pharm. Sci., 83 (1994) 1508-1510.
2. Coldman M.F., Kalinovsky T„ Poulsen B.J., Br. J. Dermatol., 85 (1971) 457- 461.
179

P-64
LEVELS OF DIOXIN-LIKE PCB CONGENERS IN ADIPOSE TISSUE OF TURKISH
MEN
M. Keski Donmez \ I. Cok >, M.H. Satiroglu 2 , B. Aydinuraz 3 , B. Henkelmann 4 , J. Kotalik 4 , K.W.
Schramm 4
1 Department of Toxicology, Faculty of Pharmacy, Gazi University, 06330 Hipodrom, Ankara,
Turkey, 2 Department of Obstetrics and Gynecology, School of Medicine, Ankara University,
06590 Cebeci, Ankara, Turkey, ^Gen-Art Women Health, IVF & Reproductive Biotechnology,
Cinnah Caddesi No: 47, 06680, Cankaya, Ankara, Turkey, 4 GSF - National Research Center for
Environment and Health, Institute of Ecological Chemistry, Ingolstaedter Landstrasse 1, 85764
Neuherberg, Germany
The Stockholm Convention on Persistent Organic Pollutants (POPs) was adopted by 125
countries including Turkey, on 22 and 23 May 2001. The convention addresses the
production, use, import, release of by-products, stockpile management and disposal of an initial 12
POPs, the so called "Dirty Dozen". Polychlorinated biphenyls (PCBs) are one of the important
members of POPs and have been used worldwide since 1929 and despite restrictions on their
production, use and disposal which have been force for many years, they continue to persist in the
environment. Because of the bioaccumulation and toxicity, usage of the PCBs for different purpose
have been restricted or banned since the begining of the 70's in most of the countries. A number of
Polychlorinated biphenyls congeners (PCBs) show "dioxinlike" toxicity. These PCBs are assigned
with a Toxic Equivalency Factor (TEF) that relates their toxicity to that of 2,3,7,8-
tetrachlorodibenzo-p-dioxin (TCDD) and are to be evaluated as dioxins. In Turkey, after 1993
PCBs are restricted for use only in closed system and banned in 1996 by the Regulation on
Dangerous Chemicals. Although this has been banned quite late, the studies addressing the PCB
exposure of general population is very limited. Dioxin-like PCBs values analyzed in this study is
important in terms of processing the first values from the Turkish population. Furthermore, if the
results of this study are considered as a pre evaluating of exposure to Polychlorinated Dibenzo-p-
Dioxins (PCDDs) and Polychlorinated Dibenzofuranes (PCDFs) the importance will increase. The
levels of dioxin-like (PCBs) were determined in 23 Turkish men living in Ankara, Turkey, in 2004.
Samples were analyzed for WHO-12 congeners (non-ortho: 77, 81, 126, 169 and mono-ortho: 105,
114, 118, 123, 156, 157, 167 and 189) using high resolution gas chromatography/high resolution
mass spectrometry (HRGC/HRMS). Concentrations ofTEQs in males were found between 1.6-24.5
pg/g, fat wt in this study. The results suggest that human background contamination by dioxin-like
PCBs is lower than that generally found in industrialized countries.
1

P-65
THE ROLE OF ANTIOXIDANT SUPPLEMENTATION IN OCCUPATIONAL
EXPOSURE TO WASTE ANAESTHETIC GASES
E. Ozcagli 1 . S. Sarda 1 , S. Izde 2 , O. Kanbak 2 , E. Kadioglu'
'Depaitment of Toxicology, Faculty of Pharmacy, Gazi University, Ankara, Turkey, 2 Ataturk
Training and Research Hospital, Ministry of Health, Ankara, Turkey
Objectives: Although the genotoxicity related to waste anaesthetic gases are controversial, a
consistent number of observations have provided evidence for an increased level of DNA strand
breaks. The goal of this research was to investigate this hypothesis and to estimate the
genoprotective role of antioxidant supplementation in technical anaesthesiology staff working in
operating theaters. Methods: Heparinized venous blood samples were collected from 17 exposed
technical anaesthesiology staff (mean age: 34.3 ± 3.5 years) and non-exposed control group (mean
age: 32.2 ± 3.4 years) and examined in the alkaline comet assay for the DNA strand breakage.
Vitamin E (300 mg/day) plus vitamin C (500 mg/day) were supplemented to the technical
anaesthesiology staff for 12 weeks and blood samples were retaken and evaluated by comet assay.
Results: The DNA breakage observed in the lymphocytes of the technical anaesthesiology staff was
21.5 ± 5.0; as calculated by total comet score (TCS). This score was significantly higher (p

PROTECTIVE EFFECT OF GINKGO BILOBA AGAINST NAPHTHALENE-INDUCED
OXIDATIVE STRESS IN MICE
A. Tozan 1 . O. $ehirli 2 , G.Z. Omurtag 1 , Cetinel 3 , G. Contuk 3 , N. Gedik 4 , G. §ener 2
P-68
'Marmara University School of Pharmacy, Departments of'Pharmaceutical Toxicology,
2 Pharmacology and, School of Medicine, department of Histology- Embryology, 4 Kasimpasa
Military Hospital, Division of Biochemistry, Istanbul / Turkey
Regarding the mechanisms of naphthalene toxicity, several hypotheses have been put forward,
among which oxidative stress and depletion of glutathione are suggested. In this study we aimed to
investigate the possible protective effect of Ginkgo biloba (EGb), an antioxidant agent, against
naphthalene toxicity in mice. BALB/c mice of either sex 25-30 g were divided into four groups
each consisting of 10 animals and were administered naphthalene in a dose of 100 mg/kg i.p for 30
days with either saline (Napht group) or Ginkgo biloba in a dose of 150 mg/kg (Napht + EGb
group). In other mice, saline (control group) or Ginkgo biloba (150 mg/kg, EGb group) was injected
for 30 days, following corn oil (vehicle of naphthalene) injection. At the end of the experiment,
following decapitation, lung, liver and kidney tissue samples were taken for histological
examination or determination of malondialdehyde (MDA), glutathione (GSH), myeloperoxidase
(MPO) activity and collagen contents. Aspartate aminotransferase (AST), alanine aminotransferase
(ALT), blood urea nitrogen and creatinine levels and lactate dehydrogenase (LDH) activity were
measured in the serum samples, while TNF-a was assayed in plasma samples. Naphthalene
administration caused a significant decrease in tissue GSH which was accompanied with significant
increases in tissue MDA and collagen levels and MPO activity. Moreover the pro-inflammatory
mediators (TNF-a), LDH activity, AST, ALT, creatinine and BUN levels were significantly
increased in the naphthalene group. On the other hand, Ginkgo biloba treatment reversed all these
biochemical indices as well as histopathological alterations induced by naphthalene. Oxidative
mechanisms play an important role in naphthalene-induced tissue damage, and Ginkgo biloba, by
inhibiting neutrophil infiltration, balancing oxidant-antioxidant status, and regulating the generation
of inflammatory mediators, ameliorates oxidative organ injury due to naphthalene toxicity.
1

P-65
THE ROLE OF ANTIOXIDANT SUPPLEMENTATION IN OCCUPATIONAL
EXPOSURE TO WASTE ANAESTHETIC GASES
E. Ozcagli 1 , S. §arda§>, S. izde 2 , O. Kanbak 2 , E. Kadioglu 1
'Department of Toxicology, Faculty of Pharmacy, Gazi University, Ankara, Turkey, 2 Ataturk
Training and Research Hospital, Ministry of Health, Ankara, Turkey
Objectives: Although the genotoxicity related to waste anaesthetic gases are controversial, a
consistent number of observations have provided evidence for an increased level of DNA strand
breaks. The goal of this research was to investigate this hypothesis and to estimate the
genoprotective role of antioxidant supplementation in technical anaesthesiology staff working in
operating theaters. Methods: Heparinized venous blood samples were collected from 17 exposed
technical anaesthesiology staff (mean age: 34.3 ± 3.5 years) and non-exposed control group (mean
age: 32.2 ± 3.4 years) and examined in the alkaline comet assay for the DNA strand breakage.
Vitamin E (300 mg/day) plus vitamin C (500 mg/day) were supplemented to the technical
anaesthesiology staff for 12 weeks and blood samples were retaken and evaluated by comet assay.
Results: The DNA breakage observed in the lymphocytes of the technical anaesthesiology staff was
21.5 ± 5.0; as calculated by total comet score (TCS). This score was significantly higher (p

P-66
GLUTATHIONE PEROXIDASE (GPX1) POLYMORPHISM IN A TURKISH
POPULATION
O. Sakalh. Y. Duydu, H.S. SUzen,
Department of Toxicology, Faculty of Pharmacy, Department of Toxicology, Tandogan-06100,
Ankara, Turkey
Reactive oxygen species (ROS) are produced as a consequence of aerobic metabolism. Oxidative
stress may occur if there is excessive production of ROS due to exposure to toxic agent or
pathological processes or when defense mechanisms are insufficient. Effect of oxidative stress has
been postulated in many conditions, including atherosclerosis, inflammatory condition, certain
cancer and the process of aging. Growing body of evidence suggest that human genetic variation in
oxidative stress-related genes may have an importance in the development of such diseases. Human
glutathione peroxidase 1 (hGPXl) is a selenium-dependent enzyme that protects cells against
oxidative damage by reducing hydrogen peroxide and a wide range of organic peroxides with
reduced glutathione.
A single nucleotide polymorphism (SNP) in the GPX1 gene has been reported at nucleotide 593, C
to T substitution which causes a proline (Pro) to leucine (Leu) substitution at codon 198
(Prol98Leu). It was shown that the Leu-containing allele was less responsive to the stimulation of
GPX1 enzyme activity. The Leu allele frequency varies by ethnic groups.
The aim of this study was to establish the frequency of polymorphic GPX1 gene in a Turkish
population. Genomic DNA was extracted from 55 healthy individuals and polymerase chain
reaction (PCR) and restriction fragment length polymorphism (RFLP) technique was used to
identify distribution of GPX1 alleles. The results showed that 23 (41.8%) individuals were
homozygous wild-type (Pro/Pro), 23 (41.8%) were heterozygous (Pro/Leu), and 9 (16.4%) were
homozygous mutant (Leu/Leu). Our preliminary findings indicate that the codon 198 variant allele
ofGPXl in Turkish population is similar to Europeans.
1

P-67
GINKGO BILOBA EXTRACT PROTECTS AGAINST MERCURY (II)-INDUCED
OXIDATIVE TISSUE DAMAGE IN RATS
G. $ener \ O. §ehirli\ A. Tozan 2 , A. Velioglu Oviin 3 , N. Gedik 4 , G.Z. Omurtag 2
Marmara University 'School of Pharmacy, Departments of Pharmacology and 2 Toxicology,
'Vocational School of Health Related Professions, 4 Kasimpasa Military Hospital, Division of
Biochemistry; Istanbul / Turkey
Mercury (II) is a highly toxic metal which induces oxidative stress in the body. In this study we
aimed to investigate the possible protective effect of Ginkgo biloba (EGb), an antioxidant agent,
against experimental mercury toxicity in rat model. Following a single dose of 5 mg/kg mercuric
chloride (HgC^; Hg group) either saline or EGb (150 mg/kg) was administered for 5 days. After
decapitation of the rats trunk blood was obtained and the tissue samples from the brain, lung, liver,
and kidney were taken for the determination of malondialdehyde (MDA) and glutathione (GSH)
levels, myeloperoxidase (MPO) activity and collagen contents. Formation of reactive oxygen
species in the tissue samples was monitored by chemiluminescence (CL) technique. BUN, creatinin,
ALT, and AST levels and tumor necrosis factor-cc (TNF-a)and lactate dehydrogenase (LDH)
activity were assayed in serum samples. The results revealed that HgCh induced oxidative damage
caused significant decrease in GSH level, significant increase in MDA level, MPO activity and
collagen content of the tissues. Treatment of rats with EGb significantly increased the GSH level
and decreased the MDA level, MPO activity, and collagen contents. Similarly, serum ALT, AST
and BUN levels, as well as LDH and TNF-a, were elevated in the Hg group as compared to control
group. On the other hand, EGb treatment reversed all these biochemical indices. Our results
implicate that mercury-induced oxidative damage in brain, lung, liver, and kidney tissues are
protected by Ginkgo biloba extract, with its antioxidant effects.
183

PROTECTIVE EFFECT OF GINKGO BILOBA AGAINST NAPHTHALENE-INDUCED
OXIDATIVE STRESS IN MICE
A. Tozan 1 . 0. Sehirli 2 , G.Z. Omurtag 1 , getinel 3 , G. Contuk 3 , N. Gedik 4 , G. §ener 2
P-68
'Marmara University School of Pharmacy, Departments of 'Pharmaceutical Toxicology,
2 3 4
Pharmacology and, School of Medicine, Department of Histology- Embryology, Kasimpasa
Military Hospital, Division of Biochemistry, Istanbul / Turkey
Regarding the mechanisms of naphthalene toxicity, several hypotheses have been put forward,
among which oxidative stress and depletion of glutathione are suggested. In this study we aimed to
investigate the possible protective effect of Ginkgo biloba (EGb), an antioxidant agent, against
naphthalene toxicity in mice. BALB/c mice of either sex 25-30 g were divided into four groups
each consisting of 10 animals and were administered naphthalene in a dose of 100 mg/kg i.p for 30
days with either saline (Napht group) or Ginkgo biloba in a dose of 150 mg/kg (Napht + EGb
group). In other mice, saline (control group) or Ginkgo biloba (150 mg/kg, EGb group) was injected
for 30 days, following corn oil (vehicle of naphthalene) injection. At the end of the experiment,
following decapitation, lung, liver and kidney tissue samples were taken for histological
examination or determination of malondialdehyde (MDA), glutathione (GSH), myeloperoxidase
(MPO) activity and collagen contents. Aspartate aminotransferase (AST), alanine aminotransferase
(ALT), blood urea nitrogen and creatinine levels and lactate dehydrogenase (LDH) activity were
measured in the serum samples, while TNF-a was assayed in plasma samples. Naphthalene
administration caused a significant decrease in tissue GSH which was accompanied with significant
increases in tissue MDA and collagen levels and MPO activity. Moreover the pro-inflammatory
mediators (TNF-a), LDH activity, AST, ALT, creatinine and BUN levels were significantly
increased in the naphthalene group. On the other hand, Ginkgo biloba treatment reversed all these
biochemical indices as well as histopathological alterations induced by naphthalene. Oxidative
mechanisms play an important role in naphthalene-induced tissue damage, and Ginkgo biloba, by
inhibiting neutrophil infiltration, balancing oxidant-antioxidant status, and regulating the generation
of inflammatory mediators, ameliorates oxidative organ injury due to naphthalene toxicity.
18

P-69
DETERMINATION OF DIACETOXYSCIRPENOL IN FOOD SPECIMENS BY HPLC
G.Z. Omurtag, A. Tozan
Department of Pharmaceutical Toxicology, Faculty of Pharmacy, Marmara University, Haydarpasa
34668, Istanbul, Turkey.
The trichothecene (TC) mycotoxins, as a group of naturally occurring contaminants of cereal and
pulse products they have been detected in fungal contaminated foods. Diacetoxyscirpenol (DAS,
anguidine) is a chemical compound which is synthesized by various species of the fungal genus
Fusarium such as Fusarium poae, F. semitectum, F. moniliforme, F. oxysporum, on foods like
cereal, pulse and their products. The occurrence of this mycotoxin depends on such factors as
temperature, humidity, food processing methods, and type of food product. DAS which is capable
of growing on agricultural products before harvest and during storage was also seen on foodstuffs.
Contamination with DAS causes a number of illnesses in man and animals such as decrease in food
consumption (anorexia), depression or inhibition on immun system function and haematoxicity. In
our study, DAS was investigated with processed the cereal and pulse products which are provided
from markets and street bazaars. DAS was detected using high performance liquid chromatography
(HPLC) with diode array detector (DAD). DAS-suspicious two specimens were analysed by gas
chromatography-mass spectrometry (GC-MS). HPLC and GC-MS were used because of their
facility in detection. In this study, DAS analysis was carried out in 69 processed cereal and 16 pulse
samples. Out of 85 samples, 53 of them were purchased from markets and 32 of them were from
street bazaars existing in Istanbul. DAS was not detected in cereal and pulse products. Cereal and
pulse products which were also exported are very important in our food chain and in our country's
economy. Therefore, foodstuffs are needed to be controlled and analysed during food process. All
mycotoxin analyses for the entire food chain have a lot of importance for healthy people and
healthy generations.
(The results presented in this study were obtained during an investigation supported by the
Scientific Research Projects Commission of Marmara University; project number SAG-
044/230804.)
18

PROTECTIVE EFFECT OF RESVERATROL AGAINST NAPHTHALENE-INDUCED
OXIDATIVE STRESS IN MICE
O. Sehirli 1 , A. Tozan 2 , G.Z. Omurtag 2 , §. Cetinel 3 , G. Contuk 3 , N. Gedik 4 , G. §ener'
P-70
'Marmara University School of Pharmacy, Departments of Pharmacology, Pharmaceutical
Toxicology, School of Medicine, department of Histology- Embryology, 4 Kasimpasa Military
Hospital, Division of Biochemistry, Istanbul / Turkey
Regarding the mechanisms of naphthalene toxicity, several hypotheses have been put forward,
among which oxidative stress and depletion of glutathione are suggested. This investigation
elucidates the role of free radicals in naphthalene-induced toxicity and the protection by resveratrol.
BALB/c mice of either sex 25-30 g were divided into four groups each consisting of 10 animals and
were administered naphthalene in a dose of 100 mg/kg i.p for 30 days with either saline (Napht
group) or resveratrol in a dose of 30 mg/kg (Napht + RVT group). In other mice, saline (control
group) or resveratrol (30 mg/kg, RVT group) was injected for 30 days, following corn oil (vehicle
of naphthalene) injection. At the end of the experiment, following decapitation, lung, liver and
kidney tissue samples were taken for histological examination or determination of malondialdehyde
(MDA), glutathione (GSH), myeloperoxidase (MPO) activity and collagen contents. Aspartate
aminotransferase (AST), alanine aminotransferase (ALT), blood urea nitrogen and creatinine levels
and lactate dehydrogenase (LDH) activity were measured in the serum samples, while TNF-a, IL-P,
IL-6 and total antioxidant capacity (AOC) were assayed in plasma samples. Naphthalene
administration caused a significant decrease in tissue GSH and plasma AOC, which was
accompanied with significant increases in tissue MDA and collagen levels and MPO activity.
Moreover the pro-inflammatory mediators (TNF-a, IL-|3, IL-6), LDH activity, AST, ALT,
creatinine and BUN levels were significantly increased in the naphthalene group. On the other hand,
resveratrol treatment reversed all these biochemical indices as well as histopathological alterations
induced by naphthalene. Oxidative mechanisms play an important role in naphthalene-induced
tissue damage, and resveratrol, by inhibiting neutrophil infiltration, balancing oxidant-antioxidant
status, and regulating the generation of inflammatory mediators, ameliorates oxidative organ injury
due to naphthalene toxicity.
18

DETERMINATION OF 7-ETHOXYRESORUFIN O-DEETHYLASE (EROD) INDUCTION
IN MULLET AS AN INDICATOR OF SEA POLLUTION IN ALIAGA BAY
O. K. Ulutas'. A. Sen 2 , B. Tutuncu 2 , i. £ok
P-71
1
Department of Toxicology, Faculty of Pharmacy, Gazi University, 06330 Hipodrom, Ankara,
2
Turkey, Biology Department, Faculty of Arts & Sciences, Pamukkale University, Kinikli Campus,
Kinikli 20017 Denizli-Turkey
Today, as a result of wide range of industrial and human activities, aquatic environment have been
increasingly polluted by numerous chemicals and xenobiotics. This pollution is a threat to the health
of organisms inhibiting the seas, as well as to human consumers of such organisms. Particularly as a
consequence of industrial activities today's aquatic environment continuously and increasingly
polluted by persistent organic pollutants (POPs) and also other chemicals and physical agents such
as pesticides, polychlorinated biphenyls (PCBs), metals, polycyclic aromatic hydrocarbons (PAHs),
radionuclides. Induction of cytochrome P4501A and of its monooxygenase activities, namely
arylhydrocarbon hydroxylase (AHH) and 7-ethoxyresorufin O-deethylase (EROD) activities, in fish
by these chemicals are the most widely used biochemical measurements in biomonitoring of
environmental contaminants. Aliaga Bay, at the west coast of Aegean Sea nearby Izmir, hosting a
wide range of industrial activities such as oil-refinery, paper factory, ship-breaking. There is no
national documentation or research on determination of pollution resulted from industrial activities
in this area. Thus obtained results from this study would be the first information regarding Aliaga
Bay. In this study, the degree of induction of cytochrome P4501A-associated EROD activity and
immunochemical detection of cytochrome P450 1A1 in leaping mullet (Liza saliens) was used as
biomarker for assessment of PAH/PCB type organic pollutants in Aliaga Bay. Mullet caught from
different section of the Bay, had approximately 52 times more EROD activity (2389.25 ± 3326.20
pmol/min/mg prot) than the feral fish sampled from clean reference site (46,33 ± 16,18
pmol/min/mg prot) near Fofa. The results of this study indicated that Aliaga Bay is highly
contaminated with PAH and/or PCB type organic pollutants.
18

P-72
EVALUATION OF EXCISION REPAIRABLE DNA LESIONS INDUCED BY LEAD
AND/OR ALA IN HUMAN LYMPHOCYTES BY USING ARA-C/CBMN ASSAY
A. Ustiindag. Y. Duydu
Ankara University, Faculty of Pharmacy, Department of Pharmaceutical Toxicology,
06100, Tandogan-Ankara - Turkey
From the previous studies it is known that lead has potential of genotoxic effects in humans by
indirect mechanisms. Genotoxic potential of lead exposure was partly attributed to the formation of
the highly reactive oxygen metabolites (ROM) in the blood. But lead ions have no ability to
generate ROM. Thus, indirect mechanism for genotoxic effect of lead is supposed to accumulation
of ALA in lead-induced DNA damage. In the present study we investigated the ability of lead and
ALA to induce excision repairable DNA lesions by using ARA-C/CBMN test method. In this assay,
N-methyl-N-nitrosourea (MNU) was used as a positive control which is a mutagen and known to
induce excision repair. According to our results, ALA significantly (p

P-73
DNA DAMAGE IN PATIENTS UNDERGOING HYPERBARIC OXYGEN THERAPY
A. Ustiindag '. A. Aydin 2 , A. Eken 2 , K. Diindar 3 , G. Uzun 3 , Y. Duydu 1
'Department of Toxicology, Faculty of Pharmacy, Ankara University, Tandodan, Ankara 06100,
Turkey, 2 Department of Pharmaceutical Toxicology, Giilhane Military Medical Academy, Ankara,
Turkey, Submarine Diseases and Centre of Hyperbaric Oxygen Treatment, Giilhane Military
Medical Academy, Ankara, Turkey
Hyperbaric oxygen (HBO) therapy is a useful method for treatment of various clinical conditions;
however, it can also cause generation of reactive oxygen species and induction of DNA damage.
The aim of the present study was to evaluate the sister chromatid exchange (SCE) frequencies in
lymphocytes from patients undergoing hyperbaric oxygen therapy (HBOT) and to determine the
sensitivity of lymphocytes from those patients to SCE induction by Mitomycin C (MMC). A
statistically significant induction in mean SCE/cell (p

P-74
THE BIOTRANSFORMATION OF TRICLOSAN BY CEPHALOSPORIUMAPHIDICOLA
S.Yilmazer, KYildirim
Sakarya University, Faculty of Arts and Sciences, Chemistry Department, 54187, Sakarya, Turkey
Triclosan is a synthetic chemical which has been added to personal health care products, fabrics,
carpets and even plastic toys as an antibacterial agent for 30 years'. Triclosan is not metabolised in
human body^ and it is an unchanged chemical accumulated in the environment due to its stable
structure 3 . The studies on the possible effects of triclosan in the environment have shown that this
chemical has harmful effects especially on algae 4 , fish 3 , and nitrite oxidizing bacteria^, and it may
behave as a pseudoestrogen^. Some studies have also shown that the worldwide and careless triclosan
usage will make triclosan a useless chemical soon due to the increasing resistance against One
aim of this study was to get harmless new metabolites via the biotransformation of triclosan by
Cephalosporium aphidicola the fungus. Another aim of this study was to compare the triclosan
metabolism in human with that of in Cephalosporium aphidicola since this fungus is a member of
moulds which are videly used in the microbial modelling of mammalian xenobiotics metabolism^. The
incubation of triclosan with Cephalosporium aphidicola for 7 days only gave one metabolite. The
results of the research supported by spectroscopic techniques suggested that the metabolite was the
starting material. This result showed that the metabolism of triclosan in human and Cephalosporium
aphidicola is performed in the same way.
1. H. N. Bharga and A. Patricia, American Journal of Infection Control, 24, 209-218, 1996.
2. M. Adolfsson-Erici, M. Pettersson, J. Parkkonen and J. Sturve, Chemosphere, 46, 1485- 1489,
2002.
3. S. R. Mueller, H. P. Singer and S. Canonica, Abstracts of Papers of the American Chemical
Society, 219, 624, 2000.
4. B. A. Wilson, V. H. Smith, F. Denoyelles and C. K. Larive, Environmental Science and
Technology, 37, 1713-1719, 2003.
5. S. N. Dokianakis, M. E. Kornaros and G. Lyberatos, Water Science and Technology, 50, 341-
346, 2004.
6. C. M. Foran, E. R. Bennett and W. H. Benson, Marine Environmental Research, 50, 153-156,
2000.
7. E. C. Cole, R. M. Addison, J. R. Rubino, K. E. Leese, P. D. Dulaney, M. S. Newell, J. Wilkins,
D. J. Gaber, T. Wineinger and D. A. Criger, Journal of Applied Microbiology, 95, 664-676,
2003.
8. A. J. McBain, A. H. Rickard and P. Gilbert, Journal of Industrial Microbiology and
Biotechnology, 29, 326-330, 2002.
9. E. A. Abourashed, A. M. Clark and C. D. Hufford, Current Medicinal Chemistry, 6, 359-374,
1999.
1

P-75
THE CAFFEINE CONTENT IN BOTH GREEN TEA AND BLACK TEA
MANUFACTURED FROM CAMELLIA SINENSIS GROWN IN ANATOLIA
E. Sarer, A.C. Agca
Ankara University, Faculty of Pharmacy, Department of Pharmacognosy, Tandogan, 06100,
Ankara, Turkey
Camellia sinensis (L.) O.Kuntze is an evergreen shrub of the Theaceae family. It grows widely in
temperate regions in Asia and is also cultivated in the rest of most countries because this plant is the
source of mostly consumed beverage in the world, called tea.Tea is derived from the leaves of
Camellia sinensis and classified in to green tea, oolong tea and black tea. This classification
depends on the degree of fermentation and manifacturing process. It is known that the content of
caffeine and polyphenols in the leaves of the plant which characterizes the taste of the tea infusion,
varies according to the different processing methods and geographical conditions . So in this study,
we wish to report the caffeine content in both green tea and black tea manifactured from Camellia
sinensis grown in Anatolia as 1.63% and 1.09%, respectively for the first time.
1

P-76
HPLC METHOD FOR THE ANALYSIS OF OLEUROPEIN IN
OLEA EUROPAEA L.
C. Altinvav. M. L. Altun
Department of Pharmacognosy, Faculty of Pharmacy, University of Ankara , 06100, Tandogan
Ankara, Turkey
Genus Olea L. is represented by 67 species. Olea europaea L. and two varieties of this species
(Olea europaea L. var. europaea Zhukovsky and Olea europaea L. var. sylvestris (Miller) Lehr.)
are growing in Turkey and are found mostly in West and South Anatolia. In this study a simple and
sensitive method for separation and determination of oleuropein in the leaves and the branches of
the natural (Olea europaea L. var.sylvestris) and cultivated (Olea europaea L. var.europaea)
varieties has been developed. Oleuropein was separated using a Nucleosil 100-5 Cig (5 pm, 250 x
4.6 mm; GL Sciences Inc.) column by isocratic elution with flow rate 1.0 ml/min. The mobile phase
composition was Water-Acetonitrile-Formic acid (84.6:15:0.4) (v/v/v). Spectrophotometric
detection was carried out at 240 nm. The linear range of detection for oleuropein was between 100-
400 pg/ ml. The oleuropein content of cultivated O.europaea (var. europaea) (Balikesir-Edremit
samples) was found to be 3.506 % for leaves and 1.438 % for branches. For natural O.europaea
(var. sylvestris) (Osmaniye samples), these values were found as 5.197 % and 1.463 %,
respectively. The oleuropein content of cultivated O.europaea (var. europaea) (Konya samples)
was found to be 4.020 % for leaves and 1.097 % for branches.
1

P-77
ANTIOXIDANT ACTIVITY OF OLEA EUROPAEA L.
A.Guvenc'.C. Altinvav 2 . M. L. Altun 2 ,
'Department of Pharmaceutical Botany, Faculty of Pharmacy, University of Ankara, 06100,
Tandogan Ankara, Turkey, department of Pharmacognosy, Faculty of Pharmacy, University of
Ankara , 06100, Tandogan Ankara, Turkey
Genus Olea L. is represented by 67 species . Olea europaea L. and two varieties of this species
(Olea europaea L. var. europaea Zhukovsky and Olea europaea L. var. sylvestris (Miller) Lehr.)
are growing in Turkey and are found mostly in West and South Anatolia. The leaves of the plant
are used as hypoglycemic and hypotensive. In this study the antioxidant activity of the freeze- dried
extracts obtained from leaves of Olea europaea L. were investigated. The antioxidant activities
were studied by two different tecniques: Qualitative DPPH (l,l-diphenyl-2-picrylhydrazyl radical)
assay to detect the free radical scavenging activity and the TBA- assay to detect liposome lipid
peroxidation. The qualitative DPPH assay was made by TLC of the extract using DPPH as the spray
reagent. The lipid peroxidation was initiated in liposomes obtained from bovine brain extracts by
addition of ascorbic acid and iron source and was measured spectrophotometrically with TBA test.
Prophyllgallate is used as a positive control.
13

HPLC METHOD FOR THE ANALYSIS OF SALICIN AND CHLOROGENIC ACID OF
VIBURNUM OPULUS L. AND VIBURNUMLANTANA L.
P-78
M. L. Altun, B. Sever Yilmaz
Department of Pharmacognosy, Faculty of Pharmacy, University of Ankara, 06100, Tandogan
Ankara, Turkey
A simple and sensitive method for separation and determination of salicin and chlorogenic acid has
been developed. Salicin and chlorogenic acid were separated using a Exsil ODS column (150 x
4.6mm, 5pm) by isocratic elution with flow rate 0.5 ml/min. The mobile phase composition was
bidistilled water, tetrahydrofuran and ortho-phosphoric acid (97.7: 1.8: 0.5) (v/v/v).
Spectrophotometric detection was carried out at 270 nm. The linear range of detection for salicin
and chlorogenic acid were between 17.775- 474 and 17.808- 474.8 |ig/ ml, respectively. The
method described was suitable for the determination of salicin and chlorogenic acid in the leaves,
branches and fruits of Viburnum opulus L. and Viburnum lantana L. It was observed that V. opulus
fruit sample has the highest salicin content (1.266), while V.lantana leaves (0.390) have the lowest
salicin content as w/w (%) in our case. On the other hand, the chlorogenic acid contents have the
highest in V.opulus fruits (1.240) and V.lantana fruits (0.010) have the lowest content as w/w (%) in
our case. This study is the first application of High Performance Liquid Chromatography (HPLC)
method to the determination of salicin and chlorogenic acid content of Viburnum opulus L. and
Viburnum lantana L. in Turkey.
1

P-79
HEPATOPROTECTIVE AND HYPOGLYCEMIC ACTIVITY OF VIBURNUM OPULUS L.
M.L. Altun 1 . H. Ozbek 2 , B. Sever Yilmaz 1 ,G. Saltan gitoglu 1 , I. Bayram 3 , N. Cengiz 4 ,
Altinyay 1
1 Department of Pharmacognosy, Faculty of Pharmacy, University of Ankara, 06100 Tandogan
Ankara, Turkey, 2 Department of Pharmacology, Faculty of Medicine, University ofYuzuncu Yil,
65300 Van, Turkey, 3 Department of Pathology, Faculty of Medicine, University ofYuzuncu Yil,
65300 Van and 4 Department of Histology, Faculty of Medicine, University of YtiziincU Yil, 65300
Van
Water extract of Viburnum opulus L. (VO) (Caprifoliaceae) was investigated for hypoglycemic
activity in diabetic mice and for hepatoprotective activity on carbon tetrachloride (CCl 4 )-induced
hepatotoxicity in rats. Biochemical parameters of hepatic damage such as serum aspartate
aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and bilirubin
concentrations were determined. CC1 4 (0.8 mL/kg i.p for 7 days) treatment increased the serum
AST, ALT, ALP and bilirubin levels significantly as compared to controls. Treatment of animals
with VO (100 mg/kg, i.p.) +CC1 4 (0.8 mL/kg i.p.) (for 7 days) lowered the levels of ALT and AST
but was not significant. It is observed that ALP levels higher than normal values. The results of
biochemical tests were also confirmed by histopathological examination. VO showed a few
balooning degeneration, apoptosis, centrilobular necrosis in the liver tissue and bridging necrosis
similar to CCl 4 -treated group but not as much as it. VO together with CC1 4 treatment showed many
ballooning degeneration, apoptosis, centrilobular necrosis in the liver tissue and bridging necrosis
similar to CC1 4 treated group but not as much as it. For the evaluation of hypoglycemic activity of
VO, glibenclamide was used as the reference agent. But, VO has not hypoglycemic effect on healty
and diabetic mice. The present study reveals that the water extract of Viburnum opulus had slight
hepatoprotective effect on carbon tetrachloride-induced acute liver toxicity in rats and no
hypoglycemic activity in mice.
1

THE EFFECTS OF (+) CATECHIN AND RUTIN AGAINST OXIDATIVE DNA DAMAGE
P-80
S. Aydin 1 , A.A. Ba§aran 2 , N. Ba§aran'
'Department of Toxicology, Faculty of Pharmacy, University of Hacettepe, Ankara, Turkey,
department of Pharmacognosy, Faculty of Pharmacy, University of Hacettepe, Ankara, Turkey
Phenolic phytochemicals are a large group of substances that have been regarded as possible
antioxidants. However the full chemical properties and the effects of phenolic phytochemicals as
antioxidants in protecting DNA against oxidative damage have not been completely examined since
they have suggested having both antioxidant and prooxidant activities. Paliurus spina-christii
(Rhamnaceae) is used as traditional herbal medicine in Anatolia. The methanolic extract of Paliurus
spina-christii was found rich in phenolic compounds. The main compound of its n-buthanolic
extract was rutin. Another polyphenols compound (+) catechin was isolated from the methanolic
extract by fractionating with ethyl acetate and purified by chromatographic methods. The structures
of the compounds were identified by chemical and spectral methods and kept after lyophilised. In
the present study, the modulating effects of (+) catechin and rutin against the oxidative DNA
damage induced by H2O2 in human lymphocytes was investigated by new single cell gel
electrophoresis or COMET assay which is relatively simple and sensitive method for measuring
DNA strand-breakage, alkali-labile abasic sites, or intermediates in base or nucleotide-excision
repair. Briefly, after incubating human lymphocytes with pi amounts of (+) catechin and rutin at
concentrations 50, 100, and 200 pM, in DMSO or PBS with or without 0,1 mM H 2 0 2 for 30
minutes at 37°C, the cells were embedded in agarose, lysed and electrophoresed and then stained
with a fluorescent DNA binding dye, before DNA damage was visualized using fluorescence
microscopy. At the concentration of 50, 100, and 200 pM rutin alone have not induced DNA strand
breakage, but at the concentration of 200 pM (+) catechin alone induced DNA damage. At the
concentrations both (+) catechin and rutin significantly reduced DNA strand breakage in human
lymphocytes, when the lymphocytes incubated with 0,1 mM H 2 0 2 (p

P-81
PHYTOCHEMICAL SCREENING OF THE BIOACTIVE EXTRACT FROM
LA UNEAE ARBRESCENS
N. Belboukhari, A. Cheriti
Phytochemistry & Organic Synthesis Laboratory, Univesity of Bechar, Algeria
In this work, launea arborescens (Asteraceae) was selected for phytochemical investigation. The
important activity of the methanol extract detected in the biological and chemical screening
performed previously and the lack of studies concerning the genus prompted this choice. The aerial
parts methanol extract of Launea arborescens (Asteraceae) displayed several activities: amongst
these, the antifungal activities against Candida albicans and Sacharomyces cerevisiae and the
antibacterial activity against E. coli, S.aureus, P.aeriginisa and Klebseila entrecocus. The extracts
presenting the most interesting activities are then selected for activity-guided phytochemical
investigation, in order to identify the active compounds. Moreover, the isolation and
characterization of further molecules presenting original chemical structures and a potential
therapeutic interest is performed.
1. A.Cheriti, N. Belboukhari, S. Hacini, & D.E1 Abed , 2005. Seminaire international sur la
valorisation des plantes medicinales dans les zones arides, Ouargla, p-12.
2. A. Cheriti, N. Belboukhari, and S. Hacini, 2004. J. Pharm. Res.;3(2),51
3. N. Belboukhari and A. Cheriti, 2006. Pakistan J. Biological. Sc;.9(l), pp-1,2 .
1

P-82
ANALYTICAL RESEARCH ON ECHIUM VULGARE L. SEED OIL
M.V. Codreanu 1 , V. Istudor 2 , N. Dociu 3 , M. Dinu'
'Department of Pharmaceutical Botany and 2 Department of Pharmacognosy, Phytochemistry,
Phytotherapy, Faculty of Pharmacy, "Carol Davila" University of Medicine and Pharmacy,
Bucharest, Romania, 3 Biotehnos, Bucharest, Romania
Echium vulgare L. (blueweed, Borraginaceae) seed fatty oil was physico-chemically analysed
in order to establish its importance as potential source of two essential fatty acids, gammalinolenic
and stearidonic acid. These vegetal compounds, with limited occurrence in nature,
have recently discovered therapeutic properties (anti-inflammatory, hypocholesterolemic and
protective for cardiovascular system). An appreciable fatty oil content of seeds (35,20-39,38%)
and physico-chemical characteristics of the oil (refractive index, relative density, acid and
iodine indices) were established. The chemical composition of the oil (fatty acids, phytosterols
and monoglycerides profiles) was determined using a gas chromatographic-mass
spectrometrical method. Blueweed seed oil was found to contain all the essential fatty acids, in
substantial percentages: gamma-linolenic (10,04%), stearidonic (15,01%), alpha-linolenic
(39,72%) and linoleic acid (14,86%). Monolinolein (0,41%) and beta-sitosterol (8,72%) were
identified too. Seeds of Echium vulgare L. from Romanian spontaneous flora could be one of
the richest source for gamma-linolenic and stearidonic acids, two polyunsaturated compounds
of therapeutic interest.
1. Christie W.W. - Gas Chromatography and Lipids. A Practical Guide, The Oily Press Ltd.,
Ayr, Scotland, 1989, 64-79, 161-166, 212-215;
2. Cisowschi W. et al. Acta Chromatographica, 2001, 11, 215-223;
3. Erdemoglu N. et al., European Journal of Lipids. Science and Technology, 2004,106(3),
160-164;
4. Fan YY„ Chapkin RS, Journal ofNutrition, 1998, 128 (9), 1411-1414;
5. James M., Ursin V., Cleland L. Am.J.Clin.Nutr., 2003, 77, 1140-1145;
6. European Pharmacopoeia, the 5 th edition, Council of Europe, Strasbourg, 2005, 110- 113,
27-129.
18

P-83
ANTIBACTERIAL ACTIVITY AND CHARACTERIZATION OF VOLATILE
CONSTITUENTS OF TWO ENDEMIC PHLOMIS SPECIES AGAINST FOOD
PATHOGENS
1 2 1 3 1
B. Demirci , K. Giiven , F. Demirci , M.Y. Dadandi , K.H.C. Baser
1 2
Department of Pharmacognosy, Faculty of Pharmacy, Department of Biology, Faculty of Science
3
and Letters, Anadolu University, 26470-Eskisehir, Department of Biology, Faculty of Science and
Letters, Erciyes University, 38039-Kayseri, Turkey
Plant extracts and essential oils constitute a moderate natural resource of antimicrobial compound
mixtures since centuries. Essential oils and components are used as natural antibacterials in food
systems, as well as to prevent the growth of food borne bacteria resulting in extension of the shelf
life of the processed food. The family Lamiaceae hosts a wide variety of essential oil bearing plants.
Although Phlomis species are not essential oil rich plants, traditional uses such as tonic, stimulant
and stomachic have been reported. In this study, the essential oils of two endemic species: Phlomis
russeliana (Sims.) Bentham and P. grandiflora H.S. Thompson var. grandiflora were obtained by
hydrodistillation from the aerial parts. Subsequently, the essential oils were analyzed by gas
chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). As a result, the main
constituents of P. russeliana essential oil were identified as sesquiterpenes, namely b-caryophyllene
(22.6%), germacrene-D (15.1%), and caryophyllene oxide (8.1%). The analysis of P. grandiflora
var. grandiflora oil also showed sesquiterpenes such as b-eudesmol (42.0%) and a-eudesmol
(16.1%) as major constituents. Furthermore, the essential oils were tested against common food
borne bacteria such as Aeromonas hydrophila, Bacillus cereus, Escherichia coli 0157:H7, Listeria
monocytogenes, Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhimurium,
Yersinia enterocolitica, and the anaerobic pathogen Clostridium perfringens using the microbroth
dilution technique. When compared with standard antimicrobials weak to moderate (125->1000
|ig/mL) minimum inhibitory concentrations (MIC) were observed.
1

P-84
TRACE ELEMENTS AND METALS IN TURKISH LICORICE
(GLYCYRRHIZA GLABRA L.) BY ATOMIC ABSORPTION SPECTROMETRY
1,2 2
F. Demirci . S. Cevheroglu
'Anadolu University, Faculty of Pharmacy, Department of Pharmacognosy, 26470-Eskisehir,
2
Ministry ofNational Defence, Army Drug Factory, 06110-Diskapi, Ankara, Turkey.
The Fabaceae (bean family) comprises important food and medicinal plant genera worldwide.
Glycyrrhiza glabra L. well known as licorice/liquorice is one of them which grows naturally in
Turkey where the roots are collected as an important export crop. This work concerns the
quantitative analysis of trace elements, minerals or heavy metals such as Mg, Fe, Zn, Ca, Cu, Cr,
Co, Cd, Mn, Pb, Ni, and Al in various plant samples. The commercially obtained plant material
used in this study was first digested by microwave heating followed by atomic absorption
spectrometric (AAS) analysis. As a result, unwanted toxic metals such as Pb, Cd, Hg, As, Ni, and
Cu were detected below the Ph.Eur. 2002 demanded ranges (0.1-50 mg/kg), differing for each
element.

P-85
CHEMICAL COMPOSITION AND ANTIOXIDANT ACTIVITY OF CAMPANULA
ALLIA RIIFOLIA
M. U. Dumlu 1 . E. GUrkan 1 , E. Tuzlaci 2
1 Department of Pharmacognosy, Faculty of Pharmacy, Marmara University,
2 Department of
Pharmaceutical Botany, Faculty of Pharmacy, Marmara Univesity, 34668 Istanbul, Turkey
Campanula alliariifolia synonime of C. lamiifolia (Campanulaceae) is distributed naturally in
Northern Turkey. The plant is a densely pubescent perennial herb, height is to 70 cm, growing in
Northern Turkey and nearly 95 species are found in the Turkish flora. Campanula species are used
for ear-pains in traditional folk medicine and known as "harebell" in the world. In Turkey, the
species are used for wound healing and they are known as '^an^egi" . The aim of the present
study is find out the chemical constituents and antioxidant activity of the plant. The chemical
constituents of Campanula alliariifolia Willd. are being investigated for the first time with the aid
of this paper. Five known compounds, which were quercetin-3-O-glucoside, quercetin-3-Orutinoside,
kaempferol-3-O-glucoside, lobetyolin ( 9-0-P-D-glucopyranosyl-2,10-tetradecadien-4,6-
diyne-8,14-diol) and lobetyol (2,10- tetradecadien-4,6-diyne-8,9,14-triol), were isolated from the
methanol extract. The antioxidant actvity of the chloroform and methanol extracts of the plant was
investigated with DPPH (1,1- diphenyl-picrilyhidrazyl) and Reducing power methods. The extracts
showed significant antioxidant activity according to the mentioned methods.
1

P-86
CHEMICAL AND ECOLOGICAL STUDIES ON VA CCINIUMMYRTILLLUS GROWING
IN BURSA-TURKEY
1 2 2 3
F.Z. Erdemgil , N. §anli , G. Alsancak , C. Tiirkben
Plant & Drug & Scientific Research Center (AUBiBAM), Anadolu University, Eskisehir 26470,
2 3
Turkey, Department of Chemistry, Suleyman Demirel University, Isparta, Turkey, Department of
Horticulture, Faculty of Agriculture, Uludag University, Bursa 16059, Turkey.
Vaccinium myrtillus L. (Bilberry, Blueberry; Shepherd's Grape Ericaceae family), which is also
known as "Coban Uziimii, Yaban Mersini" in Turkey, is a rich source of phenolic compounds (1,2).
These phenolics (e.g. phenolic acids and flavonoids) have beneficial effects on health as natural
antioxidants, anticarcinogens and antimicrobial agents (3). In folk medicine, V. myrtillus is used as
tonic, antiseptic, astringent and antidiabetic. The fresh fruits are edible. The leaf and the dried fruits of
V. myrtillus are used in phytotherapy and they were added to the French pharmacopoeia in 1992 (4).
This study was conducted to characterize wild blueberry populations in Uludag (Mount Olympus in
Bursa, Turkey) in 2005. Therefore, blueberry samples taken from the Northern side of Uludag
(Kirazliyayla, Sarialan, Cobankaya, Bakacak and Alafam) were investigated. Descriptions, habitats
and soil characteristics of blueberry plants were recorded. It was concluded that wild blueberry
populations generally localized under fir (Abies nordmanniana subsp. bornmuelleriana) trees at the
1250-1900 m altitudes, in the humid regions of the Northern side of Uludag. The pH of the soil was
ranged between 4.9 and 6.6 in the blueberry growing areas. As part of our continuing research on this
species, phenolic acids in the extracts of V. myrtillus L. leaves and fruits have been identified by
retention times of the standards and spectral data obtained by DAD. In this study, the phenolic
compounds were separated by using optimized condition with a Phenomenex Synergi Max RP C-12
column (6). Two different extraction methods were compared for sample preparation of HPLC
analysis of these compounds.
1. Davis, P.H., Flora of Turkey and the East Aegean Islands, Edinburgh University Press,
Vol.6, 1978.
2. Baytop, T., Turkiye'de Bitkilerle Tedavi, Nobel Tip Kitabevleri, Istanbul, 1999.
3. Hakkinen, S.H., TorrOnen, A.R., Food Res.Int. 33, 517-524 (2000).
4. Bruneton, J., Pharmacognosy, Phytochemistry, Medicinal Plants, Intercept Ltd., England,
1995.
5. Hakkinen, S., Heinonen, M., Karenlampi, S., Mykkanen, H., Ruuskanen, J., Torronen, R.,
Food Res.Int. 32, 345- 353 (1999).

ANTI-INFLAMMATORY AND ANTINOCICEPTIVE EFFECTS OF NERIUM OLEANDER
L.
N. Erdemoglu. E. Klipeli, E. Ye§ilada
Gazi University, Faculty of Pharmacy, Department of Pharmacognosy, 06330 Ankara, Turkey
P-87
Nerium oleander L. (Apocynaceae), oleander, is widely distributed in the Mediterranean region.
Since it is known as a poisonous plant, limited numbers of utilization have been recorded in Turkish
folk medicine. During our expeditions, an informant described that fresh oleander flowers was put
in alcohol in summer and kept as a home-remedy to alleviate her severe pain and paresthesia in her
legs. In a reference survey in the database program of Turkish folk medicine (TUHIB), a similar
utilization was also described by another informant, i.e. flowers kept 40 days in olive oil is applied
on joint against rheumatic pain (1). Moreover, leaves or flowers are used to stop pain or eczema,
sap obtained from fresh leaves for abscess or rheumatism were recorded. Besides, another part of
oleander, the fruits, is reported as anti-rheumatic and as a remedy for skin diseases in Saudi folk
medicine (2).
To investigate the anti-inflammatory and antinociceptive effects of Nerium oleander flowers and
leaves, extracts are prepared either by using water or ethanol. Both aqueous and ethanol extracts of
oleander flowers possess significant antinociceptive and anti-inflammatory activities as determined
by /-benzoquinone induced abdominal contractions test and carrageenan-induced hind paw edema
model, respectively, but that of EtOH extract was found to have more pronounced activity. As the
further studies are conducted on ethanol extract of flowers by employing bioassay-guided
fractionation process and five fractions (rc-hexane, chloroform, ethyl acetate, «-buthanol and
remaining aqueous) obtained. The ethyl acetate and n-buthanol were determined as the most potent
fractions in both anti-inflammatory and antinociceptive tests. Thus, further studies are in progress in
order to define the active constituents of the ethyl acetate and rc-buthanol extracts.
1. Ye§ilada, E., 2002. Biodiversity in Turkish Folk Medicine, ed. Bilge §ener, Kluwer
Academic/ Plenum Publishers, London, p.119-135.
2. Adam, S.E.I., Al-Yahya, M.A., Al-Farhan, A.H., 2001. Small Ruminant Research 40, 239-
244.
3

P-88
ENZYME INHIBITORY ACTIVITY OF LIGNANS FROM TAXUSBACCATA L.
N. Erdemoglu', B. Sener 1 , S. A. Nawaz 2 , M. I. Choudhary 2
1 9
Department of Pharmacognosy, Faculty of Pharmacy, Gazi University, 06330 Ankara, Turkey
HEJ Research Institute of Chemistry, International Center for Chemical Sciences, University of
Karachi, 75270 Karachi, Pakistan.
Genus Taxus L. (Taxaceae), yew, is widely distributed in the northern hemisphere, occurring in
Europe, North America, Eastern Asia and Asia Minor. There are eight Taxus species and two
hybrids in the world. T. baccata L. (European yew) is the single representative in Turkey. In our
previous phytochemical investigation of Taxus baccata L. growing in Turkey resulted in the
isolation of five lignans, namely lariciresinol, taxiresinol, 3'-demethylisolariciresinol-9'-
hydroxyisopropylether, isolariciresinol and 3-demethyliso lariciresinol (1,2). Continuing our studies
on the screening of plants for enzyme inhibitory activity to be used potential leading compounds in
the treatment of Alzheimer's disease (AD), we also screened lignans obtained from T. baccata. In
this presentation, these lignans were evaluated for their acetylcholinesterase, butyrylcholinesterase
and lipoxygenase inhibitory activities by in vitro spectrophotometric methods. All compounds
exhibited inhibitory potential against lipoxygenase enzyme. All of these lignans except
isolariciresinol, inhibited butyrylcholinesterase enzyme potentially. However, none of them showed
the inhibitory activity against acetylcholinesterase. These compounds may act as potential
compounds in the discovery of clinically useful agents for the treatment of the central nervous
system disorders.
1. Erdemoglu, N., Sener, B., Ozcan, Y., Ide S., J. Mol. Struc., 655(3), 459-466, 2003.
2. Erdemoglu, N., Sahin, E., Sener, B., Ide S., J. Mol. Struc., 692(1-3), 57-62, 2004.

ALKALOID PROFILE OF SEEDS AND AERIAL PARTS OF SOPHORA ALOPECUROIDES
VAR. ALOPECUROIDES AND THEIR ANTIMICROBIAL ACTIVITIES
N. Erdemoglu'. S. Ozkan 2 , N. AdigUzel 3 , F. Tosun 1
'Department of Pharmacognosy, Faculty of Pharmacy, Gazi University, 06330 Ankara, Turkey, 2
Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Gazi University, 06330
Ankara, Turkey, 3 Gazi University, Faculty of Science and Art, Department of Biology, 06500
Ankara, Turkey
P-89
The plant Sophora alopecuroides L. var. alopecuroides (Leguminosae) is widely distributed
Southwest and East Asia, Greece and South Russia. It grows edges of fields, banks and rarely on
sand dunes, from sea level to 1750m. Genus Sophora has two varieties, namely var. alopecuroides
and var. tomentosa (Boiss.) Chamberlain. Among them, var. alopecuroides is more common than
the var. tomentosa in Turkey. Sophora alopecuroides var. alopecuroides is a perennial plant with
rhizomatous, oblong and 7-12-paired leaflets, cream flowers and a narrowly cylindrical lomentum
fruit (1). In this presentation, the alkaloid composition of the aerial parts and seeds of Sophora
alopecuroides var. alopecuroides (SA) was investigated by GC-MS, a powerful method for the
analysis of the quinolizidine alkaloids. Sophoridine and sophocarpine were the main alkaloids.
Besides these main alkaloids, other quinolizidine alkaloids were detected. The structure of alkaloids
was identified according to their mass fragmentation patterns that combined with library search
(Wiley library databank) and compared with authentic alkaloids.In addition, antibacterial and
antifungal activities of S. alopecuroides alkaloid extracts were tested against standard strains of the
bacteria; Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis and Staphylococcus aureus
as well as the fungi; Candida albicans and Candida krusei. The aerial parts of SA alkaloid extract
presented a significant activity against -S. aureus and B. subtilis with MIC of 62.5 pg/ml, a moderate
activity on the P. aeruginosa with MIC of 250 pg/ml, and a weak activity against E. coli with MIC
500 pg/ml. The seeds alkaloid extract of SA possessed significant activity on both P. aeruginosa
and B. subtilis with MIC of 62.5 pg/ml and moderate activity on both S. aureus and E. coli. In the
anti-yeast assay, the aerial parts alkaloid extract of SA displayed significant activity against C.
krusei yeast tested at concentrations of 62.5 pg/ml. Both alkaloid extracts of SA showed weak
activity on C. albicans while aerial parts alkaloid extract of SA displayed moderate activity against
C. krusei at MICs of 250 pg/ml.
1. Chamberlain DF, Sophora L. in "Flora of Turkey and the East Aegean Islands", In: P.H.
Davis (ed.), Vol. 3, Edinburgh University Press, Edinburgh, 1970, 11-12.

P-90
GC-MS ANALYSIS AND ANTIMICROBIAL ACTIVITY OF ALKALOID EXTRACT
FROM GENISTA SANDRASICA
N. Erdemoglu 1 , S. Ozkan 2 , F. Tosun 1
1 Department ofPhannacognosy, Faculty of Pharmacy, Gazi University, 06330 Ankara, Turkey, 2
Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Gazi University, 06330
Ankara, Turkey
The genus Genista L. includes chiefly deciduous shrubs or small trees in the Mediterranean area
and Western Asia. There were thirteen Genista (Fabaceae) species in Turkish Flora, among these
species G. aucheri, G. burdurensis, G. sandrasica, G. involucrata and G. vuralii are endemic [1-3].
Genista sandrasica Hartvig & Strid., prostrate shrublet with several spreading stems, grows on
Sandras Mountain of Southwest Anatolia [2], In this presentation, we report the alkaloid profile of
the aerial parts of Genista sandrasica which was studied by capillary GC-MS. Anagyrine was the
major alkaloid. Besides this main alkaloid, /J-Isosparteine, 13-methoxylupanine, 11-oxo-cytisine,
cytisine, 5,6-dehydrolupanine, lupanine, Y-formylcytisine, baptifoline and 13a-cwcinnamoyloxylupanine
were detected as the minor alkaloids. The structure of alkaloids was
determined according to their mass fragmentation patterns which combined with library search
(Wiley library databank) and compared with authentic alkaloids. In addition, antibacterial and
antifungal activities of Genista sandrasica alkaloid extract were tested against standard strains of
the bacteria; Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis and Staphylococcus
aureus as well as the fungi; Candida albicans and Candida krusei. The alkaloid extract of G.
sandrasica showed significant activity on B. subtilis and S. aureus with MIC of 31.25 pg/ml and
62.5 pg/ml respectively, a moderate activity on the gram-negative P. aeruginosa and E. coli with
MIC of 125 pg/ml. In the yeast assay, the alkaloid extract possessed moderate activity against C.
albicans and C. krusei.
1. Gibbs PE, Genista L. in "Flora of Turkey and the East Aegean Islands", Davis PH (ed.),
Vol. 3, Edinburgh University Press, Edinburgh, 1970, pp.24-32.
2. Davis PH, Mill RR, Tan K, Genista L. in "Flora of Turkey and the East Aegean Islands", In:
Davis PH, Mill RR, Tan K (eds.), Vol. 10, Edinburgh University Press, Edinburgh, 1988,
pp.113.
3. Duran A, Dural H, Ann. Bot. Fennici., 40, 113-6, 2003.

P-91
ANTI-INFLAMMATORY, ANTINOCICEPTIVE AND ANTIOXIDANT ACTIVITIES OF
ARCTIUM MINUS (HILL) BERNH. SSP. MINUS
N. Erdemoglu 1 . N. N. Turan 2 , E. Kiipeli 1 , B. §ener', N. Abacioglu 2
1 Department of Pharmacognosy, Faculty of Pharmacy, Gazi University, 06330 Ankara, Turkey,
department of Pharmacology, Faculty of Pharmacy, Gazi University, 06330 Ankara, Turkey
Arctium minus (Hill) Bernh. ssp. minus is widely distributed in Northern and Inner Anatolia, which
is called "kocaot, kokarot, kabalak, biiytikkabalak, acikabalak". In Turkish folk medicine, Arctium
minus ssp. minus leaves are used to alleviate rheumatic pain or to against fever in sunstroke,
externally (1,2). The anti-inflammatory and antinociceptive activities of ethanolic and aqueous
extracts from the aerial part of Arctium minus ssp. minus were evaluated. Through the results only
the ethanolic extract exhibited a dose-dependent anti-inflammatory activity on carrageenan-induced
hind paw edema model in mice without inducing any gastric damage, whereas the aqueous extract
was found to be inactive in the same assay. The ethanolic extract displayed a significant
antinociceptive activity. However, the aqueous extract of the plant did not show any perceptible
antinociceptive effect on p-benzoquinone induced abdominal contractions in mice. These
experimental results have supported the folkloric utilization of the Arctium minus ssp. minus as
remedy. Furthermore, the antioxidant power of extracts from Arctium minus ssp. minus has been
evaluated by using l,l-diphenyl-2-picrylhydrazyl (DPPH) and flow injection analysis-luminol
chemiluminescence (FIA-CL). DPPH scavenger capacity of both extracts of the plant was
compared with the known antioxidative substances such as ter-butyl-l-hydroxytoluene (BHT),
ascorbic acid, quercetin and gallic acid. Although both extracts were shown to possess significant
DPPH radical-scavenging activity, that of aqueous extract was found to have more pronounced
activity. In FIA-CL system, inhibitory effect of the extracts on H 2 0 2 - or HOCI-luminol induced
peak chemiluminescence was investigated and compared to ascorbic acid. Both aqueous and
ethanolic extract were significantly reduced H 2 0 2 and HOCI-luminol chemiluminescence as
ascorbic acid-luminol chemiluminescence. The aqueous extract of plant was shown to possess a
significant scavenger activity against H 2 0 2 but ethanolic extract of plant was much more potent
antioxidant activity against HOCI-luminol chemiluminescence than H 2 0 2 -luminol
chemiluminescence. According to the results of these studies, it was concluded that Arctium minus
ssp. minus contains potent natural antioxidants capacity.
1. Kupicha FK, Arctium L. in "Flora of Turkey and the East Aegean Islands", Davis PH (ed.),
Vol.5, Edinburgh University Press, Edinburgh, 1975, 354-356.
2. Fujita T, Sezik E, Tabata M, Yeilada E, Honda G, Takeda Y, Tanaka T, Takaishi Y,
Economic Botany 49, 406-422, 1995.

P-92
ISOLATION OF AN ANTI-ULCEREGENIC FLAVONOID FROM EQUISETUM
PALUSTRE L. THROUGH BIOASSAY-GUIDED FRACTIONATION PROCEDURES IN
RATS
j. Giirbtiz'. E.Yeilada 2 , S. Ito 3
'Department of Pharmacognosy, Faculty of Pharmacy, Gazi University 06330 Etiler, Ankara,
Turkey; department of Pharmacognosy, Faculty of Pharmacy, Yeditepe University 34755
Kayidagi, Kadikoy, Istanbul, Turkey and 3 Tokyo Medical and Dental University, Inst, of
Biomaterials and Bioeng., Dept. Molec. Des., Div. B., Tokyo, Japan.
In Turkey, there are seven Equisetum species (Equisetaceae) and have been reported to be used for
various therapeutical purposes in folk medicine; i.e. as diuretic to pass kidney stone and for weightloss,
and against gastric pains or for wound healing. In our previous studies, a significant antiulcerogenic
activity of the extracts from Equisetum palustre (EP) was reported against ethanolinduced
gastric lesions in rats [1]. In addition, 80% ethanol extract showed more pronounced effect,
and this extract was separated into three fractions with successive solvent extractions (chloroform,
n-butanol/H 2 0, and remaining water fractions). All fractions showed significant activity but those
of n-butanol/H20 and remaining water fractions were found more prominent against ethanolinduced
ulcer model [2]. Further study is aimed to isolate its active anti-ulcerogenic component(s)
through bioassay-guided fractionation and isolation procedures, and then elucidate the chemical
structure by *H-,
13 C- and 2D-NMR spectroscopic techniques. Bioassay guided fractionation
yielded a flavonoid, kaempferol-3-0-D-glycopyranosyl-3-0-D-glycopyranoside (Figure 1), as the
active component which possessed remarkable activity on ethanol-induced experimental ulcer
model in rats.
2' 3'
Figure 1: Kaempferol-3-0-D-glycopyranosyl-3-0-D-glycopyranoside.
This study is financially supported by Turkish Technical and Scientific Research Council
(TUBITAK) Project No. SBAG-2564
1. Giirbiiz, i., Ustiin, O., Ye§ilada, E., Sezik, E., Akyiirek, N. Journal of Ethnopharmacology,
83,241-244, 2002.
2. Giirbiiz, L, Yeilada, E. 7 th International Symposium on Pharmaceutical Sciences (7 th
ISOPS), Ankara-Turkey, June 24-27, 2003.
8

P-93
FREE RADICAL SCAVENGER ACTIVITY OF LINUM FLAVUM SSP: SCABRINERVE
A. Aydin 1 , B. Konuklugil, 2 A. Sayal 1 , A. Eken 1 , Q. Saglik 2
'Giilhane Military Medical Academy, Department of Pharmaceutical Sciences, Ankara, Turkey,
department of Pharmacognosy, Faculty of Pharmacy, Ankara University, 06100 Tandogan, Ankara,
Turkey
The genus Linum contains about 200 species mainly annual or perennial herbs with some small
shrubs, and they are distributed all over the world in a very wide variety of habitats. The genus
Linum is represented by 39 species in the Flora of Turkey and East Aegean Islands. Twentyfour taxa
of these are endemic (1,2). Aryltetralins (podophyllotoxin types) are the main lignan constituents of
the species from Sec. Syllinum (3-7). 6-methoxypodophyllotoxin is found to be the major lignan in
this section. Lignans are phenolic compounds that are very wide spread in the plant kingdom and
show a wide variety of biological activities: antitumour, anti-HIV, immunosuppressive,
hipolipidemic, antifungal, phytoestrogen^ and antiasthmatic activities (7,9). In this study we aimed
to investigate the free radical scavenger activity of the main phenolic component of L. flavum
ssp.scabrinerve, which is endemic in Turkey and belongs to section Syllinum. In our study, we used
the The water extract of Linum species has been investigated for stable free radical DPPH' scavenger
activity DPPH' scavenger ability of extract has been studied with different concentrations and
compared with the same concentrations of known antioxidants such as BHT, Vitamin C, Gallic acid
and Quercetin. We found that extracts had DPPH" scavenger activity. However this results are lower
than the controls but we will continue to study radical scavenger activity of L. flavum ssp.
scabrinerve.with others antioxidant activity measurement methods. As a result, we can conclude
that L. flavum ssp. scabrinerve have a radical scavenger activity(8.5 mg /ml) in vitro. But we do not
know the fate of these extracts in vivo. Further in vivo studies are needed to make an exact decision
about radical scavenger activity of this Linum extract.
Financial support by University_of Ankara, Biotechnology Institute (2002-K-120-130-2) is gratefully
acknowledged.
1. Flora of Turkey and the East Islands, P.H. Davis,Vol 2. University Press Edinburg,1967.
2. Flora of Turkey and the East Aegean Islands,A. Gtiner, N. Ozhatay,T. Ekim, K.H.C. Baer.
University Press Edinburg(supp. 2) Vol 11 2000.
3. Konuklugil, B., (1997) Fitoterapia, 68(2): 183-184.
4. Konuklugil, B., Fitoterapia, 67(4): 379-381 .(1996).
5. Konuklugil, B., Biochemical Systematics and Ecology, 25(1): 75.(1997)
6. Konuklugil, B., Biochemical Systematics and Ecology, 26(1) 795-796.(1998).
7. Ward RS (1999) Lignans, Neolignans and Related Compounds. Nat. Prod. Rep. Vol:16 75- 96.
8. Charlton, J. M. (1998) Antiviral Activity of Lignans, J. Nat. Prod. Vol: 61 1447-1451.
9. Thompson LU, Seidl M M, Rickard SE, Orcheson L J, Fon HHS(1996) Nutr. Cancer Vol: 26
159-165.

P-94
ANTIOXIDANT PROPERTIES AND COMPOSITION OF SALVIA VIRGATA JACQ.
M. Kosar, F. Goger, K.H.C. Baer
Faculty of Pharmacy, Department of Pharmacognosy, Anadolu University, 26470 Eski§ehir, Turkey
Antioxidant activities and phenolic compositions of active fractions of Salvia virgata Jacq.
(Lamiaceae) from Turkey, were examined. S. virgata herb was extracted with different solvents in an
order of increasing polarity such as hexane, ethyl acetate, methanol, 50% methanol. Water extract
was also prepared from S. virgata by reflux. All solvent fractions were investigated for their total
phenolic contents, qualitative-quantitative compositions (by HPLC-PDA analysis), iron(III)
reductive activities, free radical scavenging activities (using DPPH*) and effect upon linoleic acid
peroxidation activities, also the peroxidation level was determined by TBA method. The results of
activity tests given as IC 5 o values were estimated from non-linear algorithm and compared with
standards via. butylated hydroxytoluene, ascorbic acid, gallic acid. Polar fractions were found more
active among the others in free radical activity system whereas non-polar fractions protected the
peroxidation of linoleic acid. Rosmarinic acid, caffeic acid, lutelin-O-glycoside were detected in the
extracts as in all Salvia species.
1

P-95
ANTIHYPERCHOLESTEROLAEMIC AND ANTIOXIDANT ACTIVITY ASSESSMENT
OF SOME PLANTS USED AS REMEDY IN TURKISH FOLK MEDICINE
G. Avci 1 , E. Ktipeli 2 , A. Eryavuz 3 , E. Ye§ilada 4 ,1. Kiii^ukkurt'
department of Biochemistry, Faculty of Veterinary Medicine, Afyon Kocatepe University, Afyon,
department of Pharmacognosy, Faculty of Pharmacy, Gazi University, Hipodrom 6330, Ankara,
department of Physiology, Faculty of Veterinary Medicine, Afyon Kocatepe University, Afyon,
department of Pharmacognosy, Faculty of Pharmacy, Yeditepe University, Istanbul
Ethanolic and aqueous extracts from five plant species used in Turkish traditional medicine were
evaluated for in vivo hypercholesterolemic and antioxidant activities; Agrostemma githago L.,
Potentilla reptans L., Thymbra spicata var. spiccita L., Urtica dioica L. and Viscum album var.
album L. We assayed the effects of the administration of plant extracts on serum total cholesterol,
triglyceride, HDL-C, LDL-C, glucose, AST and ALT concentrations in mice fed with cholesterolrich
diet. In addition, plasma TAA, MDA and NO x levels in the same animals were assayed. All the
aqueous plant extracts did not affect the serum cholesterol concentration. However, the ethanolic
extracts of Agrostemma githago, Thymbra spicata and Viscum album decreased the serum
cholesterol concentration in the mice fed with high cholesterol diet without inducing any gastric
damage. The ethanolic extracts of Thymbra spicata, Viscum album, Potentilla reptans and Urtica
dioica and the aqueous extract of Agrostemma githago increased the serum HDL concentration,
whereas the ethanolic extracts of Agrostemma githago, Thymbra spicata, Viscum album and Urtica
dioica decreased the serum LDL-C concentration. Thymbra spicata and Viscum album were
observed to decrease the serum triglyceride concentration. Among the plant extracts studied the
ethanolic extracts of Thymbra spicata significantly decreased the MDA level in mice. The ethanolic
extract of Potentilla reptans increased in NO x . None of these plants showed statistically prominent
activity on plasma TAA. Results of the present study indicated that the ethanolic extracts of
Agrostemma githago, Thymbra spicata and Viscum album showed potent hypocholesteroleamic
activity in the mice fed with a diet contained high-cholesterol.
1

P-96
EVALUATION OF SOME PLANTS USED IN TURKISH FOLK MEDICINE AGAINST
PARASITIC INFECTIONS FOR THEIR IN VIVO ANTHELMINTIC ACTIVITY
E. Kozan 1 , E. Kiipeli 2 , E. Ye§ilada 2
'Department of Parasitology, Faculty of Veterinary Medicine, Afyon Kocatepe University,Afyon,
Turkey,
2 Department of Pharmacognosy, Faculty of Pharmacy, Gazi University, Etiler 06330,
Ankara, Turkey
Ethanolic and aqueous extracts obtained from nine plant species from seven families selected
depending on their use in Turkish folk medicine, including Citrillus lanatus (Thunb.) Matsum.
(seed), Jasminum fruticans L. (branches), Juniperus drupacea Labill. (fruits), Juniperus nana L.
(fruit and leaves), Juniperus oxcycedrus L (fruit and leaves), Mentha longifolia L. (herba), Pinus
nigra ssp. pallasiana (Lamb.) Richt. (fruits), Plantago lanceolata L. (leaves), and Zea mays L. (seed)
were evaluated for their in vivo anthelmintic activity. Among the plant extracts studied, both
ethanolic and aqueous extracts of Jasminum fruticans, Mentha longifolia and Pinus nigra ssp.
pallasiana, the aqueous extracts of Zea mays, the ethanolic extracts of Citrillus lanatus, Juniperus
drupacea (fruit), Juniperus oxcycedrus and Plantago lanceolata displayed significant anthelminthic
activity against pinworms, Syphacia obvelata and Aspiculuris tetraptera, in mice. Rest of the
extracts from plants did not show any remarkable anthelmintic activity. The results were considered
significant at p

P-97
EVALUATION OF ANALGESIC AND ANTI-INFLAMMATORY ACTIVITIES ON
JUGLANSREGIA L.
E. Kiipeli, N. Erdemoglu, E. Yeilada
Gazi University, Faculty of Pharmacy, Department of Pharmacognosy, 06330 Ankara, Turkey
Juglans regia L. (Juglandaceae) leaves have been used mostly in worldwide traditional medicines.
In Turkish folk medicine, leaves are frequently used to alleviate fever in heat stroke or rheumatic
pain, externally (1). In our ongoing project on medicinal plants used in Turkish traditional medicine
for the treatment of rheumatism and related inflammatory diseases, the objective of this study was
to elucidate traditional use of J. regia. The ethanolic and aqueous extracts prepared from the leaves
of J. regia were tested in mice for anti-inflammatory activity using carrageenan-induced hind paw
edema model and for antinociceptive activity using /-benzoquinone induced abdominal
contractions. Both aqueous and ethanolic extracts (EtOH) showed significant antinociceptive
activity without inducing any gastric damage, but the effect of later extract was much more
pronounced than the former as well as that of aspirin. EtOH extract also showed a potent antiinflammatory
activity as potent as indomethacin. Therefore, the EtOH extract was fractionated
through subsequent solvent extractions in increasing polarity. Each fraction was administered to
experimental animals in doses depending on their ratio in the original EtOH extract. The ethyl
acetate fraction possess significant activity for anti-inflammatory and analgesic activities, while n-
hexane, chloroform, n-buthanol and remaining aqueous fractions were found totally inactive.
Further studies are necessary to assess the potential clinical use of Juglans regia, or its extract or
active principles, as analgesic and anti-inflammatory.
1. Fujita, T., Sezik, E„ Tabata, M., Yeilada, E., Honda, G., Takeda, Y., Tanaka, T„ Takaishi,
Y., 1995. Economic Botany 49, 406-422.
13

P-98
FIVE NEW TRITERPENE SAPONINS FROM ERYNGIUM CAMPESTRE
M. Kartal 1 , A.-C. Mitaine-Offer 2 , T. Paululat 3 , M. Abu-Asaker 1 , H. Wagner 4 ,
M.-A. Lacaille-Dubois 2
1 Department of Pharmacognosy, Faculty of Pharmacy, Ankara University, 06100 Tandogan-
Ankara, Turkey, 2 Laboratoire de Pharmacognosie, Unite de Molecules d'Interet Biologique,
UPRES-EA 3660, Faculte de Pharmacie, Universite de Bourgogne, 7, Bd. Jeanne D'Arc, BP 87900,
21079 Dijon Cedex, France, J Siegen University, Adolf-Reichwein-Str. 2, 57068 Siegen, Germany
and 4 Center of Pharmaresearch, Pharmaceutical Biology, Munich University, 81377 Munich,
Germany
Eryngium genus (Apiaceae) is represented by 317 species, subspecies and varieties. 1 ^Eryngium
campestre L. grows in most parts of Europe and Northern Africa and has been introduced into
North America. In the Flora of Turkey and East Aegean Islands, 2) Eryngium genus is represented by
23 species and E. campestre is known in Turkish folk-medicine as "Bogadikeni". Infusions of the
aerial and root parts are used as an antitussive, diuretic, appetizer, stimulant and aphrodisiac. 3 Only
small amounts of saponins 4,5 were isolated from the roots mainly Rl- and Al-barrigenol
glycosides. We describe in this paper the further phytochemical investigations of a «-BuOH-soluble
fraction of the MeOH extract of the roots of E. campestre. Five new triterpene saponins 1-5 were
isolated by successive chromatographic steps over Silica gel and their structures were elucidated
mainly by NMR analysis, including ID and 2D NMR ('H-'H COSY, TOCSY, NOESY, HSQC,
HMBC), and mass spectroscopy. They were characterized as 3-0-«-L-rhamnopyranosyl-(iri2)-^-
D-glucuronopyranosyl-22-0-^,7S-dimethylacryloyl-Al-barrigenol (1), 3-O-a-L-rhamnopyranosyl-
(iri2)-^-D-glucuronopyranosyl-22-0-angeloyl-Rl -barrigenol (2), 3-O-a-L-rhamnopyranosyl-
(iri2)-^-D-glucuronopyranosyl-21-(9-acetyl-22-(9-angeloyl-Rl-barrigenol (3), 3-O-a-Lrhamnopyranosyl-(in2)-/i-D-glucuronopyranosyl-21
-0-acetyl-22-0-/^dimethylacryloyl-Rl -
barrigenol (4), and 3-0-a-L-rhamnopyranosyl-(in2)-/-D-glucuronopyranosyl-22-O-angeloyl-28-
O-acetyl-Rl-barrigenol (5).
1. Worz A., Stuttgarter Beitr. Naturk. Ser. A, 1999, 596, 1-48.
2. Giiner A., Ozhatay N., Ekim T., Baser K.H.C. "Flora of Turkey and the East Aegean
Islands", Vol.11, Edinburgh University Press, Edinburgh, 2000, pp. 136-138.
3. Baytop T. "Tiirkiye'de Bitkilerle Tedavi-Gecjmisten Bugiine (Therapy with Medicinal Plants
in Turkey-Past and Present)", 2nd ed., Nobel Tip Basimevi, Istanbul, 1999, p. 169.
4. Hiller K., Linzer B. Pharmazie 1966, 21, 245-250.
5. Kartal M., Mitaine-Offer A.-C., Abu-Asaker M., Miyamoto T., Calis I., Wagner H.,
Lacaille-Dubois M.-A. Chem. Pharm. Bull. 2005, 53, 1318-1320.
1

P-99
BIOACTIVE STEROIDAL SAPONINS FROM SMILAXMEDICA
M. Sautour', T. Miyamoto 2 , M.-A. Lacaille-Dubois'
1 Laboratoire de Pharmacognosie, Unite de Molecules d'lnteret Biologique, UMIB UPRES- EA
3660, Faculte de Pharmacie, Universite de Bourgogne, 7, Bd. Jeanne d'Arc, BP 87900, 21079 Dijon
Cedex, France, 2 Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka 812-
8582,Japan.
The genus Smilax contains 350 species, which are widely distributed, mainly in tropical regions of
East Asia, and South and North America. Several Smilax species have already been studied
chemically and found to contain steroidal saponins. Previous phytochemical studies on the
methanolic extract of the roots of Smilax medica Schlecht. et Cham, have led to the isolation of
three new steroidal antifungal saponins." A further detailed investigation of the same extract has
resulted in the isolation of two new spirostanol glycosides (1-2) together with the known smilagenin
3-O-yS-D-glucopyranoside (3). 2) The n-BuOH-soluble fraction of the Me0H-H 2 0 (7:3) extract of the
rhizomes of S. medica was purified by precipitation with diethyl ether to give a crude saponin
mixture. This mixture was submitted to multiple chromatographic steps involving vacuum-liquid
chromatography (VLC) on reversed-phase Cig silica gel and medium-pressure liquid
chromatography (MPLC) on normal silica gel to yield compounds 1-3. Their structures were
elucidated mainly by ID- and 2D-NMR experiments (COSY, TOCSY, NOESY, HSQC and
HMBC), and by FABMS. They were characterized as (257)-5/-spirostan-3/-ol 3-0-/3-Dglucopyranosyl-(l—>-6)-/-D-glucopyranoside
(1) and (25i)-5y3-spirostan-3/-ol 3-(9-y5-Dglucopyranosyl-(l-^6)-[y0-D-glucopyranosyl-(l->2)]-[or-L-rhamnopyranosyl-(l->4)]-/-Dglucopyranoside
(2). 1 and 2 exhibited antifungal activity against the human pathogenic yeasts
Candida albicans, C. glabrata and C. tropicalis (MICs between 6.25 and 50 pg/ml) whereas 3 was
inactive.
1. Sautour M., Miyamoto T., Lacaille-Dubois M.A. J. Nat. Prod. 2005, 68, 1489-93.
2. Sautour M., Miyamoto T., Lacaille-Dubois M.A. Planta Med. 2006, in press.
1

P-100
GLUCURONIDE TRITERPENE SAPONINS FROM BERSAMA ENGLERIANA
A. L. Tapondjou 13 , T. Miyamoto 2 , M.-A. Lacaille-Dubois 1
1 Laboratoire de Pharmacognosie, Unite de Molecules d'Interet Biologique, UMIB UPRES- EA
3660, Faculte de Pharmacie, Universite de Bourgogne, BP 87900, F-21079, Dijon Cedex, France, 2
Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka 812-8582, Japan, J
Laboratoire de Chimie Appliquee et Environnementale, Faculte des Sciences, Universite de
Dschang, BP 183, Dschang, Cameroun.
As part of our continuing search for new bioactive triterpene saponins from Cameroonian medicinal
plants", we have investigated the stem bark of the tree Bersama engleriana Gurke (Melianthaceae)
collected in the Western highlands of Cameroon. Some Bersama species are generally used for their
antitumor 2 ', spamolytic"', cardiotonic 4 ', antibacterial 5 ' and antiviral 6 ' activities and also to treat
leprosy 7 '.The chemical constituents and biological activity of B. engleriana have not been
previously investigated and we present here the isolation and structural elucidation of five new
triterpene saponins (1-5). Their structures were established mainly by means of spectroscopic
methods (ID and 2D NMR) as well as FAB-, HRESI-mass spectrometry as 3-C4P-Dglucopyranosyl-(l->2)-P-D-glucuronopyranosyl]-28-0-[P-D-glucopyranosyl]-betulinic
acid (1), 3-
0-[P-D-glucopyranosyl-(l->2)-[P-D-galactopyranosyl-(l-^3)]-P-D-glucuronopyranosyl]-oleanolic
acid (2), 3-0-[p-D-glucopyranosyl-(l-^3)"P-D-glucuronopyranosyl]-28-C>-[P-D-xylopyranosyl-
(l->6)-P-D-glucopyranosyl]-oleanolic acid (3), 3-0-[P-D-galactopyranosyl-(l-»3)-P-Dglucuronopyranosyl]-28-0-[P-D-glucopyranosyl-(l->4)-P-D-glucopyranosyl]-oleanolic
acid (4),
and 3-0-[P-D-glucopyranosyl-(l->3)-P"D-galactopyranosyl-(l->3)-P-D-glucuronopyranosyl]-28-
0-[P-D-xylopyranosyl-(l->6)-P-D-glucopyranosyl]-oleanolic acid (5).
1. Tapondjou A.L., Lontsi D., Sondengam B.L., Shaheen F., Choudhary M.I., Atta-ur-Rahman,
Heerden F.R., Park H.J., Lee K.T. J. Nat. Prod. 2003, 66, 1266-1269.
2. Bowen I.H., Jackson B.P., Motawe H.M.I. Planta Med. 1985, 6, 48-487.
3. Makonnen E., Hagos E., Phytother. Res. 1993, 7, 211-212.
4. Vanhaelen M., Indeherberg J., Bauduin, H. J. Pharm. Sci. 1972, 61,1165-1167.
5. Taniguchi M„ Kubo I. J. Nat. Prod. 1993, 56, 1539-1546.
6. Asres A., Bucar F., Kartnig T., Witvrouw M., Pannecouque C., De Clercq E., Phytother.
Res. 2001, 15, 62-69.
7. Hutchings A., Scott A.H., Lewis G., Cunningham A. in Zulu medicinal plants-An inventory,
University ofNatal Press, Piertermaritzburg 1996, 190-191.
1

P-101
ANTIBACTERIAL ACTIVITY AND COMPOSITION OF ESSENTIAL OILS OF THYMUS
PUBESCENS BOISS & KOTSCHY CHEMOTYPE CITRONELLOL
H. Nazemiyeh 1 , F. Lotfipour 1 , A. Delazar 1 , S. Asnaashari 1 , S. Bamdad 1 , A.H.Talebpour 2
'Faculty of Pharmacy and Drug Applied Research Center, Tabriz University of Medical Sciences,
Tabriz, Iran, 2 Research Center for Agriculture and Natural Resources of East Azerbaijan.
Within the genus Thymus, chemical polymorphism of the essential oil is a widespread phenomenon.
According to the results of numerous researches a great variation has been demonstrated in the
quantity of main components within various thyme oils. Thymus pubescens grows wildly in North
and North West of Iran, in spite of this fact few investigations have been performed on this taxon.
In this study, the composition of the essential oils obtained from flowering aerial parts of two
populations of Thymus pubescens were studied by GC-MS and the results compared with published
data. The oils were Characterized by high amounts of citronellol (42.62%),geranyl acetate
(14.01%), geraniol (13.1%), nerolidol (6.59%),citronellyl acetate (3.91%), neral (1.83%) and betacaryophyllene
(1.57%) while the others reported carvacrol, thymol and para-cymene as major
compounds. These findings showed a clear chemical polymorphism whitin the Thymus pubecsens.
The essential oils were also screened for their inhibitory effects against four strains of gramnegative
bacteria namely E. coli, Pseudomonas aeruginosa, Salmonella Typhimurium and Serratia
marcescens and six strains of gram-positive bacteria namely Staphylococcus aureus, Micrococcus
luteus, Staphylococcus epidermidis, Streptococcus pneumonia, Bacillus anthracis and
Staphylococcus saprophyticus using filter paper disc methods. The results indicated a moderate
antibacterial effect more marked against gram (+) strains especially Staphylococcus epidermidis. To
obtain the Minimum Inhibitory Concentrations (MIC) of the essential oils against Staphylococcus
epidermidis, macro tube dilution method was conducted. The MIC of the essential oils was found to
range of 0.210 v/v (dissolved in hexane) against Staphylococcus epidermidis. Amikacin, hexane as
well as microbial inoculum were used as standard antibiotic, negative and positive controls
respectivily. All the experiments were performed triplicates.
1

P-102
ANTIVIRAL AND ANTIMICROBIAL ASSESSMENT OF SOME SELECTED
FLAVONOIDS
B. Ozfelik 1 ,1. Orhan 2 . G. Toker 2
'Department of Pharmacognosy, Faculty of Pharmacy, Gazi University, 06330, Ankara, Turkey,
department of Pharmaceutical Microbiology, Faculty of Pharmacy, Gazi University, 06330,
Ankara, Turkey
In the current study, the results of antibacterial, antifungal, and antiviral activity tests of four
flavonoid derivatives; scandenone (1), tiliroside (2), quercetin-3,7-0-a-L-dirhamnoside (3), and
kaempferol 3,7-O-a-L-dirhamnoside (4) are presented. Antibacterial and antifungal activities of
these compounds were tested against Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis,
Klebsiella pneumoniae, Acinetobacter baumannii, Staphylococcus aureus, Bacillus subtilis, and
Enterococcus faecalis, as well as the fungus; Candida albicans by a microdilution method. On the
other hand, both DNA virus Herpes simplex (HSV) and RNA virus Parainfluenza (PI-3) were
employed for antiviral assessment of the compounds using Madin-Darby bovine kidney and Vero
cell lines. According to our data, all of the compounds tested were found to be quite active against
S. aureus and E. faecalis with MIC values of 0.5 pg/ml, followed by E. coli (2 pg/ml), K.
pneumoniae (4 pg/ml), A. baumannii (8 pg/ml), and B. subtilis (8 pg/ml), while they inhibited C.
albicans at 1 pg/ml as potent as ketoconazole. However, only compound 3 displayed antiviral effect
towards PI-3 at the range of 8-32 wg/ml of inhibitory concentration for cytopathogenic effect
(CPE).
18

ANTIMICROBIAL ACTIONS AND FATTY ACID PROFILES OF THE SEED OILS OF
THREE URTICA SPECIES
I. Orhan'. U. Koca 2 , B. 6z

P-104
ANTIBACTERIAL ACTIVITY OF VERBASCUM OBTUSIFOLIUM HUB.-MOR. AND V.
INULIFOLIUM HUB.-MOR.
B. Ozbilgin'. S. Guzel', N. Delialioglu 2 , G. Kokdil', G. Emekda 2
1
Department of Pharmacognosy, Faculty of Pharmacy, Mersin University, Yeniehir
2
Campus, 33169 Mersin, Turkey, Department of Microbiology, Faculty of
Medicine, Mersin University, Yeni§ehir Campus, 33169 Mersin, Turkey
The genus Verbascum belonging to the family Scrophulariaceae, comprises more than 350 species
(1). Flowers and leaves of some Verbascum species are used in traditional medicine for the
treatment of various diseases and particularly as anticough, antiinflammatory and antiviral remedy
(2). In pyhtochemical studies, mucillagine, iridoids, flavonoids, saponins, phenylethanoids, lignans,
steroids and alkaloids have been reported from Verbascum species (2-4). There are 233 Verbascum
species in Turkey, 185 species being endemic (5-7). Some species of this genus have been used as
expectorant, mucolytic, sedative, diuretic and constipate in Turkish folk medicine (8). Verbascum
obtusifolium Hub.-Mor. and V. inulifolium Hub.-Mor. are endemic species growing wild in South
Anatolia. In this study, the dichloromethane and methanol extracts obtained from aerial parts of
these species were tested against three Gram-positive (Staphylococcus aureus ATCC 25923,
Enterococcus faecalis ATCC 29212 and Bacillus subtilis ATCC 6633) and two Gram-negative
(Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853) bacteria. The antibacterial
activity was determined by macrodilution method. The dichloromethane extracts were effective
against S. aureus, E. faecalis and E.coli whereas the methanol extracts were effective against S.
aureus. The methanol extract obtained from flowers of V.obtusifolium showed antibacterial activity
against B.subtilis and E.coli.
1. Heywood, VH, Flowering Plants of the World, Oxford University Pres, Oxford, p.243, 1979
2. Turker AU, Giirel, E, Phytother.Res. 19, 733-739, (2005).
3. Abou Gazar H, Tasdemir D, Ireland CM, Call I, Biochemical Systematics and Ecology. 31,
433-436 (2003).
4. Akdemir Z, Tatli I, Bedir E, Khan IA, Turk J Chem 28, 227-234 (2004).
5. Huber-Morath, A., Verbascum. In: Davis, P.H. (Ed.), Flora of Turkey and the East Aegean
Islands, Vol.6, University Press, Edinburgh, pp. 458-603 (1978).
6. Davis PH, Mill RR, Tan K.(Eds.) Flora of Turkey and The East Aegean Islands
(Suplement), Vol.10, University Press, Edinburgh, pp.190-192 (1988).
7. Giiner A, Ozhatay N, Tuna E, Baf)er K.H.C.(Eds.) Flora of Turkey and The East Aegean
Islands (Suplement 2) University Press, Edinburgh, Vol 11, p. 193 (2000)
8. Baytop T, Turkiye'de Bitkiler ile Tedavi (Ge9mite ve Bugun), 2. Baski, Nobel Tip
Kitabevleri, Istanbul (1999).

PHYTOCHEMICAL STUDIES ON THE AERIAL PARTS OF RUBIA PEREGRINA L.
U. Ozgen 1 . S. Toper 2 , C. Kazaz 3 , H. Se9en 3 , M. Cokun 4
1 Atatiirk University, Faculty of Pharmacy, Department of Pharmacognosy, 25240 Erzurum,
Turkey, J Atatiirk University, Faculty of Education, 25240 Erzurum, Turkey, 3 Atatiirk University,
Faculty of Arts and Science, Department of Chemistry, 25240 Erzurum,Turkey, 4 Ankara
University, Faculty of Pharmacy,Department of Pharmaceutical Botany, 06100 Tandogan,
Ankara,Turkey
P-105
Rubia peregrina (Rubiaceae) grows in Northwest Anatolia in Turkey (1). The underground parts of
R. peregrina have been used as a dye in Anatolia (2). In this study, phytochemical studies were
performed on aerial parts of Rubia peregrina. The powdered aerial parts of R. peregrina were
extracted with methanol. The concentrated extract was suspended in water:methanol mixture (9:1)
and partitioned with chloroform and ethyl acetate. The isolation of the compounds were carried out
using several and repeated chromatographic techniques from chloroform, ethyl acetate and aqueous
phases that partitioned from methanolic extract obtained from plant. The chromatographic studies
on aqueous phases gave two compounds: One flavonol glycoside and one iridoid glycoside. The
structures of the compounds were elucidated by means of spectral analysis (1H-NMR, 13C-NMR).
The studies on the aqueo phase are still in progress.
1. Ehrendorfer F. and Schobeck-Temesy E. (1982) Rubia L., "Flora of Turkey and the East
Aegean Islands", Vol. 7, pp. 857-861, University Press, Edinburgh (edited by P.H. Davis)
2. Baytop, T. (1999) "Therapy with Medicinal Plants in Turkey (Past and Present)", 2nd ed.
pp. 193, Nobel Tip Kitabevleri, Istanbul.
1

INVESTIGATION OF ANTIHYPERGLYCEMIC EFFECT OF PEGANUM HARMALA
SEEDS (ZYGOPHYLLACEAE) ON STREPTOZOTOCIN- DIABETIC RATS
D. Dicle 1 , H. Sagmanhgil Ozdemir' , A. Kul 2 , F. Ozgokfe 2
'Department of Pharmacology, Faculty of Medicine, University of YiizlincU Yil, 65200, Van-
TUrkiye, 2 Department of Biochemistry, Arts and Sciences Faculty, University of YuztincU Yil,
65200, Van-Turkiye
This study was planned to research the acute toxicity (LD 5 o) and the antihyperglycemic effect of the
extract prepared from P. harmala L. seeds, harmine and harmaline alkaloids fixed to be present in
the extract in the GC-MS analysis of seeds extract on the streptozotocin- diabetic rats. At the
beginning, in order to investigate LD 50 dose of P. harmala L. seeds, 64 male-female Swiss-albino
mice were separated into the groups of eight equally as four male and four female in each group.
The water extract of P. Harmala L. was given to the mice and the death rates of them were
observed during 72 hours to determine LD 50 dose of P. harmala L. seeds using probit analysis
according to Log 10. After that, seventy-two male-female waster albino rats (120-250 g) divided
into twelve groups equally (n=6, three male and three female). In order to investigate
antihyperglycemic effects of the extracts, they were given to the animals at four different non-toxic
doses determined according to LD50 calculation. Streptozotocin (STZ) solution was freshly prepared
in citrate buffer (0.1 M, pH.4.5) and was given to the rats as single dose (55 mg / kg body weight)
intraperitoneally. Some of experimental groups were formed as the control (distilled water), diabetic
control (distilled water), glibenclamide control (5 mg/kg oral) and insulin control (6 IU/kg i.p). The
others were formed as the treatment groups and they were given water extract of P .harmala L.
seeds at four different doses (0.75, 1.00, 1.25, 1.50 g/kg) by single oral. Finally, the last groups
were treated with oral administration of the harmine (50 and 100 mg/kg) and harmaline (125 and
250 mg/kg) at two different doses. After one week of STZ injection, the fasting blood glucose
levels of the animals were measured (0 th hour) and the drug and extract were applied to them. The
measurements were repeated at 60 th , 120 th and 180 th hours following the application. The data were
statistically compared using variance analysis and Duncan test and also one-way variance analysis
and Dunnett test and they were accepted significant at the level of p

PHYTOCHEMICAL STUDIES ON THE AERIAL PARTS OF RUBIA PEREGRIN A L.
U. Ozgen'. S. Toper 2 , C. Kazaz 3 , H. Sesen 3 , M. Cokun 4
1 Atatiirk University, Faculty of Pharmacy, Department of Pharmacognosy, 25240 Erzurum,
Turkey, 2 AtatUrk University, Faculty of Education, 25240 Erzurum, Turkey, 3 Ataturk University,
Faculty of Arts and Science, Department of Chemistry, 25240 Erzurum,Turkey, 4 Ankara
University, Faculty of Pharmacy,Department of Pharmaceutical Botany, 06100 Tandogan,
Ankara,Turkey
Rubia peregrina (Rubiaceae) grows in Northwest Anatolia in Turkey (1). The underground parts of
R. peregrina have been used as a dye in Anatolia (2). In this study, phytochemical studies were
performed on aerial parts of Rubia peregrina. The powdered aerial parts of R. peregrina were
extracted with methanol. The concentrated extract was suspended in water:methanol mixture (9:1)
and partitioned with chloroform and ethyl acetate. The isolation of the compounds were carried out
using several and repeated chromatographic techniques from chloroform, ethyl acetate and aqueous
phases that partitioned from methanolic extract obtained from plant. The chromatographic studies
on aqueous phases gave two compounds: One flavonol glycoside and one iridoid glycoside. The
structures of the compounds were elucidated by means of spectral analysis (1H-NMR, 13C-NMR).
The studies on the aqueo phase are still in progress.
1. Ehrendorfer F. and Schobeck-Temesy E. (1982) Rubia L., "Flora of Turkey and the East
Aegean Islands", Vol. 7, pp. 857-861, University Press, Edinburgh (edited by P.H. Davis)
2. Baytop, T. (1999) "Therapy with Medicinal Plants in Turkey (Past and Present)", 2nd ed.
pp. 193, Nobel Tip Kitabevleri, Istanbul.
P-
1

DETERMINATION OF PHENOLIC ACIDS, FLAVONOIDS, FLAVONS AND RADICAL
SCAVENGING ACTIVITIES OF CERTAIN HYPERICUM SPECIES 1
P-
N. Oztiirk'. M. Tun9el 2 ,1. Potoglu Erkara 3
'Department of Pharmacognosy, Faculty of Pharmacy, University of Anadolu, 26470 Eskiehir,
Turkey, department of Analytical Chemistry, Faculty of Pharmacy, University of Anadolu,
26470 Eskiehir, Turkey, department of Biology, Faculty of Science and Arts, University of
Eskiehir Osmangazi, 26480, Turkey.
Hypericum (Hypericaceae) species are widely distributed plants in the world and exhibit radical
scavenging activity. Their antioxidant potentials arise from their polyphenolic compounds, i.e.
flavonoids and phenolic acids. In the present study, active chemicals and antioxidant activities of
Hypericum montbretii Spach., H. origanifolium Willd. and H. perforatum L. were investigated.
Having different polarities, methanol, ethyl acetate and water were used as solvents to extract from
flowers and leaves of the plants, whose phenolic acid contents were determined. The determination
of phenolic acids in the relevant Hypericum species was achieved by using a RP-HPLC method
with photodiode array detections and they were separated on a 3pm Cis column in a 42-minutes
gradient analysis with internal standard and quantification was performed by the rate of peak
normalizations. Free radical-scavenging capacities were evaluated with the test of 2,2-diphenyl-lpicryhydrazyl
radical (DPPH*), spectrophotometrically. Using spectrophotometrical techniques,
total phenolic, flavonoid and flavonols contents in the plant extracts were analyzed by Folin-
Ciocalteu, AICI3 and AlCh+Na-acetate methods, respectively. The results were calculated, in turn,
equivalent to gallic acid and rutin. A correlation between radical scavenging capacities of extracts
with the total phenolic compoun content was observed.
This study was supported by Research Fund of Anadolu University (Project no: 30353)

ANTIOXIDANT POTENTIAL OF BARAKOL EXTRACTED FROM THE LEAVES OF
CASSIA SIAMEA LAMK.
W. Reanmongkol', S. Subhadhirasakul', H. Watanabe 2
'Faculty of Pharmaceutical Sciences, Prince of Songkla University, Hat Yai, Songkhla, Thailand,
institute of Natural Medicine, Toyama Medical and Pharmaceutical University, Toyama, Japan
In order to determine the antioxidant potential of barakol, a chromone compound named barakol
extracted from the leaves of Cassia siamea Lamk., we investigated the antioxidative capacity of
barakol using free radical scavenging assays and lipid peroxidation assay. The young leaves of
Cassia siamea were extracted with aqueous acetic acid solution, cooled and filtered. The filtrate
were basified with ammonia solution. The basic solution was extracted with chloroform. The
chloroform extracts were concentrated under reduced pressure and extracted with aqueous acetic
acid solution to give barakol. The antioxidative capacity of barakol were examined using
scavenging of superoxide anion radical and hydroxyl radical assays. The lipid peroxidation was also
determined. Barakol at the concentration of 200 pM, scavenged the superoxide radical generation
by more than 90% on superoxide radical scavenging assay. At the concentration of 400 pM, barakol
inhibited hydroxyl radical generation by 80%. In the lipid peroxidation assay, at the concentration
of 100 pM, barakol showed potent inhibition of malondialdehyde formation by 95% in brain
homogenates. These results suggest that barakol possesses scavenging free radicals and lipid
peroxidation inhibition, which may be due to the scavenging of the hydroxyl radical.
P-
3

INVESTIGATION OF ANTIHYPERGLYCEMIC EFFECT OF PEGANUM HARMALA
SEEDS (ZYGOPHYLLACEAE) ON STREPTOZOTOCIN- DIABETIC RATS
D. Dicle 1 , H. Sagmanhgil Ozdemir 1 , A. Kul 2 , F. Ozgok9e 2
'Department of Pharmacology, Faculty of Medicine, University of YuziincU Yd, 65200, Van-
Turkiye, 2 Department of Biochemistry, Arts and Sciences Faculty, University of YuziincU Yd,
65200, Van-Tiirkiye
This study was planned to research the acute toxicity (LD50) and the antihyperglycemic effect of the
extract prepared from P. harmala L. seeds, harmine and harmaline alkaloids fixed to be present in
the extract in the GC-MS analysis of seeds extract on the streptozotocin- diabetic rats. At the
beginning, in order to investigate LD 50 dose of P. harmala L. seeds, 64 male-female Swiss-albino
mice were separated into the groups of eight equally as four male and four female in each group.
The water extract of P. Harmala L. was given to the mice and the death rates of them were
observed during 72 hours to determine LD 50 dose of P. harmala L. seeds using probit analysis
according to Log 10. After that, seventy-two male-female waster albino rats (120-250 g) divided
into twelve groups equally (n=6, three male and three female). In order to investigate
antihyperglycemic effects of the extracts, they were given to the animals at four different non-toxic
doses determined according to LD 50 calculation. Streptozotocin (STZ) solution was freshly prepared
in citrate buffer (0.1 M, pH.4.5) and was given to the rats as single dose (55 mg / kg body weight)
intraperitoneally. Some of experimental groups were formed as the control (distilled water), diabetic
control (distilled water), glibenclamide control (5 mg/kg oral) and insulin control (6 IU/kg i.p). The
others were formed as the treatment groups and they were given water extract of P .harmala L.
seeds at four different doses (0.75, 1.00, 1.25, 1.50 g/kg) by single oral. Finally, the last groups
were treated with oral administration of the harmine (50 and 100 mg/kg) and harmaline (125 and
250 mg/kg) at two different doses. After one week of STZ injection, the fasting blood glucose
levels of the animals were measured (0 th hour) and the drug and extract were applied to them. The
measurements were repeated at 60 th , 120 th and 180 th hours following the application. The data were
statistically compared using variance analysis and Duncan test and also one-way variance analysis
and Dunnett test and they were accepted significant at the level of p

P-
ANALGESIC AND ANTI-INFLAMMATORY ACTIVITIES OF VIBURNUM LANTANA L.
B. Sever Yilmaz 1 , G. Saltan Citoglu'. M. L.Altun 1 , H. Ozbek 2
'Department of Pharmacognosy, Faculty of Pharmacy, University of Ankara, 06100 Tandogan
Ankara, Turkey, department of Pharmacology, Faculty of Medicine, University of Yiiziincu Yil,
65300 Van, Turkey.
Water extract of Viburnum lantana L. (VL) leaf was investigated for analgesic and antiinflammatory
activities in mice and rats. The tail flick test, acetic acid-induced writhing test and the
carrageenan-induced rat paw oedema test were used to determine of these effects. Our findings
show that VL cause dose related inhibition in the acetic acid-induced abdominal streching in mice.
When compared to aspirin, VL had inhibited chemical painful at 100 mg/kg similarly but this
extract had exhibit stronger analgesic activity than aspirin at 200 mg/kg dose. VL showed powerful
analgesic activity which was formed by thermal painful in 100 mg/kg dose. The evaluation of antiinflammatory
activity of VL was not found significant difference at 100 mg/kg and 200 mg/kg
doses. As a result, VL had detected slight anti-inflamatory activity compared to indomethacin. The
LD 5 O of VO was determined as 2.169 g/kg.

P-110
FREE RADICAL SCAVENGING EFFECTS OF VIBURNUM OPULUS L. AND
VIBURNUMLANTANA L. SPECIES GROWING IN TURKEY
G. Saltan Citoglu', M.L. Altun 1 , B. Sever Yilmaz 1 , G. Genfaslan 2 , T. Qoban 2
'Department of Pharmacognosy, Faculty of Pharmacy, University of Ankara, 06100 Tandogan
Ankara, Turkey, department of Toxicology, Faculty of Pharmacy, University of Ankara, 06100
Tandogan Ankara, Turkey.
The radical scavenging activity of Viburnum opulus L. and Viburnum lantana L. branch, fruit and
leaf water extracts were investigated by the 1, l'-diphenyl-2-picryhydrazyl (DPPH) scavenging and
superoxide anion scavenging methods. In this study water extracts prepared separately from the
fruits, branches and leaves of Viburnum lantana and Viburnum opulus species. The branch extracts
of V. lantana and V. opulus inhibited superoxide anion in a concentration dependent manner. The
fruit extracts of V. lantana did not show any scavenging effect on superoxide anion formation. V.
lantana leaf extracts showed moderate scavenger effect on superoxide anion formation whereas
V.opulus leaf extracts showed strong scavenging effect on superoxide anion in higher concentration.
All tested extracts exhibited scavenging effect on DPPH radical but with various potencies (7-94%).
When compared with Vitamin E (a -tocopherol), V opulus fruit extract and V. lantana extract
showed the highest DPPH radical scavenging activity with IC 5 o value 5.7 pg /ml and 35 pg /ml,
respectively.
226

p-111
IN VITRO EVALUATION OF ANTIOXIDANT ACTIVITY OF ESSENTIAL OIL AND
VARIOUS EXTRACTS OF ZHUMERIA MAJDAE
F. Sharififar, M. Hosseini, F. Hosseininajad
Faculty of Pharmacy, Kerman University of Medical Science, Kerman, Islamic Republic of Iran.
Zhumeria majdae known in persian as "Mohrkhosh" is an endemic plant to Hormozgan province in
Iran and has been used as carminative and for dysmenorrhea in traditional medicine. At present
study antioxidant and free radical scavenging activity of the essential oil and petroleum ether,
chloroform and methanol extracts of the plant were evaluated using two separate methods,
inhibition of free radical 2,2-diphenyl-l-picrylhydrazyl (DPPH) and ammonium thiocyanate
systems. The fractions of the essential oil and methanol extract were able to reduce the stable free
radical 2,2-diphenyl-l-picrylhydrazyl (DPPH) with an IC50 of 21.7 ± 1.58 and 16.2 ± 1.61 pg/ml
respectively in comparing to BHA (18.2 ± 1.94 pg/ml). Inhibition values of linoleic oxidation were
calculated to be 84.3% for the methanol extract. The essential oil to be showed more inhibition
(91.4% ± 2.5), similar to the synthetic antioxidant of BHA (97.8 ± 2.94). The chemical composition
of hvdrodistilled essential oils of Z.majdae was analyzed by GC/MS. A total of 36 compounds
representing 95.8% of the oil were identified: linalool (52.9%), camphor (26.15%) and limonene
(4.17%) were the main components comprising 83.22% of the oil. Separation and identification of
most active fractions of the essential oil and methanol extract will be reported in congress.
22

P-
GALLOYL QUERCETIN AND OTHER QUERCETIN DERIVATIVES FROM GERANIUM
TUBEROSUM SSP. TUBEROSUM
D. Sohretoglu, M.K. Sakar
Department of Pharmacognosy, Faculty of Pharmacy, University of Hacettepe, 06100, Sihhiye,
Ankara, Turkey
The genus Geranium (Geraniaceae) is represented by 33 species in the flora of Turkey (1). The
aerial parts of Geranium (cranesbill) species are known as "Turnagagasi" and used as tonic,
diuretic, antidiabetic, antidiarrheal and antihemorrhoidal as well as a home remedy to treat gastrict
disorders and to promote the healing of wounds (2). These species contain tannins, flavonoids,
anthocyanins and essential oils (3,4,5). We have investigated EtOAc soluble part of the methanolic
extract of G. tuberosum L. ssp. tuberosum L. By employing a combination of chromatographic
methods were isolated six flavanoids: quercetin, quercetin-3-0-P-galactopyranoside, quercetin-3-0-
P-glucopyranoside, rutin, quercetin-3-0-P-(2"-0-galloyl-glucopyranoside) and quercetin-3-0-P-(2"-
O-galloyl-galactopyranoside). The structures of the compouds were elucidated by ID ('H, 13 C) and
2D-NMR techniques (COSY, HMBC, HMQC).
1. P.H. Davis, "Geranium"in the Flora of Turkey and East Aegean Islands, ed.P.H. Davis,
Vol.2, University Press, Edinburg, 1966, pp441-474.
2. T. Baytop, "Theraphy with medicinal plants in Turkey (Past and Present)"2 nd Nobel Tip
Kitabevleri Ltd., Istanbul, 1966.
3. R. Hegnauer, "Chemotaxonomie der Pflanzen" Vol. VIII, Birkhauser Verlag, Basel, Berlin,
511-516, 1989.
4. Z.§.Akdemir, i.i. Tatli, I. Saracoglu, U.B. ismailoglu, i. §ahin-Erdemli, i.
Polyphenolic compounds from Geranium pratense and their free radical scavenging
activities, Phytochemistry, 56, 189-193, 2001.
5. D. Ercil, M. Kaloga, O. A. Radtke, M.K. Sakar, A.F. Kiderlen, H. Kolodziej, O-Galloiyl
Flavonoids from Geranium pyrenaicum and their in vitro antileishmanial activity, Turkish
Journal of Chemistry, 29, 437-443, 2005.
8

P-
ROLE OF L-ARGININE-NO PATHWAY ON DIURNAL AND GENDER VARIATION OF
NEBIVOLOL ANTINOCICEPTION
N. Senyer, E. Aypar. N. Abacioglu
Department of Pharmacology, Faculty of Pharmacy, Gazi University, 06330 Etiler Ankara Turkey
In our previous studies we demonstrated the antinociceptive effect of nebivolol on mouse PBQinduced
algesia model. In this study we examined the role of L-arginine-NO pathway on gender
differences and diurnal rhythmicity of nebivolol antinociception. Experiments performed on male
and female mice synchronized to 12:12 h light-dark at two different times of the day(09:00 and
21:00). Animals were subcutaneously treated with saline (O.lml/lOg), nebivolol (0.5 mg/kg),
morphine(ED50=0.13 mg/kg), L-NAME(75 mg/kg), L-arginine(2.0 mg/kg), nebivolol+morphine,
nebivolol +L-NAME, nebivolol+L-arginine, morphine+L-NAME, morphine+L-arginine,
nebivolol+morphine+L-NAME or nebivolol +morphine+Larginine 15 minutes before PBQ(2.5
mg/kg i.p) injection. After PBQ administration number of writhes were counted. Results were
shown as normalized (arcsin transformation) % antinociception values. Parametric and
nonparametric ANOVA tests were used for statistical analysis. Results of this study showed that
there were no gender and day-night variation in nebivolol antinociception on selected time points.
The role of L-arginine-NO pathway on nebivolol antinociception has been demonstrated in female
09:00 group.

P-116
EXPERIMENTAL PHARMACOLOGICAL RESEARCHES REGARDING THE ANTI-
INFLAMMATORY EFFECT OF THE STATINES HMG-COA REDUCTASE INHIBITORS
AND OF THEOPHYLLINE
A.N. Cristea, L.I. Turculet, C. Zbarcea
Dept. of Pharmacology and Clinical Pharmacy, Faculty ofPharmacy, University of Medicine and
Pharmacy « Carol Davila », Bucharest, 020956, Romania, 6 Traian Vuia Street
Clinical postmarketing studies showed that statines HMG-CoA reductase inhibitors produce a
spectacular decrease of the cardiovascular morbidity and mortality at patients with or without high
levels of the cholesterol (1). The laboratory investigations demonstrated that the therapy with
hypocholesterolemiant statines HMG-CoA reductase inhibitors reduces the CRP (marker of
inflammation) levels (2). This data generated the hypothesis that statines HMG-CoA reductase
inhibitors have their own anti-inflammatory activity, with similar importance as the
hypocholesterolemiant effect, in the treatment of atherosclerosis, as inflammatory disease secondary
to dislipidemia. On the other hand, studies with low dose of theophylline on voluntary patients with
atopic asthma demonstrated for theophylline an anti-inflammatory type effect (3), usefull in the
therapy of asthma, considered today an inflammatory disease. Because in the clinical studies the
anti-inflammatory effect is difficult to differentiate from the main action of the statines HMG-CoA
reductase inhibitors and theophylline (hypocolesterolemiant and bronhodilatating, respectively), in
this paper we proposed to bring a contribution to the knowledge of the anti-inflammatory effect.
The experimental model used was the classic oedema of the rat paw, induced by dextran, 0.2 ml/10
g, 0,6% solution, administrated beneath the plantar surface and measured with an Ugo Basile
plethysmometer. We choosed the inflammation induced by dextran because it has many similitudes
to the inflammation from different human diseases. We worked on groups of eight Wistar rats each,
of 120-140 g. The investigated doses were 5, 10, 15 mg/kg p.o. for simvastatin and 25, 50, 100
mg/kg p.o. for theophylline. The reference substance was phenylbutazone, in dose of 100 mg/kg,
p.o. International regulations of bioethics for researches on laboratory animals has been respected.
The statistic analyzed results showed for simvastatin: the anti-inflammatory effect at 45 minutes
(maximum inflammatory effect of dextran) for the dose of 5 mg/kg p.o. were almost equal (38,5%)
to the effect of phenylbutazone at 100 mg/kg p.o. (33,55%) and for the dose of 10 mg/kg p.o.
almost double (60,52%). The anti-inflammatory effect of simvastatin lasts longer than that of
phenylbutazone. For theophylline the anti-inflammatory effect at 45 minutes was 34,33%, 59,57%
and 41,25% for the doses 25, 50, respectively 100 mg/kg p.o. comparing with the effect of 24,38%
of phenylbutazone at 100 mg/kg. The effect lasts longer for theophylline. The results of our
research complete other experimental and clinical studies and support the anti-inflammatory effect
of the statines HMG-CoA reductase inhibitors and of theophylline.
1. Sacks F.M. et al. The N. Engl. J. of Med. 1996; 14: 1001-9.
2. Verschuren K.R. et al. Blood 2004.
3. Sullivan P., Bekir S., Jaffar Z„ Page C., Jeffery P., Costello J. The Lancet. 1994; 343: 1006-
8.
3

P-113
ROLE OF L-ARGININE-NO PATHWAY ON DIURNAL AND GENDER VARIATION OF
NEBIVOLOL ANTINOCICEPTION
N. Senyer, E. Aypar, N. Abacioglu
Depanment of Pharmacology, Faculty of Pharmacy, Gazi University, 06330 Etiler Ankara Turkey
In our previous studies we demonstrated the antinociceptive effect of nebivolol on mouse PBQinduced
algesia model. In this study we examined the role of L-arginine-NO pathway on gender
differences and diurnal rhythmicity of nebivolol antinociception. Experiments performed on male
and female mice synchronized to 12:12 h light-dark at two different times of the day(09:00 and
21:00). Animals were subcutaneously treated with saline (O.lml/lOg), nebivolol (0.5 mg/kg),
morphine(ED50=0.13 mg/kg), L-NAME(75 mg/kg), L-arginine(2.0 mg/kg), nebivolol+morphine,
nebivolol +L-NAME, nebivolol+L-arginine, morphine+L-NAME, morphine+L-arginine,
nebivolol+morphine+L-NAME or nebivolol +morphine+Larginine 15 minutes before PBQ(2.5
mg/kg i.p) injection. After PBQ administration number of writhes were counted. Results were
shown as normalized (arcsin transformation) % antinociception values. Parametric and
nonparametric ANOVA tests were used for statistical analysis. Results of this study showed that
there were no gender and day-night variation in nebivolol antinociception on selected time points.
The role of L-arginine-NO pathway on nebivolol antinociception has been demonstrated in female
09:00 group.

P-114
ANALGESIC ACTIVITY OF PUNIC A GRAN A TUM L. FRUIT RIND EXTRACT
P.P. Can'. U. Demir', N. Oztiirk; 2 , Y. Ozturk
1
Anadolu University, Faculty of Pharmacy, Department of Pharmacology, 26470 Eskiehir, Turkey,
Anadolu University, Faculty of Pharmacy, Department of Pharmacognosy, 26470 Eskiehir,
Turkey
Punica granatum (Pomegranate) is a small tree from Punicaceae family. As well as the
consumption of the fruits as a meal; its juice, pericarp and bark (from tree and roots) preparations
and plant itself have been traditionally used to cure several disorders such as colitis, oxyuriasis,
leucorrhea, menorrhagia, ulcer, hepatic damage, snakebite, paralysis and rectocele. Further, it has
been reported that external applications of these preparations are used for headache treatment in
folk medicine. The rind of the fruit is antihelminthic, useful in diarrhea, dysentery and ulcer. But to
the best of our knowledge, there have been no reports related to analgesic activity of the fruit rind
extract. The present study was aimed to evaluate the analgesic activity of the methanolic extract of
Punica granatum fruit rind. A dose of 100 mg/kg (p.o) test compound was used for the present
study. Analgesic activity of the extract was measured by tail-clip and tail-immersion tests.
Morphine (2 mg/kg) was used as standard. In tail clip tests, 100 mg/kg doses of the extract showed
analgesic activity. In contrast, no analgesic activity was observed in "tail-immersion" test. Being
important for the development of new analgesic drugs, this is the first report for the analgesic
activity of pomegranate rind extract. However, further studies are necessary.
3

EXPERIMENTAL PHARMACOLOGICAL RESEARCH ABOUT A POTENTIAL
ANXIOGENIC ACTION AND MOOD TONUS DECREASE INDUCED BY ISONIAZID
A. N. Cristea, C. D. Marineci, C. Chirita, V. Butoescu
Dept. of. Pharmacology and Clinical Pharmacy, Faculty of Pharmacy, University of Medicine and
Pharmacy "Carol Davila", 6 Traian Vuia Street, Bucharest, 020956, Romania
P-115
The topic of this study was inspired by a clinical case of a depressed young woman which
committed suicide. The depressed mood was gradually settled down, after the discontinuity of a 6
months tuberculostatic treatment with isoniazid (INH), orally given. The patient described that,
during the INH intake, she was feeling CNS stimulation effects, with hyperactivity. Starting from
the details of this clinical case and known CNS stimulatory pharmacological profile of INH (until
convulsions, at high doses), we have hypothesized a possible and probable drug-induced etiology of
this severe depression, associated with acute anxiety attacks. We considered that INH could be a
starting factor of an anxiogenic mechanism and/or of the onset of a depressed mood after the
discontinuity of an INH intensive treatment. The reason could be the depletion of the
neurotransmitters involved in the mood tonus maintenance, as consequence of prolonged CNS
stimulation induced by INH, which is an IMAO. To verify this hypothesis, we gave acute, subacute
and chronic treatment of INH to mice, orally or intraperitoneally (i.p.). We tested: the CNS
stimulatory effect (in Autotrack-type actometer, hole board, inclined plane and platform tests), the
anxiogenic effect (in suspended cross-labyrinth test) and mood tonus depression (forced swimming
test). Experimental INH used doses were small, medium and big ones (10, 25, 50, 100 and 200
mg/kg), according to the fact that we couldn't extend the period of mice administration as long as
the period of a chronic human tuberculostatic INH treatment. The measurements were made both
during the drug administration and after its interruption. We worked on 18-20 g white mice,
grouped in 10 animals/group for acute treatment or 20 animals/group for subacute and chronic
treatment. The bioethics rules of vivid laboratory animals research were followed. The results
showed that INH produced CNS stimulation, anxiety and mood tonus depression. These effects
were statistically significant and they differed (from p

P-116
EXPERIMENTAL PHARMACOLOGICAL RESEARCHES REGARDING THE ANTI-
INFLAMMATORY EFFECT OF THE STATINES HMG-COA REDUCTASE INHIBITORS
AND OF THEOPHYLLINE
A. N. Cristea, L.I. Turculet, C. Zbarcea
Dept. of Pharmacology and Clinical Pharmacy, Faculty of Pharmacy, University of Medicine and
Pharmacy « Carol Davila », Bucharest, 020956, Romania, 6 Traian Vuia Street
Clinical postmarketing studies showed that statines HMG-CoA reductase inhibitors produce a
spectacular decrease of the cardiovascular morbidity and mortality at patients with or without high
levels of the cholesterol (1). The laboratory investigations demonstrated that the therapy with
hypocholesterolemiant statines HMG-CoA reductase inhibitors reduces the CRP (marker of
inflammation) levels (2). This data generated the hypothesis that statines HMG-CoA reductase
inhibitors have their own anti-inflammatory activity, with similar importance as the
hypocholesterolemiant effect, in the treatment of atherosclerosis, as inflammatory disease secondary
to dislipidemia. On the other hand, studies with low dose of theophylline on voluntary patients with
atopic asthma demonstrated for theophylline an anti-inflammatory type effect (3), usefull in the
therapy of asthma, considered today an inflammatory disease. Because in the clinical studies the
anti-inflammatory effect is difficult to differentiate from the main action of the statines HMG-CoA
reductase inhibitors and theophylline (hypocolesterolemiant and bronhodilatating, respectively), in
this paper we proposed to bring a contribution to the knowledge of the anti-inflammatory effect.
The experimental model used was the classic oedema of the rat paw, induced by dextran, 0.2 ml/10
g, 0,6% solution, administrated beneath the plantar surface and measured with an Ugo Basile
plethysmometer. We choosed the inflammation induced by dextran because it has many similitudes
to the inflammation from different human diseases. We worked on groups of eight Wistar rats each,
of 120-140 g. The investigated doses were 5, 10, 15 mg/kg p.o. for simvastatin and 25, 50, 100
mg/kg p.o. for theophylline. The reference substance was phenylbutazone, in dose of 100 mg/kg,
p.o. International regulations of bioethics for researches on laboratory animals has been respected.
The statistic analyzed results showed for simvastatin: the anti-inflammatory effect at 45 minutes
(maximum inflammatory effect of dextran) for the dose of 5 mg/kg p.o. were almost equal (38,5%)
to the effect of phenylbutazone at 100 mg/kg p.o. (33,55%) and for the dose of 10 mg/kg p.o.
almost double (60,52%). The anti-inflammatory effect of simvastatin lasts longer than that of
phenylbutazone. For theophylline the anti-inflammatory effect at 45 minutes was 34,33%, 59,57%
and 41,25% for the doses 25, 50, respectively 100 mg/kg p.o. comparing with the effect of 24,38%
of phenylbutazone at 100 mg/kg. The effect lasts longer for theophylline. The results of our
research complete other experimental and clinical studies and support the anti-inflammatory effect
of the statines HMG-CoA reductase inhibitors and of theophylline.
1. Sacks F.M. etal. TheN. Engl. J. of Med. 1996; 14: 1001-9.
2. Verschuren K.R. et al. Blood 2004.
3. Sullivan P., Bekir S., Jaffar Z., Page C., Jeffery P., Costello J. The Lancet. 1994; 343: 1006-
8.
3

P-117
STUDY ON THE AORTA NO AND HISTOPATHOLOGIES OF AORTA AND PANCREAS
OF THE DIABETIC RATS TREATED WITH BENFLUOREX AND/OR ASCORBIC ACID
B. Gonul'. C- Ozer 1 , L. Memi 2 , O. Ekinci 2
' Department of Physiology and 2 Department of Pathology, Faculty of Medicine, University of Gazi,
06500, Besevler, Ankara, Turkey
Streptozotocin (STZ) is a chemical which is used for creating diabetes on experimental animals.
Vitamin C (AA) economy may become deranged in diabetic animals and humans. AA dissolves in
water and takes part in the water compartment of the cell and acts as an antioxidant in this
environment. It was demonstrated that AA status can influence some selected indices of Stz diabetes in
rats. Benfluorex-Bfx, l-(3-trifluoromethylphenyl)-2(2-benzoyl-oxyethyl)-aminopropane) is an
antihyperlipidemic agent. Benfluorex treatment in middle-aged rats reverses the insulin resistance.
Reducing in NO tissue levels in diabetics can be explained with increase in catabolism or decrease in
formation. In this study the stz diabetic rats treated by benfluorex and/or ascorbic acid and these
animals" aorta NOx levels, aorta and pancreas histopathology were determined. Male wistar albino rats
were used. Experimental groups: (6 animals in each group) 1. untreated controls, 2. AA administered
controls, 3. Diabetic controls, 4. Bfx treated diabetics, 5. AA treated diabetics, 6. Bfx+ AA treated
diabetics. Controls were received the similar treatment with the experimental groups. Experimental
diabetes produced by ip injection of single 45 mg/kg dose of streptozotocin (Stz). Experimental
diabetes demonstrated by increasing blood glucose levels in the 2nd and 25th days of experiment.
After 21 days treatment by benfluorex (Bfx, 50 mg/kg/day, ig) and/or ascorbic acid (AA,
20mg/kg/day, ig). The animals killed by decapitation while the animals were under anesthesia. The
pancreas and aort tissues were removed quickly, a piece of aorta used for NOx determination. NOx
levels were detected by VCl 3 +Griess reactants and ELISA reader. Other pieces fixed in the formalin
solution and embedded in paraffin, then 4 micron thick sections were obtained and stained with
hematoxylen and eosin(H&E). These slides were examined by light microscopy. The glucose levels
and the body weights of animals observed during the experimental period. The results compared by
Anova variance analysis and Mann Whitney U tests. Body weigth and blood glucose levels of Bfx+AA
treated diabetic animals were similar to control animals' levels. Specimens of arcus aorta from all
groups showed no pathological change. In slides of pancreas from groups 1, 3, 4 and 6 Langerhans
islets were found to be in normal size and number without inflammatory or fibrotic alterations. Also,
exocrine component, ductal and vascular structures showed no abnormality.In group 2, there was
diffuse atrophy in Langerhans islets observed as reduction in both islet size and number. No
inflammation or fibrosis was present. The exocrine pancreas, ductuli and vessels showed no significant
pathological change. In group 5, atrophic changes in Langerhans islets were focal with most of the
endocrine component being normal. Diabetic rats' NOx levels decreased. In group 6 NOx levels
significantly increased.
Conclusion; Bfx+AA treatment has beneficial effects on the diabetic condition of rats. Thus this
combined therapy may also be useful for diabetic patients.
2

P-118
MELOXICAM, A CYCLOOXYGENASE-2 INHIBITOR, STIMULATES
HEMATOPOIESIS IN GAMMA-IRRADIATED MICE WHEN ADMINISTERED EITHER
PRIOR OR AFTER IRRADIATION
M. Hofer, M. Pospisil, *V. Znojil, J. Hola, A. Vacek, L. Weiterova, D. Streitova, A. Kozubi'k
Laboratory of Experimental Hematology, Institute of Biophysics, Academy of Sciences of the
Czech Republic, Kralovopolska 135, CZ-612 65 Brno, Czech Republic; institute of Pathological
Physiology, Medical Faculty, Masaryk University, Komenskeho nam. 2, CZ-662 43 Brno, Czech
Republic
Previous studies have revealed that non-selective inhibition of cyclooxygenases by classical nonsteroidal
antiinflammatory drugs (NSAIDs) supports radiation-suppressed hematopoiesis in mice.
However, administration of classical NSAIDS is frequently accompanied by undesirable
gastrointestinal effects which have been found in our experiments to aggravate the acute intestinal
radiation syndrome after the exposure of mice to high radiation doses. The experiments whose
results are presented here were aimed at testing whether the positive effects on hematopoiesis of
classical NSAIDs would be retained if cyclooxygenase-2-selective inhibitor meloxicam was
administered to the experimental animals. Meloxicam was given either in a single dose one hour
before irradiation with the dose of 6.5 Gy of gamma-rays or in four daily doses administered on
days 3, 4, 5, and 6 after irradiation with the dose of 4 Gy. In comparison with saline-treated
controls, significantly higher numbers of hematopoietic progenitor cells for granulocytes and
macrophages (GM-CFC) and for erythrocytes (BFU-E), as well as those of the peripheral blood
cells were found in meloxicam-treated mice. These findings suggest that meloxicam should be
further studied as a part of the treatment regimen for myelosuppression of various origin in the man.
3

P-119
THE INVESTIGATION OF ANTINOCICEPTIVE ACTIVITY OF MEMANTINE WITH P-
BENZOQUINONE WRITHING TEST IN DAY-AND NIGHT RHYTHM
P. K1I19, E. Aypar, N.N. Turan, N. Abacioglu
Department of Pharmacology, Faculty of Pharmacy, University of Gazi, 06330, Etiler, Ankara,
Turkey.
The N-methyl-D-aspartate (NMDA) receptor has long been considered as an important target for
the treatment of chronic pain. An NMDA receptor antagonist, Memantine, inhibits the development
of opioid tolerance and is essentially devoid of side effects at doses within the therapeutic range.
The aims of this study were firstly, if memantine has antinociceptive effect on mice PBQ induced
algesia model (diurnal) and compare this effect with morphine antinociception. Secondly, to
investigate the role of NO pathway. Experiments performed on male mice synchronized to 12:12 h
light-dark at two different times of the day (09:00-21:00). Animals were subcutaneously treated
with saline (O.lml/lOg) memantine (O.Olmg/kg), morphine (ED 50 =0.13 mg/kg), L-NAME (75
mg/kg), L-arginine (2.0mg/kg), memantine+morphine, memantine+L-arginine, memantine+L-
NAME, morphine+L-NAME, morphine+L-arginine, memantine+morphine+ L-NAME,
memantine+morphine+ L-arginine 15 minutes before PBQ (2.5 mg/kg, i.p.) injection. After PBQ
administration numbers of writhes were counted. Results were shown as normalized (arcsin
transformation) % antinociception values. Parametric ANOVA tests were used for statistical
analysis. We have observed that there were significant differences day-night variations in
memantine antinociception on selected time points. Memantine was abolished analgesic activity of
L-arginine and L-NAME when they were co-administration. It is likely that these results will need
advance research.
3

PHENOTYPING CONTRIBUTION TO SUCCESSFUL PERFORMANCE OF CLINICAL
BIOEQUIVALENCE STUDY OF NITRENDIPINE FORMULATIONS
J. Kopecky 1 . J. Zoulova 1 , J. Pastera 1 , K. Macek 2 , J. Chladek 1 , P. Anzenbacher 1 , J. Kvetina 1
P-120
'institute of Experimental Biopharmaceutics, Joint Research Centre of PRO.MED.CS Praha a.s. &
Academy of Sciences of the Czech Republic, Heyrovskeho 1207, CZ-50003 Hradec Kralove, Czech
Republic and 2 Teaching Hospital, Sokolska 581, CZ-50005 Hradec Kralove, Czech Republic
According to EMEA-CPMP Note for guidance on the investigation of bioavailability and
bioequivalence [1], "the bioavailability and bioequivalence studies with drugs known to be subject
to major genetic polymorphism could be performed in panels of subjects of known phenotype or
genotype for the polymorphism in question". Calcium channel blocker nitrendipine - used in the
treatment of hypertension - is highly metabolised by cytochrome P450 (CYP) 3A4 [2]. Hepatic and
small bowel CYP3A4 content and activity is known to vary 10-20-fold among individuals. Though
such a wide interindividual variability has been explained by ethnic and/or diet differences [2],
newer investigations make also genetic contribution evident [3]. Our aim was to develop a simple
method for sorting out the extreme metabolisers (both rapid and slow) of nitrendipine and to use
this method in the bioequivalence study of two nitrendipine formulations. To realise this task, the
peak plasma concentrations (c max ) of nitrendipine found after a single dose (20 mg) administration
of nitrendipine tablet (Baypress) were evaluated. In a pilot study with 18 healthy volunteers, the
Cmax acquired values from 1.4 to 51.3 ng/ml (median 6.95 ng/ml). As the time to reach c max (WO
was usually 1 or 2 hours in the pilot study, we decided to include into the pivotal bioequivalence
study of two nitrendipine formulations only volunteers who showed: a) 1 or 2 hours after "the
diagnostic (20 mg single dose) nitrendipine administration", the nitrendipine plasma concentrations
in the range 5-40 ng/ml (i.e. less than one order), and b) neither in 1 hour nor in 2 hours after the
administration these concentrations exceeding 40 ng/ml. Of 45 examined volunteers, 9 were
extreme metabolisers (8 rapid and 1 slow). Remaining 36 healthy volunteers entered the
bioequivalence study (open, randomized, cross-over) which proved bioequivalence of two
compared oral tablet nitrendipine formulations conclusively (the 90% confidence intervals for
AUCs and c max lie within an interval of 0.80-1.25). For the detailed results of this study see our
further abstract/poster at this Symposium (Zoulova J., Kopecky J. et al.: The relationship between
pharmacokinetics in human volunteers and in vitro dissolution test on the model of two oral
nitrendipine formulations). We conclude that performing a bioequivalence study after phenotyping
selection of healthy volunteers contributed to the successful evidence of bioequivalence of
nitrendipine formulations.
1. EMEA - CPMP: Note for guidance on the investigation of bioavailability and
bioequivalence (CPMP/EWP/QWP/1401/98). London, 26 July 2001,
www.emea.eu.int/pdfs/human/ewp/140198en.pdf
2. Dresser G.K. et al. Clin. Pharmacokinet. 38:41, 2000
3. Ozdemir V. et al. Pharmacogenetics 10:373, 2000
3

P-121
INHIBITION OF INDUCIBLE NITRIC OXIDE SYNTHASE RESTORES ATTENUATION
OF END OTHELIUM-DEPENDENT AND ENDOTHELIUM-INDEPENDENT
RELAXATIONS IN RAT SUPERIOR MESENTERIC ARTERY EXPOSED TO
ENDOTOXIN
E. Ozveren, B. Korkmaz, C.K. Buharalioglu, B. Tunctan
Department of Pharmacology, Faculty of Pharmacy, Mersin University, 33169, Mersin, Turkey
We have previously shown that increased production of vasodilator arachidonic acid products
bycPLA2a/COX-2 pathway rather than prostacyclin and nitric oxide (NO) contributes to reversal
ofendotoxin (ET)-induced vascular hyporeactivity by inhibition of calmodulin kinase II (CaMKII)
in ratsuperior mesenteric artery. There are conflicting results concerning the effect of ET on the
endotheliumdependentrelaxations in mesenteric arteries isolated from endotoxemic rats and the
arteries incubatedwith ET in vitro. Although increase or no change in endothelium-independent
relaxations has also beenshown in the mesenteric arteries isolated from endotoxemic rats, there is
no study in the arteries incubatedwith ET in vitro. This study was conducted to determine whether
ET impairs the endothelium-dependentand endothelium-independent relaxations in the rat isolated
superior mesenteric artery with endothelium/^ vitro. Contribution of inducible NO synthase (iNOS),
85 kDa cytosolic phospholipase A2a (cPLA2a),inducible cyclooxygenase (COX-2) and CaMKII to
the ET-induced impairment of relaxation responseswere also investigated. Incubation with ET (100
pg/ml) for 4 h significantly reduced relaxations to anendothelium-dependent vasodilator,
acetylcholine (ACh) (10 pM), and an endothelium-independentvasodilator, glyceryl trinitrate
(GTN) (10 pM) in the norepinephrine (0.001-1000 pM)-contracted rings.The ET-induced decrease
in ACh- and GTN-evoked relaxations were prevented by a selective iNOSinhibitor, phenylene-1,3-
bis[ethane-2-isothiourea] dihydrobromide (1,3-PBIT) at 0.1 pg/ml concentration. 1,3-PBIT at 1 and
10 pg/ml concentrations in the presence of ET did not change the ACh- and
GTNinducedrelaxations. A selective cPLA2« inhibitor, methyl arachidonyl fluorophosphonate (0.1,
0.3 and 1 pg/ml), and a selective COX-2 inhibitor, l,4-diamino-2,3-dicyano-l,4-bis[2-
aminophenylthio]butadien(DFU) (0.01, 0.1 and 1 pg/ml), and selective CaMKII inhibitor, N-[2-(N-
(4-chlorocynnamil)-Nmethylaminomethyl)phenyl]-N-[2-hydroxiethyl]-4-methoxybenzensulphonamide
(KN-93) (0.1, 0.3 andl pg/ml), had no effect on the ET-induced decrease in the AChand
GTN-evoked relaxations. These datasuggest that the ET-induced attenuation of endotheliumdependent
and endothelium-independentrelaxations in the rat superior mesenteric artery may be
related to the increased activity of iNOS ratherthan the cPLA2«, COX-2 and CaMKII.
This study was supported by Mersin University Research Foundation.
3

THE EFFECTS OF BENFLUOREX AND/OR ASCORBIC ACID ADMINISTRATION ON
THE KIDNEY OXIDANT PROCESSES IN DIABETIC RATS
P-122
C. Ozer. B. Gonul
Faculty of Medicine, Department of Physiology, University of Gazi, Ankara, 06500, Turkey
Streptozotocin (STZ) is an agent used in creating experimental diabetes. Benfluorex-Bfx is an
antihyperlipidemic agent. Increased oxidant stress plays an important role in ethiopathogenesis of
chronic complications of diabetes. It is expected that support with vitamin C (Ascorbic Acid-AA) in
diabetics could be beneficial in terms of the control of oxidizing events. The aim of this study, to
search the effects of AA and/or Bfx treatment on the oxidant and antioxidant processes in the
kidney of diabetic rats. Wistar Albino rats were separated 6 groups: 1-Control, 2-AA, 3-Diabetes
(D), 4- D+AA, 5. D+Bfx, 6. D+Bfx +AA. Rats were treated with single dose Stz (45mg/kg, i.p.) for
induction of diabetes. After 48 hours, rats, fasting blood glucose levels over 200mg/100ml, were
included in the diabetes groups. After 21 days treatment by Bfx (50 mg/kg/day ,ig) and/or AA
(20mg/kg/day,ig) the animals were sacrificed under anesthesia. Kidneys were removed quickly and
malondialdehyde (MDA) and glutathione (GSH) levels were determined. ANOVA and Mann-
Whitney U tests were used for statistical analysis. Diabetic rats' MDA levels increased while GSH
levels decreased compared to controls. Bfx and AA alone decreased the MDA levels and increased
the GSH levels in kidney tissue. Bfx was more effective on these parameters than AA treatment.
MDA and GSH levels were similar with controls in Bfx + AA treated group. According to our
results Bfx + AA treatment has beneficial effects on diabetic rats' kidney. This combined therapy
may be considered as a useful treatment for diabetic patients.
Table : The effects of Bfx and/or AA administration on the kidney MDA and GSH levels
Control
(a)
AA
(b)
Diabetes
(c)
D+AA
(d)
D+Bfx
(e)
D+Bfx+AA
(f)
MDA
(nmol/g)
33.6 ±2.4 29 ± 1.3 85 ±4 67 ± 4.4 39.1±2.4 31.1 + 2
GSH 8.3 ±0.9 10.2 ± 1.2 3.2 ±0.7 4.2 ±0.8 8.1 ± 1 10 ±0.7
(jimol/g)
Each value is the mean ± SD.
MDA : p

P-
GASTROPROTECTIVE EFFECT AND CYTOTOXICITY OF LABDENAMIDES
J.A. Rodriguez 1 , C. Theoduloz 1 , R. Izquierdo 2 , T. Yanez 1 , L. Astudillo 2 , G. Schmeda-Hirschmann 2
1 Departamento de Ciencias Basicas Biomedicas, Facultad de Ciencias de la Salud, Universidad de Talca,
Casilla 747, Talca, Chile, 2 Laboratorio de Quimica de Productos Naturales, Instituto de Qui'mica de
Recursos Naturales, Universidad de Talca, Casilla 747, Talca, Chile
The gastroprotective effect of the resin obtained from the large South American tree Araucaria araucana
has been recently reported as well as the comparative activity of natural and hemisynthetic labdane
diterpenes in the HCl/EtOH gastric lesions model in mice. 15-Hydroxyimbricatolal, 15-
acetoxyimbricatolal, 15-acetoxylabd-8(17)-en-19-oic acid and the corresponding methyl ester proved to be
the most active derivatives. Following our studies on the gastroprotective effect of terpenoids, different
aromatic amides at C-19 from 15-acetoxylabd-8(17)-en-19-oic acid and the isomer 15-acetoxylabd-8,9-en-
19 oic acid were prepared. The derivatives comprised the benzyl-, />anisidyl-, toluidyl-, p-yodophenyl-,
2,4-dimethylphenyl-, 2,4-dimethoxyphenyl-, 2-bromo-, 3-bromo- and 2-hydroxy-5-chlorophenyl amides.
The gastroprotective effect of the compounds was assessed in the HCl/EtOH gastric lesions model in mice
and the cytotoxicity of the products was determined using MRC-5 fibroblasts and AGS gastric cells. The
gastroprotective effect of the benzylamides belonging to the 8,9- and 8,17-en labdane was assessed at 12.5,
25 and 50 mg/kg. In the 8,9-en derivative a statistically significant gastroprotective effect was observed
starting at 12.5 mg/kg, reducing the gastric lesions by 51%, while the effect elicited by the 8,17-en
compound was significantly different to the control (67% reduction of lesions) at 25 mg/kg. The result ol
this experiment allowed us to use the 25 mg/kg dose for the comparison of the different amides prepared.
In the 8,17-en series, the best gastroprotective effect was elicited by the benzyl- and 3-bromophenylamide
reducing gastric lesions by 68 and 76%, respectively while the /-yodophenyl- and 2-bromophenylamide
reduced the lesions by 57-59%. In the 8,9-en labdane derivatives, the highest effect was provided by the
benzyl- and 3-bromophenylamide from 15-acetoxylabd-8(17)-en-19-oic acid as well as the benzyl- and p-
toluidylamide of 15-acetoxylabd-8,9-en-19 oic acid. The p-toluidyl-, benzyl- and 2-hydroxy-5-
chlorophenylamide reduced lesions by 77, 69 and 63%, respectively. A 50% reduction of ulcers was
elicited by the p-yodophenyl- and 2,4-dimethoxyphenylamide. In the cytotoxicity studies, compounds
displayed low toxicity with IC 5 o >1000 pM. As a whole, the amides showed better gastroprotective effect
than the parent diterpenes, with much lower cytotoxicity. Highest cytotoxicity towards AGS cells was
observed for the 2-bromophenyl- and 2-hydroxy-5-chlorophenyl amides in both diterpene series, with IC5c
in the range of 14-34 pM towards AGS cells and fibroblasts (IC 50 10-37 pM).
Supported by FONDECYT, Grant N° 1030583 and the Programa de Productos Bioactivos, Universidad de
Talca. Rafael Izquierdo thank the Gobierno Regional Grant for the Memoria de Titulacion. We are grateful
to Prof. Dr. Antonio Palenzuela, Universidad de La Laguna, Tenerife, Spain, for recording the mass
spectra.

P-124
A COMPARATIVE STUDY OF ANTIOXIDATIVE PROPERTIES OF SOME MEDICINAL
PLANTS GROWING IN TURKEY
I. i. Tatli Cankava 1 . F. Kovoh 2 , A. Sezgin 1 , O. Mumcu 1 , M. Martin 3 , S. Sahbaz 4 , F. Bailleul 4 , N.
Ezer 1
'Hacettepe University, Faculty of Pharmacy, Dept. of Pharmaceutical Botany, Sihhiye, 06100
Ankara-Tiirkiye, 2 Universite de Lille 2, Faculte de Pharmacie, Laboratoire de Pharmacologic et de
Pharmacie Clinique, B.P. 83 59006 Lille cedex, France,
3 Departement de recherche sur les
Lipoproteines et l'Atherosclerose INSERM UR 545, Institut Pasteur de Lille et Faculte de
Pharmacie, Universite de Lille 2, B.P. 245, 59019 Lille cedex, France, 4 Universite de Lille 2,
Faculte de Pharmacie, Laboratoire de Pharmacognosie, B.P. 83 59006 Lille cedex, France
Excessive production of free radicals is known to induce oxidative damage in cells and causes several
serious diseases (1). This is why in recent time a great attention is devoted to the research of new effective
antiradical and antioxidative substances. Due to the negative effects of some synthetic antioxidants, there is
an increased tendency for their replacement with natural ones (2). In an ethnopharmacological screening of
selected medicinal plants (Hypericum orientale L., Helichrysum plicatum Dc. subsp. plicatum, Centaurea
drabifolia Sm. subsp. drabifolia, Centaurea drabifolia Sm. subsp. detonsa (Bornm.) Wagenitz, Achillea
wilhelmsii C. Koch, Rubus canescens Dc. var. canescens), methanolic extracts from Turkish medicinal
plants were assayed in vitro to detect freeradical scavenging activity.The antioxidant activities were studied
by three different techniques: 1. Qualitative and quantitative freeradical scavenging assays using DPPH. For
the qualitative assay, extracts (10 pl/spot) and reference (green tea) were applied on silica gel 60 F254 TLC
plate, developed with appropriate eluents, dried out and stained with DPPH solution in methanol. For
quantitative studies, calibration curves were obtained with extracts in 5 different concentrations (10, 5, 2.5,
1.25, 0.625 mg/ml). The decrease in absorbance was followed at 517 nm (DPPH), until the reaction reached
a plateau (steady state). The radical scavenging effects were evaluated by the decrease in absorbance at
maximum absorbance and expressed as inhibition percentage (IP). The methanol extracts exhibited a strong
dose-dependent inhibition of DPPH activity which was similar to green tea. Rubus canescens var. canescens
extract was the most active with IP 94 % towards DPPH. 2. The ability of these species to scavenging
superoxide anions was determined in the hypoxanthine-xanthine oxidase system using cell-free experiments.
Control experiments were performed to find out if the scavenging of superoxide radical was in fact done by
inhibiting xanthine oxidase. These extracts (each 10 mg/ml concentratiosn) show inhibitory activity on the
enzyme. Besides these, they were able to scavenge the hydrogenperoxide radical. 3. These extracts were also
investigated by the non-enzymatic lipid peroxidation assay for their ability to act as radical scavenging
agents. The results will evaluate that the methanolic extracts exhibited Cu +2 -induced peroxidation of
phospholipid liposomes in a dose-related manner (conc. 10, 5, 2.5, 1.25 and 0.625 mg/ml). These antiradical
properties of selected species which are mostly due to the presence of phenolic compounds in methanolic
extracts support the traditional use of these species for the treatment of hemorrhoids, diarrhea and abdominal
pains.
1. Mahakunakorn, P. et al. (2004). Biol. Pharm. Bull. 27: 38-46.
2. Cuendet, M. et al. (1997). Helv. Chim. Acta 80: 1144-1152.

GASTROPROTECTIVE AND CYTOTOXIC EFFECT OF SEMISYNTHETIC FERRUGINOL
DERIVATIVES
C. Theoduloz', C. Areche 2 , G. Schmeda-Hirschmann 2 ,1. Razmilic 2 , T. Yanez 1 , J A. Rodriguez 1
1 Departamento de Ciencias Basicas Biomedicas, Facultad de Ciencias de la Salud, Universidad de Talca,
Casilla 747, Talca, Chile, Laboratorio de Quimica de Productos Naturales, Instituto de Quimica de
Recursos Naturales, Universidad de Talca, Casilla 747, Talca, Chile
Dehydroabietic acid derivatives have been shown to display gastroprotective effect in animal models.
Ferruginol is an abietane diterpene occurring in wood and bark of the native gymnosperm Prnmnopitys
andina (Poepp. Ex Endl.) de Laub (Podocarpaceae). A dose-response study of ferruginol in the HCl-EtOH
induced gastric lesions in mice showed that ferruginol inhibited gastric lesions at 25 mg/kg similar to
lansoprazole at 20 mg/kg. To disclose some structure-activity in this diterpene, some 17 semisynthetic
ferruginol derivatives were prepared and their gastroprotective activity was assessed on experimentally
induced gastric lesions in mice and cytotoxicity in human lung fibroblasts (MRC-5) and human epithelial
gastric (AGS) cells. The highest gastroprotective effect was provided by the compounds 1, 7, 8, 16 and 17
being as active as lansoprazole at 20 mg/kg. Compounds 6, 14 and 15 did not displayed anti-ulcer activity.
No relation was observed between lipophilicity values and the gastroprotective effect. Highest cytotoxicity
towards AGS cells was observed for compounds 2, 4, 6, 7, 8, 15, and 17 (IC 5 o in the range 18-31 |^M)
while compounds 9-14, that exhibited the highest lipophilicity values, and 16 were the less cytotoxic
showing IC5o values > 1000 pM. Compounds 2, 6, 7, and 15 were most active against fibroblasts (IC50 in
the range 19-24 pM). Compounds 9-14, 16 and 18 showed low cytotoxicity (IC50 values > 1000 pM). The
best activity/cytotoxicity ratio was found for the derivative 16 with a lesion index comparable to
lansoprazole at 20 mg/kg and a cytotoxicity > 1000 pM towards MRC-5 and AGS cells, respectively. In
conclusion, many derivatives showed a better gastroprotective effect/cytotoxicity ratio than the parent
compound ferruginol. These promising results encourage further pharmacological studies of these
compounds as potential gastroprotective drugs.
Supported by FONDECYT Grant N° 1060841 and Programa de Investigation en Productos Bioactivos,
Universidad de Talca. C. Areche thanks the Universidad de Talca for a Doctoral Grant.
P-1
241

THE RELATIONSHIP BETWEEN PHARMACOKINETICS IN HUMAN VOLUNTEERS
AND IN VITRO DISSOLUTION TEST ON THE MODEL OF TWO ORAL NITRENDIPINE
FORMULATIONS
J. Zoulova', J. Kopecky 1 , J. Pastera', Z. Svoboda', J. Chladek', K. Macek 2 , J. Kvetina'
1 Institute of Experimental Biopharmaceutics, Joint Research Centre of PRO.MED.CS Praha a.s. &
Academy of Sciences of the Czech Republic, Hradec Kralove, 2 Faculty Hospital, Hradec Kralove,
Czech Republic
The aim was to investigate the relationship between dissolution rate and bioavailability on the
model of oral nitrendipine dosage forms with immediate release. Nitrendipine is insoluble in water,
after oral administration is rapidly and nearly completely absorbed (about 90 %), but due to a high
effect of the first pass through the liver - CYP3A4 - the systemic availability is only 20-30 % [1].
Nitrendipine belongs to the class II (low solubility, high permeability) in the Biopharmaceutical
Classification System (BCS) [2], Two oral nitrendipine tablet formulations A and B were compared
in the dissolution test (paddle metod, paddle-shaft wobbing at 50 rpm, 2% lauryl sulfate medium)
and bioequivalence study (open, randomized, cross-over). Formulations were administered in a
single oral dose of 20 mg with a wash-out period of 7-10 days to 36 healthy volunteers (20 male, 16
female, 18-47 years). Blood samples were obtained 0-36 hours after the administration. The
determination of plasma nitrendipine concentrations was performed by gas chromatography. The
dissolution profiles were different between formulations. The similarity factor f 2 = 44 (f 2 value
between 50 and 100 suggests that the two dissolution profiles are similar). In contrast,
bioequivalence of the formulations was proven by statistical tests according to Schuirmann [3] in
bioequivalence range 80-125 % for all tested parameters:
Parameter Nitrendipine A Nitrendipine B Point estimate 94 % confid. interval
AUCO-xxj [ng.h/ml] 62.8+38.5 60.2+32.7 103 % 93.9-112.1
AUCo-» t
[ng.h/ml] 59.0+36.0 56.2+31.0 103 % 94.7-112.9
Cmax [ng/ml] 19.02+12.30 16.61+9.48 111 % 98.7-124.3
tmax [h] 1.91±1.08 1.99+0.75 -0.25 h -0.42 - 0.0 h
tl/2 [h] 6.37±4.44 6.56+4.83
- -
Conclusion: the dissolution rate is not a limiting factor for bioavailability of orally administered
drugs with physicochemical properties similar to nitrendipine (class II in BCS).
1. Drugs 33: 123-155 (1987)
2. Pharm. Res. 12: 413-420 (1995)
3. J. Pharmacokin. Biopharm. 15: 657-680 (1987)
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242

ABSTRACTS OF
POSTER PRESENTATIONS
-II-

P-1
SIMULTANEOUS DETERMINATION OF QUINAPRIL AND
HYDROCHLOROTHIAZIDE IN TABLETS BY RATIO SPECTRA DERIVATIVE
SPECTROPHOTOMETRY AND CHEMOMETRIC METHODS
S. Altinoz'. E. Din 2
'Hacettepe University, Faculty of Pharmacy, Department of Analytical Chemistry
06100 Ankara, Turkey, department of Analytical Chemistry, Faculty of Pharmacy, Ankara
University, 06100, Tandogan, Ankara, Turkey
Simultaneous determination of quinapril (QA) and hydrochlorothiazide (HCT) in tablets were
accomplished by ratio spectra first order derivative spectrophotometry (graphical method) and
chemometric method (numerical method). Both methods do not require any chemical separation
step. In the application of two analytical methods, the absorption spectra in the working range of
4.0-20.0 pg/mL QA and 2.5-12.5 pg/mL HCT were plotted in the wavelength range of 210-350 nm.
In the graphical approach, the absorption spectra of QA and its binary mixtures in the selected
spectral range of 210 - 280 nm were divided by the standard spectrum of lOpg/mL HCT and their
absorption spectra were obtained. In the similar way, the ratio spectra of HCT in the wavelength
region of 210-350 nm were also obtained by using the standard spectrum of 12 pg/mL QA. First
derivative of the ratio spectra obtained in the above steps were calculated by AX=5 nm interval for
both drugs. Calibration equation were obtained by measuring the ratio spectra derivative amplitudes
of the minima at 219.9 nm for QA and 283.2 nm for HCT in the above mentioned spectral ranges
for each drug. In the numerical method, the critical wavelengths corresponding to maximum points
at 213.0 nm for QA and 220.0 nm for HCT in the zero-order absorption spectra were selected to
construct the least squares calibration (CLS). Both graphical and numerical methods developed in
this study were completely validated and applied to the quantitative analysis of tablets containing
QA and HCT. The results obtained from the developed methods were compared with each other as
well as those obtained by classical derivative spectrophotometry given in the literature and the
difference was not observed statistically significant.
245

P-18
THE DETERMINATION OF AMOXICILLIN BY FLOW INJECTION ANALYSIS, UV-
SPECTROPHOTOMETRIC AND TITRIMETRIC METHODS
G. Altiokka
Department of Analytical Chemistry, Faculty of Pharmacy, Anadolu University, 26470, Eskijehir,
Turkey
Extended spectrum penicillins- ampicillin and amoxicillin have been used in antibacterial therapy
for many years. Amoxicillin is an orally absorbed, semi-synthetic broad-spectrum antimicrobial
drug. A direct determination of amoxicillin using flow injection analysis (FIA) with UV- detection,
potentiometry and conductometry and its application to the pharmaceutical tablets are described in
this study. The best carrier solvent system was found to be consisting of 10 percent MeOH (v/v)
solution at pH 9.0. A flow rate of 1 mL.min" 1 was pumped and active material was detected at 228
nm. The limit of detection (LOD) and limit of quantification (LOQ) for FIA were calculated to be
3.3xl0" 7 M (S/N=3) and l.OxlO' 7 M (S/N=10), respectively. We studied two pharmaceuticals
containing amoxicillin. In the analysis of tablets, the RSD values were found to be 1.36, 1.21 and
1.18 (for AMOKLAVIN®), 1.10, 1.16, 1.14(for REMOXIL®) for FIA, potentiometric and
conductometric methods, respectively.
246

VOLTAMMETRIC METHODS FOR ANALYTICAL DETERMINATION OF
NABUMETONE IN PHARMACEUTICAL DOSAGE FORMS, HUMAN SERUM AND
URINE
Y.Altun'. B.Dogan 2 , S.A.Ozkan", B.Uslu 2
1
Gazi University, Gazi Faculty of Education, Department of Chemistry, 06500, Teknikokullar,
2
Ankara, Turkey, Ankara University, Faculty of Pharmacy, Department of Analytical Chemistry,
06100. Tandogan, Ankara, Turkey
Nabumetone, naphthyl alkanone designated chemically as 4- (6-methoxy- 2-naphthalenyl)-2-
butanone, is a nonsteroidal anti-inflammatory drug that exhibits anti-inflammatory, analgesic and
antipyretic properties in pharmacologic studies. To our knowledge, no information about the
electrochemical redox properties of nabumetone and its analytical application has appeared in the
literature. This work throws a more deep light upon the electrochemical behavior and elucidation of
the electrode reaction pathway of nabumetone at the glassy carbon electrode. Validated
voltammetric procedures are also described for the trace determination of the nabumetone in bulk
form, pharmaceutical formulation, human serum and urine, without the need for sample
pretreatment or time-consuming extraction or evaporation steps prior to the drug analysis. The
electrochemical oxidation of nabumetone was investigated by cyclic, linear sweep, differential
pulse (DPV) and square wave (OSWV) voltammetry using glassy carbon electrode. The oxidation
of nabumetone was irreversible and exhibited diffusion controlled process depending on pH. The
dependence of intensities of currents and potentials on pH, concentration, scan rate, nature of the
buffer was investigated. For analytical purposes, very well resolved diffusion controlled
voltammetric peaks were obtained at pH 3.7 acetate buffer. The peak current was proportional to
the concentration of nabumetone in the range 1x10"^ - 8x10"^ M with a slope of 7.44xl0 4 pA/M (r:
0.999) for DPV and 9.05x10 4 pA/M (r: 0.999) for OSWV. Validation parameters of the methods
showed it to be accurate, precise and linear over the concentration range of analysis with minimum
detectability of 2.55xl0" 7 M for DPV and 2.85xl0' 7 M for OSWV. The proposed methods were
successfully applied to the determination of nabumetone in pharmaceutical dosage form, human
serum and urine samples.
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247

POSTVALIDATION AND EVALUATION OF THE PERFORMANCES OF A CAPTOPRIL
BIOANALYTICAL METHOD IN A BIOEQUIVALENCE STUDY
V. Anuta, G. Ionica, E. Gutu, C. Mircioiu
University of Medicine & Pharmacy "Carol Davila", Faculty of Pharmacy, Bucharest-Romania
Post validation of a bioanalytical method in bioequivalence studies is essentially a
"pharmacokinetic and pharmacoeconomic validation". A bioanalytical method for determination
of captopril in plasma samples was developped in the laboratory, based on derivatization of
captopril by "capture" of free -SH groups of captopril with monobromobimane. Critical aspect of
the method concerns high instability of captopril both in vivo and ex-vivo", following dimerization
of SH groups as well as capture of aminacids from plasma. A first consequence of these
phenomena is the very short half-time of captopril and difficulties in evaluting its pharmacokinetics
for longer periods. Limit of quantification and and length of the sampling interval were finally
choiced such as to assure the determination of at least 80 % of the Area Under Curve (AUC), as is
required by bioequivalence rules.
P-1
600
500
400
300
200
100
0
mean ( n= 24) plasma levels of captopril
—
2 4 6 8 10
time (h)
This postfactum evaluation showed that an interval of 4 hours for sampling interval, instead of 8
h as was planned in the development phase, was sufficient ( as can be observed, with naked eye
from fig. 1) for assuring experimental determination of more than 95 % of AUC. It is also to take
into consideration the modification of estimated im following modification of sampling interval.
248

GC-NPD METHOD DEVELOPMENT AND VALIDATION FOR DETERMINATION OF
PRILOCAINE HC1 IN PHARMACEUTICAL PREPARATIONS
A. Atila', Y. Kadioglu', A.Temizer 2
'Department of Analytical Chemistry, Faculty of Pharmacy, Ataturk University, 25240, Erzurum,
Turkey, department of Analytical Chemistry, Faculty of Pharmacy, Hacettepe University, 06100,
Ankara, Turkey
Local anesthetics are drugs which are used to reversibily block nerve function. They prevent
conduction of electrical impulses by the membranes of nerve and muscle when these drugs applied
in sufficient concentration at the site of action. Prilocaine [2-( Propil amino )-o-propionotoluidine
mono hydrochloride] has been used succesfully to alleviate pain associated with medical procedures
and extensively metabolized by the liver [1,2]. A simple, rapid, precise, accurate and specific GC-
NPD method for the directly determination of prilocaine in pharmaceutical preparation (citanest)
was developed and validated.Lidocaine was used to be internal standard. The retention time of
prilocaine and lidocaine in GC-NPD method using ultra colomn 2 (methyl phneyl silicone) were
6.80 min and 7.10 min, respectively. Split injection was used and the carrier gas was helium at a
flow-rate of 0.7 ml min" 1 . Helium (9 ml min -1 ), hydrogen (4ml min" 1 ), synthetic air (60 ml min" 1 )
were used as auxiliary gases for nitrogen phosphorus detector. The injector and detector
temperatures were 280°C. The temperature of the GC oven was as follows, initial temperature 90°C
, final temperature 300°C, hold 7 ml/min, ramp rate 20°C min" 1 Calibration curves were prepared
over the concentration ranges 40 to 1000 ng ml" 1 . The precision of this method was less than 2 %
calculated as the relative standard deviation (RSD) accuracy better than 3 % (n=6) . LOD and LOQ
values were 30 ng ml" 1 and 40 ng ml" 1 , respectively. The developed method was applied directly and
easily to the analysis of pharmaceutical preparation. R.S.D. were found to be %7.42. In general, by
its simplicitiy, rapidity and sensitivity, GC-NPD method can be used for the routine quality control
analysis of the investigated drug in pharmaceutical preparations.
1. J.Klein, D. Fernandes, M. Gazarian, G. Kent, G. Koren, Journal of Chromatography B, 655,
1994,83-88
2. A.S. Gross, A.Nicolay, A. Eschalier, Journal of Chromatography B, 728,1999,107-115
P-1
249

THE DETERMINATION OF TIOCONAZOLE BY FLOW INJECTION ANALYSIS, UV-
SPECTROPHOTOMETRIC AND TITRIMETRIC METHODS
Z. Atkoar
Department of Analytical Chemistry, Faculty of Pharmacy, Anadolu University, 26470, Eski§ehir,
Turkey
Tioconazole is a member of the imidazole class compounds and is a medicine that has antifungal
effect. The ability of quantity determination of tioconazole with UV-spectrophotometric,
potentiometric, conductometric and FIA methods was researched in this study. For the FIA, the best
carrier solvent was found to be consisting of 10 percent methanol (v/v) solution. A flow rate of 1
mL.min" 1 was pumped and analyte was detected at 219 nm. The calibration equation of tioconazole
was linear in the range of l.OxlO" 5 - 5.0xl0" 5 mol.L" 1 . The limit of detection (LOD) and limit of
quantification (LOQ) for FIA was calculated 4.2xl0" 7 mol.L" 1 and 1.7xl0" 7 mol.L" 1 , respectively. It
was found that UV spectrophotometric, FIA and potentiometric methods are feasible for
determination on the condition being worked with standard tioconazole. However the
conductometric method is suitable for the tioconazole when is in excess amount. The methods
which were developed were practiced to the commercial pharmaceutical formulation. Because the
amount of tioconazole in the pharmaceutics was not sufficient for the conductometric titration to be
practiced, it was stabilized that this method was not suitable for the determination of tioconazole in
the pharmaceutical preparations. In the analysis of pharmaceutical preparations, the RSD values
were found to be 1.29, 0.75 and 1.07 for FIA, potentiometric and UV-spectrophotometric methods,
respectively. The results which were handed from these methods were evaluated and it was founded
that the proposed method were useful and suitable for the determination of tioconazole in the
pharmaceutical dosage forms.
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250

SPECTROFLUORIMETRIC AND SPECTROPHOTOMETRIC DETERMINATION OF
ROPINIROLE HYDROCHLORIDE IN TABLETS
Z. Aydogmus
Department of Analytical Chemistry, Faculty of Pharmacy, Istanbul University, 34116 Beyazit,
Istanbul, Turkey
Ropinirole, 4-[2-(dipropylamino)ethyl]-l,3-dihydro-2H-indol-2-one, is active dopamine D2-
receptor agonist and has been used in the treatment of parkinson's disease. Very few
chromatographic techniques have been reported for the determination of ropinirole in biological
fluids such as HPLC method with UV detection [1], Thermo spray LC-MS/MS methods have been
applied for the determination of drug metabolites [2] and pharmacokinetics evaluation. Until now,
no spectrofluorimetric and spectrophotometric methods have been described for the determination
of ropinirole in pharmaceutical formulations.In this study, two accurate, sensitive, and simple
spectrofluorimetric and spectrophotometric methods were developed for the determination of
ropinirole hydrochloride in tablets. The spectrofluorimetric method was based on derivatization
with 7-chloro-4-nitrobenzofurazan (NBD-C1) in borate buffer. After the extraction with chloroform
the fluorescence intensity of the derivative was measured at 530 nm with excitation at 464 nm. The
color was found to be stable for at least 48 h in this solvent. The optimum conditions of the reaction
were investigated and it was found that the reaction proceeds quantitatively, pH 8.5, 70 °C in 10
min when the mole ratio of the reagent to drug was 25. The spectrophotometric method was
performed by measuring the absorbance at the wavelength of 250 nm in methanol solvent. The
linearity ranges were found to be 0.01-1.200 ug/ml and 2.5-24 ug/ml for the spectrofluorimetric and
spectrophotometric methods, respectively. The detection limits were 0.0012 ug/ml and 0.48 ug/ml
for the spectrofluorimetric and spectrophotometric methods, respectively. The proposed methods
were applied to the assay of ropinirole hydrochloride in tablets. The results of these two proposed
methods were compared statistically.
1. J. Bhatt, A. Jangid, R. Shetty, B. Shah, S. Kambli, G. Subbaiah, S. Singh, J. Pharm.
Biomed. Anal., 40, 1202-1208 (2006)
2. C.R. Blakley, J.J. Carmody, M.L. Vestal, J. Am. Chem. Soc. 102 5931-5933 (1980)
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251

HPLC SCREENING OF 1,3,4-OXADIAZOLE MODIFIED CHLOROPHENYLUREAS
AND FLUROBENZAMIDES WITH CALIXARENE-BONDED STATIONARY PHASES
G. Bazylak. A. Malak
Department of Pharmaco-Bromatology, Faculty of Pharmacy, Collegium Medicum, Nicolaus
Copernicus University, Jagiellonska 13, PL-85-067 Bydgoszcz, Poland
P-14
Retention profiles in series of the neutral and highly hydrophobic 1,3,4-oxadiazole containing
chlorophenylureas and fluorobenzamides indicating analgesic activity were determined in the
narrow-bore isocratic HPLC systems employing calix[6]arene- as well as tert-butyl calix[4]arenebonded
stationary phases. When acetonitrile + 2.65 mM phosphoric acid (55 : 45, v/v), pH 3.25,
mobile phase was applied retention of these compounds increased with diminishing of their overall
hydrophobicity according to the general preference of more polar compounds by calixarene cavity
in time of its non-specific host-guest supramolecular interactions with halogenated substances. The
size of calixarene cavity and its upper-rim substitution did not changed observed retention order,
resolution and selectivity of separation for oxadiazoles. Compare to retention on the highly-endcapped
octadecylsilica HPLC column a highly improved separation of some positional isomers of
halogenated oxadiazoles were observed on the both used calixarene-type HPLC supports. In
addition formation of the 1:1 inclusion host-guest complexes between each calixarene-type
stationary phase and oxadiazoles were studied with molecular modelling MM + and AMI methods.
The structural and energetic factors leading to the hydrogen bond stabilized inclusion complexes
between these species were considered and used for explanation of observed retention sequence and
selectivity of oxadiazoles in applied HPLC systems.
252

EFFICIENCY OF CALIXARENE-BONDED STATIONARY PHASES IN NARROW-BORE
HPLC SCREENING SCHEMES FOR BETA-AGONISTS AND BETA-BLOCKERS
G. Bazylak , A. Malak
Department of Pharmaco-Bromatology, Faculty of Pharmacy, Collegium Medicum, Nicolaus
Copernicus University, Jagiellonska 13, PL-85-067 Bydgoszcz, Poland
P-1
Regularities in retention of commonly prescribed 10 beta-agonists and 16 beta-blockers on the
calix[6]arene- as well as tert-butyl calix[4]arene-bonded stationary phases in the isocratic narrowbore
HPLC systems were studied. Decreased retention and separation efficiency of these solutes
were observed after increasing concentration of acetonitrile or methanol from 10 to 80 percent in
the acidic aqueous mobile phase. Reduced retention times were obtained on the calixarene-type
stationary phase with smallest cavity. Compare to retention on the electrostatically shielded
octadecylsilica-type HPLC packing XTerra RP-18 both studied calixarene-type packings offers
baseline separation of rather not rich-component mixtures of racemic beta-blocking drugs as
metoprolol, oxprenolol, p-oxprenolol, atenolol, propranolol, bufuralol in time not exceeding 3.0
minutes. However, the partial separation of (R)- and (S)-enantiomers of bisoprolol, atenolol,
salbutamol and fenoterol was achieved on the achiral tert-butyl calix[4]arene packing eluted at 22
°C with acetonitrile + 1.40 mM phosphoric acid (25:75), pH 3.25, and flow rate 0.5 ml/min. For
explanation of this unusual effect the results of detailed molecular modelling calculations on the
formation of supramolecular host-guest complexes between upper-rim modified cavity of some
unstable conformers of calixarene moiety and each enantiomer of mentioned drugs were presented.
Multivariate statistics procedures were also applied to evaluate the usefulness of collected here
retention data in various HPLC systems as the pharmacological similarity/diversity screening tool
of beta-adrenergic and beta-adrenolytic drugs.
253

NEW SINGLE ISOMER p-CYCLODEXTRIN DERIVATIVE AS A CHIRAL SELECTOR
IN CAPILLARY ELECTROPHORESIS
P-1
J. Boonleang 1 . R.G. Carlson 2 , J.F. Stobaugh 3
'Department of Pharmaceutical Chemistry, Prince of Songkla University, Songkhla, 90112,
Thailand, department of Chemistry, The University of Kansas, Lawrence, KS, 66045, USA,
department of Pharmaceutical Chemistry, The University of Kansas, Lawrence, KS, 66047,
USA
The specific mono-6-substituted-P-cyclodextrin derivative carrying three negative charges, mono-
[6-deoxy-6-(6-sulfooxy-5,5-bis-sulfooxymethyl-hexylthio)]-p-cyclodextrin, was synthesized in
order to increase separation window and resolution power of a chiral selector in enantiomeric
separation in capillary electrophoresis. Since this compound composes of only one single
component with appropriate number of negative charges, it might improve peak shape and
separation efficiency as well. By first synthesizing three negatively charged arm, mono-(6-
mercapto-2,2-bis-sulfooxymethyl-hexyl)sulfuric acid ester trisodium salt, this was then connected
to P-cyclodextrin by nucleophilic substitution reaction with mono-(6-tosyl)-P-cyclodextrin in dry
DMF at 70 °C overnight to give, mono-[6-deoxy-6-(6-sulfooxy-5,5-bis-sulfooxymethyl-hexylthio)]-
P-cyclodextrin , the desired product in good yield (80%). Its structure was confirmed by 'HNMR,
,3 CNMR and MS (FAB, positive). This compound can enantiomerically resolve many of the chiral
basic drugs used in this study, clenbuterol, ephedrine, epinephrine, propranolol, pseudoehedrine,
terbutaline and verapamil with baseline resolution; however, it gave partial resolution of ibuprofen,
the chiral acidic drug used in this study.
254

DETERMINATION OF TICLOPIDINE IN PHARMACEUTICAL FORMULATION BY
FLOW INJECTION ANALYSIS (FIA) WITH UV-DETECTION
N. O. Can. G. Altiokka
Department of Analytical Chemistry, Faculty of Pharmacy, Anadolu University, 26470, Eskiehir,
Turkey
A precise and accurate FIA method for the quantification of ticlopidine HC1 (TIC) in
pharmaceuticals is described. The best suitable carrier solvent is distilled water. Sample solution
(3,196 x 10" 5 M TIC) was prepared in this solvent and injected to the instrumental system at a flow
rate of 1,0 mL.min" 1 . The signals were detected by a UV detector at 214,2 nm. The calibration curve
of TIC was linear in the concentration range of 1,598 x 10" 5 M - 4,794 x 10" 5 M. The intra- and
inter-assay precision were less than 2,0 %. The method exhibited a good linearity with the
correlation coefficients. The effects of the tablet excipients were insignificant at the 95% probability
level. The calculated tablet content was 99% which is agreement with the ranges stated by
pharmacopoeias.
P-1
255

SPECTROPHOTOMETRIC METHOD FOR THE DETERMINATION OF ROPINIROLE
IN TABLETS
A. Onal, S. Caglar
Istanbul University, Faculty of Pharmacy, Department of Analytical Chemistry, 34452, Beyazit,
Istanbul - Turkey
P-18
Simple and rapid spectrophotometric procedure has been established for the quantitation of
ropinirole in pharmaceutical preparations. The method is based on the reaction between the
examined drug and bromocresol green [1] producing ion-pair complex. In the acidic buffer (pH:4) it
gives a yellow color after chloroform extraction and absorbance was measured at 412 nm. The
optimization of the reaction conditions was investigated. Beer's law is obeyed in the concentration
range 1.5-15 fag mL"'. The molar absorbtivity, Sandell sensitivity, detection and quantification
limits are also calculated. The developed method was applied successfully for the determination of
the drug in the commercial preparation and it can be recommended for the routine analysis of
ropinirole in tablets.
1. Safwan A., Raghad A. II Farmaco, 60, 771-775, 2005.
256

EXPERIMENTAL DESIGN-BASED DEVELOPMENT AND VALIDATION OF A
RAPID MICELLAR ELECTROKINETIC CHROMATOGRAPHIC METHOD FOR THE
SIMULTANEOUS DETERMINATION OF ISONIAZID AND PYRIDOXINE
HYDROCHLORIDE IN A PHARMACEUTICAL FORMULATION
E. Nemutlu, M. Celebier, B. Uyar, S. Altinoz
Hacettepe University, Faculty of Pharmacy, Department of Analytical Chemistry
06100 Ankara, Turkey
An efficient and reliable micellar electrokinetic chromatography (MEKC) method has been
developed for the simultaneous determination of isoniazid and pyridoxine hydrochloride in
pharmacutical formulations. A chemometric approach, two level full-factorial design, was used to
search for the optimum conditions of separation. Three parameters were selected for this study: the
buffer pH, the buffer concentration and SDS (Sodium Dodecyl Sulfate) concentrations. Resolutions,
peak symmetry and analysis time were established as responses. The two analytes were separated
within 6 min with the optimized conditions: 50 mM borate buffer, 25 mM SDS, pH 7.8, 30 °C, at
50 mbar 4 s injection and 30 kV. The method was validated with respect to stability, linearity range,
limit of detection and quantitation, precision, accuracy, specificity, and robustness. The developed
method was applied to the pharmaceutical preparation.
P-1
257

VOLTAMMETRIC DETERMINATION OF PHENAZOPYRIDINE HYDROCHLORIDE
IN HUMAN URINE AND TABLET DOSAGE FORMS
M. Citak'. S. Yilmaz 1 , G. Tinker', Y. Dilgin', S. Yagmur 1 , H. Erdugan 2
P-14
'Department of Chemistry, Faculty of Arts and Sciences, Canakkale Onsekiz Mart University,
17020, Canakkale, Turkey, 2 Department of Biology, Faculty of Arts & Sciences Canakkale
Onsekiz Mart University, 17020, Canakkale, Turkey
Phenazopyridine hydrochloride (PAP) exerts an analgesic effect on the mucosa of the urinary tract
and is used to provide symptomatic relief of pain in conditions such as cystitis and urethritis [1,2].
An electroanalytical method was developed for the direct quantitative determination of PAP in
spiked human urine and tablet dosage forms. The electrochemical reduction and determination of
PAP has been carried out at a carbon paste electrode in various aqueous solution in the pH range of
0.51-12.00 (Britton-Robinson, acetate, phosphate buffers and 0.5 M sulfuric acid solution) by cyclic
(CV) and Osteryoung square wave voltammetry (OSWV). To the best of our knowledge the
polarographic reduction of PAP has been investigated using a hanging mercury drop electrode
(HMDE) [1,2], but electrochemical reduction on carbon paste electrode (CPE) and voltammetric
determination have not yet been published. The best results were obtained for the quantitative
determination of PAP by OSWV method in 0.5 M sulfuric acid ( pH 0.51) at -0.056 V. The peak
current and peak potential depend on pH, so its influence and also scan rate effects were studied.
The diffusion controlled nature of the peak was established. This electroanalytical procedure
enabled to determine PAP in the concentration range 2.5x10" 8 -2.5x10" 6 M. A linear relationship was
obtained between peak current and concentration with equation of regression equal to I p (pA) =
4.78 xlO 5 C (M) - 0.037 (r = 0.996). Limit of detection and limit of quantitatification were
obtained as 7.5xl0" 9 and 2.5x10" 8 M, respectively. The proposed voltammetric technique was
validated and good recoveries were obtained in spiked urine and tablet dosage forms.
1. M.S.Suzy, Talanta 50 (1999) 133.
2. P.Surmann, P. Aswakun, Arch. Pharma. 318 (1985) 14.
258

P-14
EXTRACTIVE SPECTROPHOTOMETRIC STUDY OF RIZATRIPTAN AND ITS
DETERMINATION IN BULK AND IN PHARMACEUTICAL FORMULATIONS
E. Cicek. N. Erk
Ankara University, Faculty of Pharmacy, Department of Analytical Chemistry 06100 Tandogan -
Ankara-Turkey
Rizatriptan is a novel, selective 5-HTlB/lD receptor agonist, which is used in the treatment of
migraine headache. The main aim of this work was to investigate the reaction between rizatriptan
and sulphonphtalein dyes that are bromocresol green (BCG), bromocresol purpur (BCP) and
alizarin red (ARS). In this work, accurate, simple, rapid and sensitive three new extractive
spectrophotometric methods were developed for the determination of rizatriptan, producing ion-pair
salts in organic phases. In developing the methods, pH, time and temperature parameters were
considered and we provided optimum conditions with measurements. The proposed methods
involve formation of coloured chloroform extractable ion-pair complexes with BCG, BCP and
ARS. The complex species, which are extractable to chloroform phase were quantitatively
determined using at 618, 616, 602 nm for methods BCG, BCP and ARS, respectively. These ionpair
complexes were calculated by molar ratio methods. Linear calibration curves were obeyed over
the concentration range 0.750-2.25 pg-ml" 1 for BCG, 1.50-2.10 pg.ml" 1 for BCP and 0.90-1.65
pg.ml"' for ARS ion-pair complexes. These methods can be practise in pure form and in dosage
form successfully. The results obtained compared with the first derivative data and no difference
was found.
1. S.M. Blaih, H.H. Abdine, F.A. El-Yazbi and R. A. Shaalan; Spectroscopy Letters,33(1),91-
102(2000)
2. N. Rahman, N. A. Khan, S. N. H. Azmi; II Farmaco 5947-54 (2004)
3. Y.M. Issa, F.M. Abdel-Gawad, M.A. Abou Table and H.M. Hussein; Analytical Letters, 30(11),
2071-2084(1997)
4. A.L. El-Ansary, Y.M. Issa and W. Selim ; Analytical Letters, 32(5), 955-969(1999)
5. S. Altinoz, G. Uar, and E. Yildiz Analytical Letters, 35 (15) 2471-2485,(2002)
259

P-14
CONTINUOUS WAVELET TRANSFORM FOR THE RESOLUTION OF THE
OVERLAPPING VOLTAMMETRIC SIGNALS AND SIMULTANEOUS
DETERMINATION OF LEVODOPA AND BENSERAZIDE
S. Demircan', I. Siislu', E. Din 2 , S. Altinoz'
'Department of Analytical Chemistry, Faculty of Pharmacy, Hacettepe University, 06100, Sihhiye,
Ankara, Turkey, 2 Department of Analytical Chemistry, Faculty of Pharmacy, Ankara University,
06100, Tandogan, Ankara, Turkey
The overlapping voltammetric signals of levodopa (LD) and benserazide (BE) in their binary
mixture were processed by continuous wavelet transform (CWT) and the transformed signals
were used for the simultaneous determinations of these compounds in a pharmaceutical
formulation. The Osteryoung Square Wave Voltammograms of LD and BE in Britton Robinson
(BR, pH = 3) buffer were plotted in the potential range of -196 and 1200 mV using glassy carbon
electrode versus Ag/AgCl reference electrode. In the signal data processing, various continuous
wavelet family with different the scale parameter (a) was tested and Haar CWT method (a = 4)
was found to be the optimal wavelet transform for the signal processing. Calibration functions in
the linear dynamic range of 4.0-18.0 pg/mL for LD and 2.0-10.0 pg/mL for BE were obtained by
measuring the transformed voltammetric amplitude at 460 mV and 320 mV for LD and BE,
respectively. The validation of the Haar CWT method was carried out by analyzing the
independent set of the synthetic binary mixtures containing LD and BE. The mean recoveries and
relative standard deviations were found to be 98.9 % and 2.31 % for LD 101.6 % and 1.76 % for
BE. This proposed method was applied to the real samples consisting of LD and BE in the
commercial pharmaceutical as capsules. The experimental results obtained from the Haar CWT
method were compared with those obtained by the literature method. A good agreement was
observed for the obtained results.
260

P-14
GC-FID METHOD DEVELOPMENT AND VALIDATION FOR DETERMINATION OF A -
TOCOPHEROL (VITAMIN E) IN PHARMACEUTICAL PREPARATIONS
F. Demirkaya, Y.Kadioglu
Department of Analytical Chemistry, Faculty of Pharmacy, Ataturk University, 25240, Erzurum,
Turkey
Vitamin E is a term used to designate a family of related compounds (tocopherol and tocotrienols).
a-tocopherol (a-TC)(5,7,8 trimethyltocol) is the most biologically active form of vitamin E. It
functions as a chain-breaking antioxidant that prevents the propagation of free radical reactions and
as one of the most important fat-soluble antioxidant in biological systems [1-2]. A simple, rapid,
precise, accurate and specific GC-FID method for the directly determination of underivatized a-TC
in vitamin and multivitamin tablets was developed and validated. Separation of underivatized a-
tocopherol in pure substance and pharmaceutical preparations was performed in about 8.4 min,
using HP-5 capillary column. Splitless injection was used and the carrier gas was nitrogen at a flowrate
of 2 ml min" 1 . Nitrogen (25 ml/min), hydrogen (40 ml/min) and synthetic air (400 ml/min) were
used as auxiliary gases for the flame ionization detector. The injector and detector temperatures
were 300°C. Specificity of this method has been demonstrated by the representative chromatograms
for standard a-TC in vitamin and multivitamin tablets. Different temperature programs were
investigated for exception of matrix interference and the others vitamins in multivitamins tablets.
The end of this investigation, the best temperature program was selected for a good resolution. The
method has a wide linear over the 1-30 pg/ml of concentration range. The method was completely
validated and proven to be rugged. The precision of this method was calculated as the relative
standard deviation (R.S.D.) was less than 8 %, and accuracy (relative error ) was better than 11 % (n
= 6). Sensitivity of component on-column was in the nanogram range, from limit of detection
(LOD) to almost 300 ng ml"', thus allowing analysis of different samples of possible concentrations
of a-tocopherol. The developed method was applied to directly and easily to the analysis of the
pharmaceutical preparations. R.S.D. were found to be 0.59 % in Grandpherol® (soft gelatine
capsule; 200 I.U.), 6.59 % in Megadyn® (film tablet; 10 mg) and 1.54 % in (Supradyn® drage; 10
I.U.). This validated GC-FID, in conjunction with other methods, is potentially could be
successfully applied for routine laboratory because of its simplicity, rapidity, sensitivity, precision
and accuracy.
1. Pyka A., Sliwiok J., Chromatographic separation of tocopherols. Journal of
Chromatography, 2001,935,71 -76
2. Brigelius-Flohe R., Traber M.G., Vitamin E: function and metabolism. The FASEB
Journal,! 999,13, July, 1145-1155.
261

DETERMINATION OF CARBAMAZEPINE IN PHARMACEUTICAL PREPARATIONS
USING HPLC-DAD METHODS
F. Demirkava, Y. Kadioglu
Department of Analytical Chemistry, Faculty of Pharmacy, Atatiirk University, 25240, Erzurum,
Turkey
P-14
Carbamazepine (CBZ), 5-H-Dibenz [b.f] azepine-5-carboxomide, which has been developed as an
anticonvulsant drug, is used as an antiepileptic, antidepression and antimanic drug [1-2].
A simple, rapid, precise, accurate and specific HPLC-DAD method for the quantitative
determination of CBZ in pure forms and in pharmaceutical preparations were developed and
validated. Chromatography was conducted using a mobile phase of acetonitrile-Milli-Q grade water
(30:70 v/v) pumped at a flow-rate of 1 ml min"' through a Phenomenex Bondolone reversed-phase
Cl8 column (150 x 3.9 mm. 5 mm. USA). The injection was 10 ml and the peaks were detected
at 220 nm. The integrator attenuation was 8 and the chart speed was 0.2 cm min"'. Retention time
of CBZ was 8.2 min and the total run time for an assay was approximately 9 min. System suitability
parameters calculated under the optimized experimental conditions were; capacity factor (k ) 4.46;
symmetry factor 1.05 and column efficiency (n) 4303 plates /m. To determine the linearity of
HPLC-DAD, calibration standard solutions of CBZ were prepared. The linear ranges were found to
be 0.25-25 mg ml"'. The regression equations obtained by least square regression method were
y=449820x+l 15048 (y and x are mean peak area and concentration, respectively, r =0.999). The
limit of quantification (LOQ) and the limit of detection (LOD) were 70 ng ml"' and 50 ng ml"',
respectively. Intra-day and inter-day precision, expressed as the relative standard deviation (RSD)
was less than 7%, and accuracy (relative error ) was better than 7 %. The developed method was
applied to directly and easily to the analysis of the pharmaceutical preparations. R.S.D. were found
to be 0.90 % in Tegretol® tablet; (200 mg/tablets), 0.79 % in Karberol® tablet; (200 mg/tablets )
and 0,69 % in Karbelex® tablet; (300 mg/tablets ).
This validated HPLC-DAD method can be used for the routine quality control analysis of the
investigated drug in pharmaceutical preparations.
1. Bradley, M.K., Rene, H., Levy, R.R., Matson, R., Meldrum, B., Penry, J.K., Dreifuss F.E.,
(Eds.), Antiepileptic Drugs, New York, Third Edition, Raven Press, Ltd., 1989,505
2. Oiling, M., Mensinga, T.T., Barends, D.M., Groen, C., Lake, O.A., Meulenbent, J.,
Biopharmmaceutics and Drug Disposition, 1999, 20,19.
262

P-14
DETERMINATION OF NIFEDIPINE AND PHENYTOIN BY HPLC IN HUMAN
GINGIVAL CREVICULAR FLUID AND PLASMA
A. Dincel'. G.N. GUnctt 2 , F. Caglayan 2 , A. Bozkurt 1
'Faculty of Medicine, Department of Pharmacology, 2 Faculty of Dentistry, Department of
Periodontology, Hacettepe University, Ankara, Turkey
One of the widely known side effects of nifedipine and phenytoin is gingival overgrowth. The aim
of this study is the determination of nifedipine and phenytoin concentrations in gingival crevicular
fluid (GCF) and plasma by HPLC. Separation of nifedipine from GCF was performed by
Microsphere, C lg (100 x 4.6 mm, particle size 3 pm) analytical column and methanol, sodium
acetate (pH=4.0, 10 mM) (60:40, v/v) containing mobile phase at 0.8 ml/min. Detection of
nifedipine and nitrendipine (internal standard, 0.5 pg/ml) was performed by UV/Vis detector at 235
nm. GCF samples were extracted by using a mixture of methanol and water (50:50, v/v). Plasma
samples were extracted by using a mixture of hexane and dichloromethane (70:30, v/v). The
calibration curve for nifedipine was linear over the concentration range of 0.01-0.5 pg/ml. The
mean recovery (±SD) from GCF was 99.05±3.72 % for nifedipine at a concentration of 0.1 pg/ml
(n=6). The mean recovery (+SD) from plasma was 102.03±5.62 % for nifedipine at a concentration
of 0.1 pg/ml (n=6). Separation of phenytoin from GCF and plasma was performed by XTerra, C lg
(250 x 4.6 mm, particle size 5pm) analytical column and acetonitrile, K 2 HP0 4 (pH=5.0, 20 mM)
(30:70, v/v) containing mobile phase at 1.2 ml/min flow rate. Phenytoin and mephenytoin (as an
internal standard, 5 pg/ml) were detected by UV/Vis detector at 240 nm. GCF samples were
extracted by using a mixture of acetonitrile and water (50:50, v/v). Protein precipitation from
plasma samples was performed by using acetonitrile. The calibration curve for phenytoin was linear
over the concentration range of 0.02-5 pg/ml. The mean recovery (±SD) from GCF was
101.15±4.12 % for phenytoin at a concentration of 1 pg/ml (n=6). The mean recovery (+SD) from
plasma was 105.22±3.17 % for phenytoin at a concentration of 1 pg/ml (n=6). Consequently, this
study describes a simple, sensitive, and practical HPLC-UV/Vis method which permits
determination of nifedipine and phenytoin in human gingival crevicular fluid and plasma samples.
263

P-14
DETERMINATION OF PROPARACAINE IN HUMAN AQUEOUS HUMOR USING A
VALIDATED IIPLC-UV/VIS METHOD AND SYSTEM SUITABILITY TEST
A. Dincel', N.E. Bai 2 , H. Atila 3 , A. Bozkiirt 1
1 2
Hacettepe University, Faculty of Medicine, Department of Pharmacology, Faculty of Pharmacy,
Department of Analytical Chemistry, 3 Ankara University, Faculty of Medicine, Department of
Ophthalmology, Ankara, Turkey
Proparacaine is mainly used as a local anesthetic in ophthalmic experience, minor and cataract surgery.
The aim of this study is development of high performance liquid chromatographic (HPLC) method for
determination of proparacaine levels in aqueous humor. In addition, developed method was validated,
system suitability tests were considered and long term perfonnance of the method was ascertained.
Separation of proparacaine from aqueous humor was perfonned by Bondesil (4.6 x 250 mm, particle
size: 5 }j.m) analytical column eluted with a mobile phase containing acetonitrile, sodium dihydrogen
phosphate (pHN3.0, 20 mM) (30:70, v/v) at a flow rate of 1.2 ml/min. Proparacaine and lidocaine
(internal standard) was detected by UV/Vis detector at 220 nm. The retention times for lidocaine and
proparacaine were 5.58 and 12.01 min, respectively. After liquid-liquid extraction with cyclohexane,
human aqueous humor samples were injected to the HPLC system. Calibration curve was linear in the
range of 75-700 ng/ml. The limit of detection (LOD) and limit of quantitation (LOQ) were 25 and 75
ng/ml respectively. The recovery of proparacaine from human aqueous humor samples was 111.64 % at
100 ng/ml. In inter-assay and intra-assay precision and accuracy analysis, the relative standard deviation
was in the range of 0.96 % and 7.98 % and the relative error was in the range of 0.45 % and 11.69 % at
proparacaine concentration levels of 100, 250 and 600 ng/ml. Deliberately diverted pH, flow rate,
acetonitrile ratio and wavelength from their optimum values result in no statistically significant change
in the response of developed method. The results of different analysts were not statistically different
from each other, suggesting that the HPLC method developed in this study was robust and ruggest. For
the establishment of system suitability tests, the values of chromatographic parameters obtained from
the response of independent proparacaine standard samples (600 ng/ml) were evaluated. The variations
in all chromatographic parameters were not statistically significant over a period of 3 months. In
addition, Shewhart's quality control charts for developed HPLC method were constructed from the
response of independently prepared standard proparacaine samples with/without extraction for 3
months. All of the proparacaine responses were in the range of upper and lower control limits (mean ± 3
S.D) in both cases. Human aqueous humor samples obtained from patients topically treated with
proparacaine were successfully analysed by using the HPLC method presented. Consequently, this
study describes a sensitive, selective, accurate and precise HPLC-UV/Vis method having a long term
reliability which permits determination of proparacaine in human aqueous humor samples.
This study is partially supported by Hacettepe University, Research Center (Project No.:
03D05301001).
264

P-14
ELECTROCHEMICAL OXIDATION OF PEFLOXACIN AT BORON-DOPPED
DIAMOND ELECTRODE AND ITS DIRECT DETERMINATION IN SERUM AND
PHARMACEUTICS BY SQUARE WAVE AND DIFFERENTIAL PULSE
VOLTAMMETRY
B. Dogan, B. Uslu, SA. Ozkan
Department of Analytical Chemistry, Faculty of Pharmacy, University of Ankara, 06100, Tandogan, Ankara /
Turkey
Pefloxacin Mesylate Dihydrate is a member of fluoroquinolone group antibacterial drug. It is
clinically used as the antibiotic of first choice for general bacterial infectious diseases, and its
efficacies are highly appreciated. There is no written report dealing with electrochemical studies,
oxidation mechanism and analytical assay from pharmaceuticals or biological media of this
compound by voltammetric techniques. Electrochemical methods have proved to be highly sensitive
and selective for the determination of organic molecules including drugs in pharmaceutical
formulations and human body fluids. Boron-doped diamond electrodes have attracted much recent
attention for electrochemical determination. Its features include low voltammetric background
current, a wide potential window, low adsorption of organic molecules and high electrochemical
stability. In this study the electrooxidation of pefloxacin at boron-doped diamond and glassy carbon
electrodes was carried out using cyclic, differential pulse (DPV) and square wave (SWV)
voltammetric techniques. The dependence of current intensities and potentials on pH, scan rate,
nature of the buffer, concentration were investigated with both electrodes. According to the linear
relationship between the peak current and the concentration, differential pulse (DPV) and square
wave (SWV) voltammetric methods for pefloxacin assay in pharmaceutical dosage forms and
biological fluids were developed using boron-doped diamond electrode. The pH dependence
studies were performed in the pH range of 1.5 - 12.00 in various buffer solutions. The oxidation of
pefloxacin was irreversible and exhibited a diffusion controlled process. The slope of the log Ip -
log v linear plot was 0.49 indicating the diffusion control in 0.5 M H2SO4. For analytical purposes a
sharp peak was obtained at +1.20 V (vs Ag/AgCl) by differential pulse voltammetry (DPV) and
+1.24 V (vs Ag/AgCl) by square wave voltammetry in 0.5 M H2SO4 acid at boron-doped diamond
electrode. At concentrations lower than about 2x10" 4 M the peak current obtained by DPV and SWV
is linearly related to the concentration and is suitable for quantitative determination in tablets and
human serum. The linear response was obtained in the ranges of 2x10" 6 - 2x10" 4 M for supporting
electrolyte and spiked serum sample. The detection limit was found as 4.12xl0" 7 M and 4.65x10" 7 M
for DPV, 1.53xl0~ 7 M and 5.77xl0" 7 M for SWV techniques for supporting electrolyte and spiked
serum sample, respectively. The repeatability of the methods was found as 0.49 and 0.37 % for peak
currents and 0.45 and 0.14 % for peak potentials for DPV and SWV, respectively. Precision and
accuracy of the developed method was checked by recovery studies. No electroactive interferences
from the excipients and endogenous substances were found in the pharmaceutical dosage forms and
in the biological samples, respectively.
265

P-14
BINARY COMPLEXES OF ASPARTIC ACID AND VALINE WITH Cu(II)
IN AQUEOUS SOLUTIONS: STABILITY AND THERMODYNAMIC PARAMETERS
A. S. Ba§tug, N. Yars Ozarslan, S. Ekinci Goz
Department of Basic Pharmaceutical Sciences, Faculty of Pharmacy, Marmara University,
Haydarpasa 34668, istanbul, Turkey
The studies on complex formation reactions between metal ions and biologically important ligands
have received considerable attention for a long time. In this work, the stability constants of the 1:1
binary complexes of Cu(II) with aspartic acid and valine and the protonation constants of these
ligands were determined potentiometrically at a constant ionic strength of /= 0.10 mol L" 1
(NaC10 4 ) in aqueous solutions at 5.0, 20.0 and 35.0°C. UV-VIS spectroscopic studies were also
0 o
performed in order to confirm the complex formation. The thermodynamic parameters AGf, A//f
and ASf were reported for the complex formation reactions. The enthalpy change of Cu(II) with
aspartic acid complexation is found to be positive. The driving force is entropy for this reaction.
The enthalpy change of Cu(II) with valine complexation is found to be negative. The driving forces
are entropy and enthalpy for this reaction. The contribution of enthalpy to the decrease in free
energy is 51 %.
266

P-14
SIMULTANEOUS DETERMINATION OF PARACETAMOL, CAFFEINE
AND PROPYPHENAZONE IN TERNARY MIXTURES BY CAPILLARY
ELECTROPHORESIS
D. Emre. N. Ozaltin
Hacettepe University Faculty of Pharmacy Department of Analytical Chemistry, 06100, Sihhiye,
Ankara, Turkey
In this study, a new micellar electrokinetic capillary chromatography method was developed to
analyze pharmaceutical preparations containing ternary combination of paracetamol (PAR),
caffeine (KAF) and propyphenazone (PRO), simultaneously by capillary electrophoresis. Best
results were obtained by the use of 20 mM pH 9.0 borate buffer containing 30 mM sodium
dodecylsulphate as the background electrolyte. Diflunisal (DiF) was used as internal standard. The
separation was performed through a fused silica capillary (50 |im internal diameter, 44 cm total
length, 35.5 cm effective length) at 25°C with the application of 3 seconds of hydrodynamic
injection at 50 mbar pressure and a potential of 29 kV. Detection wavelength was 200 nm. Under
these conditions, the migration times were found to be 5.17 min for PAR, 5.51 min for KAF,
7.20 min for DIF, and 9.37 min for PRO. Linearity ranges for the method were determined as 2-200
|ig mL" 1 for PAR and KAF and 3-200 pg mL" 1 for PRO. Limit of detections were found as 0.6
(ig mL" 1 for PAR and KAF and 0.8 pg mL" 1 for PRO. According to the validation study, it was
proved that the developed method was accurate, precise, sensitive, selective, rugged and robust.
Three pharmaceutical preparations, which are produced by different drug companies in Turkey,
were analyzed by the developed method. One of the same preparations was also analyzed by the
derivative ratio spectrophotometric method reported in literature. No significant differences were
found statistically between the results obtained from developed method and the method reported in
literature.
267

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ELECTROCHEMICAL BIOSENSOR TECHNOLOGY COMBINED WITH
MAGNETIC PARTICLES
A. Erdem 1 , H. Karadeniz 1 , F. Sayar 2 , G. Giiven 2 , M. Ozsoz 1 , E. Pikin 2
'Ege University, Faculty of Pharmacy, Analytical Chemistry Department, Bornova, Izmir, Turkey,
2 Hacettepe University, Faculty of Engineering, Chem. Eng. Dept and Bioengin Div., and
TUBITAK-Biyomedtek: Center for Biomedical Technologies, Beytepe, Ankara, Turkey
After discovery of electroactivity in nucleic acids at the beginning of the sixties (1), many
electrochemical approaches have been developed for analyzing or quantification of nucleic acids
and its interactions (2-5). A new kind of affinity biosensors as "DNA Biosensor" or "Genosensor"
based on nucleic acid recognition processes have been rapidly developed towards the goal of simple
and low-cost point-of-care detection of specific nucleic acid sequences related with genetic and
infectious diseases. Genosensors could be applied to produce credit card-sized sensor arrays for
clinical applications such as detection of pathogenic bacteria, tumors, and genetic disease, or for
forensics (2,3,6). The ultimate goal is to design DNA biosensors as preparing a basis for the future
DNA microarray systems allowing to combine them with magnetic particles (7-12), or modify them
with nanomaterials such as nanoparticles and nanotubes (13-15). Recently, there has been an
interest in magnetic particles in order to design of new electrochemical biosensor approaches (7-
12) resulting in efficient magnetic separation. The development of electrochemical biosensor
technology based on the magnetic assay in combination with magnetic particles in different size
from pin to nm to detect DNA hybridization is the goal of this study. A disposable graphite sensor,
pencil graphite electrode (PGE) and differential pulse voltammetry (DPV) was used for measurment
of guanine oxidation signal observed at +1.0 V after DNA hybridization with its biotinylated probe
attached to streptavidin coated magnetic particles. The use of PGE brings some important
advantages such as being easy to use (single-use) and portable, which are crucial properties of
devices for DNA chip technology. In combination of magnetic hybridization surfaces with singleuse
transducers, the label-free electrical detection eliminates the needs for external indicators or any
advanced surface modification, and thus results in a greatly simplified protocol.
1. E. Palecek, Nature, 1960, 188, 656.
2. J. Wang, Nucl. Acids Res., 2000, 28, 3011.
3. E. Palecek, M. Fojta, Anal. Chem., 2001, 73, 75A.
4. M. I. Pividori, A. Merkoci, S. Alegret, Biosensors andBioelectronics, 2000,15, 291.
5. A. Erdem, M. Ozsoz, Electroanal., 2002, 14, 965.
6. A. Erdem, M. I. Pividori, M. Del Valle, S. Alegret, J. Electroanal Chem., 2004, 567, 29.
7. J. Wang, A.-N.Kawde, A. Erdem, M. Salazar, Analyst, 2001, 126, 2020.
8. E. Palecek, R. Kizek, L. Havran, S. Billova, M. Fojta, Anal. Chim. Acta, 2002, 469, 73.
9. J. Wang, D.Xu, A. Erdem, R. Polsky and M. Salazar, Talanta, 2002, 56, 931.
10. J. Wang, G.-U. Flechsig, A. Erdem, O. Korbut, P. Grundler, Electroanalysis, 2004,16, 928
11. A.Erdem, D. Ozkan Ariksoysal, H. Karadeniz, P. Kara, A. Sengonul, A.A. Sayiner, M. Ozsoz, Electrochem.
Comm. 2005, 7, 815
12. A. Erdem, M.I. Pividori, A. Lermo, A. Bonanni, M. Del Valle, S. Alegret, Sens. Actuators. B. Chem. 2006,
114, 591.
13. J. Wang, J. Li, A.J. Baca, J. Hu, F. Zhou, W. Yan, D.-W. Pang, Anal. Chem., 2003, 75, 3941.
14. J. Wang, Anal. Chim. Acta, 2003, 500, 247.
15. S.E. Baker, W. Cai, T.L. Lasseter, K.P. Weidkamp, R.J. Hamers, Nano Lett., 2002, 2, 1413.
268

UV SPECTROPHOTOMETRIC STUDY FOR THE QUANTITATIVE DETERMINATION OF
MOEXIPRIL HYDROCHLORIDE AND HYDROCHLOROTHIAZIDE
T. Av$ar, E. C'sek, N. Erk
Department of Analytical Chemistry, Faculty of Pharmacy, University of Ankara, 06100 Tandogan-
Ankara- Turkey
Moexipril hydrochloride (MO) and hydrochlorothiazide (HY), a new combination, is indicated in
the treatment and management of edema and hypertension. In this present study, two simple,
accurate, rapid and sensitive spectrophotometric methods have been developed for the quantitative
determination of MO and HY in bulk and in laboratory-prepared mixture. First spectrophotometric
method, first derivative UV spectrophotometry, depends on the measurement of the analytical
signals at 241.5 nm for MO and at 282.8 nm for HY in the first derivative spectra of the mixture in
methanol. Linear regression equations in the first derivative spectrophotometric method calculated
by the least square regression method were Y : -1.32 C + 3.0 10" 4 for MO and Y: -3.57 C +
1.5 10" 3 for HY (where Y is analytical signals in the first derivative spectra , and C is the
concentration) in laboratory-prepared mixtures, and linear correlation coefficient were found
0.9980 for MO and 0.9987 for HY. In the method, the relative standard deviations (RSD) were
found as 1.07 % for MO and 1.59 % for HY. The recovery test was performed from laboratoryprepared
mixture. The mean recoveries were found as 97.9 % for MO and 101.2 % for HY. In the
other method, ratio spectra first derivative spectrophotometry based on the measurement of ratio
spectra first derivative amplitudes at 242.4 nm for MO and at 274.5 nm for HY. In this method, the
representative linear equations were established as Y : - 98.02 C+ 4.15 10" 2 for MO and Y: -26.71
C + 8.0 10~ 3 for HY (where Y is analytical signals in the ratio spectra first derivative spectra, and
C is the concentration) in laboratory-prepared mixtures with the correlation coefficients of 0.9975
and 0.9954 respectively. Both methods showed good linearity in the ranges 1.50-13.0 pg-ml" 1 for
MO and 0.50 -11.0 pg.ml" 1 for HY. In this method, the relative standard deviations (RSD) were
calculated as 1.34 % and 1.60 % for for MO and HY respectively. The recovery test was performed
from laboratory-prepared mixture and the recovery averages were found as 99.8 % for MO and 99.9
% for HY. The described methods could be applied to commercial pharmaceutical dosage forms.
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269

DETERMINATION OF PINDOLOL IN TABLET BY HEAVY ATOM INDUCED ROOM
TEMPERATURE PHOSPHORESCENCE
N. Ertap, E. §atana, N. G. Goger
Gazi University, Faculty of Pharmacy, 06330 Etiler Ankara, Turkey
Pindolol is a beta P-adrenoceptor antagonist drug. Heavy atom induced-room temperature
phosphorescence (HAI-RTP) technique is described for the determination of pindolol in tablet. The
phosphorescence signal was monitored in the presence of potassium iodide as heavy atom reagent
and sodium sulphite as deoxygenating agent. The excitation and emission wavelengths were 280
and 442 nm, respectively. The calibration curve was linear between l.OxlO" 6 and 6.3x 10" M
pindolol in aqueous solution. The limit of detection and limit of quantification was found as 2.6x10"
7 and 8.7xl0" 7 M respectively. The precision in terms of relative standard deviation at 3.0xl0" 6 M
concentration level was found as 1.4 %. The method has been applied to determination of pindolol
in tablet.
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270

P-1
DERIVATIVE SPECTROMETRY: A NEW WAY TO ASSESS THE FREE-RADICAL
SCAVENGING POWER OF ANTIOXIDANTS
D. Fompeydie, J.- P. Gramond, P. Levillain
Department of Analytical Chemistry, Faculty of Pharmacy, University Rene Descartes, 4 avenue de
l'Observatoire 752070 Paris cedex 06, France
Second derivative spectrometry is applied to the determination in-vitro of the free radical
scavenging activity of various chemicals and beverages by measuring the decrease of the
absorbance at 517 nm given by a stable free-radical: diphenyl picryl hydrazyl (D.P.P.H.) in
presence of an antioxidant. By the way of derivation, one can eliminate the influence of colored or
troubled matter, which is not the case by using HPLC with visible spectrometry detection as
recently published. Moreover, HPLC is solvent and time consuming. The limit of detection was
determined at lpM/L. The linearity is far better than that obtained by HPLC mainly at lower
concentrations. The antioxidant power is measured versus that of trolox used to establish the
standard curve. The results show that resveratrol has an antioxidant power intermediate between
that of rutin and ascorbic acid. Of the beverages tested coffee has the greatest antioxidant power.
These results are in accordance with those obtained by other methods using more sophisticated
instruments.
271

P-14
QUALITY CONTROL OF SULFAMETHOXAZOLE AND TRIMETHOPRIM IN
COMMERCIAL TABLETS USING LC- MS (ION TRAP)
L. Gene'- 2 . N. §anli 3
'Anadolu University, Faculty of Pharmacy, Department of Pharmaceutical Technology, 26470,
Eskiehir, Turkiye, 2 Plant, Drug and Scientific Researches Center of Anadolu University
(AUBiBAM), 26470, Eski§ehir, Turkiye, 3 Siileyman Demirel University, Faculty of Science and
Arts, Department of Chemistry, 32260, Isparta, Turkiye;
Sulphonamides have been used in the treatment of acute, uncomplicated urinary-tract infections,
particularly those caused by Echerichia coli and some other bacterial infections. Sulfamethoxazole
is also used often in combination with diaminopyrimidine potentiators such as pyrimethamine or
trimethoprim, in the treatment of protozal infections, (malaria and toxoplazmosis). There are a lot of
pharmaceutical alternative product of Co-trimoxazole tablets in Turkish drug markets. For the
quality control of these formulations, weight deviation, hardness, friability, diameter-height ratio,
content uniformity of the active substance and in vitro dissolution technique were performed.
Dissolution tests were performed according to the paddle method described in USP 26, Apparatus
II. The rotating speed was 75 rpm and the temperature was 37±0.5°C. Dissolution studies were
carried out in 900 mL 0.1 M hydrochloric acid. Reversed phase liquid chromatography coupled
with mass spectrometry, LC-MS (ESI ion trap), has been frequently used because of its reliability,
sensitivity and accuracy for many applications. Chromatography was carried out on a Zorbax
Eclipse XDB C8 analytical column (4,6x150 mm; 5; Agillent Tech.) and Zorbax SB CI8 1-Pack
analytical column (2.1x30 mm; 3.5; Agillent Tech.). The column performance was tested and
mobile phase conditions and peak shapes were investigated. The method developed was validated
for the analysis of sulfamethoxazole and trimethoprim in commercial tablets.
272

P-1
DETERMINATION OF ANTHRALIN AND SALICYLIC ACID IN CREAM
FORMULATIONS BY SYNCHRONOUS FLUORESCENCE SPECTROMETRY
N.G. Goger. N. Erta§
Gazi University, Faculty of Pharmacy, Department of Analytical Chemistry, 06330 Ankara/
Turkey
Anthralin (l,8-dihydroxy-9-anthrone) is an efficient drug for the topical treatment of psoriasis. This
work describes a simple, rapid, selective, and sensitive synchronous fluorescence spectrometry for
simultaneous determination of anthralin and salycilic acid in pharmaceutical creams without any
time-consuming extraction steps prior to analysis Throughout the study methanol was used as
solvent and pH of the cream samples was adjusted to 8.0 using borate buffer. The synchronous
fluorescence spectrum was obtained over the range of 200-600 nm and AA, values were 140 nm for
anthralin and 110 nm for salycilic acid. Quantitative determinations were realized using the
emmission intensities at 386 nm for anthralin and 295 nm for salicylic acid. The lineer calibration
graphs were drawn over the ranges of 8.7xl0" 6 -2.61xl0" 5 M for anthralin and 7.41xl0" 8 - 2.96xl0" 7
M for salicylic acid. The proposed method was precise with RSD values, less than % 1.5. The
obtained LOD (S/N=3), 3.02X10 -7 -4.17X10' 9 M and LOQ (S/N=10), l.OlxlO -6 -1.38xl0" 8 M
values for anthralin and salicylic acid; respectively show sensitivity of the method.
273

SPECTRAL, ANALYTICAL, THERMAL AND ANTIMICROBIAL STUDIES OF A
NOVEL SODIUM 2-[4(2-HYDROXY-3-IZOPROPYLAMINOPROPOXY) PHENYL]
ACETAMIDE (ATENOLOL) DITHIO CARBAMATE AND ITS CADMIUM COMPLEX
A. Golcti, P. Yavuz
Kahramanmara SiitfU imam University, Faculty of Science and Arts, Department of Chemistry,
46100, Kahramanmara, Turkey.
Atenolol dithiocarbamate (ADTC) and its complex with Cd(II) have been synthesized. These newly
synthesized products have been characterized by elemental analyses (C, H, N and S), thermal
[thermogravimetry (TG) and differential thermal analyses (DTA)] as well as by spectral [UV, IR
and NMR ('H)] studies. The stability constants (P) in dimethyl sulfoxide (DMSO) of metal
complexes of ADTC have been determined by UV-Vis data. The antimicrobial activities of the
cadmium complex has been screened in vitro against the organisms Bacillus megaterium DSM 32,
Bacillus brevis NRS, Yersinia enterecolilica CMC 120, Micrococcus luteus La 2971, Pseudomonas
aeruginosa ATCC 27853, Enterococcus feacalis ATTC 15753 and Kluyveromyces marxianus
ADH1, the yeast cultures Candida tropicalis FMC 23, Candida albicans ATCC 10231 and
Kluyveromyces fragilis NRRL 2415.
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274

P-1
APPLICATION OF ELIMINATION VOLTAMMETRY FOR MICROANALYSIS OF
OLIGONUCLEOTIDES AND NUCLEIC ACIDS BASES
F. Jelen 1 , L. Trnkova 2 , R. Mikelova 2 , A. Kourilova 1 , S. Hason 1 , E. Palecek 1
'Institute of Biophysics, Academy of Sciences of the Czech Republic, Kralovopolska 135, 612 65
Brno Czech Republic, department of Theoretical and Physical Chemistry, Faculty of Science,
Masaryk University, Kotlarska 2, 611 37 Brno, Czech Republic
Elimination voltammetry with linear scan (EVLS) enables elimination of selected voltammetric
current components contributing to a total current [1]. In comparison to usual voltammetric methods
the EVLS is capable to resolve overlapping signals, specifically by using the elimination function,
which eliminates the charging and kinetic currents, and conserves the diffusion current. In the case
of adsorbed electroactive species this elimination function yields a well readable peak-counter peak
signal [2-4], which can be utilized in the adsorptive stripping (AdS) or adsorptive transfer stripping
(AdTS). The elimination procedure was applied to the analysis of nucleic acids and short synthetic
homo- and hetero-deoxyoligonucleotides (ODNs) containing adenine (A), cytosine (C) and guanine
(G). The EVLS results showed that the elimination voltammetric signal of C is separated from the
signal of A, and the elimination signals of A and C reflect their sequences in ODN molecules. The
detection is based on the adsorptive stripping voltammetric measurements in connection with
hanging mercury drop electrode (HMDE) or carbon paste electrode (CPE) [5], ODNs determination
was performed with released nucleic acid bases in presence of copper ions [6, 7]. Our results show
that the EVLS in connection with the AdS procedure is a useful tool for both qualitative and
quantitative microanalysis of ODNs.
1. Dracka O., J., Electroanal. Chem. 402 (1996) 19.
2. Trnkova L., Kizek, R., Dracka, O., Electroanalysis. 12 (2000) 905.
3. Trnkova L., Jelen F., Postbieglova I., Electroanalysis. 15, (2003) 1529.
4. Trnkova L., J. Electroanal. Chem., 582 (2005) 258.
5. Palecek E., F. Jelen, in (Palecek E., Scheller F., Wang J., Eds.) Perspectives in
Bioanalysysis. Vol. 1 Electrochemistry of nucleic acids and proteins. Towards
electrochemical sensors for genomics and proteomics, Elesevier, New York, 2005, pp. 74-
173.
6. Farias P.A.M., Wagener A.D.R., Castro A.A., Anal. Lett., 34 (2001) 1295-1310
7. Jelen F., Kourilova A., Pecinka P., Palecek E., Bioelectrochemistry, 63 (2004) 249-252
This work was supported by the grants INCHEMBIOL MSM0021622412 and BIO-ANAL-MED
LC06035 from the Ministry of Education, Youth and Sports and the grant A100040602 from the
Grant Agency of the Academy of Sciences of the Czech Republic.
275

P-18
DEVELOPMENT OF DISPOSABLE GENOSENSOR USED FOR ELECTROCHEMICAL
DETECTION OF DNA AND DRUG-DNA INTERACTIONS
H. Karadeniz, A. Erdem, M. Ozsoz
Department of Analytical Chemistry, Faculty of Pharmacy, University of Ege, 35100, Bornova,
Izmir, Turkey
The use of nucleic acid technologies has significantly improved preparation and diagnostic
procedures in life sciences. Nucleic acid layers combined with electrochemical or optical
transducers produce a new kind of affinity biosensors as DNA Biosensor (gene-based biosensor;
genosensor) for small molecular weight molecules. Electrochemical DNA biosensors are attractive
devices for converting the hybridization event into an analytical signal for obtaining sequencespecific
information in connection with clinical, environmental and forensic investigations, or drug-
DNA interactions (1-5). The electrochemical detection of DNA as double stranded DNA (dsDNA)
or single stranded DNA (ssDNA) immobilized onto a disposable graphite sensor; pencil graphite
electrode (PGE) by dip-coating procedure was performed by using differential pulse voltammetry
technique (DPV) as measuring the oxidation signal of guanine at about +1.0 V vs Ag/AgCl
reference electrode. The measurements were performed using AUTOLAB-PGSTAT 30
electrochemical analysis system with GPES 4.8 software package (Eco Chemie, The Netherlands)
and also, PalmSens PC vs 1.50 (Palm Instruments BV, The Netherlands). The effect of some
experimental conditions was studied based on guanine signal; such as, different concentrations of
dsDNA or ssDNA immobilized onto PGE surfaces and different immobilization time of dsDNA
onto surfaces. The antibiotic Mitomycin C (MC) [laS-(laa,8p,8aa,8ba)]-6-amino-8-
[[(aminocarbonyl)oxy]methyl)-l,la,2,8,8a,8b-hexahydro-8a-methoxy-5 methylazirino [2',3':3,4]
pyrrolo[l,2-a]-indole-4,7-dione is an antitumor agent used in clinical chemoteraphy against a broad
spectrum of solid tumors. MC has sitotoxic character and this molecule also give a damage to
normal human cells (6). For electrochemical monitoring of interaction between drug, MC and
dsDNA, both oxidation signals of guanine and MC were measured before/after interaction. As a
result of the interaction of MC with dsDNA, there was observed a great decrease at both oxidation
signals of guanine and MC. As a conclusion, the electrochemical detection of dsDNA and ssDNA
was performed using this disposable genosensor. Additionally, the electrochemical detection of MC
interaction with dsDNA was done successfully by using faster, more sensitive and less laborious
technique with the advantages of this genosensor. The detennination of interaction between DNA
and DNA-targetted molecules would be valuable in the design of the molecule-specific
electrochemical biosensor for application in diagnosis tests and in the development of drugs for the
chemoteraphy.
1. E. Palecek, M. Fojta, Anal. Chem., 73, 74A-83A (2001).
2. A. Erdem, M. Ozsoz, Electroanalysis, 14, 965-974 (2002).
3. J. Wang, A.N Kawde, A. Erdem, M. Salazar, Analyst 126 (11) 2020-2024, (2001).
4. H. Karadeniz, B. Gulmez, F. Sahinci, A. Erdem, G.I. Irem Kaya, N. Unver, B. Kivcak, M.
Ozsoz, J. Pharm. Biomed. Anal., 33, 295-302, (2003).
5. A. Erdem, B. Kosmider, R. Osiecka, E. Zyner, J. Ochocki, M. Ozsoz, J. Pharm. Biomed.
Anal., 38 (4), 645-652, (2005).
6. J. Cummings, V. Spanswick, J. Smyth, Eur J Cancer, 31A, 1928-1933 (1995).
276

P-1
SEPARATION OF CITALOPRAM AND DESMETHYLCITALOPRAM BY
CAPILLARY ZONE ELECTROPHORESIS
S. Karakaya, K.Cakir, E. §atana, N. Erta, N. G.Goger
Gazi University, Faculty of Pharmacy, Department of Analytical Chemistry, 06330 Ankara /
Turkey
A capillary zone electrophoretic (CZE) method was developed for separation of citalopram and its
pharmacologically active metabolite, desmethylcitalopram. Separation was accomplished in a 75
pm i.d, fused silica capillary with an effective length 56 cm. A solution of 10% acetonitrile in 25
mM phosphate buffer was used as background and sample solution. The pH of the solutions was
adjusted to 9.3 throughout the study. During analysis the instrument was operated at 20 kV positive
polarity generating a current level of approximately 135 pA. All samples were introduced by
hydrodynamic injection for 5 seconds at 50 mbar pressure and 200 nm was used as detection
wavelength. The standard and sample solutions were fdtered through 0.20 pm syringe fdters. The
migration times were 4.70 and 5.02 minutes for citalopram and desmethylcitalopram, respectively.
The proposed method was precise with RSD values, 1.2 and 2% for citalopram and
desmethylcitalopram, respectively. The limit of detection and limit of quantification were obtained
as 1.7xl0" 5 M and 3.9xlO" 5 M for citalopram.
277

OPTIMISATION OF A REVERSED-PHASE HIGH PERFORMANCE LIQUID
CHROMATOGRAPHIC METHOD FOR ANALYSIS OF MONOTERPENE GLYCOSIDES
IN PAEONIA MASCULA
S. Kovunoglu'. Demircan 2 ,1. Call 3 , S. Kir 2
'Department of Basic of Pharmaceutical Sciences, Faculty of Pharmacy, Hacettepe University,
06100, Sihhiye, Ankara, Turkey, department of Analytical Chemistry, Faculty of Pharmacy,
Hacettepe University, 06100, Sihhiye, Ankara, Turkey, department of Pharmacognosy, Faculty of
Pharmacy, Hacettepe University, 06100, Sihhiye, Ankara, Turkey
Paeonia, the largest genus in the family Paeoniacea, is represented by 7 species in the flora of
Turkey. The methanolic extract of the dried roots of Paeonia mascula, an Oriental medicinal herb,
have been reported to contain these cardiovascular protective monoterpene glycosides. Crude
extract was subjected to open column chromatography on normal silica-gel using dichloromethanemethanole-water
mixtures as eluents to yield fractions. The structure elucidation of the compounds
were achieved by combination of one and two dimensional NMR techniques and mass
spectrometry. As a continuation of our work on the monoterpene glycosides, a qualitative reversedphase
high performance liquid chromatographic (HPLC) method for the determination of the
monoterpene glycosides have been developed. The developed method was performed by HPLC
coupled with diode array detector. The method is based on the use of a Nucleosil 100-5 C )8 (5 pm,
250 x 4.6 mm) column with a mobile phase composed of acetonitrile-water. A wavelength of 230
nm for diode array detection was selected. The method was optimized by changing the
chromatographic parameters such as organic solvent ratio and pH of mobile phase. The effect of
buffer type and concentration were also investigated. Optimal conditions were selected by
calculating capacity and tailing factors of each peaks and by identifying the resolution for the
separation. Low retention time, symmetric peak, high peak area and high resolution were obtained
by optimized method. The developed method will be usefull for determination of monoterpene
glycosides in Paeonia mascula after all validation parameters have been performed.
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278

P-1
RESEARCH ON THE SOME 2,4-DI- AND 2,3,4-TRISUBSTITUTED BENZIMIDAZO[l,2-
A] PYRIMIDINES HAVING POTENTIAL ANTICANCER ACTIVITY
A. Merip', H. Karadeniz 2 , A. Erdem 2 , M. Ozsoz 2
1 Anadolu Univ., Faculty of Pharmacy, Dep. of Pharmaceutical Chemistry, 26470, Eskisehir,
Turkey, 2 Ege Univ., Faculty of Pharmacy, Analytical Chemistry Department, 35100, Bornova,
Izmir, Turkey
Nucleic acids offer the analytical chemist a powerful tool in the recognition and monitoring of
many important compounds [1-4]. A recent active area of research is to explore the nature and
dynamics of binding small molecules to biomacromolecules. The design of site-and conformationspesific
reagents provide new studies for the rational drug design [1-7]. Binding of small molecules
to deoxyribonucleic acid (DNA) occur through primarily in three modes: electrostatic interactions
with the negative-charged nucleic sugar-phosphate structure, binding interactions with two grooves
of DNA double helix and intercalation between the stacked base pairs of native DNA [5], The some
2,4-di- and 2,3,4-trisubstituted benzimidazo[l,2-a]pyrimidine derivatives (1-10) were previously
synthesized and tested for IC50 values against 5RP7 and F2408 cell lines (Figure) [8]. The result of
IC50 evaluations had been suggested that the existence and abundance of methyl substituent on the
structure increase the cytotoxic activity. Consequently, the substances abundantly bearing methyl
RI
N
//
N
R 2
//
N
-R^
Ri R2 R 3
Comp
OH H Me 1
OH H Ph 2
OH H n-Pr 3
OH Ph Me 4
OH Me Me 5
Ph H Me 6
Ph H Ph 7
Me H Me 8
Me Me Me 9
subtituent had been found promising as
potential tineoplastic activity [8],
The detection of interaction between
some of these compounds and double
stranded DNA (dsDNA) was also
studied electrochemically based on the
difference at oxidation signals of
electroactive DNA bases; guanine or
adenine, and also the selected ones of
these compounds. The determination of interaction between the DNA targetted molecules and DNA
would be valuable in the design of sequence-specific DNA binding molecules for application in
chemotherapy and in the development of tools for the point-of-care tests and diagnosis in genetics.
An understanding of the structural orientations, kinetics and thermodynamics associated with these
complexes is pivotal to design and development of novel "next-generation" chemotherapeutic
agents.
1. L.B. Mc Gown, M. Joseph, J. Pitner, G. Vonk, C. Linn, Anal. Chem, 1995, 67,663A .
2. J. Wang, Nucl. Acids Res., 2000, 28, 3011.
3. E. Palecek, M. Fojta, Anal. Chem, 2001, 73, 75A.
4. A. Erdem, M. Ozsoz, Electroanalysis, 2002, 14, 965.
5. C. Xia, S. Guoli, J. Jianhui, Y. Ruqin, Anal. Lett, 1999, 32,717.
6. H. Karadeniz, B. Gulmez, F. Sahinci, A. Erdem, G.I. Irem Kaya, N. Unver, B. Kivcak, M. Ozsoz, J. Pharm.
Biomed. Anal, 2003, 33, 295.
7. A. Erdem, B. Kosmider, R. Osiecka, E. Zyner, J. Ochocki, M. Ozsoz, J. Pharm. Biomed. Anal, 2005, 38(4),
645
8. A. Meri, Z. incesu, A. Karayel, S. Ozbey, Bioorg. Med. Chem. Lett, 2006, submitted.
279

P-1
PHARMACOGENETICS AND ITS IMPACT ON PHARMACY EDUCATION
D.O. Unal
Department of Molecular Biology and Genetics, Bogazici University, Bebek, 34342, Istanbul,
Turkey
Pharmacogenetics became an established science in 1950's and Vogel published the word
'pharmacogenetics' in 1959. Pharmacogenetics is the study of how genes influence an individual's
response to drugs. Pharmacogenetic researches have gained enormous momentum, with recent
development in molecular genetics. Research in pharmacogenetics is currently developing in two
main directions: 1. identifying specific genes and gene products associated with various diseases,
which may act as new drug targets and 2. identifying genes and allelic variants of genes that affect
our response to drugs. Single nucleotide polymorphisms (snips) are variations in a single DNA base
located at a specific position in the genome. This can be used to map genes related to diseases.
Proteins encoded by specific genes associated with various diseases are expected to become targets
for new drugs. Polymorphism in any genes, including genes encoding drug receptors, drug
transporters and cell signaling pathways can be important determinants of response to drugs. To
understand the genetic variation of person pharmacogenetic tests must be applied but
pharmaceutical testing is currently used in only limited number of research hospitals. But first, legal
and ethical questions related to knowledge of a person's genes have to be resolved. Innovative
approaches to drug discovery and high throughput screening technologies are giving a new
direction to pharmaceutical sciences. To meet the needs of this new area, pharmaceutical education
has to set new priorities to keep pace related to genomic technologies and develop new education
program for both undergraduate and graduate curricula. Educators and pharmacy school members
have the responsibility of deciding how and to what extent new technologies will be used in the
education program. New educational programs in pharmacy must include the principles and
applications of these new technologies to enable the student to understand the basic aspects of new
drug development and molecular mechanism of drug action on biological systems. Well educated
pharmacist in genomic technologies may have an opportunity as a valuable scientist in the health
care system.
Pharmacogenomics will become an important component in pharmacy and medicine in the coming
decade.
280

P-1
USE OF CARBON NANOTUBES IN BIOSENSOR SYSTEM FOR SIGNAL
ENHANCEMENT
P.O. Ariksoysal' M. Ozsoz, P. Kara Kadayifcilar, G. Yalcin, S. Cavdar, B. Meric
Department.of Analytical Chemistry, Faculty of Pharmacy, Ege University, 35100 Bornova-Izmir,
Turkey
Quick advances in nanotechnology field have provided a diversity of nanoscale materials with
highly controlled and unique optical, electrical, magnetic, or catalytic properties [1-3]. Researchers
have recently begun to borrow these nanomaterials and apply them to different kinds of applications
especially for diagnosis of disease [4], Identifying infectious organisms, quantitating gene
expression, and sequencing genomic DNA on chips all rely on the detection of nucleic acid
hybridization. Electrochemical biosensors, which couple the inherent specificity of DNA recognition
reactions with the high sensitivity of physical transducers, hold great promise for sequence-specific
detection. Detecting specific DNA sequences have been the ultimate goal of many electrochemical
genosensor systems [5-11]. Recently, carbon nanotubes received enormous interest because of their
high surface area, high electrical conductivity, high chemical stability and significant mechanical
strength [12-15], These characteristics allowed their use for different novel applications. In
particular, researchers have focused on DNA modified nanotubes and nanoparticles because of their
unique characteristics such as applicable for practical diagnosis tests [16,17], Optical and some of
electrochemical genosensors based on nanotubes have already been studied extensively and
optimized in detail, but electrochemical ones are still waiting for surprising new advances. Here, we
describe a novel electrochemical DNA biosensor using carbon nanotubes modified carbon paste
(CPE) electrode. The signal enhancement based on the oxidation signal of guanine was measured in
order to understand the role of carbon nanotubes for immobilization of DNA onto CPE. The carbon
paste electrode was prepared with the mixture of carbon paste and carbon nanotubes. Addition of
carbon nanotubes in CPE showed significant signal enhancement in the voltammetric signal of
guanine oxidation compared to the signal obtained from bare carbon paste electrode. These results
represent applicable and sensitive DNA detection for clinical genetic analysis.
1. Daniel, M. C„ and Astruc, D„ Chem. Rev. (2004) 104 (1), 293.
2. Khairutdinov, R. F, Colloid J.(1997) 59 (5), 535.
3. Hicks, J. F, et al, J. Am. Chem. Soc.(2002) 124 (44), 13322.
4. Ying Wang, Zhiyong Tang, and Nicholas A. Kotov, Nanotoday, (2005), pp.20.
5. Millan, K„ Saraulo, A, Mikkelsen, S.R, Anal. Chem., (1994) 66, 2943.
6. Ozsoz, M, Erdem, A, Kerman, K, Ozkan, D, Tugrul, B, Topcuoglu, N, Ekren, H, Taylan, M, Anal.Chem.,
(2003), 75,2181.
7. Wang, J.; Nucleic Acids Res. (2000), 25, 3011.
8. Erdem, A.; Kerman, K; Meric, B.; Akarca, U. S.; Ozsoz, M. Electroanalysis. (1999), 10, 586.
9. Ozkan Ariksoysal , D, Karadeniz, H, Erdem, A, Sengonul, A, Sayiner, A.A, Ozsoz, M, Anal Chem.,
(2005) 77 : 4908.
10. Palecek, E.; Fojta, M.; Anal Chem. (2001), 73, 74A.
11. Lucarelli, F„ Palchetti, I, Marrazza, G„ Mascini, M„ Talanta, 56 (2002) 949.
12. Ajayan, P.M., Chem. Rev., 99 (1999) 1787.
13. Ajayan, P.M., Iijima, S, Nature, 361 (1993) 333.
14. Baughman, R.H, Zakhidov, A, de Heer, W.A., Science, 297 (2002) 787.
15. Iijima, S„ Ichihashi, T„ Nature, 363 (1993) 603.
16. Shipway, A. N, Katz, E, Willner, I, ChemPhysChem., (2000), 1, 18-52.
17. 1'aunescu, T, Rajh, T, Wiederrecht, G, Master, J, Vogt, S„ Stojicevic, N„ Protic, M, Lai, B„ Oryhon, J. ,
Thurnauer, M„ Woloschak, G„ Nature Materials,(2003), 10.10380/nmat875,1-4.
281

P-14
A SPECTROPHOTOMETRIC METHOD FOR THE DETERMINATION OF
AMLODIPINE BESYLATE IN HUMAN PLASMA
M. Oztiirk. Y. Kadioglu
Department of Analytical Chemistry, Faculty of Pharmacy, Ataturk University, 25240, Erzurum,
Turkey
Amlodipine besylate is a dihydropyridine type long acting calcium channel blocker with slow onset
of vasodialatory action. The use of this important life saving drug is approved for the treatment of
variant and stable angina and hypertension. The purpose of this study is to develop and validate a
UV-Spectrophotometric method for the determination of amlodipine besylate in human plasma. The
sample preparation for the plasma assay involves precipitation of plasma proteins with diethyl ether
and hexan. Spectrophotometrically, amlodipine besylate was determined by measuring of the
absorbance values at 360 nm. Beer's Law was obeyed in the concentration range 2.0-17.0 pg mL" 1 .
A typical calibration curve had the regression equation of y=0.0114x+0.0204 with a correlation
coefficient (r) of 0.9973 (n=6). Limit of detection (LOD) has been determined as 1.5 pg mL" 1 , limit
of quantification (LOQ) as 2 pg mL" 1 . Absolute recovery, precision and accuracy assays were
carried out. The results were good; the mean absolute recovery values range from 80.25 % to 92.48
in all the concentrations studied, the relative standard deviations were less than 2 % and for all
concentrations of this compound the accuracy was higher than %94.5. Repeatability is given as
intra-day and inter-day precision and accuracy where evaluated by analyzing three different
concentration of amplodipine. Accuracy of the method was checked for different days at three
concentration levels at 5.75, 10.75, 14.5 mg ml" 1 . Due to the results of the present study, the
developed spectrophotometric method is concluded as accurate, sensitive, precise and reproducible
range over the concentration range examined.
282

SIMULTANEOUS SPECTROPHOTOMETRIC DETERMINATION OF ATENOLOL AND
CHLORTALIDONE IN A PHARMACEUTICAL PREPARATION USING PRINCIPAL
COMPONENT REGRESSION METHOD
I.M. Palabiyik, F. Onur
Department of Analytical Chemistry, Faculty of Pharmacy, University of Ankara, 06100,
Tandogan, Ankara, Turkey
The combination of atenolol and chlortalidone is widely used in the treatment of hypertension. In
this study, a spectrophotometric method is described for the simultaneous determination of atenolol
and chlortalidone in their combination. The obtained data were evaluated by using principal
component regression (PCR) method. The concentration data matrix were prepared by using the
synthetic mixtures containing these drugs in 0.1 M NaOH. The absorbance data matrix
corresponding to the concentration data matrix was obtained by the measurements of absorbances in
the range 240 - 290 nm in the intervals with AX = 2 nm at 26 wavelengths in their zero - order
spectra, then, calibration or regression was established by using the absorbance data matrix and
concentration data matrix for the prediction of the unknown concentrations of atenolol and
chlortalidone in their mixture. The procedure did not require any separation step. The linear range
was found to be 10 - 200 pg/mL for atenolol and 6-20 pg/mL for chlortalidone. Mean recoveries
and relative standard deviations for this method were found to be 99.98 % and 0.70 % for
chlortalidone and 100.15 % and 1.01 % for atenolol respectively in the synthetic mixtures of title
drugs. To select the number of factors, a cross-validation method, leaving out one sample at a time
was employed using training sets. In the method; fourteen factors for both atenolol and
chlortalidone in their mixture were found optimum for the determinations. We found the prediction
error sum of squares (PRESS) and root-mean squares (RMS) minimum with these factors. This
method was successfully applied to a tablet marketed in Turkey and the results were compared
statistically with an HPLC method.
P-1
283

P-1
SIMULTANEOUS DETERMINATION OF SOME BINARY MIXTURES USING
MULTIVARIATE SPECTROPHOTOMETRY AND DERIVATIVE RATIO
SPECTROPHOTOMETRY
H. Salem
Analytical Chemistry Department, Faculty of Pharmacy, Minia University, Minia, Egypt
This work is concerned with the simultaneous determination of four binary mixtures; sulperide with
mebeverine hydrochloride (Mix. I), diloxanide furoate with metronidazole (Mix. II), hyoscine
butylbromide with dipyrone (Mix. Ill) and cinnarizine with piracetam (Mix. IV) without previous
separation by three different methods. The first method is the use of multivariate
spectrophotometric calibration for the simultaneous determination of four binary mixtures, in which
the components of the four mixtures show a considerable degree of spectral overlapping. The
resolution of the studied binary mixtures has been accomplished by using partial least squares (PLS)
regression analysis. Although the components show an important degree of spectral overlap, they
have been simultaneously determined with high accuracy, with no interference from pharmaceutical
dosage forms excipients. A comparison is presented with the related multivariate method of
classical least squares (CLS) analysis, which is shown to yield less reliable results due to severe
spectra overlap presented by the studied compounds. The second method is the application of
derivative ratio spectrophotometry, in which the ratio spectrum was obtained by dividing the
absorption spectrum of the mixture by that of one of the components. The proposed procedures
were successfully applied for the simultaneous determination of the cited drugs in laboratory
prepared mixtures and in commercial pharmaceutical preparations. The validity of the proposed
methods was assessed by applying the standard addition technique where the percentage recovery
of the added standard was found to be 99.15±1.354, 98.97±0.988, 99.57±1.087 and 99.80±0.879 for
mixtures I, II, III and IV, respectively, using multivariate spectrophotometric method and
99.15±1.354, 98.97±0.988, 99.57±1.087 and 99.80±0.879 for mixtures I, II, III and IV, respectively,
using derivative ratio method. The proposed procedures are rapid, simple, require no preliminary
separation steps and can, therefore, be used for routine analysis of the studied drugs in quality
control laboratories.
284

SPECTROFLUORIMETRIC STUDY OF THE CHARGE-TRANSFER COMPLEXATION
OF CERTAIN FLUOROQUINOLONES WITH 2,3,5,6-TETRAFLUORO-P-
BEZOQUINONE
D. Geffken 1 , H. Salem 2
1 Institute of Pharmacy, Pharmaceutical Chemistry Department, University of Hamburg,
Bundesstrasse 45, 29146 Hamburg, Germany, 2 Analytical Chemistry Department, Faculty of
Pharmacy, Minia University, Minia, Egypt.
P-167
A highly sensitive spectrofluorimetric method was developed for the first time, for the analysis of
ten fluoroquinolones (FQs) antibacterials, namely amifloxacin (AMI), ciprofloxacin (CIP),
difloxacin (DIF), enoxacin (ENO), enrofloxacin (ENR), lomefloxacin (LOM), levofloxacin (LEV),
norfloxacin (NOR), ofloxacin (OFL) and pefloxacin (PEF) in their pharmaceutical dosage forms or
in biological fluids through charge transfer (CT) complex formation with fluoranil (TFQ). The TFQ
was found to react with these drugs to produce stable complexes, and the fluorescence intensity of
the complexes was greatly enhanced. The formation of such complexes was also confirmed by both
infrared and ultraviolet-visible measurements.The different experimental parameters that affect the
fluorescence intensity were carefully studied. At the optimum reaction conditions, the drug-TFQ
complexes showed excitation maxima ranging from 270 to 285 nm and emission maxima ranging
from 450 to 460 nm. Rectilinear calibration graphs were obtained in the concentration range 0.02 to
3.1 pg ml" 1 for the studied drugs. The method has been successfully applied to determine their
pharmaceutical dosage forms with good precision and accuracy compared to official and reported
methods as revealed by t- and F-tests. They also applied for the determination of studied drugs in
human urine samples.
285

SIMULTANEOUS DETERMINATION OF TRIMETHOPRIM AND
SULFAMETHOXAZOLE IN HUMAN PLASMA SAMPLES BY HIGH PERFORMANCE
LIQUID CHROMATOGRAPHY
E. Sayar 1 - 2 . S. ^ahin 1 , Y. Ozkan 3 , A. Sava§er 3 , S. Cevheroglu 2 , A. A. Hincal 1
P-18
•Hacettepe University, Faculty of Pharmacy, 06100-Ankara, Turkey; 2 Ministry of Defence, Army
Drug Factory, 06310-Ankara, Turkey, 3 Gulhane Military Medical Academy, 06018-Ankara, Turkey
Co-trimoxazole is a bacteriostatic antibiotic combination of trimethoprim (TMP) and
sulfamethoxazole (SMZ), in the ratio of 1 to 5, used in the treatment of a variety of bacterial
infections. Several methods such as microbiological, gas-liquid chromatography, mass
spectrophotometry and high performance liquid chromatography (HPLC) have been used for
determination of TMP and SMZ in pharmaceutical or biological samples. Of these, HPLC has been
the most widely used method for determination of these compounds. The aim of the present study
was to develop an HPLC method for simultaneous determination of TMP and SMZ in plasma
samples using antipyrine as the internal standard. The compounds (TMP and SMZ) in plasma
samples were determined after liquid-liquid extraction. Before analysis, plasma samples were
deproteinized with acetonitrile, defatted with hexane, and then extracted with dichloromethane.
Separation of TMP and SMZ was performed at 25°C on a Cg column (4.6 x 250 mm; 5 pm) with
max plot detection technique, using a mobile phase (pH 6.2) consisted of potassium hydrogen
phosphate (7.5g), acetonitrile (400 ml), methanol (100 ml), water (qs 1000 ml) which was delivered
at a flow rate of 1 ml/min. The proposed method was validated as to specificity, precision, linearity,
accuracy, sensitivity, detection limits and stability. The calibration curve was characterized by its
regression coefficient, slope, and intercept. These compounds were well separated with retention
times of 5 min for TMP, 7 min for antipyrine and 9 min for SMZ. The proposed method was linear
in the range of 10-5000 ng/ml for TMP and 50-50000 ng/ml for SMZ. The detection limits were 10
ng/ml for TMP and 50 ng/ml for SMZ. The results of the validation studies indicated that the
reverse phase HPLC method developed for analysis of TMP and SMZ is rapid, sensitive and
precise. This method was successfully applied to the determination of these compounds in plasma
samples obtained from the bioequivalence study of Co-trimoxazole tablets manufactured by Army
Drug Factory.
286

P-1
PHOTOSTABILITY OF SOME IMIDAZOLINE DERIVATIVES IN SOLID STATE
B. Marciniec, M. Ogrodowczyk, K. Dettlaff, M. Stawny
Department of Pharmaceutical Chemistry, Karol Marcinkowski University of Medical Sciences,
6 Grunwaldzka, 60-780 Poznan, Poland
Antazoline and naphazoline being derivatives of 2- imidazoline belong to the most often applied
drugs for alleviation of the first symptoms of allergy. They are components of many simple or
complex preparations such as eye drops, nose drops, gels, nose aerosols, and antazoline is also
applied in the form of injections. Stability of the therapeutic substance depends on the form of drug,
conditions of its production and storage. There is always a risk that as a result of chemical
transformations, including photodegradation, the therapeutic effect of the drug can be weakened due
to the formation of products either pharmacologically inactive or even toxic. Photostability of 2-
imidazoline derivatives in solutions has been studied from the early 1970s [1, 2], This study was
performed on two imidazoline derivatives: antazoline and naphazoline in solid state. According to
the Polish Pharmacopoeia VI [3] antazoline and naphazoline are the substances that must be
protected against light so determination of their photostability seems of significance. The process of
photodegradation was conducted according to the ICH requirements [4], with a xenon lamp of 1500
W and emitting radiation in the range 320-800 nm as a source of light. Samples of 0.1 g of the
therapeutic substances studied were placed in glass and plastic cuvettes and irradiated for 5 days.
After this time the samples were subjected to the qualitative and quantitative analysis by the
organoleptic method (form, colour, solubility) and spectrophotometric methods (UV and IR). The
products of photolysis were observed by the thin layer chromatography (TLC) and the
chromatographic - spectrophotometric method (TLC/UV). The results have shown that antazoline
and naphazoline hydrochlorides in solid phase meet the ICH requirements of photostability as they
do not show photodegradation changes even in drastic experimental conditions of the maximum
dose of visible irradiation (6 min lux • h). Therefore, both derivatives in the solid phase can be
concluded to be characterised by high photostability and do not require protection against light, in
contrast to these substances in solution for which the protection against light is recommended.
1. Rozanska M, Wojcik Z.: Biul. Inst. Lekow 15, 43-49, 1968
2. Sortino S, Cosa G, Scaiano J.C.: New J. Chem. 24, 159-163, 2000
3. Pharmacopeia Polonica, Editio VI, PTFarm, Warsaw, 2002
4. ICH, Stability Testing of New Drug Substances and Products, International Conference on
Harmonization IFPMA, Geneva, 1993
287

THE INFLUENCE OF RADIATION STERILIZATION ON THIAMPHENICOL PART I
1 1 2
B. Marciniec , M. Stawny and M. Kozak
l
Department of Pharmaceutical Chemistry, Karol Marcinkowski University of Medical Sciences,
2
6 Grunwaldzka, 60-780 Poznan, Poland, Department of Macromolecular Physics, Faculty of
Physics, Adam Mickiewicz University, 85 Umultowska, 61-614 Poznan, Poland
Thiamphenicol (D (+) - threo - 2 - (dichloroacetamido) - 1 - [p - (methylsulfonyl)phenyl] - 1,3 -
propanodiol) is an antibiotic of a wide spectrum of antibacterial activity. This substance is a
synthetic derivative of chloramphenicol, an antibiotic known for many years and obtained from
Streptomyces venezuele [1]. Thanks to the modification of the chemical structure of
chloramphenicol involving a replacement of the nitric group of the phenyl ring by a
methylsulphonic substituent, thiamphenicol is much safer in use (gives fewer undesirable side
effects) and is used not only for topical but also for general applications [2, 3], Thiamphenicol used
as injections or eye drops must be sterilised, most conveniently by ionising radiation. However,
prior to the application of irradiation sterilisation the effect of this procedure must be carefully
established as irradiation may induce physicochemical changes altering the pharmacological
activity [4, 5], The study was undertaken to check the effect of irradiation on physicochemical
properties of thiamphenicol in solid phase. Thiamphenicol was subjected to e-beam irradiation in
the doses of 25, 100 and 400 kGy, and the character and direction of possible changes were detected
by organoleptic analysis (form, colour, solubility) and a number of analytical methods: UV, IR
spectrophotometry, TLC chromatography, SEM observations, X-ray diffraction, polarimetry and
thermal (DSC). No significant changes in the properties of thiamphenicol irradiated with a dose of
25 kGy were detected. Irradiation with higher doses (100 and 400 kGy) did not cause changes in the
IR spectrum but resulted in a small increase in the intensity at maximum wavelength of the UV line,
a change in the colour, the appearance of the products of radiolysis (TLC), changes in the XRD
pattern, in SEM image, in the melting point value and optical rotation. Linear correlations were
found between the size of the irradiation dose and the decrease in the melting point and melting
enthalpy, characterised by the coefficients r= 0.9831 and r = 0.9622, respectively. A correlation was
also found between the dose and the changes in the optical rotation detected by the polarimetric
method; the rotation angle decreased from -21.8 to -16.5 optical rotation= f(dose), characterised
by r= 0.9998. The results have shown that ionising irradiation causes changes in the
physicochemical properties of thiamphenicol, but they appear after irradiation with doses much
higher than that of 25 kGy, which is a standard dose applied for sterilisation.
1. J. Ehrlich, Q. R. Bartz, R. M. Smith, D. A. Joslyn, P. R. Burkholder, Science, 106 (1947), 417
2. M. Bergeret, N. Boutros, J. Raymond, Medecine et Maladies Infectieuses, 33 (2003), 276
3. J. A. Turton, A. C. Havard, S. Robinson, D. E. Holt, C. M. Andrews, R. Fagg, T. C. Williams,
Food. Chem. Toxicol., 38 (2000), 925
4. W. Bogl, Radiat. Phys. Chem., 25 (1985), 425
5. T. Pronce, B. Tilquin, J. Chim. Phys., 94 (1997), 390
P-1
288

OPTIMISATION, VALIDATION AND DETERMINATION OF ZAFIRLUKAST IN
HUMAN SPIKED PLASMA BY A REVERSED PHASE HIGH - PERFORMANCE LIQUID
CHROMATOGRAPHY
P-1
j. Siislii, S. Altinoz
Department of Analytical Chemistry, Faculty of Pharmacy, Hacettepe University, 06100, Sihhiye,
Ankara, Turkey
Zafirlukast (ZAF) is a selective and competitive orally administered inhibitor of the cysteinyl
leukotrienes LTC4, LTD4 and LTE 4 in human airways. ZAF is currently indicated for the
prophylaxis and chronic treatment asthma in adults and children aged > 7 years. In this study, a
simple and sensitive reversed phase high - performance liquid chromatographic method was
developed for the determination of zafirlukast in spiked human plasma. Analysis was carried out on
Nucleosil Ci8 100 A (150 mm x 4.6 mm, i.d, 5pm) column with a mixture of acetonitrile : acetate
buffer (pH 3.0) (70 : 30, v/v) as the mobile phase, at a flow rate of 0.8 mL min"'. The peak was
detected by a ultraviolet detector set at a wavelength of 240 nm. Piribedil was used as an internal
standard. The retention times were about 3.9 min for piribedil and 5.8 min for zafirlukast. The
method was suitably validated with respect to specificity, linearity, limit of detection and
quantitation, accuracy, precision, recovery and stability. The calibration range was linear from
49.69 - 437.50 ng mL" 1 in spiked plasma. The limit of detection (LOD) value was 19.23 ng mL" 1
(S/N = 3), with RSD of 11.11 % and the limit of quantitation (LOQ) value was 49.69 ng mL" 1 with
RSD of 9.25 % for developed method. The relative recovery was 98.42 ± 0.53 % with RSD of 1.33
% at level 254.78 ng mL" 1 of zafirlukast concentration. The developed method was successfully
applied to the spiked human plasma with high recovered, good reproducibility, selective extraction
procedure and short analysis time.
289

A REVERSED PHASE HIGH - PERFORMANCE LIQUID CHROMATOGRAPHIC AND
CAPILLARY ZONE ELECTROPHORETIC METHODS FOR THE DETERMINATION
OF ZAFIRLUKAST IN PHARMACEUTICAL FORMULATIONS
P-1
I. Siislii, §. Demircan, S. Altinoz, S. Kir
Department of Analytical Chemistry, Faculty of Pharmacy, Hacettepe University, 06100, Sihhiye,
Ankara, Turkey
Zafirlukast is a competitive and selective leukotriene receptor antagonist, which is indicated for the
prophylaxis and treatment of mild to moderate persistent and chronic asthma. Simple, rapid,
sensitive and reliable a reversed phase high - performance liquid chromatographic (HPLC) and a
capillary zone electrophoretic (CZE) methods were developed and validated for the determination
of ZAF in pharmaceutical formulations. The experimental and instrumental parameters were
investigated and optimized for the zafirlukast determination. HPLC analysis was carried out on
Nucleosil C]g 100 A (150 mm x 4.6 mm, i.d, 5pm) column with a mixture of acetonitrile : acetate
buffer (pH 3.0) (70 : 30, v/v) as the mobile phase, at a flow rate of 0.8 mL min" 1 . CZE analysis was
carried out in a fused silica capillary (i.d. 50 pm, total length 80.5 cm and effective length 72.0 cm)
with 50 mM borate buffer at pH 8.5 as the running buffer, at applied voltage of 30 kV and the
samples were injected hydrodynamically for 3 s at 50 mbar. Piribedil was used as internal standard
in these methods. The ultraviolet detector was used in HPLC method and diode array detector was
used in CZE method. These detectors were set at a wavelength of 240 nm. The linear calibration
ranges were 25.0 - 900.0 ng mL" 1 and 2.0 - 80.0 pg mL" 1 and the detection limits (LOD) were 10.0
ng mL" 1 and 0.75 pg mL" 1 for HPLC and CZE methods, respectively. These methods showed good
sensitivity, accuracy, precision, selectivity, robustness and ruggedness. The proposed methods were
successfully applied for the determination of zafirlukast in its pharmaceutical formulations. The
results obtained by the proposed methods were compared with the electrochemical method reported
in the literature and statistical analysis showed no significant difference between the proposed
methods.
290

P-1
SPECTROFLUOROMETRIC DETERMINATION OF CITALOPRAM HBr
IN TABLETS
E. Satana, N. Erta§, N. G. Goger
Gazi University, Faculty of Pharmacy, Department of Analytical Chemistry
06330 Ankara / Turkey
Citalopram, (l-[3-(dimethylamino) propyl]-l-(4-fluorophenyl)-l,3-dihydroisobenzofuran-5-
carbonitrilmonohydrobromide) a selective serotonin reuptake inhibitor, is used as an antidepressant
agent. In this study a spectrofluorometric method was developed for the determination of
Citalopram HBr in pharmaceutical formulations, tablets. Calibration curves were linear over the
range of 5.0xl0" 7 - 2.5xl0" 6 M at excitation wavelength 240 and emission wavelength 300 nm. The
limit of detection (LOD) and limit of quantification (LOQ) were 7.9x10" 9 , 2.7x10' 8 M, respectively.
The proposed method was applied to determination of Citalopram in tablets. The results for
Citalopram in tablets were in agreement with the labeled quantities. The proposed method was
accurate with 100.5-101.2% recovery values and precise with 1.2 % RSD values.
291

P-14
DETERMINATION OF CITALOPRAM USING FLOW INJECTION -SOLID PHASE
EXTRACTION (SPE) WITH SPECTROFLUOROMETRIC DETECTION
E. Satan a, N. Erta$, N.G. Goger
Department of Analytical Chemistry, Faculty of Pharmacy, University of Gazi, 06330, Ankara,
Turkey
Citalopram, a selective serotonin reuptake inhibitor, is used as an antidepressant agent. In this study
an on-line solid phase extraction (SPE) method with spectrofluorometric detection was developed.
SPE cartridge contains 200 mg CI8 with a particle size of 60 (im. The best solvent system was
found to be consisting of ethanol and 0.1 M acetic acid (40:60, v/v) for elution. The optimised
values of injection volume and flow rate were, 1 ml and 2.5 ml min" 1 , respectively. The excitation
and emission wavelengths were 240 and 300 nm, respectively. The linearity was investigated in the
concentration range of l.OxlO" 7 -1.5xl0" 6 M in aqueous solution and 2.5xl0" 7 -1.2xl0" 6 M in serum.
The LOD and LOQ values were found to be 1.6xl0" 8 and 5.2x20" 8 M for tablets and 1.7xl0" 8 and
5.5x10" M in serum, respectively. The proposed method has been applied successfully for the
determination of the Citalopram HBr in pharmaceutical tablets and also in spiked human serum.
The recovery values of the method were 99.5-100.6% for tablets and 98.8-100.9% for serum.
292

COLORIMETRIC AND ATOMIC ABSORPTION SPECTROMETRY DETERMINATION
OF MUCOLYTIC DRUG AMBROXOL THROUGH ION PAIR COMPLEX FORMATION
WITH IRON (III) AND THIOCYANATE
A. Levent, Z. Senttirk
Yuziincii Yil University, Faculty of Science and Letters, Department of Analytical Chemistry,
65080 Van, Turkey
By the use of ion pair formation in analytical chemistry, which permits the combination of different
analytical techniques such as extraction and spectrometry, the increase in the sensitivity and
selectivity of the determination is achieved. Analysis by extraction of ion pairs using molecular
spectrophotometry has been known for many years. However, the use of charged metal complexes
allows the use of indirect atomic absorption spectrometric (AAS) applications for the determination
of organic compounds in recent years. Ambroxol is a compound with potent mucoliytic activity, for
which it is used as an expectorant and bronchosecretolytic in therapeutics. It is administered as a
hydrochloride salt using mostly oral formulations.
P-1
Br
The purpose of this work is to study the application of ion pair complex formation to develop a
simple spectrophotometric and AAS method for the determination of Ambroxol in pharmaceutical
formulations. These procedures depend upon the reaction of iron (III) metal ion with the drug in the
presence of thiocyanate ion in the pH range 1-3.5 to form stable ion pair complex which is
extractable into chloroform. The red coloured complex was determined either colorimetrically at
510 nm or by indirect AAS via the determination of the iron content in the formed complex. The
optimum experimental conditions for pH, concentrations of Fe 3+ and SCN", shaking time, phase
ratio, and the number of extractions were determined. Under the proposed conditions, linearity was
obeyed in the concentration range 4.1xl0" 6 M-5.7xl0" 5 M using colorimetric and AAS methods,
respectively. The proposed methods were applied for the determination of Ambroxol in pure and
tablet dosage form. The results obtained were compared with those obtained by applying the high
performance liquid chromatographic method with UV detection.
293

ELECTROCHEMICAL BEHAVIOUR OF PURINE DERIVATIVE ACYCLOVIR AND ITS
VOLTAMMETRIC DETERMINATION IN AQUEOUS AND MICELLAR MEDIA
P. Talay Pinar, Z. Senttirk
Yuziincii Yil University, Faculty of Science and Letters, Department of Analytical Chemistry,
65080 Van, Turkey
Substituted purines represent an important category of compounds actively studied as potential
therapeutics against viral infection. Because they constitute the essential components of biologically
important compounds such as polynucleic acids, and because their electrochemical and enzymatic
oxidations follow similar mechanistic pathways, the knowledge of voltammetric behavior of these
compounds is of biological interest. It is therefore important to examine the electrochemistry of
acyclovir, guanine analogue that is used extensively in the treatment of skin infections caused by
herpes simplex and varicella zoster virus. The aim of the present work is to investigate the
electrochemical properties of acyclovir in the pH and scan rate range 1.8-12.0 and 5-5000 mV s" 1 ,
respectively, at a glassy carbon electrode in aqueous and micellar media by cyclic and differential
pulse voltammetry. It can be concluded that in aqueous medium the electrochemical oxidation of
acyclovir involves irreversible pH-dependent four-electron and four-proton transfer process through
the formation of intermediate 8-oxoacyclovir. The investigation of the redox mechanism of
acyclovir was also carried out in micellar system. In case of sodium lauryl sulfate as anionic
surfactant, existence of several oxidation steps, depending on the potential scan range, implied a
complex and interesting oxidation pathway of acyclovir, resulting not only in formation of 8-
oxoacyclovir, but also in formation of several other intermediates and products. For analytical
purposes, a very resolved diffusion controlled voltammetric peaks were obtained in 0.2 M sulphuric
acid solution with addition of 10" 3 M sodium lauryl sulfate using differential pulse mode, with a
detection limit of 1.26xl0" 6 M. By developing method based on glassy carbon modified electrode in
micellar medium, it was possible to improve the sensitivity of determination of acyclovir in
pharmaceutical tablet forms with a mean recovery and its relative standard deviation of 99.9 % and
1.6 %, respectively.
P-1
294

P-1
SPECTROFLUORIMETRIC DETERMINATION OF FLUOROQUINOLONES IN
PHARMACEUTICAL PREPARATIONS
S. Tatar Ulu
Department of Analytical Chemistry, Faculty of Pharmacy, Istanbul University, Beyazit, Istanbul,
Turkey
Quinolones constitute a large class of synthetic antimicrobial agents that are highly effective in the
treatment of many types of infectious diseases, particularly those caused by bacteria. New
quinolones are continually being developed as bacterial species develop resistance to existing
quinolones [1]. Simple, rapid, and highly sensitive spectrofluorimetric method for the determination
of four fluoroquinolone drugs in tablets has been developed. The method is based on derivatization
with 4-chloro-7-nitrobenzofurazan (NBD-C1) and monitoring the fluorescence of the formed
fluoroquinolones-NBD derivatives at emission 535 nm and excitation 464 nm. The reaction
conditions were studied and optimized. The linear ranges and limits of detection are 23.5-500
ng/mL, 7.0 ng/mL for ciprofloxacin, 28.5-700 ng/mL, 8.5 ng/mL for enoxacin, 29.5-800 ng/mL, 9.2
ng/mL for norfloxacin, 33.5-1000 ng/mL, 9.98 ng/mL for moxifloxacin, respectively. The mean
recovery was about 99.46 % for these fluoroquinolones. Intra-day and inter-day RSD and RME
values at three different concentrations were determined. The results demonstrate that the method
has linearity, acceptable precision/accuracy and robust. The method is highly sensitive and specific.
The results obtained are in good agreement with those obtained by the official and reference
method.
1. Andriole V (2000) The Quinolones,Third Edition 517
295

P-18
DETERMINATION OF PARACETAMOL AND 4-AMINOPHENOL BY REVERSED
PHASE ION-PAIRING HIGH PERFORMANCE LIQUID CHROMATOGRAPHY
1 1 2 1
E. Ucakturk , A Ye§ilada , N. Basci , K. Ulubayram
1 2
Department of Basic Pharmaceutical Sciences, Department of Analytical Chemistry, Faculty of
Pharmacy, Hacettepe University, 06100 Sihhiye Ankara, Turkey
Paracetamol (acetaminophen) is a valuable non-steroidal drug with analgesic, and antipyretic
activities. A simple, reliable and convenient chromatographic method for determination of
paracetamol (P) and its main impurity; 4-aminophenol (4-AP), has been developed and validated by
using reversed phase ion-pairing HPLC. In order to optimize the chromatographic conditions, the
influence of different parameters (organic solvent, pH and ion pairing reagent concentration) on
capacity factor, resolution and peak symmetry were systematically investigated. The analysis was
performed in an isocratic mode on a reversed phase Nucleosil 100-5 C18 column (5 pm, 250 x 4.6
mm, i.d.) with diode array detector (DAD) detection. Isocratic mixture of 50 mM ammonium
acetate buffer/methanol 80/20 (v/v) containing 15 mM tributylamine (TBA) (pH 7) was found as a
suitable mobile phase composition for the determination of P and 4-AP. Under optimized
conditions, correlation coefficients for calibration curves, in the ranges 5-125 ng/mL for P and 0.4-5
pg/mL for 4-AP, were both found as 0.9999. The limit of detection (S/N=3) values were 0.05
pg/mL and 0.2 pg/mL for P and 4-AP, respectively. The limit of quantification (S/N=10) was 0.2
pg/mL for P and 0.4 pg/mL for 4-AP. The proposed chromatographic method was successfully
applied to the analysis of commercially available paracetamol tablets with a recovery of 99 %.
The method developed in this study is selective, accurate, precise, and reproducible according to
the evaluation of validation parameters and can be applied for the analysis of commercially
available paracetamol tablets.
296

DETERMINATION OF RISEDRONATE SODIUM IN PHARMACEUTICAL
PREPARATIONS BY FIRST ORDER DERIVATIVE UV SPECTROPHOTOMETRIC
METHOD
G. Ugurlu. N. Ozaltm
Department of Analytical Chemistry, Faculty of Pharmacy, University of Hacettepe, Sihhiye,
Ankara, Turkey
First derivative UV spectrophotometric method was developed for the analysis of risedronate
sodium which is used in osteoporosis treatment [l].The determinations of risedronate sodium in
biological samples including HPLC [2-4] have been reported in literature. In this study a simple,
rapid, reliable (and validated) first order derivative UV- spectrophotometric method was developed
for the determination of risedronate sodium in pharmaceutical preparations. The UV spectrum of
excipients gave absorption at 262 nm where standard solution gave maximum absorption. This
situation caused interference. Therefore first derivative UV spectrum was selected to prevent
interference with excipients. 258 nm was chosen for measurement. The solutions of the standards
and pharmaceutical samples were prepared in water. The linearity range of the method was 0.6-120
pg mL" 1 . The limit of detection was 0.2 pg mL' 1 . The method was validated and applied to the
determination of risedronate sodium in pharmaceutical preparation (tablet contains 5 mg risedronate
sodium). It was concluded that the developed method was accurate, sensitive, precise, rugged and
useful for the quality control of risedronate sodium in pharmaceutical preparations.
P-1
1. Borah, B., Dufresne, T.E., Chmielewski, P. A., Johnson, T. D., Chines, A., Manhart, M. D.
Bone. 2004, 34 (4), 736-746
2. Jia J.H., Li W„ Zhao K. Journal of Chromatography. B. 2006, 562, 171-175
3. Vallano, P. T., Shugarts, S. B„ Kline, W. F„ WoolfE. J., Matuszewski, B.K. Journal of
Chromatography. B. 2003, 794, 1, 23-33
4. Usui, T., Watanabe, T., Higuchi, S. J. Chromatogr. 1992, 584, 213-220.
297

P-18
DETERMINATION OF ROSUVASTATIN CALCIUM IN PHARMACEUTICAL
PREPARATIONS BY UV SPECTROPHOTOMETRIC METHOD
B. Uyar, M. Celebier, S. Altinoz
Department of Analytical Chemistry, Faculty of Pharmacy, University of Hacettepe, Sihhiye,
Ankara, Turkey
Rosuvastatin Calcium is a synthetic lipid lowering agent which is used in hypercholesterolemia. It
is a selective and competitive inhibitor of HMG-CoA reductase [1]. The determinations of
Rosuvastatin Calcium from pharmaceutical preparations including HPLC [2-3] and HPTLC [4],
from biological samples with HPLC [5-6] have been reported in literature. In this study a simple,
rapid and reliable UV spectrophotometric method was developed for the determination of
Rosuvastatin Calcium in pharmaceutical preparations. The solutions of standard and pharmaceutical
samples were prepared in MeOH. 243 nm was chosen for measuring the absorbances of
Rosuvastatin Calcium solution. The developed method was validated with respect to specificity,
linearity range, limit of detection and quantitation, accuracy, precision and ruggedness. The
linearity range of the method was 1-60 pg mL" 1 . The limit of detection was 0,33 pg mL" 1 . The
developed and validated method was applied to the determination of Rosuvastatin Calcium in
pharmaceutical preparations. There were no interference with the excipients.
1. Cheng J.W.M., Clinical Therapeutics, 2004, 26(9), 1368-1387.
2. Mehta T.N, Patel A.K, Kulkarni G.M. and Suubbaiah G, Journal of AOAC International,
2005, 88 (4), 1142-7.
3. Hull C.K, Penman A.D., Smith C.K. and Martin P.D., Journal ofChrom. B, 2002, 772, 219-
228.
4. Sane R.T, Kamat S.S, Menon S.N., Inamdar S.R. and Mote M.R, JPC-Modern TLC. 18
(103), 194-198.
5. Hull C.K, Martin P.D, Warwick M.J. and Thomas E, Journal of Pharm. Biomed. Anal,
2004, 35, 609-614.
6. Trivedi R.K, Kallem R.R, Mullangi R. and Srinivas N.R, Journal of Pharm. Biomed. Anal.
2005, 39, 661-669.
298

P-18
ARTIFICIAL NEURAL NETWORK FOR THE SIMULTANEOUS QUANTITATIVE
PREDICTION OF HYDROCHLOROTHIAZIDE AND LOSARTAN POTASSIUM IN
TABLETS
E. Din. O. Usttindag
Ankara University, Faculty of Pharmacy, Department of Analytical Chemistry, 06100, Ankara,
Turkey
Artificial neural network (ANN) calibration was applied to the quantitative prediction of
hydrochlorothiazide (HCT) and losartan potassium (LST) in tablets without using chemical
separation, extraction and graphical treatment of absorption spectra. In the application of ANN
method, the concentration set of 84 mixtures containing HCT and LST in the concentration range of
0-40 (J.g/ mL within methanol were prepared and then the absorption spectra of the concentration set
were plotted in the spectral range of 200-300 nm. The absorption values (x-block) corresponding to
the concentration set (y-block) were obtained by using the measurements at the 40-wavelenght
points in the above spectral range. The deviation from Lambert-Beer law at high concentration
limits was observed in the non-linear dynamic ranges corresponding to 0-40 pg/mL for both drugs.
In this case, the ANN chemometric calibration was computed by using the non-linear relationship
between concentration set (x-block) and their corresponding absorption data (y-block). In order to
determine the optimal ANN calibration model with different neuron sizes, various topological
networks were tried and finally a training network 40 neurons in the input layer, 20 neurons in to
hidden layers and two output for the calibration and prediction steps were found to be suitable for
the construction of ANN calibration for the simultaneous quantitative prediction of HCT and LST
in commercial tablet formulation. The validation of non-linear ANN approach was carried out by
analyzing the synthetic mixtures of two analyzed drugs. Mean recovery results and relative standard
deviations were found between 100.0 % and 0.26 for HCT and 100.1 % and 0.23 for LST. The
ANN approach was applied to real samples and the experimental results obtained were observed in
a good agreement with the literature HPLC method.
299

P-18
ELECTROCHEMICAL BIOSENSOR FOR THE INTERACTION OF
DNA AND ANTITUMORAL AND CYTOTOXIC CHALCONE DERIVATES
G. Yal9in, M. Cizmecioglu, P. Kara Kadayifcilar, D. Ariksoysal, S. Cavdar, B. Meric, O. Sogut, M.
Ozsoz
Ege University Faculty of Pharmacy, Dept. of Analytical Chemistry,
DNA biosensor technologies are currently under intense investigation owing to their great promise for rapid
and low-cost detection of specific DNA sequences in human viral and bacterial nucleic acids (1). A recent
active area of research to explore is the nature and dynamics of binding of small molecules to
biomacromolecules such as DNA. The design of site-and conformation-specific reagents provides new
studies for the rational drug design (2-3). The interaction of DNA with other molecules is an important
fundamental issue in life sciences. The methods based on DNA interactions have great importance for
understanding the action mechanisms of some anti-tumor and anti-viral drugs and some carcinogenic
molecules and to design new DNA-targeted drugs and also to screen these drugs in vitro. The interactions of
some anticancer drugs with DNA have been studied by a variety of techniques (4-5) and in recent years,
there is a growing interest in the electrochemical investigations of interactions between anticancer drugs and
other DNA-targeted molecules and DNA (6).The interactions of some anticancer agents with DNA have
been investigated by a variety of techniques ant there is a groving interest in the electrochemical methods for
the determination of anticancer agents (7-8). In this study, the detection of interaction between antitumoral,
antioxidant and cytotoxic chalcone derivates and DNA was performed by monitoring the differences
between guanine oxidation signals at a pencil graphite electrode (PGE) by using fish sperm dsDNA and
synthetic oligonucleotides. The electrochemical procedure for the detection of interaction is based on two
methods namely (a) interaction at electrode surface and (b) interaction at solution phase (9-10). It was shown
that after interaction of the antitumoral chalcone derivates with the base pairs in dsDNA, the voltammetric
signal of guanine greatly decreased. After the interaction of these chalcone derivates with oligonucleotides, a
higher decrease in the oxidation signals of guanine was observed under the same conditions. The interaction
of chalcone derivates with dsDNA in solution-phase was also investigated and the results were compared
with the ones obtained by surface immobilized dsDNA. As a result, the oxidation signals of guanine were
used for detecting the interaction mechanism of chalcone derivatives with the DNA surface. Detecting the
voltammetric behavior of several drugs that interact with DNA would be valuable in the design of sequencespecific
DNA binding molecules for application in chemotherapy and in the development of biotechnological
tools for the point-of-care tests based on DNA. Progress in this laboratory is towards the goal of determining
the voltammetric behavior of newly synthesized drugs with DNA, thus introducing the electrochemical
methods to solve the drug - DNA interaction mechanisms.
1. J. Wang, Nucl. Acids Res. 2000, 28, 3011.
2. S.R. Mikkelsen, Electroanalytical 8 (1996) 15.
3. J. Wang, A. N. Kawde, E. Sahlin, Analyst 2000, 125, 5.
4. K. Kerman, B. Meric, D. Ozkan, P. Kara, A. Erdem, M. Ozsoz, Anal. Chim. Acta, 2001, 450, 45.
5. E. Palecek, M. Fojta, Anal. Chem, 2001,73 74A.
6. A. Erdem, K. Kerman, B. Meric, M. Ozsoz, Electroanalysis, 2001, 13 219.
7. A. Erdem, M. Ozsoz, Electroanalysis 2002,14, 965.
8. M. Ozsoz, A. Erdem, P. Kara, K. Kerman, D. Ozkan, Electroanal, 2003, 15, 613.
9. J. Wang, M. Ozsoz, X. Cai, G. Rivas, H. Shiraishi, D.H. Grant, M. Chicarro, J.R. Fernandes, E. Palecek,
Bioelectrochem. Bioenerg, 1998,45 33.
10. A. Erdem, M. Ozsoz, Anal. Chim. Acta 437 (2001) 107.
300

P-18
DETERMINATION OF INDIGOTINE AND TARTRAZINE IN A SOFT DRINK BY HIGH
PERFORMANCE LIQUID CHROMATOGRAPHY
G. Yalcin 1 . S. Seyhan 2
'Marmara University Faculty of Pharmacy, Analytical Chemistry Department 34668 Haydarpasa
* 9 • *
Istanbul, Marmara University, Science Institute, Goztepe, Istanbul
A quantification method was developed for Indigotine and Tartrazine by high performance liquid
chromatography using diode array detector. The chromatographic column was Phenomenex, Luna,
ODS-2 RP- CI8(2) (5p m, 4.6 x250 m m i.d.), mobile phase was 6 mM tetrabutylamonium
hydrogensulfate aqueous solution - acetonitrile (57 : 43), the diod array detector was Agilent 1100
Model. The wavelength was 610 nm for Indigotine; 427 nm for Tartrazine (band width: 4nm), Flow
rate was 1 mL.min"', injection volume was 50 pL, pressure was 138 bar. The linearity was obtained
in the concentration range of 2.35-35.25 x 10" 6 mol.L" 1 , y = 561.Olx + 5.5978 (r 2 = 0.9999); the
limit of detection was determined as 5.3 x 10~ 7 mol.L" 1 and the limit of quantification was
determined as 1.76 x 10" 6 mol.L"' for Indigotine. The linearity was obtained in the concentration
range of 7.1-71.4 x 10" 6 mol.L" 1 , y = 561.01x + 5.5978 (r 2 = 0.9997); the limit of detection was
determined as 2.05 x 10" 6 mol.L"'and the limit of quantification was determined as 6.82 x 10" 6
mol.L" 1 for Tartrazine. The recovery was 81.49 % for Indigotine; 84.83 % for Tartrazine. The
amount of Indigotine was found as 56.00 mg / kg, the amount of Tartrazine was found as 80.72 mg
/ kg in a commercial soft drink.
301

DEVELOPMENT AND VALIDATION OF A GRADIENT HPLC METHOD FOR THE
SIMULTANEOUS DETERMINATION OF ROSIGLITAZONE AND METFORMIN IN
HUMAN PLASMA
P-184
C. Yardimci'. N. Ozaltin 1 , A. Giirlek 2 , M. I ildak 2 , A. Harmanci 2
'Department of Analytical Chemistry, Faculty of Pharmacy, Hacettepe University, 06100, Sihhiye,
Ankara, Turkey, 2 Division of Endocrinology, Department of Internal Medicine, Faculty of
Medicine, Hacettepe University, 06100, Sihhiye, Ankara, Turkey
Rosiglitazone (R), one of the newly available members of the thiazolidinedione family, and
metformin (M), a biguanide, are effective antihyperglycaemic agents with different modes of
action. The combination of R with M offers a rational therapeutic approach to the treatment of type
2 diabetes. A gradient reversed-phase HPLC method was developed and validated for the
simultaneous determination of rosiglitazone and metformin in human plasma. The analysis was
performed on a Ace 5 phenyl column (250 x 4.6 mm i.d., 5 pm) using a gradient method starting
with mobile phase composed of acetonitrile:5 mM sodium acetate pH 5.5 (75:25, v/v). The flow
rate was 1 mL/min. UV detection was performed at 245 nm and verapamil was used as internal
standard. The total run time was less than 10 min. Sample preparation included a simple protein
precipitation with acetonitrile. Validation experiments were performed to demonstrate stability,
specificity, linearity, accuracy, precision and ruggedness. The limit of quantification was 100
ng/mL for R, 250 ng/mL for M. The extraction recoveries were 100.02-105.0 % for R and 105.64-
103.88 % for M. The applicability of the method was demonstrated by the analysis of plasma
obtained from patients following administration of R and M.
302

PIROXICAM RELEASE FROM ACRYLATE-BASED pH-SENSITIVE HYDROGELS
S.Yarimkaya, H. Basan
P-185
Department of Analytical Chemistry, Faculty of Pharmacy, Gazi University, 06330, Etiler, Ankara
A non-steroidal anti-inflammatory agent, piroxicam (PX), was incorporated into the pH-sensitive
monolithic systems prepared by copolymerization of 2-hydroxyethyl methacrylate (HEMA) with
sodium/ammonium acrylates in the presence of crosslinking agent and redox initiator. Drug loading
was performed before polymerization and crosslinking processes. PX containing polymeric discs
were cut into pieces ranging from 12 to 25 mg and compressed in the form of tablet. In vitro release
studies were carried out in simulated gastric fluid for 1 h and followed by simulated intestinal fluid
at 37°C. The release rate of PX was controlled by changing the composition of the polymeric
matrix. Results showed that, in the low pH of the stomach, swelling degree of the acrylate-based
hydrogels was low and released amount of the PX was in the range of 3.9 to 8.2%, depending upon
the composition of the acrylate hydrogels. But, in the intestine, release rates were ranging from 85.4
to 97.9%. The controlled delivery of PX to the small intestine rather than to the stomach would
reduce the severity of the side effects such as gastric irritation and ulceration.
30

P-186
DETERMINATION OF LEFLUNOMIDE IN PHARMACEUTICAL TABLETS
BY FLOW-INJECTION ANALYSIS
D. Yeniceli, D. Dogrukol-Ak, M. Tuncel
Department of Analytical Chemistry, Faculty of Pharmacy, Anadolu University,
26470, Eskisehir, Turkey
A flow injection analysis (FIA) of leflunomide using UV-detection is described, in this study. The
most suitable carrier solvent was found to be an aqueous solution of EtOH (25 %, v/v).
Leflunomide was determined at the optimum conditions such as flow rate of 0.8 mL.min" 1 and
detection wavelength of 260 nm. The method has been validated and linearity was examined in the
range of 2.8xl0" 6 -l.lxl0- 4 M. The limit of detection (LOD) and quantitation (LOQ) were calculated
to be 1.60xl0" 7 M (S/N = 3.3) and 4.8xl0" 7 M (S/N = 10), respectively. The application of the
proposed method has been performed in pharmaceutical tablets of leflunomide and very good
results were obtained. The results were compared with those obtained from UV-spectrophotometry.
Statistically insignificant difference was found between the methods. As a result, the FIA method
for the determination of leflunomide in pharmaceutical tablets can be proposed as precise, accurate,
sensitive and cheap method for routine analysis laboratories.
30

P-187
COMPARISON OF ZERO- AND FIRST-ORDER DERIVATIVE
SPECTROPHOTOMETRIC METHODS FOR DETERMINATION OF INSULIN IN
PHARMACEUTICAL PREPARATIONS
B. Yilrnaz, Y. Kadioglu
Department of Analytical Chemistry, Faculty of Pharmacy, Ataturk University, 25240, Erzurum,
Turkey
Insulin is the most important regulatory hormone in the control of glucose homeostasis consisting
of 51 amino acids shared between two intramolecular chains and with a molecular weight of 5800
g/mol [1], The aim of this study is to develop and validate zero- and first-order derivative
spectrophotometric methods for the determination of insulin in pharmaceutical preparations. The
solutions of standard of insulin were prepared in 0.01 M HC1 solution and its maximum absorption
wavelength for zero- and first-order derivative spectrophotometric methods are at 276 and 280 nm,
respectively. For calibration curves, insulin solutions containing 5, 10, 20, 30, 40, 50, 60, 70 pgml
were prepared by diluting with 0.01 M HC1 of the standard insulin solution (100 pg ml ) for zeroand
first-order derivative spectrophotometric methods. Developed zero- and first-order derivative
spectrophotometric methods in this study are accurate, sensitive, precise, reproducible and can be
directly and easily applied to Actrapid HM injectable form as pharmaceutical preparation.
Statistical analysis (Student t-test) of the obtained results showed no significant difference between
the proposed two methods.
1. Y.W. Chien, Drug. Dev. Ind. Pharm. 22 (1996)753
30

P-188
SIMULTANEOUS DETERMINATION OF METOPROLOL, PROPRANOLOL AND
PHENOL RED IN SAMPLES FROM RAT IN SITU INTESTINAL PERFUSION STUDIES
P. Zakeri-Milani 1 , H. Valizadeh 1 - 2 , Y. Azarmi 12 , M. Barzegar Jalali 1 , H. Tajerzadeh 3 , Z.
Islambulchilar 1
1 Department of Pharmaceutics, Faculty of Pharmacy, Tabriz University of Medical Sciences,
Tabriz, Iran, 2 Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran,
J Department of Pharmaceutics, Faculty of Pharmacy, Tehran University of Medical Sciences,
Tehran, Iran.
Single-pass intestinal perfusion technique (SPIP) is the most used classic technique employed in the
study of intestinal absorption of compounds in which a non-absorbable marker such as phenol red is
used to correct the water flux. A simple and rapid reversed-phase high performance liquid
chromatographic method with UV detection at 227 nm was developed for simultaneous quantitation
of propranolol and metoprolol along with phenol red for in-situ permeability studies. The mobile
phase was a mixture of 55% methanol, 45% of 0.05 M KH 2 P0 4 aqueous solution (adjusted to pH 6)
and 0.2 % (v/v) triethylamine. Analysis was run at a flow rate of 1 ml/min with a 9 min run time.
The calibration curves were linear for all three compounds (r > 0.999) across the concentration
range of 7.5-125 (ig/ml with a limit of detection of 4.24, 2.18 and 8.57 ng/ml and limit of
quantification of 14, 7.2 and 28.3 ng/ml for metoprolol, propranolol and phenol red respectively.
The coefficient of variation for intra-assay and inter-assay precision was less than 8% and the
accuracy was between 93.6-107%. Using the SPIP technique and the suggested HPLC method for
sample analysis, the mean values of 0.49 e -4 (±0.19) cm/sec and 0.32 e -4 (± 0.09) cm/sec were
obtained for propranolol and metoprolol intestinal permeability coefficients respectively.
30

P-189
QSAR OF SOME ANTIMICROBIAL l,4-BENZOXAZINE-3-ON DERIVATIVES
AGAINST C. KRUSEI
S. Alper-Hayta, I. Yildiz, E. Aki-Sener, O. Temiz-Arpaci, I. Yalfin
Ankara University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry
A group of l,4-benzoxazine-3-ones was isolated from maize, wheat and rye several years ago. 2,4-
Dihydroxy-l,4-benzoxazine-3-one, DIBOA and its methoxy derivative DIMBOA have been shown to
inhibit germination of spores of the phytopathogenic fungi. They play an important role in the chemical
defense of cereals against deleterious pests such as insects, pathogenic fungi and bacteria. These molecules
are present naturally in the plants as glucosides from which the aglycones are released rapidly by enzymatic
hydrolysis after physical and biological injury of the plants and they exhibit antifungal, antibacterial and
insecticide properties [1-3]. In this study, the QSAR analysis of a set of previously synthesized compounds
[4-5] tested for growth inhibitory activity against Candida krusei, was performed by using the computerassisted
multiple regression procedure. Regression analysis and calculations were run on a PC using the
BILIN[6] statistical program package. The activity contributions for substituent effects of these compounds
were determined from the correlation equation (Eq. 1) for predictions of the lead optimization.
R
R= H, CH 3 ,C 2 H 5
Ri=H, CH 3 , CI, COOC2H5
R 2 =H, N0 2
log l/c = +0.0499 (±0.039) B, (R)
+0.169 (±0.053) ct ( ri)
+0.0569 (±0.020) I X( R2)
Eq. 1
n =16 R 2 =0.942 s =0.018 F =31.471 Q 2 =0.790 S PRE ss =0.024
As a result of this study, this equation (Eq.l) was found to be the best fit for the predictions according to the
examined validation test results. QSAR analysis revealed that the substitution of position R and Ri were
more significant than the position R2 for the tested antifungal activity. The QSAR was developed based on
the a values of the substituents at 6. position, where C is the minimum concentration of compound that
inhibited growth of C. krusei. From this relationship, we see that an electron-withdrawing substituents of 6-
position of benzoxazine favor inhibition of growth. The positive coefficients of B/ parameter indicated that
the width of the substituent at position 4 was conducive for the activity. Additionally, a nitro group at
position 7 of the fused ring moderately helped to inhibit growth. We could say that width substituent R is
important for the antifungal activity against C. krusei. These observations could guide us to design further
new lead antifungal compounds.
1. A.I. Virtanen, P. K. Hietala, Acta Chem. Scand. 14(2) (1960) 499-502.
2. C. L. Tipton, J. A. Klun, R. R. Husted, M. D. Pierson, Biochemistry 6 (1967) 2866-2870.
3. J. A. Klun, C. L. Tipton, J. F. Robinson, D. L. Ostreem, M. Beroza, Agric. Food Chem. 18(4) (1970) 663-665.
4. I. Yalcin, B. P. Tekiner, I. Yildiz-Oren, O. Temiz-Arpaci, E. Sener-Aki, N. Altanlar, Indian J. Chem. 42B
(2003) 905-909.
5. S. Alper-Hayta, E. Aki-Sener, B. Tekiner-Gulbas, O. Temiz-Arpaci, I. Yildiz, I. Yalcin, N. Altanlar, Eur. J.
Med. Chem. (in press).
6. H. Kubinyi, "Quantitative models. In QSAR: Hansch Analysis and Related Approaches" 1993, Vol 1;
Mannhold, R, Krogsgaard-Larsen, P, Timmerman, H, VCH Verlagsgesellschaft mbH:Weinheim, pp 57-89.
30

P-190
MELATONIN-RETINOIDS AS POTENT ANTIOXIDANTS
Z. Ates-Alagoz', T. Coban 2 , E. Buyiikbingol'
Ankara University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry 1 , Department
of Toxicology 2 , Tandogan 06100, Ankara, Turkey
A number of retinoid related compounds represent classes of antioxidative and proapoptotic agents
with promising potential in the treatment of neoplastic diseases. Indeed, the synthetic retinoid amide
fenretinide [N-(4 hydroxyphenyl) retinamide] induces apoptosis of cancer cells and acts as a
chemotherapeutic drug in cancer therapy. In the present work, as a continuation of our studies on
retinoid-type of compounds, the synthesis of melatonin retinamide derivatives was studied as a
novel series of melatonin retinoids, using the condensation reaction sequence involving
tetrahydrotetramethylnaphthalene carboxylic acid and appropriate melatonin-type moieties. Despite
of the weak DPPH inhibition activity pattern of the syntetized compounds, some of them showed a
strong inhibition on lipid peroxidation (IVa-b, Va and Vlla-c, 88%, 96%, 90%, 94%, 93% and 86%,
respectively at 10" 4 M concentration) when the melatonin (85% at 10" 4 M concentration) was used as
a reference compound.
IV (a-c)
1. N. Takahashi, K. Tamagawa, Y. Kubo, T. Fukui, H. Wakabayashi and T. Honda, Bioorg. Med.
Chem. 11, 3255-3260 (2003).
2. S. Stole, Life Sciences 65, 1943 (1999).
3.R. J. Reiter, Progress in Neurobiology 56, 359(1998).
4. Z. Ates-Alagoz, Z. Buyukbingol, E. Buyukbingol, Die Pharmazie, 60, 643-647 (2005).
5. Z. Ates-Alagoz, E.Buyukbingol, Heterocyclic Communications,!, 455 (2001).
30

P-191
DNA-BINDING ACTIVITY OF SOME METAL COMPLEXES
K. Benkli 1 , G. Ulufam 2 , N. Beynek 2 , Z. Seller 3 , G. Akalin 3 , G. Turan 1
'Department of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Anadolu, 26470,
Eski§ehir, Turkey, department of Chemistry, Faculty of Science, University of Trakya, 22030,
Edirne, Turkey, department of Biochemistry, Faculty of Pharmacy, University of Anadolu, 26470,
Eski§ehir, Turkey,
In this research, metal-ion controlled synthesis of some complexes in the presence of Zn(II), Cd(II),
Hg(II), Pb(II) was studied. 2,2'-bipyridyne-6,6'-dicarboxylaIdehyde and appropriate metal salt were
dissolved in MeOH, the mixture were refluxed. In MeOH, 2,6-bis(2-
aminothiophenoxymethyl)pyridine was added dropwise with stirring. The mixture then was
refluxed for more two hours and filtered hot. The solvent of the reaction mixture was reduced to
13
half its original volume and then the mixture was placed in a refrigirator to induce crystallisation " .
The crystals were filtered and dried. The structure of the compounds were elucidated by IR, 'H-
NMR, MASS and Elemental Analyses. The mitocondrial activity of U20S cells after exposure to
complexes was determined by colorometric assay which detects the conversion of 3-(4,5-dimethylthiazollvl-2)-2,5-diphenyltetrazolium
bromide (MTT) to formazon 4 . DNA-binding activity of all
compounds was also investigated by using plasmid DNA pUC18 purified from E. Coli 5 .
1. N.F. Curtis, J. Chem. Soc, 4409-17, (1960).
2. N. Beynek, M. McPartlin, B. P. Murphy, I. J. Scowen, Polyhedron, 17, 2137, (1998).
3. T. Adatia, N. Beynek, B. P.Murphy, Polyhedron, 14, 335, (1995).
4. C. M. Che, J. S. Huang, Coord. Chem. Rev, 242, 97-113, (2003).
5. T. Mosmann, Journal of Immunological Methods. 65, 55-63, (1983).
6. J. J. Criado, J. L. Manzano, E. Rodriguez-Fernandez, J. Of Inorganic Biochemistry, 96,
311-320, (2003).
30

P-192
DETERMINATION AND INVESTIGATION OF ELECTROCHEMICAL BEHAVIOUR OF 2-
PHENYLINDOLE DERIVATIVES: DISCUSSION ON POSSIBLE MECHANISTIC
PATHWAYS
S.P.Bozkava 1 . B.Dogan 2 , S.SUzen 1 , D.Nebioglu 1 , S.A.Ozkan 2
1 2
Department of Pharmaceutical Chemistry, Department of Analytical Chemistry, Ankara University,
Faculty of Pharmacy, 06100, Tandogan, Ankara, Turkey.
Indole derivatives constitute an important class of therapeutical agents in medicinal chemistry including
anticancer, antioxidant, antirheumatoidal and anti-HIV. Some of 2-phenylindole (2PI) sulfamates are known
inhibitors of steroid sulfatase with antiproliferative activity in breast cancer cells also 2PI with sulfur containing
side chain also shows significant in vivo antineoplastic and antiestrogenic acitivity. In this study, some 2-
phenylindole (2PI) derivatives were synthesized and investigated electroanalytically by voltammetric
determination. Due to the existing resemblance between electrochemical and biological reactions it can be
assumed that the oxidation mechanisms taking place at the electrode and in the body, share similar principles. 2-
phenylindole (2PI) derivatives were prepared by Fischer indole synthesis.
Table 1. Electroanalytically evaluated 2PI derivatives
No R, R 2 R 3 R4 R 5 R«
1 -H -H -H -H -H -H
2 -H -H -H -H -CI -H
3 -H -H -H -CI -CI -H
4 -H -H -H -H -NH 2 -H
5 -H -H -OH -H -CH 3 -H
6 -H -H -H -N0 2 -CI -H
7 -N0 2 -N0 2 -H -H -H -H
No previous electrochemical data were available concerning the oxidative or reductive behaviour of 2PI
derivatives. All 2PI derivatives were electrochemically oxidized in a broad pH range (1.8-12) using a glassy
carbon disc electrode. In addition to the electrooxidative behaviour, compounds 6 and 7 also showed
electroreductive behaviour over a broad pH range due to their aromatic nitro groups. Current-voltage curves of
all 2PI derivatives showed one well defined oxidation peak and one wave at pH less than 7.0. All 2PI
derivatives were electrochemically reduced at the indole ring in a broad pH range (1.8-10) using a HMDE.
Compound 6 and 7 showed additional reduction due to their nitro groups. All the voltammograms of 2PI
derivatives showed oxidation steps on nitrogen atom in the indole ring, which electroactive in both acidic and
basic media leading finally to hydroxylation of benzene ring. Possible oxidation mechanism of 2PI derivatives
on nitrogen atom in indole ring is shown in scheme. (Quoted from Radi et al.)
-(H+, c")
H+ 2H20
iXVR - 2(£ -- h+) fx\
In this study possible oxidation / reduction products of some 2PI derivatives that are relevant to metabolism
products of these molecules were characterised electrochemically and possible mechanistic pathways have been
discussed.
30

BIOCHEMICAL RESPONSES OF CHEMICAL COMPOUNDS TO FREE RADICALS
S.P. Bozkava', S. Olgen 1 , T. Qoban 2 , D. Nebioglu 1
P-193
'Department of Pharmaceutical Chemistry, 2 Department of Pharmaceutical Toxicology, University of
Ankara. Faculty of Pharmacy, 06100, Tandogan, Ankara, Turkey.
Protein kinases are considered as the major target for activation by tumor promoting and progression agents
and play an important role in carcinogenesis. In addition, protein kinases can be activated by oxidative stres
and inhibited by antioxidants. A series of thioindolines and indolin-2-ones have been reported in the
literature (S. Olgen, IL Farmaco 60, 497-506, 2005) as inhibitors of the protein kinases, were also showed
important antioxidant effects. It was found that the tested indoline derivatives showed preventive
antioxidative action probably similar to superoxide dismutase (SOD) enzyme in the system generating
superoxide radicals (0~ 2 ) and also showed protective action against deoxyribose degradation obtained by
hydroxyl radicals (HO) and reacted with peroxide (ROO) radicals. These findings suggest that the
compounds can protect against redox stimulation of cellular protein kinases. Several indole-2 and 3-
carboxamides are proposed to be selective cyclooxygenase-2 inhibitors. It was reported that that
cyclooxygenase increase the reactive oxygen species (ROS) production, thus the enzyme can participate in
oxidative stress. The broad spectrum of the observed antioxidant activity of the indole2- and 3-carboxamides
suggested us that they can scavenge oxygen free radicals, and some of them also scavenge the 0 2 produced
by the cyclooxygenases. In the basis of these findings we aimed to synthesize novel N-substituted indole-2
and 3-carboxamide derivatives in order to identify the possible antioxidant activity of the compounds and to
guide perspectively the design of new analogues. In this study, we synthesized 1-benzyl and substituted
benzyl carboxylic acids by the method of Murakami et al. (Synthesis, 738-740, 1984). Indole-2 and 3-
carboxamides were prepared using SOCl 2 as a carboxyl activator according to the method described by
Olgen et al (Pharmazie, 57, 238-242, 2002).
2nd Location
No R R, R 2
1 H H H
2 H H F
3 H H CI
4 H CI CI
5 H F F
6 F H H
7 F H F
8 F H CI
9 F CI CI
10 F F F
3rd Location
No R R, R 2
1 H H H
2 H H F
3 H H CI
4 H CI CI
5 H F F
6 F H H
7 F H F
8 F H CI
9 F CI CI
10 F F F
Synthesized N-substituted indole-2 and 3-carboxamide derivivatives have been assayed for their antioxidant
acitivity followed by compared with a-tocopherol. The structure-activity relationship of these indole
derivatives were also evaluated by comparing with those synthesized N-substituted indole amides
previously.
3

P-194
SYNTHESIS OF ELACRIDAR ANALOG AS CHEMOSENSITIZER IN
MULTIDRUG RESISTANCE IN CANCER
V.D. Dandekar. V.S. Velingkar
K.M.Kundanani College of Pharmacy, Mumbai, India
Elacridar exhibits Potent, selective chemosensitizing activity with low toxic potential. [1] The
present work involves the synthesis of novel elacridar analog.
OMe
H
Synthesis of Elacridar analog with structure 4-[2-(6',7'-dimethoxytetrahydroisoquinoline-2-
yl)ethyl]acridone-4-carboxanilide involved alkylation of 6,7-dimethoxy tetrahydroisoquinoline with
2-(4-nitrophenyl) ethyl bromide to obtain an intermediate with structure 6', 7'-dihydroxy-2[2-(4-
nitrophenyl) ethyl] tetrahydroisoquinoline which was then methylated to 6', 7'-dimethoxy-2[2-(4-
nitrophenyl) ethyl] tetrahydroisoquinoline. The nitro group of dimethoxy compound was then
reduced to corresponding amine by using ammonium sulfide (Yield=78.35%) or tin and
hydrochloric acid (Yield=57.8%) to obtain 4-[2-(6',7'-dimethoxytetrahydroisoquinoli-2-yl) ethyl]
benzenamine. The amine and acridone-4—carboxylic acid were then condensed by using DCC to
get the final Elacridar analog (yield=53.8%, M.P.=173°C). The intermediates involved were
synthesized using alternate methods than that reported in literature [2], Progress of all reactions was
monitored by thin layer chromatography. The compounds were characterized by infrared and 'H
NMR spectroscopy. Reaction steps were optimized with respect to yield and reaction time. The
work involved optimization of yield and reduction of reaction steps, use of alternate
catalyst/reducing agents and effective cost reduction
1. Dodic, N., DumaitreJB., Daugan,A., Pianetti,P., J. Med. Chem., 38, 2148 (1995)
2. Lednicer, D., Mitscher, L., "Polycyclic fused Heterocycles", in The organic chemistry of
Drug Resistance, Wiley-Interscience Publication, Vol. 6, 197 (1929)
32

P-195
SYNTHESIS AND ANTIMICROBIAL ACTIVITY OF SOME [l(2H)-PHTHALAZINON-2-
YL]-ACETYL/PROPYLTHIOSEMICARBAZIDES
D. S. Dogruer'. T. Onkol 1 , L. Uzun 1 , S. Adak 1 , S. Ozkan 2 , M. F. Sahin 1
'Gazi University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, Ankara, Turkey,
2 Gazi University, Faculty of Pharmacy, Department of Microbiology, Ankara, Turkey.
Many compounds bearing thiosemicarbazide structure have been reported to have antimicrobial
activity. These observation prompted us to synthesize new twelve acetylthiosemicarbazide,
propylthiosemicarbazide derivatives which bind at two position of phthalazinone ring. Structures of
the synthesized compounds have been confirmed by IR, 'H-NMR and elemental analysis.
n
R
1 Phenyl
1 Benzyl
1 Phenethyl
1 4-Chlorophenyl
1 4-Metoxyphenyl
1 4-Methylphenyl
i II n
2 Phenyl
V^^/N—(CH 2 )n—C—NH—NH—C—NH—R 2 Benzyl
0 2 Phenethyl
2 4-Chlorophenyl
2 4-Metoxyphenyl
2 4-Methylphenyl
The synthesized compounds will be tested against two Gram (+) bacteria (S. aureus, B. subtilis),
two Gram (-) bacteria ( P. aeruginosa, E.coli) and two yeast-like fungus such as C. albicans and C.
Parapsilosis by using disc diffusion method. Ciprofloxacin and Ketoconazole will be used as
control agents.
3

P-196
SYNTHESIS AND ANALGESIC AND ANTI-INFLAMMATORY ACTIVITIES OF ETHYL
(6-SUBSTITUTED-3(2//)-PYRIDAZINONE-2-YL) ACETATE DERIVATIVES
Y. Diindar 1 . M. Gok9e ! , E. KUpeli 2 , M. F. §ahin', N. Noyanalpan 1
'Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Gazi University, 06330,
Hipodrom-Ankara-Turkey, department of Pharmacognosy, Faculty of Pharmacy, Gazi University,
06330, Hipodrom-Ankara-Turkey.
The main objective in current pain research is to develop improved NSAI analgesics which devoid
of gastrointestinal (GI) side effects. In the course of a search for new molecules with analgesic
activity, we previously reported the synthesis of 6-substituted 3(2/^)-pyridazinone 1 derivatives
possessing analgesic and anti-inflammatory properties. Some compounds possessed significant
analgesic effects in the phenylbenzoquinone-induced writhing test (PBQ test). The derivatives
exhibiting highest activity had no gastric ulcerogenic effect and no toxicity even at 200 mg/kg
doses. In the present study, using 6-substituted 3(2/f)-pyridazinone derivatives as starting materials,
we synthesized the title compounds, ethyl (6-substituted-3(2//)-pyridazinone-2-yl) acetate 2
derivatives. Analgesic and anti-inflammatory effects of ethyl (6-substituted-3(2//)-pyridazinone-2-
yl) acetate 2 derivatives were evaluated in the phenylbenzoquinone-induced writhing test (PBQ test)
and carrageenan-induced paw edema method, respectively. Side effects of the compounds were
examined on gastric mucosa. None of the compounds showed gastric ulcerogenic effect compared
with reference nonsteroidal anti-inflammatory drugs (NSAIDs). A significant relation has been
observed between anti-inflammatory effect and the substituents. A short study implies that the
modification of the chemical group at the position 6 of 3(27f)-pyridazinone system influences
analgesic and anti-inflammatory activities. The structures of these new pyridazinone derivatives
were confirmed by their IR, 'H-NMR spectra and elemental analysis.
Comp.
R
o
2a 4-(Phenyl)piperazine
w
2b 4-(Benzyl)piperidine
2 3
/
C—OCH0CH3
HnC 2c 4-[(4-Chloro)phenyl]piperazine
\ 2d 4-[(2-Ethoxy)phenyl]piperazine
N—N 2e 4-[(2-Fluoro)phenyl]piperazine
2f 4-(2,3-Xysyl)piperazine
V" 2g 4-[(3-Trifluoromethyl)phenyl]piperazine
2
2h 4-(2-piridyl)piperazine
3

SYNTHESIS AND IN VITRO ANTIBACTERIAL ACTIVITY AGAINST PSEUDOMONAS
AERUGINOSA OF SOME NOVEL 2-PHENYL BENZOXAZOLES
T. Ertan 1 .1. Yildiz 1 , B. Tekiner-Giilba 1 , K. Bolelli 1 , O. Temiz-Arpaci 1 , S. Ozkan 2 ,
U. Abbasoglu 2
P-197
'Ankara University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, 06100,
Tandogan, Ankara, Turkey, 2 Gazi University, Faculty of Pharmacy, Department of Pharmaceutical
Microbiology, 06330, Etiler, Ankara, Turkey
A number of cases of multidrug resistant bacterial infections is increasing at an alarming rate. As
well as the clinicians have become reliant on vancomycin as the antibiotic for serius infections
resistant to traditional agents', there is still need for the new class of antibacterial agents.
Benzoxazole ring is the structural isoster of heterocyclics in naturally occuring nucleotides such as
adenine and guanine and takes place in the molecular structure of calcimycin, an antibiotic isolated
from a strain Streptomyces chartreusis 2 . Substituted benzoxazoles are found to be associated with
various chemotherapeutical activities such as antitumor, antiviral and antimicrobial activities 3 " 6 . In
this study, several novel 2-(p-substitutedphenyl)-5(or6)-nitrobenzoxazol derivatives were
synthesized by condensing of appropriate o-aminophenols and suitable acids in the presence of
polyphoshoric acid in order to investigate their antibacterial activity against Pseudomonas
aeruginosa. Chemical structures of the compounds were elucidated by using IR, 'H-NMR and Mass
spectroscopic analyses. The minimum inhibitory concentrations (MIC) of the newly synthesized
compounds were determined against Pseudomonas aeruginosa ATCC 10145 as Gram-negative
bacteria by using twofold serial dilution technique and were compared to standard drugs, ampicillin
and ofloxacine, The synthesized compounds were found to inhibit the in vitro growth of screened
microorganisms, showing the MIC values between 31.25 - >250 pg/ml.
Ri:H, C(CH 3 ) 3 , C 2 H 5 , F, Br
R 2 : H, N0 2
R 3 : H, N0 2
1. S.K. Fridkin, R.P. Gaynes, Clin. Chest. Med. 20 (1999) 303-316.
2. B.J. Abbot, D.S. Fukuda, D.E. Dorman, J.L. Occolowitz, M. Debeno, I. Farhner,
Antimicrob. Agents. Chemother. 16 (1983) 808.
3. A. Akbay, I. Oren, O. Temiz, E. Aki-Sener, I. Yalcin Arzneim.-Forsch. /Drug Research, 53
(2003) 266-271.
4. I.Yildiz, I. Yalcin, E. Aki-Sener, N. Ucarturk, Eur. J. Med. Chem.,39 (2004) 291-298.
5. I. Yildiz-Oren, B. Tekiner, I. Yalcin, O. Temiz-Arpaci, E. Aki-Sener, N. Altanlar, Arch.
Pharm., 337 (2004) 402-410.
6. O.Temiz-Arpaci, B. Tekiner-Gulbas, I. Yildiz, E. Aki-Sener, I. Yalcin, Bioorg. & Med.
Chem., 13 (2005) 6354-6359.
3

P-198
HETARYL SUBSTITUTED PYRAZOLO [3,4-Z>]QINOLINONES BY ONE-STEP
CYCLOCONDENSATION REACTIONS
M. K. Giimus. E. Kirpi, H. K. Beker, S. Kaban
Yildiz Technical University , Faculty of Science and Arts, Department of Chemistry, Organic
Chemistry, Davutpasa Campus, 34210, Merter-Istanbul, Turkiye
The research on the pyrazole[3,4-£]quinolinones is of current interest due to their exceptional
properties as calcium antagonists 1 " 3 and as powerful arteriolar vasadilators 4 ' 5 and have been
evaluated for enzymatic inhibitory activitiy 6 . We have been studying an efficient method for the
synthesis of various fused heterocyclic compounds having hetaryl-substituted pyrazole[3,4-
Z]quinolinone systems. In all cases the reaction gave a single product and structures were elucidated
by spectroscopic methods (IR, 'H- and 13 C NMR).
CHO
H3C
NH,
II \
/
H3C CH3
NH,
CHO
Ar-N
ci
Ar = _ Q or -^^c Het: Quinolin-4-yl or Quinolin-8-yl
1. F. Bossert and W. Vater, Med. Res. Rev., 9, 291 (1989).
2. D. I. Trigle, D. A. Langs, and R. A. Janis, Med. Res. Rev., 9, 123 (1989).
3. A. Fleckenstein and G. Grun, Arzneim. Forsch., 22, 334 (1972).
4. M. R. Bell and J. H. Ackerman, US Patent 4,920,128; Chem. Abstr., 113, 178015b, 1990
5. R. Alajarin, J. Alvarez-Builla, J. J. Vaquero, C. Sunkel, J. Fau, P. Statkov, and J. Sanz,
Tetrahedron Asymmetry, 4, 617 (1993).
6. F. Gatta, M. Pomponi and M. Marta, J. Hetereocyclic Chem., 28, 1301 (1991).
3

P-199
CYCLOCONDENSATION OF 2-AMINOBENZIMIDAZOLE OR 3-AMINO-l,2,4-TRIAZOLE
WITH DIMEDONE AND VARIOUS HETARYL CARBOXALDEHYDES
H. K. Beker, M. K. Gumus, E.Kirpi, S. Kaban
Yildiz Technical University , Faculty of Science and Arts, Department of Chemistry, Organic
Chemistry, Davutpasa Campus, 34080, Merter-Istanbul, Turkiye
It has been reported that condensed heterocyclic systems with partially or completely reduced
pyrimidine rings are of interest since they include valuable pharmacological properties as effective
coronary vasodilators 1,2 and calcium channel antagonists 3 ' 4 . In this research we have examined the onepot
cyclocondensation reaction of some amino-substituted benzimidazole and 1,2,4-triazole with
dimedone and various quinolinecarboxaldehydes, respectively. The structure of compounds was
elucidated by using IR, 'H- and 13 C NMR spectral data.
+
H3C
CH3
1. R. Alajarin, J. Alvarez-Builla, J. J. Vaquero, C. Sunkel, J. Fau, P. Statkov, and J. Sanz,
Tetrahedron Asymmetry, 4, 617 (1993).
2. Y. Tsuda, T. Mishina, and M. Obata, Jpn. Pat, 61227584; Chem. Abstr., 106, 176416
(1987).
3. F. Bossert and W. Vater, Med. Res. Rev., 9, 291 (1989).
4. D. I. Trigle, D. A. Langs, and R. A. Janis, Med. Res. Rev., 9, 123 (1989).
3

P-200
SYNTHESIS OF NEW 3,3'-BIS-l,3-THIAZOLIDIN-4-ONE DERIVATIVES
BY ONE-POT THREE-COMPONENT CONDENSATION
E. Kirpi, M. K.Gumiis. K. Beker, S. Kaban
Yildiz Technical University , Faculty of Science and Arts, Department of Chemistry, Organic
Chemistry, Davutpasa Campus, 34080, Merter-Istanbul, Turkiye
Bisthiazolidinone derivatives show different pharmacological activities depending on the
substituents and types of the moieties between the thiazolidin rings connecting the N-thiazolidine
rings . Owing to the presence of 2 and 2' equivalent stereogenic centers bisthiazolidinones can be
obtained as racemic 2R, 2'R / 2S, 2'S and 2R, 2'S-meso isomers, which usually exhibit
stereoselective pharmacological properties 2 . In this study, we report the synthesis of a new series of
bisthiazolidinones analogues.
o
Het: Pyridin-2-yl or Pyridin-3-yl
The synthesis of new 3,3'-bis-l,3-thiazolidin-4-ones was carried out by reacting properlysubstituted
pyridinecarboxaldehydes with equimolar amount of suitable diamines in the presence
of an excess of mercaptoacetic and 2-mercaptopropionic acid in toluene.
1. Benetello F, Bombieri G, Del Pra A, Orsini F, Previtera T, Vigorita M.G, Journal of
Molecular Structure, 443, 131-39, 1998.
2. Vigorita M.G, Ottana R, Monforte F, Maccari R, Trovato A, Monforte M.G, Taviano
F, Bioorganic and Medicinal Chemistry Letters, 11, 2791-94, 2001.
3

P-201
ANTIOXIDANT ACTIVITY STUDIES OF NOVEL Y-ACYL DEHYDROALANINE
DERIVATIVES
G. Gurkok 1 , T. Coban 2 , S. Siizen'
'Department of Pharmaceutical Chemistry, 2 Department of Pharmaceutical Toxicology, Ankara
University, Faculty of Pharmacy, 06100, Tandogan, Ankara, Turkey
Oxidative stress has been implicated in the development of many neurodegenerative diseases such
as Parkinson and Alzhemier's disease and also related to those diseases corresponding with aging,
artherosclerosis, rheumatoid arthritis and carcinogenesis. Olefins such as dehydroalanines have
been shown to inactivate free radicals by forming stabilized free radical adducts. Among these
molecules N-acyl dehydroalanines react with scavenge oxygen and hydroxyl radicals. This study
has surveyed the synthesis, characterization and in vitro effects on rat liver lipid peroxidation levels,
and DPPH free radical scavenging activities of some A^-acyl dehydroalanine derivatives. The results
indicated that compounds c, f and j slightly scavenged the level of DPPH radical at 10" 3 M
concentration by about 27, 46, and 56 %, respectively while compounds a, d, e, f, g, h showed the
strong inhibitory effect on the lipid peroxidation at the 10" 3 M and 10" 4 M concentrations and the
inhibition rate were in the range of 76-90 %.
DCCI / HONsu
- V
O
CH,
OH
Appropriate amine
" • V
CH,
NH CH, g HN
b HN CH 3
CH,
JZ
HN CH,
CH,
N
o
NH
\ /
3

P-202
SYNTHESIS AND BIOLOGICAL ACTIVITY OF SUBSTITUTED BENZYLISO-
THIOUREA DERIVATIVES
1 1 2 2 3
Z. Kazimierczuk . M. Chalimoniuk , A.E. Laudy , B.J. Starosciak , R. Moo-Puc , R. Cedillo-
Rivera
1
Polish Academy of Science Medical Research Center, 5 Pawinskiego St, 02-106 Warsaw, Poland,
2
Medical University, Department of Pharmaceutical Microbiology, 3 Oczki St, 02-007 Warsaw,
Poland, Unidad Interinstitutional de Investigation Medica, IMSS/UADY, Yuc, Mexico
The series of benzylisothiourea derivatives substituted onto benzene ring as well on the nitrogen
atoms was prepared by the reaction of un- and N-substituted thioureas with respective benzyl
bromides or chlorides. Additionally, two thiophene and furane isothiourea derivatives were
synthesized. The compounds were isolated as hydrochlorides or hydrobromides.
® e e
NHR 6 CI or Br
NHR7
^
NH2
NH2
2a: X = S,R = C1
2b:X = 0, R = N02
cP
lh: R 2 , R4 = N02, Ri,R 3 ,R 5 = H, Rsj = CH 3
la: R 3 = Br, Ri, 2 ,R4, 7 = H
li: R 2 , R 3 = CI, Ri, 4 -7 = H
lb:R3=N0 2) Ri, 2 , R4,7 = H
lj: R 2 , R 3 = CI, Ri, 4 -6 = H, R 7 = CH 3
lc: Ri=N0 2 , R 2 -7 — H
lk: R 2 , R 3 = CI, Ri, 4 ,5 = H, R6 j7 = CH 3
Id: R],R 3 = N0 2 , R 2 , R4. 7 = H 11: R 2 , R 3 = CI, Ri, 4 -6 = H, R 7 = C 2 H 5
le: Ri, R 3 = CI, R 2 , R4- 7 = H
lm:R 2 , R 3 = CI, R,, R 4 . 6 = H, R 7 = allyl
If: R 2 , R4 = N0 2 , Ri,R 3 ,R 5 -7 = H lo:Ri. 5 = F, R 6j 7 = H
lg: R 2 , R4 = N0 2 , Ri,R 3 ,R 5 , 6 = H, R 7 = CH 3 lp: R M =Br, R^ = H
The antibacterial and antigiardial (Giardia lambria) activity of the synthesized compounds was
studied. The most potent growth-inhibition of Gram-positive bacteria was observed for Id, If and
lp (MIC=12.5-50 mg/ml). Against Gram-negative bacteria the most active was Id (MIC= 12.5-100
mg/ml). Some of isothiourea derivatives (lc, lg, lj and 11) showed modest antiprotozoal activity
(IC50 = 1.14-1.27 mg/ml; metronidazale as control = 0.21 mg/ml). The obtained isothiouronium
salts inhibit the constitutive NO-synthase in few cases (li, lo, lp and 2a) stronger or comparable to
used as control drugs -7-nitroindazole or N-nitroarginine.
The study was supported by the Foundation for Development Diagnostics and Therapy, Warsaw,
Poland.
32

P-203
SYNTHESIS, ANTIFUNGAL AND ANTIOXIDANT SCREENING OF SOME NOVEL
BENZIMIDAZOLE DERIVATIVES
I. Kerimov', G. Ayhan-Kilcigil 1 , M.Iscan 2 , N. Altanlar 3 , B. Can-Eke 2
'Department of Pharmaceutical Chemistry, Ankara University, Faculty of Pharmacy, 06100,
Tandogan, Ankara, Turkey, 2 Department of Pharmaceutical Toxicology, Ankara University, Faculty
of Pharmacy, 06100, Tandogan, Ankara, Turkey, department of Pharmaceutical Microbiology,
Ankara University, Faculty of Pharmacy, 06100, Tandogan, Ankara, Turkey
Some novel benzimidazole derivatives were synthesized and their in vitro effects on rat liver
microsomal NADPH-dependent lipid peroxidation (LP) levels, microsomal ethoxyresorufin O-
deethylase (EROD) and antifungal activities were determined. The significant decrease in LP levels
was noted by compounds 4c (52%), 4e (58%) and 4h (43%) at 10" 3 M concentration. Compounds 4c
(100.0%), 4h (100.0%), 5c (98.0%) and 5h (100.0%) inhibited the microsomal ethoxyresorufin O-
deethylase (EROD) enzyme activity better than that of the specific inhibitor caffeine (85%). Among
these compounds, only the compounds 4b and 4h exhibited moderate activity against C.albicans
whereas the others had no remarkable effects.
5a, 5c,5e-5f,5h
* 0 e:^ f. -Q-, g: -£> h: ,-Q-.
Scheme: Synthetic route for the preparation of the compounds
32

P-204
SYNTHESIS AND ANTIMICROBIAL ACTIVITIES OF 5-[4-SUBSTITUTED PIPERAZIN-
1-YL] CARBOXYAMIDE DERIVATIVES OF NEW BENZIMIDAZOLES
C. Kus'. G. Ayhan-Kilcigil 1 , M. Tur^bilek 1 , N. Altanlar 2
'Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Ankara University, 06100-
Tandogan, Ankara, Turkey , 2 Department of Microbiology, Faculty of Pharmacy, Ankara
University, 06100-Tandogan, Ankara, Turkey
Benzimidazoles have diverse pharmacological effects and are comprised in many drugs in which
they have been widely investigated through their parasitic and antiviral activities. In previous
studies, we reported the synthesis and biological evaluation of l,2,5(6)-trisubstituted
benzimidazoles as antimicrobial agents' 2 . In this study, 2-(Substitutedphenyl)-5-[4-
substitutedpiperazin-l-yl]-carboxyamido-lH-benzimidazole derivatives were obtained by the
cyclization of 3,4-diaminobenzoic acid using Na2S205 adduct of appropriate benzaldehyde
compounds. Benzimidazolecarboxylic acids were converted to acyl chlorides with SOCI2, followed
by dehydrohalogenation of corresponding acyl chlorides and substituted piperazine to obtain the
planned derivatives. Antimicrobial activities of the synthesized compounds are currently under
investigation.
R =2,5-difluoro, 4-methoxy,
3,4-dimethoxy, 2,4-dichloro
R' = methyl, 4-fluorophenyl
1) soci2
2) R '-Nl' VjH
\ /
H
1. H. Goker, M. Tun

SYNTHESIS OF 1-ARYL-3-ETHYLAMINO-1 -PROPANONE HYDROCHLORIDES AND
EVALUATION OF THEIR ANTICONVULSANT ACTIVITIES BY MES AND scMet
TESTS
1 1 2 3 2
H. I. Gul . K. Kiiciikoglu , M. C'zmecioglu , U. Cah§ , E. Erciyas
P-205
Ataturk University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry,
2
Erzurum,Turkey, Ege University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry,
Izmir, Turkey, Hacettepe University, Faculty of Pharmacy, Department of Pharmaceutical
Chemistry, Ankara, Turkey.
Mono Mannich bases, l-Aryl-3-ethylamino-l-propanone hydrochlorides, have been synthesized
and tested for their anticonvulsant activities by MES and scMet tests, which are the indicator of
grand-mal and petite-mal epilepsy, respectively. Compounds Al, A2, A4 and A5 were new.
Anticonvulsant activity results have been shown at Table 1. Of the compounds, A3, which is
methoxy derivative, was the only compound, which did not show toxicity even at 300 mg/kg.
Other compounds synthesized have shown toxicity at 300 mg/kg after 20 minutes from the
injection. As a result, compound A3, regarding to its nontoxicity can be a candidate compound for
developing new compounds for grand mal epilepsy .
i_i
Ar C CH2CH2 N C2H5 -HCI
O
Al (C 6 H 5 ), A2 (p-CHj-QH,), A3 (p-CHjO-QR,), A4 (p-OH-C 6 H 4 ), A5 (p-Cl- C 6 H 4 ), A6 (C 4 H 3 S- (2-Thienyl))
Figure 1: l-Aryl-3-ethylamino-l-propanone hydrochlorides synthesized
Table 1: Phase I anticonvulsant screening of the synthesized compounds
Compound
MES ScMet Toxicity
number Vi hour 4 hour '/2 hour 4 hour Vi hour 4 hour
mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg
30 100 300 30 100 300 30 100 300 30 100 300 30 100 300 30 100 300
Al 1/1 1/1
*
0/1 0/1
*
1/1 1/1
*
0/1 0/1
*
0/4 0/4
*
0/2 0/2
*
A2 1/1 1/1
*
0/1 0/1
*
1/1 1/1
*
0/1 1/1
*
0/4 0/4
*
0/2 0/2
*
A3 0/1 1/1 1/1 1/1 1/1 0/1 0/1 0/1 0/1 0/1 0/1 0/1 0/4 0/4 0/4 0/2 0/2 0/2
A4 0/1 1/1
*
1/1 1/1
•
0/1 0/1
*
0/1 0/1
*
0/4 0/4
*
0/2 0/2
*
A5 1/1 1/1
*
0/1 0/1
*
0/1 0/1
*
1/1 1/1
*
0/4 0/4
*
0/2 0/2
*
A6 1/1 1/1
*
0/1 0/1
*
1/1 1/1
*
0/1 0/1
*
0/4 0/4
*
0/2 0/2
*
32

NEW 2-METHYL-0-ACYL-0XIMIN0-DIBENZ[b,e]0XEPINS SYNTHESIS AND SPECTRAL
CHARACTERIZATION
C. Limban'. A.- V. Missir 1 ,1. C. Chirita', C. Draghici 2
P-206
'Department of Pharmaceutical Chemistry, University of Medicine and Pharmacy "Carol Davila",
Bucharest, Traian Vuia 6, sect. 2, 020956, Romania, 2 The Organic Chemistry Center of Romanian
Academy "Costin C.D. Nenitescu" Bucharest, Splaiul Independentei, 202B, 77208, Romania
The present application is a continuation-in-part of our research concerning the synthesis and
characterization of new 0-acyl-oximino-dibenz[b,e]oxepins. The remarkable pharmacological
efficiency of the compounds with dibenz[b,e]oxepinic structure, such as Doxepine, known for
antidepressive action and lower side effects, and the positive results of the previous
pharmacological tests that we effectuated with some substances with the simillar structure
synthetized by us, led us to obtain new compounds from the dibenz[b,e]oxepin seria. In the case of
the previously synthesized dibenz[b,e]oxepins, the pharmacological tests revealed that some
compounds (e.g. O-benzoyl-1 l-oximino-6,1 l-dihydro-dibenz[b,e]oxepin; O-(p-methoxybenzoyl)-
1 l-oximino-6,1 l-dihydro-dibenz[b,e]oxepin; 0-(p-chlorobenzoyl)-l l-oximino-6,11-dihydro-dibenz
[b,e]oxepin) are psychoactive and others (e.g. 0-(p-fluorobenzoyl)-l l-oximino-6,11-dihydrodibenz[b,e]oxepin,
0-(p-nitrobenzoyl)-ll-oximino-6,11-dihydro-dibenz[b,e]oxepin) are sedatives
and anxiolytics. We did some chemical modeling and combined in the same molecule the
dibenz[b,e]oxepinic system and the oximinic group, double bound with the carbon from 11-position
of the dibenz[b,e]oxepinic nucleus. The synthesis of the new compounds contains three stages. In
the first stage, the 2-(4-methyl-phenoxymethyl)-benzoic acid was prepared by treating the phtalide
with potassium p-cresolate in xylene. The resulted potassium salt of 2-(4-methyl-phenoxymethyl)-
benzoic acid showed a good solubility in an aqueous solution of sodium hydroxide and was
separated from xylene. The aforementioned acid was precipitated using a mineral acid solution. The
potassium salt of p-cresol was obtained using the p-cresol and potassium hydroxide in xylene, and
the resulting water was removed by azeotropic distillation. The 2-methyl-6,l 1-dihydrodibenz[b,e]oxepin-l
l(6H)-one was synthesized in the second stage by a Friedel-Crafts cyclization
of the 2-(4-methyl-phenoxymethyl)-benzoic acid chloride in dry 1,2-dichloroethane. The
aforementioned acid chloride was obtatined by refluxing the coresponding acid with thionyl
chloride in excess, but could also be obtained by using different anhydrous solvents as reaction
medium, such as 1,2-dichloroethane. The desired ketone was prepared directly from the 2-(4-
methyl-phenoxymethyl)-benzoic acid by using various agents for anhydrization (e.g. poliphosporic
acid), but the yields were smaller. The new compounds were prepared by acylation of the 2-methyl-
1 l-hydroximino-6,1 l-dihydro-dibenz[b,e]oxepin with different benzoic acid chlorides, in dry
benzene, and in the presence of anhydrous pyridine as a proton fixator. The oxime was resulted by
treating the 2-methyl-6,l l-dihydro-dibenz[b,e]oxepin-l l(6H)-one with hydroxylamine
hydrochloride in the presence of pyridine. We established the optimal reaction conditions in
synthesizing process of the new compounds with high purity and yields. The new compounds,
which have not been mentioned in the literature concerning this domain, have been characterized by
their physical constants (melting point, solubility) and the structures were confirmed by 'H-NMR,
13 C-NMR and IR spectral methods. Following the obtaining of new compounds from
dibenz[b,e]oxepin series with potential pharmacological action, we synthesized eight new 2-methyl-
0-acyl-oximino-dibenz[b,e]oxepins. The spectral analysis confirmed the final and intermediate
compounds structures and also the synthesis that we have done.
32

P-2 07
SYNTHESIS AND ANTIMICROBIAL ACTIVITY OF SOME NEW
2, 4-TH [AZOLIDINEDIONE DERIVATIVES
O. Bozdag-Diindar 1 , A. Mente^e', N. Altanlar 2 , R. Ertan 1
Ankara University, Faculty of Pharmacy, Departments of, 'Pharmaceutical Chemistry and
2 Microbiology, Tandogan, Ankara, Turkey
The presence of a thiazolidine ring in penicillins and related derivatives was first recognized for
their occurrence in nature [1]. Indeed, heterocycles compounds which contain the thiazolidine ring
system have been reported to exhibit wide spectrum of biological activities. Depending on the
substituents, the thiazolidine ring can induce different pharmacological properties such as
antibacterial, antifungal [2], antidiabetic [3], cardiotonic [4], anticonvulsant [5], cyclooxygenase
and lipoxygenase inhibitory [6] activities. It has been known that the entrance of arylidene moieties
at different positions of the thiazolidine ring enhanced the antimicrobial activity [2, 7]. Prompted by
these investigations, in this study, we report the synthesis of some novel 2,4-thiazolidinedione
derivatives incorporating with two known bioactive heterocyclic nuclei such as thiazole and
thiazolidinedione as seen in below. Chemical structure of the synthesized compounds has been
elucidated by their IR, 'H NMR, Mass and Elementary analysis data. The synthesized compounds
were tested for their antifungal and antibacterial activities in vitro. All the compounds were found
active against used microorganisms.
O
1. Brown, F. C., Chem. Rev., 61, 463 (1961).
2. DeLima, M. C. A., Costa, D. L. B., Goes, A. J. S. et al., Pharmazie, 47, 182 (1992).
3. Cantello, B. C. C„ Cowthorne, M. A., Cottam, G. P. et al., J. Med. Chem., 37, 3977 (1994).
4. Andreani, A., Rambaldi, M., Locatelli, A. et al., Eur. J. Med. Chem., 28, 825 (1993).
5. El-Feky. S. A. H., Pharmazie, 48, 894 (1993).
6. Boschelli, D. H., Connor, D. T., Kuipers, P. J. et al., Bioorg. Med. Chem. Lett., 2, 705
(1992).
7. Labouta, I. M„ Salama, H. M., Eshba, N. H. et al., Eur. J. Med. Chem., 22, 485 (1987)
325

P-208
STUDY OF FORMATION OF ARTIFACTS UNDER DICHLOROMETHANE REACTION
WITH SOME NITROGENOUS DRUGS
A . Mohammadi 1 , M.P. Hamedani 2 , M. Amini 2 , H. Heidari'
'Department of drug and food quality control, Faculty of Pharmacy, Medical sciences university of
Tehran, Tehran Iran, department of medicinal chemistry, Faculty of Pharmacy, Medical sciences
university of Tehran, Tehran Iran.
The interactions of amines and some alkaloids with halogenated hydrocarbon solvents are well
known [1], These solvents and in particular dichloromethane are often used in the research and
development processes of the most of pharmaceutical compounds including synthesis,
identification, purification, extraction, assay, metabolic studies and etc. The information regarding
to such interactions which will have an effect on the results of the relevant processes, will improve
quality control and quality assurance purposes. In this work, we report a kinetic study which was
carried out on the quaternization reaction of Clozapine in dichloromethane under reflux condition
and room temperature at different times. The structure of clozapine - dichloromethane adduct,
clozapine chloromethochloride, was elucidated using 'H - NMR spectroscopy. In addition a HPLC
- UV method was developed and validated for the determination of intact drug. The peak of
clozapine was completely disappeared in the resultant chromatograms after treating with
dichloromethane to 10 minutes at concentrations of 1 and 5 pg/ml at room temperature.
1. A.H. Beckett and H. M. Ali, Journal of Chromatography, 177 (1979) 255 - 262.
32

P-209
RESEARCH ON ANTIDEPRESSANT DIBENZOCYCLOHEPTATRIENIC COMPOUNDS
L. Morusciag. G. M. Nitulescu, C. Stecoza, D. Nuta, A. Missir
Pharmaceutical Chemistry Department, Faculty of Pharmacy, University of Medicine and Pharmacy
"Carol Davila", Bucharest, Traian Vuia 6, 020956
This paper presents our research on the synthesis of new dibenzocycloheptatrienic compounds,
being known the antidepressant properties of some drugs (e.g. Demexiptylline) with similar
structure. Our lead compound has two biologically active structures in the same molecule: the
10,1 l-dihydro[a,d]cycloheptatrienic nucleus and a oximinic group. We used 5Hdibenzo[a,d]cyclohepten-5-one
as intermediate substance which was obtained from 10,11-dihydro-
5H-dibenzo[a,d]cyclohepten-5-one. The resulting suberenone was transformed into the
corresponding oxime, and this one was acylated with different aromatic acid chlorides giving novel
0-acyl-oximino-5H-dibenzo[a,d]cycloheptenes. The synthetized compounds were characterized
through their physical properties and their structures were confirmed by 'H-NMR and 13 C-NMR
analysis. The synthesized new 0-acyl-oximino-5H-dibenzo[a,d]cycloheptenes compounds which
were characterized by spectral analysis, showed antidepressant activity, eventually.
32

P-210
NEW DIBENZOSUBERONE N-ACYLATED OXIMES
L. Morusciag, A. Missir, I. Chirita, C. Limban
Pharmaceutical Chemistry Department, Faculty of Pharmacy, University of Medicine and Pharmacy
"Carol Davila", Bucharest, Traian Vuia 6, 020956
This paper is a continuation of our research concerning the synthesis of dibenzocycloheptadienic
compounds (1), being known their properties as antidepressant agents. These substances were
obtained by condensation of 5-oximino-10,l l-dihydro-5H-dibenzo[a,d]cycloheptene (2) with
different substituted benzoic acid chlorides for the purpose of synthesize the new dibenzosuberone
N-acylated oximes.
COOH
PPA.
\ HjNOHCI RCOCI
^OCOR
The 10,ll-dihydro-5H-dibenzo[a,d]cyclohepten-5-one (3) was obtained starting from phtalic
anhydride (4) which is reacted with phenyl-acetic acid to give benzylidenphtalide (5) which is
reduced with iodhydric acid and red phosphorus to 2-(2-phenyl-ethyl)benzoic acid (6). The 2-(2-
phenyl-ethyl)benzoic acid is cyclizated by heating with poliphosphoric acid at 170°C to give 10,11-
dihydro-5H-dibenzo[a,d]cyclohepten-5-one. The synthesized compounds were characterized
through their physical properties and their structures were confirmed by 'H-NMR,
13 C-NMR
analysis and elemental analysis.
32

P-211
SYNTHESIS AND ANTIMICROBIAL ACTIVITY OF SOME (5-CHLORO-2-
OXOBENZOTHIAZOLIN-3-YL) ACETO/PROPANO HYDRAZIDES
T. Onkol'. S. Ito 2 , B. Ozfelik 3 , B. Cakir', M.F. $ahin'
'Gazi University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, Ankara, Turkey,
institute for Medical and Dental Engineering, Tokyo Medical and Dental University, Tokyo, Japan,
3 Gazi U niversity, Faculty of Pharmacy, Department of Microbiology, Ankara, Turkey
There has been considerable interest in the chemistry of 2-(3H)-benzothiazole ring systems, a core
structure in various synthetic pharmaceuticals displaying a broad spectrum of biological activity'
such as antimycobacterial, antifungal, anticancer, antituberculosis, anticonvulsant,
antiinflammatory, and analgesic activities. Meantime, various hydrazide and hydrazone derivatives
have shown antibacterial and antifungal activities. In the present study, eleven (5-chloro-2-
oxobenzothiazolin-3-yl) aceto/propanohydrazides have been synthesized. Structure of the
compounds synthesized has been elucidated by the aid of IR and 'H-NMR spectral data and
elemental analyses.
R: H, CH3
R,:H,CI, CH3, OCH3
Figure 1
The synthesized compounds were tested against three Gram-positive (S. aureus, B. subtilis, E.
faecalis) and three Gram-negative bacteria (P. aeruginosa, E. coli, K. pnomoni). The antifungal
activities of compounds were evaluated in vitro against a yeast-like fungus such as C. albicans
using the microdilution method. Among the tested compounds, the most effective antimicrobial
activity was observed with (5-chloro-2-oxobenzothiazolin-3-yl) propanohydrazide derivatives at
concentration of 32pg/ml (MICs) against E. faecalis. Ampicilline, Ofloxasine, and Ketoconazole
were used as control agents.
32

P-212
CENTRAL NERVEOUS SYSTEM ACTIVITY OF 2-(3,5-DIMETOXY-4-
HYDROXYPHENYL)-5,6-DICHLORO-(lH)-BENZIMIDAZOLE (DPCB)
O.D. Can", Y. Ozkay 2 , U. Demir', i. Iikdag 2 , Y. Oztiirk
i
Anadolu University, Faculty of Pharmacy, Department of Pharmacology, 26470 Eskiehir, Turkey,
Anadolu University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, 26470
Eski§ehir, Turkey
It is known that synthetic chemical compounds especially lipophylic ones have various effects on
central nervous system. Benzimidazoles are the examples to such compounds in
whichbenzimidazoles have been reported several times for their potential pharmacological effects
such as analgesic, sedative , selective 5-HT4 receptor antagonists. As a benzimidazole derivative,
we have synthesized 2-(3,5-Dimetoxy-4-hydroxyphenyl)-5,6-dichloro-(lH)-benzimidazole (DPCB)
for screening its central nervous system activity. The effects of the DPCB (500 mg/kg) on
exploratory behaviour, spontaneous motor activity and motor coordination (Rota-rod performance)
were investigated in mice. Intraperitonal (i.p) administration of DPCB induced a significant
(P

ASSESSMENT OF STRUCTURES OF POLY(RIBO)NUCLEOTIDE-PHOSPHOLIPID
SELF-ASSEMBLIES AND THEIR IMPLICATIONS IN GENE TRANSFECTION AND
DNA CHROMATOGRAPHY
Y. Mana.vba§i', E. SUleymanoglu 2
'Department of Pharmacology, department of Pharmaceutical Chemistry, Gazi University,
Faculty of Pharmacy, 06330 - Ankara, Turkey,
P-213
Physicochemical features related to preparation and use of self-assemblies formed between
multilamellar and unilamellar liposomes and poly (ribo) nucleotides with various conformation and
sizes are presented. The roles of divalent metal cation or surfactant-induced adsorption, aggregation
and adhesion between single- and double-stranded polyribonucleotides and phosphatidylcholine
vesicles are emphasized. Nucleic acid condensation and compaction mediated by Mg 2+ , Ca 2+ and
both single chain and gemini surfactants were followed with regard to interfacial interaction with
lipid vesicles. Our previous and more recent microscopic, spectroscopic and microcalorimetric
measurements of liposomes and poly (ribo) nucleotides and their ternary complexes with inorganic
cations and detergents were used to build the thennodynamic model of their structural transitions.
In general, the increased thermal stability of the phospholipid bilayers is achieved by affecting their
melting transition temperature by nucleic acid induced electrostatic charge screening.
Measurements give evidence for the stabilization of polynucleotide helices upon their association
with liposomes in presence of cations with various valency. Such an induced aggregation of
vesicles either leads to heterogeneous multilamellar DNA-lipid arrangements, or to DNA-induced
bilayer destabilization and lipid fusion. In contrast, stable monodisperce complexes are formed after
precompaction of DNA with surfactant, followed by addition of vesicles. Surfactants bind to DNA
in a cooperative manner resulting in rise in sizes of the resulting DNA-surfactant complexes due to
their aggregation. The formation of these bundles is governed by both electrostatic and hydrophobic
interactions of surfactant chains, the reaction being mediated by condensed counterions, steric
hindrance or by intrinsic chain flexibility. In addition to nucleic acid-liposome formulations,
relevant structures, as seen in Langmuir-Blodgett monolayers and Black Lipid Membranes (BLM)
setups are also presented. The further employment of these transfection competent polyelectrolyte
nanostructures as improved formulations in therapeutic gene delivery trials, as well as in DNA
chromatography, is discussed.
3

SYNTHESIS AND CYTOTOXIC ACTIVITY OF PLATINUM(II) AND PLATINUM(IV)
COMPLEXES WITH 5(6)-CHLORO-2-HYDROXYMETHYLBENZIMIDAZOLE
LIGANDS AGAINST MCF-7 AND HeLa CELL LINES
S. Utku'. F. Giimti 2 , S. Giir 3 , A. Ozkul 4
P-214
'Department of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Mersin, 33169,
Mersin, Turkey, department of Pharmaceutical Chemistry, Faculty of Pharmacy, University of
Gazi, 06330 Etiler-Ankara, Turkey, department of Microbiology, Faculty of Veterinary
Medicine, University of Kocatepe, 03200, Afyonkarahisar, Turkey, department of Virology,
Faculty of Veterinary Medicine, University of Ankara, 06110, Ankara, Turkey
Cisplatin, m-diamminedichloroplatinum(II), and other platinum-based drugs such as carboplatin, cisdiammine(l,l-cyclobutanedicarboxylato)platinum(Il),
and oxaliplatin, (ra«5-i,i-cyclohexane-l,2-diamine
oxalatoplatinum(II)) are used to treat testicular tumors as well as a variety of other human solid tumors, but
most of the drugs used are intrinsically resistant and acquired resistance commonly develops during
treatment [1]. The need for cisplatin analogs which are less toxic and have broader spectra of activity led to
the synthesis of a large number of platinum complexes over the past three decades. The replacement of the
leaving chloride groups affects mainly tissue and intracelluar distribution of the cisplatin analogues. On the
other hand, the replacement of amine groups can result in different structural and formational alterations in
the target DNA, which may then affect the character of biological activities of the anologues [2], The more
hydrophobic cisplatin analogues were expected to enhance affinity of the damage-recognation proteins to the
platinated site, which could more effectively protect the mentioned proteins from excision repair. It has been
shown that increasing cytotoxicity of cisplatin analogues, in which NH 3 groups were replaced by more
hydrophobic amine ligands, was correlated with growing hydrophobicity of these analogues [3]. Some Pt(I V)
complexes have shown potential as powerful anticancer drugs. It is widely believed that reduction to Pt(II) is
essential for the anticancer activity of Pt(IV) complexes to be effected. The reduction potentials of
diam(m)ine Pt(IV) complexes are dependent on the nature of the axial and equatorial ligands, but the axial
ligands generally exert the stronger influence [4]. Although, some of Pt(IV) compounds, including iproplatin
(CHIP, JM9, cz's-dichloro-Zraws-dihidroxy bis (isopropylamine)platinum(IV)), tetraplatin
(ormaplatin,tetrachloro[(l,2-diaminocyclohexane)platinum(IV)]) and satraplatin (JM216, trans.cisbis(acetato)aminedichloro(cyclohexylamine)platinum(IV)cyclohexylamine)
have been tested in clinical
trials, there are still no Pt(IV)-based therapeutics in routine clinical use [5], In the present study, as an
extension of the previous investigation on the probable antitumor activity of platinum complexes of
benzimidazole ligands to determine the effect of axial and equatorial ligand variation on the cytotoxic
activities of the platinum complexes, a series of Pt(II) and Pt(IV) complexes with 5(6)-chloro-2-
hydroxymethylbenzimidazoles as non-leaving amine ligand and chloro, iodo, hydroxo ligands as leaving
groups was synthesized and evaluated for their in vitro cytotoxic activities on the human MCF-7 and HeLa
cell lines.
1. Jamieson, E. R„ Lippard, S. J., Chem. Rev., 99, 2467-2498 (1999).
2. Brabec, V, Kasparkova, J, Drug Resistance Updates, 8, 131-146 (2005).
3. Tallen, G, Mock, C, Gangopadhyay, S. B, Kangarloo, B, Krebs, B, Wolff, J. E. A, Anticancer
Res., 20, 445-450 (2000).
4. Hall, M. D, Hambley, T.W., Coor. Chem. Rev., 232, 49-67 (2002).
5. Markovic, M, Knezevic, N, Momcilovic, M, Grguric-Sipka, S., Harhaji, L, Trajkovic, V,
Stojkovic, M. M, Sabo, T, Miljkovic, D„ [Pt(HPxSC)Cl 3 ], Eur. J. Pharm., 517, 28-34 (2005).
32

P-215
THE IN VITRO HEPATIC MICROSOMAL METABOLISM OF 2-SUBSTITUTED-
BENZIMIDAZOLES IN RATS
1 2
O. Algul , M. Ulgen
'Mersin University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry ,33169,
Mersin,Turkey,
2 University of Marmara, Faculty of Pharmacy, Department of Pharmaceutical
Chemistry, 81010 Haydarpasa, Istanbul, Turkey
Benzimidazole and its derivatives with a potential antimicrobial, antitumor activity have
previously been synthesized in our laboratories. They were prepared according to the Phillips
method which was described earlier. Equimolar amounts of diamine (o-phenylenediamine, 3,4-
diaminotoluene, etc.) and the corresponding carboxylic acid in 5.5N hydrochloric acid were
refluxed for 2-30h. The solution was cooled in an ice bath and neutralized with sodium
bicarbonate. The resulting precipitate was fdtered off, washed several times with water and
purified by recrystallization. In the present study, the in vitro hepatic microsomal metabolism
of (I) was carried out using hepatic washed rat microsomal preparations fortified with
NADPH. The substrate (I) and its potential metabolite (II) were then separated by using a
reverse phase HPLC system consisted of a Cjg column and a mobile phase of
acetonitrilerwater (50:50) at a flow rate of 1 ml/min with UV detection at 254 nm. The
substrate (I) was incubated with rat microsomal preparations in the presence of NADPH,
extracted into DCM and finally evaporated under nitrogen. Results will be discussed in detail in the
present poster.
3

THE SYNTHESIS AND CHARACTERIZATION OF SOME NEW RUTHENIUM AND
IRON COMPOUNDS
V. Aldea, V. Uivarosi, B.Velescu
University Of Medicine And Pharmacy "Carol Davila" Bucharest, Faculty Of Pharmacy, Dept.
Inorganic Chemistry, 6-Th Traian Vuia Str., Bucharest
P-216
The paper presents the preparation method of two new compounds of ruthenium (III) and iron (III)
with ferron (7-iodo 8-hydroxy quinoline 5-sulphonic acid), having the potential chemotherapeutic
activity. The reaction was undertaken in DMSO medium at 70°C. The compounds were
precipitated following dilution with water and isolated from the reaction medium. In order to
establish the structure the compounds were analysed by using IR spectrophotometry, UV-VIS
spectrophotometry and conductivity measurements. The results supported the idea that the structure
of these new compounds.
H3OS
H3OS
DMSO
DMSO
Where Me=Fe(III), Ru(III)
3

P-217
SYNTHESIS OF (l-[3-(PIPERIDINOMETHYL)-4-HYDROXYPHENYL]-3-ARYL-2-
PROPEN-1-ONE AND EVALUATION OF THEIR ANTIFUNGAL ACTIVITIES
K.O. Yerdelen', H.I. Gul', F. Sahin 2
'Deparment of Pharmaceutical Chemistry, Faculty of Pharmacy, Ataturk University, 25240,
2
Erzurum, Turkey , Department of Genetics and Bioengineering, Faculty of Engineering and
Architecture, Yeditepe University, 34755, Kayisdagi, Istanbul Turkey.
In this study, Mannich bases with piperidine, (l-[3-(piperidinomethyl)-4-hydroxyphenyl]-3-aryl-2-
propen-1-one, B1-B5) were synthesized starting from the chalcones (l,3-diaryl-2-propen-l-one,
A1-A5). Chemical structures of the compounds have been confirmed by 'H-NMR, 13 C-NMR, IR,
and UV spectra and elemental analyses. Antifungal activities of the compounds have been tested
against 3 fungi species pathogenic in humans [Trichophyton rubrum (Hak-8), Trichophyton
mentagrophytes (Hak-9), Microsporum canis (Hak-4)] and 13 fungi species pathogenic in plants
[Sclerotinia sclerotiorwn), Sclerotinia minor, Alternaria alternate (AA-1121), Aspergillus flavus
(Hak-23), Aspergillus variecolor (IO-Balik), Fusarium acuminatum, Fusarium oxysporum (ED-10),
Fusarium solani (ED-IS), Fusarium tabacinum (ED-IT), Moniliania fructicola (FS-M),
Penicillium spp. (P-TY), Rhizopus spp. (R-27), Rhizoctonia solani (EB-ML)] at the concetration
range of 2-64 pg/ml by microdilution method using Amphotericin -B as the reference compound.
Of the compounds, Al against plant pathogens Sclerotinia sclerotioruma and Rhizoctonia solani,
B4 against plant pathogen Aspergillus variecolor, B5 against human pathogen Trichophyton
rubrum have shown 2-4 times more powerful antifungal activity compared with Amphotericin-B.
Of the compounds synthesized, B5 against human pathogenic fungi, Al and B4, against plant
pathogenic fungi can be choosen as candidate compounds for further studies to develop new
antifungal compounds.
3

SYNTHESIS OF l-[3-(DIBENZYLAMINOMETHYL)-4-HYDROXYPHENYL]-3-ARYL-2-
PROPEN-1-ONE AND EVALUATION OF THEIR CYTOTOXIC ACTIVITIES
K..O. Yerdelen'. M. Gul 2 ' 3 , H. I. Gul', O. Hanninen 3 , M. Atalay 3
1
Deparment of Pharmaceutical Chemistry, Faculty of Pharmacy, Ataturk University, 25240,
Erzurum, Turkey, Deparment of Physiology, Faculty of Medicine, Ataturk University, 25240,
Erzurum, Turkey, Deparment of Physiology, Faculty of Medicine, University of Kuopio, 1627,
Kuopio, Finland.
P-218
In this study, Mannich bases with dibenzylamine (l-[3-(dibenzylaminomethyl)-4-hydroxyphenyl]-
3-aryl-2-propen-l-one, C1-C5) were synthesized starting from the chalcones (l,3-diaryl-2-propen-
1-one, A1-A5). Chemical structures of the compounds have been confirmed by 'H-NMR, ^C-
NMR, IR, and UV spectra and elemental analyses. Cytotoxic activities of the compounds have been
tested against rat skeletal muscle derived myoblast cells (L6) and transformed human T
lymphocytes (Jurkat). Melphalan and 5-fluorouracil were also tested as reference drugs. Except C3,
all compounds have shown 1.29-5.40 times more powerful cytotoxicity than 5-FU, and the
compounds A3, C2 and C4 have shown 2.76, 1.17, 1.95 times more powerful cytotoxicity than
melphalan against L6 cells, respectively. Preparation of Mannich bases with dibenzylamine from
the chalcones increased the cytotoxicity 1.39 and 2.67 times at the compounds C2 and C4,
respectively, compared with their corresponding chalcones. While all compounds synthesized had
2.01- 6.63 times more powerful cytotoxicity than 5-FU against Jurkat cells, except C3, all other
compounds showed 1.09-1.68 times more powerful cytotoxicity than melphalan. Preparation of
Mannich bases from the chalcones increased the cytotoxicity 1.11, 1.46 times respectively at the
compounds CI and C5 compared with the corresponding chalcones against Jurkat cells.The
compounds synthesized have been found more selective against L6 cells compared with Jurkat
cells. To conclude, preparation of Mannich bases from the chalcones has been found to be a useful
modification to develop new compounds with cytotoxic activity.
3

SYNTHESIS OF l-[3-(DIBENZYLAMINOMETHYL)-4-HYDROXYPHENYL]-3-ARYL-2-
PROPEN-1 -ONE AND EVALUATION OF THEIR ANTICONVULSANT ACTIVITIES
H. I. Gul', K.O. Yerdelen'. U. Call/
Deparment of Pharmaceutical Chemistry, Faculty of Pharmacy, Ataturk University, 25240,
2
Erzurum, Turkey, Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Hacettepe
University, 06100, Sihhiye/Ankara, Turkey.
P-219
In this study, Mannich bases with dibenzylamine (l-[3-(dibenzylaminomethyl)-4-hydroxyphenyl]-
3-aryl-2-propen-l-one, C1-C5) were synthesized and the chemical structures of the compounds
have been confirmed by 'H-NMR,
13 C-NMR, IR, and UV spectra and elemental analyses.
Anticonvulsant activities of the compounds were evaluated by MES, scMet tests. Neurotoxicities of
the compounds were also evaluated by rotorod test. None of the compounds showed neurotoxicity
at the screening of anticonvulsant activity. Compound C4 at MES test, compounds C2, and C3 at
scMet test have shown anticonvulsant activity at different dose levels (30-300 mg/kg) and time
periods (1/2 h, 4 h). To conclude, of the compounds C4 against grand-mal epilepsia, C2 and C3
against petite-mal epilepsia can be choosen as candidate compounds to develop new anticonvulsant
compounds for further studies.
3

P-220
EVALUATION OF ANIONIC CHITOSAN DERIVATIVES AS ENTERIC COATING
POLYMERS FOR DICLOFENAC SODIUM TABLETS
K.M. Aiedeh. H.S. AlKhatib, M.O. Taha
Faculty of Pharmacy, University of Jordan, Amman, 11942, Jordan
To evaluate the potential of chitosan succinate and chitosan phthalate as enteric coating polymers
for diclofenac sodium tablets. The solubility of the new chitosan derivatives was evaluated in
different media to check their suitability for enteric applications. Diclofenac sodium core tablets
were coated with either derivative and drug release was evaluated according to the USP method for
delayed release (enteric) preparations. The effects of storage and elevated temperature and humidity
on drug release were evaluated too. The solubility profile of chitosan succinate and chitosan
phthalate was completely different from that of chitosan. The new derivatives showed significantly
improved solubility in basic media while their solubility in acidic media decreased in comparison to
the native polymer. Chitosan phthalate coated tablets complied with USP delayed release
preparations specifications, while chitosan succinate coated tablets released a high percentage of the
drug in the acid stage and failed to provide the needed dissolution criteria for enteric tablets.
Storage under ambient conditions as well as under elevated temperature and humidity slowed down
the release from tablets coated with these polymers. Chitosan phthalate succeded as an enteric
coating polymer for diclofenac sodium tablets while chitosan succinate did not. The enteric
behavior of chitosan phthalate was shown to be affected significantly by curing at elevated
temperatures and high humidity conditions.
3

P-221
THE EFFECT OF POLYMER TYPE AND RATIO ON THE EXTENDED
RELEASE OF ATENOLOL FROM HPMC TABLETS
E.Algin, O. inal, T. Baykara
Department of Pharmaceutical Technology, Faculty of Pharmacy, University of Ankara, 06100,
Tandogan, Ankara, Turkey
Atenolol is a water-soluble cardioselective beta-blocker agent which is widely used in the treatment
of hypertension and angina pectoris. Hydroxypropylmethylcellulose (HPMC), is a hydrophilic
polymer which used to control drug release from matrix tablet systems because of its easy
compression property and accommodation to high levels of drug. The objective of the present work
was to prepare Atenolol extended release tablet formulations by altering the type and the ratio of
HPMC in combination with a lactose based direct compression agent (DC-agent). In order to
prepare extended release tablets two viscosity grades of HPMC (Methocel® K100LV and K15M)
and a DC-agent (Pharmatose DCL11®) were used as matrix materials. Each of the tablet
formulation with total mass of 260 mg including 100 or 50 mg Atenolol were compressed in a
hydraulic press with a flat-faced punches of 8 mm diameter and a compaction pressure of 200 MPa
for 10 seconds. Drug release studies were carried out according to the method given for Delayed
Release Articles in USP XXVII and the USP XXVII-Apparatus II was used at 50 rpm rate. The
amount of the Atenolol was determined spectrophotometrically at 274 nm. Validation of
spectrophotometric analysis for determination of atenolol in dissolution media was done by
performing linearity and range, precision, accuracy and spesicifity. Results indicated that HPMC
types and ratios were found to be effective on drug release from matrix tablet formulations. Using
equal amounts of the low and high viscosity grade HPMCs as a mixture in the formulation resulted
with a discontiniously drug release profile. Formulations including low viscosity grade HPMC at
high amounts (drug: polymer: DC-agent at 10:10:6 and 5:9:12) gave more linear release profiles.
Drug release data of formulations were evaluated by mathematical models (Zero Order, First Order,
Higuchi, Hixson-Crowell and Korsmeyer-Peppas). The kinetics of Atenolol release from the
formulations were generally best fit to the Korsmeyer-Peppas kinetic model and drug relase
mechanism show Anomolous transport mechanism according to their values of n.
1. Algin,E, Kilifarslan, M, Karata, A, Yuksel, N, Baykara, T. "Effects of different direct
tabletting agents on release of Verapamil HC1 from cellulose matrix tablets" in 7 th Proceedings
of International Symposium on Pharmaceutical Sciences, ISOPS-7", Ankara, p.l 10, 2003.
2. Algin,E, Kiliarslan, M, Karata, A, Yuksel, N, Baykara, T. J. Fac. Pharm. Ankara,33(3),
125-137, 2004.
3. Lotfipour, F, Nokhodchi, A, Saeedi, M, Norouzi-Sani, S, Sharbafi, J, Siahi-Shadbad, M.R. II
Farmaco, 59, 819-825,2004.
3

EFFECT OF FORMULATION VARIABLES ON THE DRUG RELEASE STABILITY OF
POLYVINYL ACETATE/POVIDONE BASED MATRICES
H.S. AlKhatib. S. Mohesin, K. Aiedeh, Y. Bustanji
Faculty of Pharmacy, University of Jordan, Amman, 11942, Jordan
P-222
To evaluate the effect of formulation variables on the drug release stability from tablets based on a
commercially available extended release matrix material, Kollidon SR®, upon heat treatment and
accelerated stability testing. Extended release chlorpheniramine maleate tablets were formulated
using Kollidon SR® as a matrix forming material. The formulation variables evlauted in these
tablets were the type and level of the additional fdler excipients (Lactose, dibasic calcium
phosphate and microcrystalline cellulose), particle size of lactose as a fdler excipient as well as the
level of colloidal silicon dioxide as an anti-aging agent. Tablets were subjected to dry heat treatment
at 60°C over 4 days as well as accelerated stability testing under elevated temperature and humidity
conditions (40°C/75%RH) for three months. Drug release from the different formulations was
evaluated according to the USP method and the release profiles were fitted to the Higuchi square
root equation to determine the apparent rate constant. The effect of the dry heat or the heat/humidity
treatment on the dissolution properties of the Kollidon SR® based matrices was evaluated through
the percentage change in the apparent release rate constant. Drug release from Kollidon SR® based
matrices was found to be sensitive to the type and level of the added filler excipient. A linear
relationship between the filler content and the apparent drug release rate was observed for lactose
while an exponential relationship was observed for the other two fillers. Drug release was found to
depend significantly on the type and level of the filler excipient. The effect of lactose particle size
on drug release stability was minimal. Colloidal silicon dioxide exerted a significant anti-againg
effect that was proportional to its content in the matrix tablets. Formulation aspects affect the drug
release from Kollidon SR® based matrices significantly. Proper use of added excipients may
enhance dry heat / heat-humidity behavior of these matrix tablets. Colloidal silicon dioxide showed
efficacy as an antiaging agent in Kollidon SR® based matrices.
30

P-223
EFFECT OF POLYETHYLENE GLYCOL AND SODIUM LAURYL SULPHATE ON THE
COMPACTION CHARACTERISTICS OF EUDRAGIT
M.O. Emeje, O.O. Kunle
Department of Pharmaceutical Technology and Raw Materials Development National Institute for
Pharmaceutical Research and Development (NIPRD) Idu, P.M.B.21 Garki - Abuja, Nigeria
A study of the compaction characteristics of Eudragit L-100 in the presence and in the absence of
two commonly used additives, polyethylene glycol 6000 (PEG 6000) and sodium lauryl sulphate
(SLS) was carried out. Eudragit granules, with and without the additives were prepared separately
by the wet granulation method and compacts were made at varying compression pressures.
Compaction characteristics using Kawakita and Heckel analysis revealed a concentration dependent
effect of the additives on the compressibility of Eudragit granules, with 2.5% PEG 6000 producing
the largest effect and 5.0% PEG 6000, the least. Irrespective of the concentration of PEG 6000, the
yield value increased, while SLS either had no effect or decreased the yield value of Eudragit
granules. The highest yield value of 21.69KN was produced by formulations containing 2.5% PEG
6000. The results of this study showed that PEG 6000 and SLS were found to affect the
deformation characteristics of Eudragit L-100. The extent and nature of the effect depended on both
the type and concentration of the additive used. SLS was found to increase the deformation of
Eudragit more than PEG 6000. The results of this study show that additives such as PEG 6000 and
SLS affect the compaction characteristics of Eudragit L-100, and this could affect its retardant
behavior.
3

EVALUATING THE EFFECT OF DONOR COMPARTMENT COMPOSITION ON THE
IONTOPHORETIC DELIVERY OF PROPRANOLOL HCL THROUGH HUMAN SKIN
D. Hassanzadeh, F. Monajemzadeh
P-224
Depatment of Pharmaceutics, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz,
Iran
Transdermal delivery of drugs using Iontophoretic methods improves the low absorption of high
molecular weight and hydrophilic compounds and enhances the amount of drug absorbed. The
purpose of this study was to evaluate the receptor compartment composition on the Iontophoretic
delivery of Propranolol HCL through human skin. Anodal ionthophoresis was used to evaluate the
propranolol transport through Human female abdominal skins .The epidermal membranes were
mounted between two glass half-cells with the stratum corneum facing the donor compartment. The
transport studies were carried out using HEPES buffer sopiked with [-^H] Propranolol HCL or
[^H20]. Drug concentration in the receptor compartment was measured by Liquid sentilation
technique.The transportation Value (t0 was calculated and recorded. The results indicated that the
drug transportation Value decreased as the NaCl concentration increased in the Donor compartment
and reached the plateau at the concentration of 70 millimole of NaCl in the donor compartment.
Thus in anodal Ionthophoresis, anion concentration in the receptor compartment causes a decrease
in the Cation transportation from the donor to the receptor compartment and it is important to
consider such a conditions in in vitro studies using NaCl solutions.
32

P-225
SILDENAFIL (VIAGRA®) ENHANCES EXCITATORY SYNAPTIC TRANSMISSION IN
RAT HIPPOCAMPUS
1* 2* 2 2 2
S. Ilbasmis Tamer , N. Wijayawardhane , K. Parameshwaran , S. Uthaythas , M. Dhanasekaran ,
2 1
V. Suppiramaniam , T. Degim
*Equally contributed, Department of Pharmaceutical Technology, Faculty of Pharmacy, Gazi
2
University, Ankara, Turkey, Department of Pharmacal Sciences, Harrison School of Pharmacy,
Auburn University, Auburn, AL 36849, USA.
Sildenafd citrate has been used as therapy for erectile dysfunctions. Recent studies support that
sildenafil improves acquisition and retention of memory in rodents. In the central nervous system,
nitric oxide (NO) is believed to function as a retrograde messenger following glutamatergic N-
methyl-D-aspartate (NMDA) neurotransmission by stimulating soluble guanylyl cyclase (sGC).
Stimulation of sGC increases cGMP levels, resulting in further release of glutamate. This may
contribute to hippocampal long-term potentiation (LTP), a cellular substrate of memory, mediated
by enhancement of glutamatergic neurotransmission in the hippocampus. This study was aimed to
investigate whether sildenafd enhances the glutamatergic neurotransmission in the hippocampus.
Electrophysiological techniques were used to test the hippocampal functions that correlate with
learning and memory performances. One month old Sprague-Dawley rat offspring were treated with
sildenafd citrate (lOmg/kg) or saline for 6 days using the gavage technique. After the termination of
treatment brains were removed and 400 (im thick slices were prepared using standard techniques.
Stimulations were delivered at CA3 area and field excitatory post synaptic potentials (fEPSPs) were
recorded from CA1 area. Input/output curves, paired-pulse facilitation plots were generated. LTP
was induced by theta burst stimulation. Our study showed that the slices from sildenafil exposed
rats had significantly greater responses for the same amount of stimulus, compared to controls.
According to the preliminary data obtained, significant paired-pulse facilitation and LTP were also
noted in the animals treated with sildenafil citrate. The results indicate that sildenafil citrate may
improve learning and memory by enhancing glutamatergic excitatory synaptic transmission in the
hippocampus. Further studies need to be carried out to elucidate the underlying mechanisms of
sildenafil as a memory enhancer.
3

P-226
EVALUATION OF PHYSICOCHEMICAL PROPERTIES OF INDOMETHACIN
LIQUISOLID COMPACTS
1,2 1 2 1,3
Y. Javadzadeh , M.R. Siahi , S. Asnaashari , A. Nokhodchi
i
Department of Pharmaceutics, Faculty of Pharmacy-Tabriz University of Medical Sciences-Iran,
2 3
Drug applied research center, Tabriz University of Medical Sciences-Iran, Medway School of
Pharmacy, Central Ave, University of Kent and Greenwich, Chatham, Kent, ME4 4TB, England
The potential of liquisolid systems to improve the dissolution properties of water-insoluble agent
(indomethacin) was investigated. In this study, physicochemical properties of indomethacin
liquisolid tablets, effect of aging and type of the vehicle was also investigated. To this end, several
liquisolid tablets formulations containing various ratios of drug: solvent and different vehicles were
prepared. X-ray crystallography and differential scanning calorimetry (DSC) were used for
evaluation of physicochemical properties of indomethacin. Liquisolid formulations exhibited
significantly higher drug dissolution rates, in different dissolution media, compared to compacts
prepared by the direct compression technique. The results showed that enhanced dissolution rate of
indomethacin liquisolid tablets was due to an increase in wetting properties and surface area of drug
available for dissolution. In order to investigate the effect of aging on the hardness and dissolution
rate of liquisolid compacts, the formulations were stored at 25°C/75% relative humidity for a period
of 12 months. The results showed that aging had no significant effect on dissolution profile of
liquisolid tablets. Liquisolid compacts containing propylene glycol as vehicle produced higher
dissolution rate in comparison with other liquisolid compacts containing PEG 400 or polysorbate 80
with the same concentration. The results of DSC and X-ray crystallography did not show any
changes in crystallinity of the drug and interaction between indimethacin and exipients (Avicel and
silica) during the process.
3

P-227
EVALUATION ON CAPABILITY OF LIQUISOLID TECHNIQUE IN PREPARATION OF
SUSTAIN RELEASE FORMULATION OF PROPRANOLOL HYDROCHLORIDE
1,2 , 2 1,3
Y. Javadzadeh , L. Musaalrezaee , S. Asnaashari , A. Nokhodchi
'Department of Pharmaceutics, Faculty of Pharmacy-Tabriz University of Medical Sciences-Iran,
2 3
Drug applied research center, Tabriz University of Medical Sciences-Iran, Medway School of
Pharmacy, Central Ave, University of Kent and Greenwich, Chatham, Kent, ME4 4TB, England
Sustained release systems due to some advantages have special place in drug delivery field. There
are some methods for preparing sustained release systems which among them controlling of drug
dissolution is one of the best technique due to its simplicity and low cost. There are several
techniques for this mean that each one has own problems that made them useless in industrial field.
Liquisolid technique is new method that almost was used for enhancing dissolution rate of some
drugs. Using this system for preparing sustained release systems is new idea. It is claimed that such
systems could release drugs according to zero order kinetics such as osmotic pump systems. Then in
this research a new method was developed to retard a water soluble drug such as propranolol
hydrochloride using a simple and cheap method which is capable of producing in industrial scale.
For this mean Eudragit RS and RL and silica were selected as carrier and coating material
respectively. Different concentrations of drug in liquid medication were prepared. According to the
load factors, different tablets were compressed and dissolution test was done in two different media.
X-ray crystallography and DSC were used for evaluating of formation of any complex between
drug and excipients or any crystallinity changes during the process. Tablets were kept in laboratory
temperature for assessing the aging effect on hardness and dissolution profile of systems. The
results showed that liquisolid systems are able to retard the drug in such way that some formulation
released only 60 % of drug during 8 hr. These systems showed better retardation in compare with
matrix system. Except early minutes, zero order release was obtained. Aging had no effect on
hardness and dissolution profile of drug. X-ray crystallography and DSC ruled out any change in
crystallinity or complex formation during the process.
3

DEVELOPMENT OF AN HPLC METHOD FOR DETERMINATION OF TERBUTALINE
SULPHATE IN HEPATIC PERFUSION SAMPLES
Y. Karabey, S. Sahin, A. A. Hincal
Hacettepe University, Faculty of Pharmacy, Department of Pharmaceutical Technology, Division of
Biopharmaceutics and Pharmacokinetics, 06100 Ankara, Turkey
Terbutaline sulphate (TBS) is a synthetic P-adenoreceptor stimulant that is mainly used for the
treatment of bronchial asthma, chronic bronchitis and emphysema. Although variety of HPLC
methods have been extensively used to quantify TBS amount in pharmaceutical dosage forms, these
methods differ with respect to HPLC columns, detection methods and mobile phase components.
The aim of this study was to develop an HPLC method to be used for determination of TBS in the
hepatic outflow samples obtained from the isolated perfused rat liver preparation. Before analysis,
the hepatic perfusate samples containing TBS were filtered through a 0.45 pm filter, and then
injected into the HPLC system at a volume of 20 pi. Separation of TBS was performed at ambient
temperature on C ]8 column (25 cm x 4.6 mm, 10 pm) with UV detection at 270 nm, using a mobile
phase (pH 4.0) consisted of 150 mM ammonium acetate and methanol (90:10, v/v) which was
delivered at a flow rate of 2 ml/min. The proposed method was validated as to specificity, precision
(repeatability and reproducibility), linearity, accuracy, sensitivity, and stability. All standard
solutions for the calibration curve were prepared using blank perfusate obtained from the perfused
rat liver preparation. The calibration curve was characterized by its regression coefficient, slope,
intercept and lover limit of quantification. Under the conditions used, TBS was well separated from
the matrix components with a retention time of 6.24 min. The proposed method was linear in the
range of 1-120 pg/ml (r = 0.9999) with a lower limit of quantification of 1 pg/ml. The results of the
validation studies with low relative standard deviation values (e.g. less than 2% for repeatability and
reproducibility) indicated that the reverse phase HPLC method developed for analysis of TBS was
simple, rapid, sensitive and precise, and it can be successfully applied to the determination of this
compound in hepatic outflow samples.
P-
3

P-
EVALUATION ON CAPABILITY OF LIQUISOLID TECHNIQUE IN PREPARATION OF
SUSTAIN RELEASE FORMULATION OF PROPRANOLOL HYDROCHLORIDE
1,2 , 2 1,3
Y. Javadzadeh , L. Musaalrezaee , S. Asnaashari , A. Nokhodchi
Department of Pharmaceutics, Faculty of Pharmacy-Tabriz University of Medical Sciences-Iran,
2 3
Drug applied research center, Tabriz University of Medical Sciences-Iran, Medway School of
Pharmacy, Central Ave, University of Kent and Greenwich, Chatham, Kent, ME4 4TB, England
Sustained release systems due to some advantages have special place in drug delivery field. There
are some methods for preparing sustained release systems which among them controlling of drug
dissolution is one of the best technique due to its simplicity and low cost. There are several
techniques for this mean that each one has own problems that made them useless in industrial field.
Liquisolid technique is new method that almost was used for enhancing dissolution rate of some
drugs. Using this system for preparing sustained release systems is new idea. It is claimed that such
systems could release drugs according to zero order kinetics such as osmotic pump systems. Then in
this research a new method was developed to retard a water soluble drug such as propranolol
hydrochloride using a simple and cheap method which is capable of producing in industrial scale.
For this mean Eudragit RS and RL and silica were selected as carrier and coating material
respectively. Different concentrations of drug in liquid medication were prepared. According to the
load factors, different tablets were compressed and dissolution test was done in two different media.
X-ray crystallography and DSC were used for evaluating of formation of any complex between
drug and excipients or any crystallinity changes during the process. Tablets were kept in laboratory
temperature for assessing the aging effect on hardness and dissolution profile of systems. The
results showed that liquisolid systems are able to retard the drug in such way that some formulation
released only 60 % of drug during 8 hr. These systems showed better retardation in compare with
matrix system. Except early minutes, zero order release was obtained. Aging had no effect on
hardness and dissolution profile of drug. X-ray crystallography and DSC ruled out any change in
crystallinity or complex formation during the process.
3

P-
ENHANCING DISSOLUTION RATE OF HIGH DOSE WATER-INSOLUBLE DRUG
(CARBAMAZEPINE) USING LIQUISOLID TECHNIQUE
1,2 , 2 1,3
Y. Javadzadeh , B. Jafari Navimi Pour , S. Asnaashari , A. Nokhodchi
i
Department of Pharmaceutics, Faculty of Pharmacy-Tabriz University of Medical Sciences-Iran,
2 3
Drug applied research center, Tabriz University of Medical Sciences-Iran, Medway School of
Pharmacy, Central Ave., University of Kent and Greenwich, Chatham, Kent, ME4 4TB, England
Dissolution rate of water- insoluble drugs is the major problem in pharmaceutical sciences. There
are several methods for enhancing dissolution rate of water-insoluble drugs. Among them, liquisolid
technique is a new and promising technique for achieving this goal. Capability of this technique for
enhancing dissolution rate of drugs well established. Due to using more carrier material in
formulation of liquisolid tablets, there is a limitation in formulation of high dose drugs such as
carbamazepine as a tablet. In fact this method is applicable only for low dose drugs. In this research
we developed a new method to incorporate a high dose water- insoluble drug such as
carbamazepine to its liquisolid tablets. For this mean, different materials were used in preparing of
liquid medication in different concentration of the drug. Flowability and compressibility tests were
done for assessing of them. After formulation of a tablet, dissolution test was done about
formulation in two different media. For evaluating of aging on the hardness and dissolution rate of
the tablets, formulations were kept in laboratory temperature for 4 month. X-ray crystallography
and DSC were used for evaluating of formation of any complex between drug and excipients or any
crystallinity changes during the process. The results showed that our method is able to incorporate
high amount of the drug to the formulation that had well flowability and compressibility. This
means that this method can solve the most disadvantages of the liquisolid systems. Among the
material that was incorporated in liquid medication, PVP showed better results from view point of
flowability and dissolution rate. With increasing the amount of the PVP, better dissolution rate was
obtained. The formulation that had lower silica as a coating material had better dissolution rate.
Increasing the concentration of the drug in liquid medication showed higher dissolution profile.
Aging had no effect on hardness and dissolution profile of drug. X-ray crystallography and DSC
ruled out any change in crystallinity or complex formation during the process. Then with this new
method it is possible to formulate high dose drugs as a liquisolid tablets with enhanced dissolution
rate.
3

P-
EVALUATION OF ERYTHROMYCIN TOPICAL GEL RELEASE WITH TWO
METHODS
M. Jelvehgari, M.R. Rashidi, V. Rahmani
School of Pharmacy, Tabriz University of Medical Sciences
Erythromycin is a macrolyide antibiotic which its gel formulation is indicated for topical treatment
of acne vulgaris. It has less irritation, increase tolerability and better efficacy more than another
topical preparation use in acne. First the solubility of erythromycin in different solvents (water and
alcohol) was evaluated, then formulations were prepared using gelling agent (hydroxypropyl
cellulose). The stability of gels was evaluated in three different temperatures, refrigerator, room
temperatures and 40°C oven. The in vitro release of drug was assessed using static diffusion cell
with dialysis membrane and without dialysis membrane into the plate. The concentration of drug
was analyzed by means of UV spectrophotometer at the maximum wave length of 208.6 nm. The
results showed that release of drug in two methods follow zero order release mechanism. Also our
findings showed that increasing the amount of gelling agent in two methods increase the drug
release.
3

DEVELOPMENT OF AN HPLC METHOD FOR DETERMINATION OF TERBUTALINE
SULPHATE IN HEPATIC PERFUSION SAMPLES
Y. Karabev. S. Sahin, A. A. Hincal
P-230
Hacettepe University, Faculty of Pharmacy, Department of Pharmaceutical Technology, Division of
Biopharmaceutics and Pharmacokinetics, 06100 Ankara, Turkey
Terbutaline sulphate (TBS) is a synthetic P-adenoreceptor stimulant that is mainly used for the
treatment of bronchial asthma, chronic bronchitis and emphysema. Although variety of HPLC
methods have been extensively used to quantify TBS amount in pharmaceutical dosage forms, these
methods differ with respect to HPLC columns, detection methods and mobile phase components.
The aim of this study was to develop an HPLC method to be used for determination of TBS in the
hepatic outflow samples obtained from the isolated perfused rat liver preparation. Before analysis,
the hepatic perfusate samples containing TBS were filtered through a 0.45 pm filter, and then
injected into the HPLC system at a volume of 20 pi. Separation of TBS was performed at ambient
temperature on C ]8 column (25 cm x 4.6 mm, 10 pm) with UV detection at 270 nm, using a mobile
phase (pH 4.0) consisted of 150 mM ammonium acetate and methanol (90:10, v/v) which was
delivered at a flow rate of 2 ml/min. The proposed method was validated as to specificity, precision
(repeatability and reproducibility), linearity, accuracy, sensitivity, and stability. All standard
solutions for the calibration curve were prepared using blank perfusate obtained from the perfused
rat liver preparation. The calibration curve was characterized by its regression coefficient, slope,
intercept and lover limit of quantification. Under the conditions used, TBS was well separated from
the matrix components with a retention time of 6.24 min. The proposed method was linear in the
range of 1-120 pg/ml (r = 0.9999) with a lower limit of quantification of 1 pg/ml. The results of the
validation studies with low relative standard deviation values (e.g. less than 2% for repeatability and
reproducibility) indicated that the reverse phase HPLC method developed for analysis of TBS was
simple, rapid, sensitive and precise, and it can be successfully applied to the determination of this
compound in hepatic outflow samples.
348

P-
BIOEQUIVALENCE OF CEFDINIR DRY SUSPENSION AFTER SINGLE ORAL
ADMINISTRATION IN THAI HEALTHY VOLUNTEERS
S. Kasiwong', C. Ratanajamit 2 , D. Faroongsarng 3 , N. Phadoongsombut 4 , W. Jintapakorn 5
'Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences, Prince of Songkla
University, Thailand, department of Clinical Pharmacy, Faculty of Pharmaceutical Sciences,
Prince of Songkla University, Thailand, department of Pharmaceutical Technology, Faculty of
Pharmaceutical Sciences, Prince of Songkla University, Thailand, department of Internal
Medicine, Faculty of Medicine, Prince of Songkla University, Thailand.
Cefdinir is the third generation cephalosporin and widely used for both gram positive and gram
negative antibacterial. Cefdinir dry suspension is very useful for children patients; however its price
is quite high for most Thai patients. Thai local company has tried to develop this formulation and
prove the bioavailability with the original formulation. This study was to compare pharmacokinetic
parameters and bioequivalence of cefdinir dry suspensions in 16 Thai male volunteers. The method
was designed as a double-blind randomized, single dose, crossover study with 7 days washout
period. The 2 brands of cefdinir dry suspensions were Samnir® (Siam Bheasach Co Ltd, Thailand)
as test and Omnicef® (Warner Lambert, USA) as reference. Blood samples were collected at 0, 0.5,
0.75, 1, 2, 3, 4, 5, 6, 8, 10, 12 and 24 hours after 2xl25-mg/5 ml oral administration. Plasma
samples were determined by FIPLC, reverse phase (CI8), with UV detector and lower limit of
quantitation at 0.1 pg/ml. Pharmacokinetic parameters such as C max , AUC 0 .oc, T MAX and TI/ 2 were
calculated by WINNOLIN program and the logarithmic difference of ratio of AUC and C max were
evaluated by statistical ANOVA test. The pharmacokinetic data of two brands were 2.23±0.74 and
2.04+0.66 pg/ml for C max , 12.22+4.21 and 11.33+3.38 pg.ml/hr for AUC 0 .oc, 3.38+1.09 and
3.50+0.97 hour for T max and 2 hours for TI/ 2 , of test and reference, respectively. For statistical
comparison, the 90% confidence intervals of test/reference ratios, were 97.8-120.4% for C max and
94.3-117.9% for AUC 0 .oc, which were in the range of 80 -125%. For the results, it is concluded that
the two brands of cefdinir dry suspensions are bioequivalent for both rate and extent of drug
absorption.
3

P-234
AN IMPROVEMENT OF PHYSICOMECHANICAL PROPERTIES OF
CARBAMAZEPINE CRYSTALS
M. Maghsoodi. A. Nokhodchi, D. Hassanzadeh
Department of Pharmaceutics, School of Pharmacy, Tabriz Medical Sciences University, Tabriz
51664, Iran
In order to improve particle properties, new processes combining granulation and crystallization are
being developed. This work deals with the spherical crystallization process by the quasi-emulsion
mechanism applied to carbamazepine, a pharmaceutical drug. The aim of the present study was to
produce of spherical grains made of small crystals of a drug that have adequate properties for direct
compression when manufacturing tablets. Crabamazepine was' crystallized under different
conditions and the obtained spherical crystals were examined in terms of flow properties, particle
size analysis, compression and dissolution behaviors. Physical characteristics of the crystals were
studied for the morphology of crystals using scanning electron microscope, for the identification of
polymorphism by x-ray powder diffraction and for thermodynamic properties using differential
scanning calorimetry. The results showed that the agglomerates produced at 5 °C under stirring rate
of 300 rpm had superior flow than other agglomerates. Further more the results suggest that
agglomerates flow and pack smoothly from the hopper into the die and that tablets formed from
agglomerates attain uniformity in weight due to spherical shape of the treated samples. The results
showed that, generally, the treated carbamazepine samples (agglomerated forms) possessed superior
mechanical and better dissolution rate characteristics to untreated crystals. The results of DSC and
x-ray showed that untreated sample and agglomerates were form III and form I of carbamazepine
respectively.
32

P-231
BIOEQUIVALENCE OF CEFDINIR DRY SUSPENSION AFTER SINGLE ORAL
ADMINISTRATION IN THAI HEALTHY VOLUNTEERS
S. Kasiwong 1 . C. Ratanajamit 2 , D. Faroongsarng 3 , N. Phadoongsombut 4 , W. Jintapakorn 3
'Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences, Prince of Songkla
University, Thailand, department of Clinical Pharmacy, Faculty of Pharmaceutical Sciences,
Prince of Songkla University, Thailand, department of Pharmaceutical Technology, Faculty of
Pharmaceutical Sciences, Prince of Songkla University, Thailand, department of Internal
Medicine, Faculty of Medicine, Prince of Songkla University, Thailand.
Cefdinir is the third generation cephalosporin and widely used for both gram positive and gram
negative antibacterial. Cefdinir dry suspension is very useful for children patients; however its price
is quite high for most Thai patients. Thai local company has tried to develop this formulation and
prove the bioavailability with the original formulation. This study was to compare pharmacokinetic
parameters and bioequivalence of cefdinir dry suspensions in 16 Thai male volunteers. The method
was designed as a double-blind randomized, single dose, crossover study with 7 days washout
period. The 2 brands of cefdinir dry suspensions were Samnir® (Siam Bheasach Co Ltd, Thailand)
as test and Omnicef® (Warner Lambert, USA) as reference. Blood samples were collected at 0, 0.5,
0.75, 1, 2, 3, 4, 5, 6, 8, 10, 12 and 24 hours after 2xl25-mg/5 ml oral administration. Plasma
samples were determined by HPLC, reverse phase (CI8), with UV detector and lower limit of
quantitation at 0.1 pg/ml. Pharmacokinetic parameters such as C max , AUC 0 .oc, Tm ax and T1/2 were
calculated by WINNOLIN program and the logarithmic difference of ratio of AUC and C max were
evaluated by statistical ANOVA test. The pharmacokinetic data of two brands were 2.23+0.74 and
2.04+0.66 pg/ml for C max , 12.22+4.21 and 11.33+3.38 pg.ml/hr for AUC 0 . K , 3.38+1.09 and
3.50+0.97 hour for T max and 2 hours for T1/2, of test and reference, respectively. For statistical
comparison, the 90% confidence intervals of test/reference ratios, were 97.8-120.4% for C max and
94.3-117.9% for AUCo-*, which were in the range of 80 -125%. For the results, it is concluded that
the two brands of cefdinir dry suspensions are bioequivalent for both rate and extent of drug
absorption.
3

P-232
FORMULATION AND CLINICAL EVALUATION OF MYRTUS MUCOADHESIVE
PASTE IN THE TREATMENT OF RECURRENT APHTHOUS STOMATITIS
1 2 3 4
P. Khazaeli , G. Chamani , M. Mehrabani , N. Mohammadi
'Pharmaceutics, School of Pharmacy, Kerman Univ. of Med. Sci, Iran, 2 Oral Medicine, Dental
School, Kerman Univ. of Med. Sci, Iran, 3 Pharmacogenozy, School of Pharmacy, Kerman Univ. of
Med. Sci, Iran, 4 Pharmacist, Iran
Aim: Recurrent aphthous stomatitis (RAS) is an oral lesion with high prevalence. Its management is
directed toward treatment of symptoms. Myrtle is an herbal drug that has been used for RAS
treatment during last years. Its essential oil has antiseptic and antimicrobial properties. One of the
best factors in the treatment of aphthous lesion is, remaining of drug in location, so, a dosage form
was tried to design that can hold the drug for a long time in location. For this purpose mucoadhesive
drug delivery system was selected. Method & Material: The essential oil was extracted from myrtle
leaf by using water in Clevenger apparatus. Oral mucoadhesive paste was prepared by
compounding of sodium carboxy methyl cellulose, pectin and gelatin in plastibase. The best
formulations were selected for clinical trial. In this double blind clinical trial, individuals were
divided into two groups, A and B that received Mocuadhesive paste containing myrtle essence and
mucoadhesive paste without drug as placebo, respectively. The size of lesions was measured by
periodontal probe at 1, 2, 6 and 10 days. Also by use of VAS scale, daily pain was measured at 10
days period. Results: The time of burning sensation relief in A, B groups were 1.64±0.74 and
4.91±1.30 days respectively, which there was significant difference between two groups (p

P-233
COMPACTION CHARACTERISTICS OF ETHYLCELLULOSE IN THE PRESENCE OF
SOME CHANNELING AGENTS
M.O. Emeje, P.O. Kunle
Department of Pharmaceutical Technology and Raw Materials Development
National Institute for Pharmaceutical Research and Development (NIPRD) Idu,
P.M.B.21 Garki - Abuja, Nigeria
The purpose of this study was to investigate the effect of some commonly used release enhancers on
the compaction characteristics of ethylcellulose. The wet granulation method of massing and
screening was used and compacts were produced by compressing granules for 60 seconds at various
compression pressures. The Heckel equation was used for the analysis of results which shows that
ethylcellulose alone showed better compressibility than formulations with additives. The
hygroscopic additives; sorbitol, and polyethylene glycol 4000 produced triphasic Heckel plots,
while the non hygroscopic additives; polyethylene glycol 10000 and mannitol produced biphasic
plots. The results show that the presence of additives reduced the degree of packing of
ethylcellulose in the die with mannitol having the most effect. The presence of additive in
ethylcellulose formulations increased the pressures at which plastic deformation of the granules
occurred. The extent of this was dependent on the type of additive, with mannitol imparting the
highest resistance to deformation of ethylcellulose granules and sorbitol the least. The study showed
that polyethylene glycol 4000, 10000, sorbitol and mannitol affect the plasticity of ethylcellulose
granulations, the extent and nature of the effect is dependent on the nature of the additive.
3

P-234
AN IMPROVEMENT OF PHYSICOMECHANICAL PROPERTIES OF
CARBAMAZEPINE CRYSTALS
M. Maghsoodi, A. Nokhodchi, D. Hassanzadeh
Department of Pharmaceutics, School of Pharmacy, Tabriz Medical Sciences University, Tabriz
51664, Iran
In order to improve particle properties, new processes combining granulation and crystallization are
being developed. This work deals with the spherical crystallization process by the quasi-emulsion
mechanism applied to carbamazepine, a pharmaceutical drug. The aim of the present study was to
produce of spherical grains made of small crystals of a drug that have adequate properties for direct
compression when manufacturing tablets. Crabamazepine was' crystallized under different
conditions and the obtained spherical crystals were examined in terms of flow properties, particle
size analysis, compression and dissolution behaviors. Physical characteristics of the crystals were
studied for the morphology of crystals using scanning electron microscope, for the identification of
polymorphism by x-ray powder diffraction and for thermodynamic properties using differential
scanning calorimetry. The results showed that the agglomerates produced at 5 °C under stirring rate
of 300 rpm had superior flow than other agglomerates. Further more the results suggest that
agglomerates flow and pack smoothly from the hopper into the die and that tablets formed from
agglomerates attain uniformity in weight due to spherical shape of the treated samples. The results
showed that, generally, the treated carbamazepine samples (agglomerated forms) possessed superior
mechanical and better dissolution rate characteristics to untreated crystals. The results of DSC and
x-ray showed that untreated sample and agglomerates were form III and form I of carbamazepine
respectively.
352

P-235
DESIGNING AND EVALUATING THE PHYSICOCHEMICAL PROPERTIES OF A
HYDROGEL BASED ELECTRODE IN IONTOPHORETIC DELIVERY DEVICES
D. Hassanzadeh, F. Monajemzadeh. A. Beihaghy
Department of Pharmaceutics, Faculty of pharmacy, Tabriz University of Medical Sciences, Tabriz,
Electrodes are the most sensitive part of the iontophoretic devices and can be classified into two
broad groups: Inert and reversible electrodes. Reversible electrodes differ from inert ones in that
they don't cause water electrolysis but they make proteionous drugs to precipitate. Because of these
drawbacks, hydrogel based electrodes have been developed recently which are more compatible
with human skin and can be prepared easily. The purpose of the present study is evaluating the
physicochemical properties of a hydrogel based electrode during direct current iontophoresis.
Different hydrogels were prepared using HPMC K4M, K15M and K100M and PVPK25, as the
polymeric agents and citrate -phosphate buffer as the dispersing medium. An ionthophoretic device
was designed and the hydrogel was placed in the donor phase which was separated from the
receptor compartment by a dialysis membrane and the current intensity was maintained constant
during the experiment (0.38 mA/cm 2 ). The pH values, voltage and conductivities were recorded at
certain time intervals up to 120 minutes and the impedance values were calculated and reported for
each experiment. All the results were a statistically analyzed by ANOVA test which was performed
using SPSS software. The results revealed that, the physicochemical properties of the hydrogels
prepared with HPMC K4M, remains almost unchanged during a 120 minute constant current
iontophoretic delivery. Thus it can be a good candidate in drug delivery using hydrogel electrodes
with low drug instability causing problems.
3

P-236
FORMULATION OF PAROMOMYCIN SULFATE NIOSOMES
M.H. Moshafi. A. Pardakhti, H. Daneshvar, M. Koohzad
Dept. of Pharmaceutics, School of Pharmacy, Kerman University of Medical Sciences
Parasitic contaminations are one of the major problems in different countries. One of these
infections is cutaneous leishmaniasis caused by Leishmania major. Different therapeutic agents
and dosage forms were used for treatment of this disease such as sodium stibogluconate,
meglumine antimonite and paromomycin. Local application of paromonycin sulfate ointment in
cutaneous leishmaniasis has produced different effects and in some cases it was not effective. It
has been proposed that weak penetration of this drug to contaminated macrophages was the main
reason of ineffectiveness of it. For overcoming this problem various penetration enhancers must be
used. One of the penetrations enhancers is vesicular drug delivery systems such as liposomes and
niosomes. In thes study the niosomal formulations of paromomycin sulfate were prepared by using
polyoxyethylene alkyl ethers (Brij 52, 58 and 92), cholesterol and sodium lauryl sulfate as a
negatively charged inducing agent. The used methods for noisome preparation were classic fdm
hydration and sonication. The size distribution, (measured by laser light diffraction method)
physical stability, morphological characterization and encapsulation efficiency of paromomycin
sulfate niosomal formulations were studied. Paromomycin sulfate concentration was measured by
a microbiologic method ( agar diffusion ) against S. epidrmidis Cholesterol molar percent increase
resulted in size reduction in Brij 52 and 58 niosomes and a conterary result in liquid state Brij 92
ones [1], Paromomycin sulfate encapsulation efficiencies were reduced after sodium lauryl sulfate
molar ratio increment, possibly to competition between these two charged water-soluble agents.
Narrow size distribution, high encapsulation efficiency good stability and uniform shapes of
niosomes were the major aspects of suprior noisome formulation choice. Microscopical
(observation) revealed round and multilamellar vesicles (MLVs) in the most cases. Topical
administration of selected formulations in patient infected with L.major will be evaluated in future
studies.
1. Devaraj GN, Parakh SR, et al. Journal of Colloid and Interface Sciences. 2002, 251: 360-
365.
3

P-237
INVESTIGATIONS ON TABLETS OF IBUPROFEN SOLID DISPERSIONS
N. O. Sahin, C. Erdogan, K. Mut
Mersin University, Faculty of Pharmacy, Department of Pharmaceutics, Yenisehir Campus, Mersin
33169, Turkey
| Ibupofen is a nonsteroidal anti-inflammatory drug, which is not soluble in water and causes
gastrointestinal problems. In order to improve the aqueous solubility of the drug and enhance the
dissolution rate, physical mixture (PM) and inclusion complex (INC) of ibuprofen with skimmed
milk were prepared employing liyophilization technique in our previous studies. Enhancement of
aqueous solubility of ibuprofen was achieved upon formation of INC. In vivo studies conducted on
rats revealed reduction of gastric disorders. In the present study, we aimed to prepare tablets of
ibuprofen solid dispersion and physical mixtures. Tablets were prepared by direct compression and
characterized by means of dimension, height, content uniformity, and hardness. In vitro dissolution
profiles of tablet formulations were also investigated according to the method given in USP XXII.
120 n
c
3 90 -
O
E 60 -
re
•o
0) (A 30
re
CC
0) »
0 H
I I
20 40
60 80
Time (minute)
100
—TB
—SD-IB
Figure 1. Comparison of release profiles of ibuprofen from tablets and solid dispersions
3

P-238
ANTIBIOTIC DELIVERY SYSTEMS FOR CYSTIC FIBROSIS PATIENTS
M. Alipour, A. Omri
Department of Chemistry and Biochemistry, Laurentian University, Sudbury, Ontario, Canada
Cystic fibrosis is the most common lethal autosomal recessive disease found in the Caucasian
population with a frequency of approximately one in 2,500 births. Chronic respiratory infections
with Pseudomonas aeruginosa are the leading cause of morbidity and mortality in such individuals.
Once Pseudomonas aeruginosa colonizes the cystic fibrosis patient's lung, it cannot be eradicated
even by the most aggressive antibiotic therapy. The aim of this work is to evaluate bactericidal
activity of liposomes-entrapped aminoglycosides against clinical isolates of Pseudomonas
aeruginosa from cystic fibrosis patients. Gentamicin, amikacin, and tobramycin were incorporated
into liposomes prepared by the modified dehydration-rehydration method. The sizes and the
encapsulation efficiencies of these vesicles were determined. The minimum inhibitory
concentrations and the time-killing curves of free and liposomal drugs against clinical isolates of
Pseudomonas aeruginosa were assessed. The average liposomal size was below 400 nm in
diameter. The encapsulation efficiency ranged from 10 ± 1% for gentamicin, to 36 ± 2% for
amikacin, and 15 ± 2% for tobramycin. The minimum inhibitory concentrations of liposomal
aminoglycosides for clinical isolates of Pseudomonas aeruginosa were lower compared to the
corresponding free drugs. The most notable difference was seen for a highly resistant strain PA-
48912-2 which displayed a minimum inhibitory concentration of 16 mg/L for liposomal amikacin
compared to 256 mg/L for the free drug. In addition, the time-Killing values for liposomal
aminoglycosides were either equivalent to or better than that of the free antibiotics. Liposomal
aminoglycosides are more potent anti-pseudomonal antibiotics with improved killing time and
prolonged antimicrobial activity and warrant further pre-clinical investigations into the use of these
formulations for the treatment of chronic pulmonary infections.
3

P-239
GENTAMICIN ACTIVITIES OF PHOTOPOLYMERIZED POLY (ETHYLENE GLYCOL)
DIACRYLATE (PEG-DA) AND 2-HYDROXYETHYL METHACRYLATE (HEMA)
HYDROGELS: IN-VITRO
S. Ozkan'. F. Ayhan 2 , U. Abbasoglu 1
' Gazi University, Faculty of Pharmacy, Department of Pharmaceutical Microbiology, Ankara,
Turkey, 2 Mugla University, Faculty of Science, Department of Chemistry, Kotekli, Mugla, Turkey
Hydrogels based on poly(ethylene glycol)-diacrylate (PEG-DA) and 2-hydroxyethyl methacrylate
(HEMA) were polymerized with crosslinking agent ethylene glycol diacrylate (EGDMA) under
mild photoinitiating conditions (0.5 wt % initiator 2,2-dimethoxy-2-phenylacetophenone (DMPA),
10 mW/cm 2 of 365 nm light and 5 min). PEG-DA and HEMA concentrations of disks which have
1±0.3 mm thickness, were 30 % and 50 % w/w and 40 % and 60 % w/w, respectively. 10 mg/ml
Gentamicin sulphate was incorporated into the hydrogel during photopolymerization and its release
kinetics was tested by spectrophotometric method at 255 nm wavelenght in phosphate buffer (pH
7.4) and citrate buffer (pH 2.2). The drug release in phosphate buffer is faster as compared to citrate
buffer. With decreasing polymer percentage, the rate of drug release is almost 75 % of the loaded
drug within 4 hours in phosphate buffer. Antimicrobial efficiency of the samples was tested by agar
diffusion method in two different bacterial cultures (,Staphylococcus aureus ATCC 25923,
Pseudomonas aeruginosa ATCC 10145). The diameter of the inhibition zone (mm) surrounding
each sample was measured after 24 h incubation of drug loaded disks onto agar plates at 37°C. Agar
diffusion test results also confirm that polymerization conditions did not adversely affect the
antimicrobial activity of gentamicin sulphate.
3

P-240
KINETIC RELEASE OF CHLORPHENIRAMINE MALEATE FROM DIFFERENT
VESICULAR, PRONIOSOMAL AND TOPICAL FORMULATIONS
1 2 2
A. Pardakhty , J. Varshosaz , S.M. Hossaini
1
Department of Pharmaceutics, Kerman University of Medical Sciences, Iran, PO Box 76175-493,
2
Department of Pharmaceutics, Isfahan University of Medical Sciences
Vesicular systems are considered very promising to overcome the permeation barrier of the skin,
especially the upper layer, i.e. stratum corneum. Furthermore, they can be used as vehicles for
controlled percutaneous drug delivery. We have reported the sorbitan monopalmitate-based
proniosomes for transdermal delivery of chlorpheniramine maleate [CPM] (1). The objective of this
study was the assessment and comparison of topical preparations effect on the release kinetic of
CPM. For proniosomal formulations, lipids (span 40 and cholesterol [Choi] or dicetylphosphate
[DCP] or egg lecithin) with alcohol (ethanol, propanol or isopropanol) and aqueous phase
containing 2% of CPM were mixed at 70°C until a clear solution was formed that on cooling
converted to a proniosomal gel. Niosomes were prepared by hand-shaking method 50°C.
Hydrophilic ointment (USP) and a w/o cream with 2% CPM also were formulated as non vesicular
preparations.The particle size and particle size distribution of niosomes prepared directly or from
proniosomes, were determined by laser-light scattering method (Mastersizer X, Malvern
Instruments, UK). For encapsulation efficiency measurement and release study, CPM was analyzed
spectrophotometrically at 261 nm. Release tests were performed by a Franz- cell using cellulose
nitrate membrane with 0.1 pm pore size at 37°C. Different formulations were compared for their
dissolution efficiencies over 6 h (DE6%). Inclusion the negatively charged DCP both in
proniosomes and niosomes increased the mean volume diameter of vesicles probably due to
increasing the surface free energy of them. Brij 52 containing niosomes also was larger than brij 72
ones (p< 0.05) apparently to higher HLB or hydrophilicity of the first surfactant. Whereas the
highest entrapment efficiency in proniosomes were 15.7±0.99% related to span 40/Chol/lecithine
with ethanol, brij 72/Chol and span 60/Chol niosomes had encapsulation efficiencies more than
40%. Our results showed that vesicles formed from different alcohols were of different size and
followed the order: ethanol> propanol> isopropanol. In vitro release studies provide valuable
information as tot the product behavior in vivo. As the results of DE60% of CPM from different
proniosomal formulations indicate, regardless of the type of alcohol, all series of vesicles containing
lecithin show lower DE60% than those with DCP (p

P-241
CREAM FORMULATION AND SPF DETERMINATION OF LAWSONIA INERMIS
EXTRACT
M. Rezaeifar,' P. Khazaeli 1 , F. Sharififar 2 , G.R. Dehghan 1
'Department of Pharmaceutics, School of Pharmacy, Kerman University of Medical Science,
Kerman, Iran, department of Pharmcogenozy, School of Pharmacy, Kerman University of Medical
Science, Kerman, Iran
The proper properties of Lawsonia inermis (Henna) in moisturizing, softening and antiinflammation
in skin, has been established since many years ago. The UV absorption spectrum from
Lawsonia extract led us to evaluate the antisolar activity of this extract; Lawson is a color material
with naphthoquinone structure which can find in Henna leaves (1). The amount of Lawson based on
different plant origins were reported between 0.2- 1% (2). There are some other components in
henna leaves such as Laxanthon I, II, Beta sitosterol glucoside, Luteolin, Luteolin-7- O- glucoside
and Acacetin- 7- O- glucoside (3). In this study fresh leaves of henna were collected at April from
Shadad and Rigan region, south east of Iran. The collected leaves were dried and so milled to 36
mesh. Maceration method was used for extraction. 15Og of powdered leaves were macerated by
using saturated solution of sodium carbonate in water for 24 hours. The aqoues phase was collected
and acidified with hydrochloric acid, so mixed with sufficient amount of chloroform and shaked
vigorously. The organic phase was collected and dehydrated with sodium sulfate. This extract was
concentrated with rotary evaporation at vacuum condition. Lawson was confirmed is this extract by
use IR spectrum. In formulation step, cream bases were formulated at different HLB values between
5 -14. The cream base with HLB value equal to 9 was selected as best one. Also, between four
methods in cream preparation, the addition of aqoues phase to oil phase containing surfactants was
selected. Then the cream bases containing Lawson (3%) were formulated. For In vitro determination
of SPF, three different concentrations from this antisolar cream were prepared in methanol. By
transmittance reading in a range of wavelengths, PF, MPF, and so SPF was calculated. The SPF
value for formulated cream containing Lawson was 3.88. The results showed that Lawson can not be
used as an effective anti- solar agent alone, but it can be used as a tanning agent and also as a
booster for antisolar activity of other active ingredients.
1- Trease, G.E, Evans W.G. "A text book of Pharmacogenozy" 1983, London, pp 171-80.
2- Latif, A; Isolation of a vit - k active compound from the leaves of Lawsonia, chemical
composition of the air- dried leaves, Indian, J.Agr, Sci, 29 (2), pp. 147-150 (1950).
3- Mahmoud, Z.f, et al; Constituent of henna leaves growing in Egypt, fitoterapia, pp. 51, 153
(1980).
3

P-242
INFLUENCE OF STABILIZERS AND pH OF EXTERNAL AQUEOUS PHASE ON THE
CHARACTERISTICS OF POLY (e-CAPROLACTONE) MICROPARTICLES PREPARED
BY A MULTIPLE EMULSION (W/OAV) SOLVENT EVAPORATION TECHNIQUE
O. Sonakin, A. Karatas, M. Kiln^arslan, T. Baykara
Department of Pharmaceutical Technology, Faculty of Pharmacy, University of Ankara, 06100,
Tandogan, Ankara, Turkey
In this study levobunolol HC1 loaded poiy(s-caprolactone) (PCL) microparticles were prepared by
the multiple emulsion (W/O/W) solvent evaporation technique and it is aimed to examine the
effects of two process factors, type and concentration of stabilizer and pH of external aqueous
phase, on the encapsulation efficiency, particle size and drug release of the microparticles. Solvent
evaporation technique based on W/O/W method has been prepared as an alternative method for
encapsulation of hydrophilic drugs as levobunolol HCI which is commonly used nonselective
ophthalmic beta-blocker. PCL is well-known polymer because of excellent tissue compatibility,
biodegradability and existing regulatory approval. The presence of a surfactant is of significant
importance to influence the colloidal stability and particle size in this method. Therefore polyvinyl
alcohol (PVA) and pluronic F68 (PF68) are used as stabilizers of the emulsion at the different
concentrations varied from 0.25 to 1.5 % (w/v) in the external aqueous phase of pH 5.5 and 12
individually. The highest encapsulation efficiency and drug release were obtained from the
microparticles prepared with the 0.5 % PVA concentration at the both pH 5.5 and 12 of external
phase. It is determined that particle sizes of microparticles decreased with increased PVA
concentration. But increasing the concentration of PVA up to 1 and 1.5 resulted in an aggregation of
the particles leading to an increase of particle size. At the higher pH of external phase the
encapsulation efficiency and particle size were higher than that of pH 5.5. Whenever the pH of
external phase exceeds the pKa of levobunolol HCI (pKa 9.5) increases in drug entrapment (from
3.78 to 30.6) were observed because of reducing in the degree of drug ionization. As for PF68,
lower encapsulation efficiency and particle sizes (3 pm) were obtained. Due to the smaller particle
sizes of these particles the drug release was higher than that of microparticles prepared by PVA. In
conclusion this study revealed that levobunolol HCI loaded microparticles could be prepared by
W/O/W solvent evaporation technique and the higher pH of external phase from the pKa of drug
and also 0.5 % PVA concentration ensure a good emulsification process and therefore leads to
smaller particles with the highest encapsulation efficiency.
This work has been supported by Management of Scientific Research Projects of Ankara University
30

P-243
ANTIMICROBIAL ACTIVITY OF HYDROPHILIC OINTMENT BASE AND NON-IONIC
CREAM BASE CONTAINING ESSENTIAL OIL FROM KAEMPFERIA GALANGA
1 2 1 2
A. Sunthornpit, S. Yuenyongsawad , D. Maneenuan , S. Kummee
'Department of Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, Prince of Songkla
2
University, Songkhla, 90112, Thailand, Department of Pharmacognosy and Pharmaceutical
Botany, Faculty of Pharmaceutical Sciences, Prince of Songkla University, Songkhla, 90112,
Thailand
The essential oil (K. galanga oil) extracted from the rhizome of Kaempferia galanga Linn.
(Zingiberaceae) has been reported to show antimicrobial activity against Staphylococci aureus,
Streptococcus faecalis, Bacillus subtilis, Samonella typhi, Shigella flexneri, Escherichia coli and
Candida albicans. The present study aimed to determined the minimum inhibition concentration
(MIC) of K. galanga oil against S. aureus and C. albicans and to evaluate an antimicrobial activity
of Hydrophilic ointment base containing 5 % w/w K. galanga oil and Non-ionic cream base
containing 5 % w/w K. galanga oil against S. aureus and C. albicans at initial and after storage for
6 months at room temperature (30° C). The fresh rhizomes of K. galanga were extracted by water
distillation.The essential oil (0.30%w/w) obtained was then collected and stored at 4 °C.
Hydrophilic ointment base (composed of stearyl alcohol, white petrolatum, sodium lauryl sulfate,
propylene glycol, methylparaben, propylparaben and purified water), Non-ionic cream base
(composed of cetostearyl alcohol, cetomacrogol 1000, white soft paraffin, liquid paraffin, propylene
glycol paraben concentrate and purified water) were prepared by separated heating of water phase
and oil phase then mixed and K. galanga oil 5 % w/w was added at 45C, The K. galangal oil
creams prepared were namely according to cream bases; Hydrophilic ointment base(cream A),Nonionic
cream base(cream B). The MIC of K. galanga oil against S. aureus and C. albicans were
determined by microdilution broth method. Antimicrobial activity of A and B at initial and after 6
month storage were assayed by the agar-cup diffusion method. Gentamycin cream, Clotrimazole
cream and Tea tree oil cream were used as positive control. The result showed that MIC of K.
galanga oil against S. aureus ATTC25923 and C. albicans were 1 mg/ml and 4 mg/ml respectively.
For antimicrobial activity, cream A kept at 30°C for 6 months showed significantly increased
inhibition zone diameter, 27.43+0.92 mm against S. aureus compared to initial, 13.17+ 0.25 mm
(p

P-244
PREPARATION AND CHARACTERIZATION OF SOLID DISPERSIONS OF
PIROXICAM WITH HYDROPHILIC CARRIERS
H. Valizadeh. P. Zakeri-Milani, A. Nokhodchi
Department of Pharmaceutics, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz,
Iran, Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
The objective of this study was to improve the dissolution rate of a poor water soluble drug,
piroxicam, by solid dispersion systems using mannitol, dextrin, myrj 52 and Eudragit® El00. Solid
dispersions were prepared by three different methods depending on the type of carrier, and
evaluation of the dispersions properties was performed using solubility measurement, dissolution
studies, Fourier-transform infrared spectroscopy and X-ray powder diffractometry. The dissolution
rate of piroxicam was markedly increased in solid dispersion of myrj 52, Eudragit® El 00 and
mannitol. Dissolution of the drug from solid dispersions prepared with dextrin was slower than
physical mixtures. Solubility studies revealed a marked increase in the solubility of piroxicam with
an increase in myrj 52 and Eudragit® El00 concentrations. Mannitol and dextrin concentrations
had no effect on the solubility of piroxicam. Data from the X-ray diffraction and FT-IR
spectroscopy showed that piroxicam was amorphous in the solid dispersions prepared with dextrin
and Eudragit® El 00. Crystallinty of piroxicam was decreased in the solid dispersions prepared with
Mannitol, but in the case of Myrj52 there was not any difference between the crystallinity of
piroxicam in physical mixtures and solid dispersions.
32

P-245
PREPARATION AND DISSOLUTION KINETICS OF MEFENAMIC ACID
MICROSPONGES
A. Yurdasiper, B. Kaynarsoy, F. Sevgi
Department of Pharmaceutical Technology, Faculty of Pharmacy, Ege University,
35100, Bornova, Izmir, Turkey
In this study, we aimed to prepare mefenamic acid (MFA) loaded microsponges by emulsionsolvent
diffusion method (1). MFA is a non steroidal anti inflammatory drug (NSAID) possessing
analgesic and antipyretic activity. Its reported half life is 2 hours. The short half life and increased
need of patient compliance, especially in the management of chronic rheumatic conditions, suggest
the need for controlled release preparations of MFA. Eudragit RS 100 was used as polymer. The
organic internal phase containing MFA, Eudragit RS 100 and triethylcitrate (TEC) was gradually
added to distilled water which contained PVA as emulsifying agent. TEC was used as a plastifier.
The mixture was stirred for 2 hours at 25 °C to remove dichloromethane from the reaction flask.
The formed microsponges were filtered and washed with distilled water before being air dried. For
the evaluation of the effect of drug:polymer ratio on the physical characteristics and release profiles
of microsponges, different weight ratios of drug to Eudragit RS 100 (1:1, 3:1, 5:1, 7:1, 9:1) were
employed. Although the total amount of polymer and emulsifier were kept constant, the volume of
external phase were changed (2). Drug release was investigated using the paddle method of the USP
XXV. Phosphate buffer (pH 7.4) were used as dissolution media (3). The amount of drug was
determined by spectrophotometry at 285nm. Dissolution profiles of the microsponges were plotted
and evaluated kinetically.
1. Comoglu T, Goniil N, Baykara, T, Int. J. Pharm. 242, 2002, 191-195
2. Jelvehgari M, Siahi-shadbad M R, Azarmi S, Martin G P, Nokhodchi, A, Int. J. Pharm,
2005,9-18
3. Kawashima Y, Iwamoto T, Niwa T, Takeuchi T, Hino, T, J. Microencap. 10, 1993, 329-340
3

P-246
POSSIBILITY OF USING EUDRAGIT® E100/ CARBOMER 940P INTERPOLYEIJ

P-247
MEMBRANE TRANSPORT OF DRUGS IN INTESTINAL TRACT OF THE RAT AND ITS
CORRELATION WITH HUMAN INTESTINAL ABSORPTION
1 1 2 1 1
P. Zakeri-Milani , H. Valizadeh , H. Tajerzadeh , Y. Azarmi , M. Barzegar-Jalali ,
Z. Islambolchilar , S. Barzegar
'Department of Pharmaceutics, Faculty of Pharmacy, Tabriz University of Medical Sciences,
2
Tabriz, Iran, Department of Pharmaceutics, Faculty of Pharmacy, Tehran University of Medical
Sciences, Tehran, Iran.
The prediction of human intestinal permeability and fraction absorbed of oral dose using single-pass
intestinal perfusion technique (SP1P) in rats was the aim of study. Permeability coefficients in
anaesthetized rats were determined for 14 compounds. Drug solution in phosphate buffered saline
(PBS) was perfused through the intestinal segment with flow rate of 0.21 ml/min and samples were
taken from outlet tubing at different time points up to 90 min. Phenol red was used as a nonabsorbable
marker to correct water flux through the segment. Drug concentrations in samples were
determined using HPLC equipment and permeability coefficients were calculated. The results
showed approximately 12.5 times difference in magnitude for rat permeability coefficients of a
series of passively absorbed compounds. These values were compared with published data for
human intestinal permeability, and a strong correlation was found between P e ff (rat) and P e ff
(human); (Peff (human) = 11-04 P e ff (rat) - 0.0003; R 2 = 0.93, P0.0001). Consequently in human
the fraction dose absorbed (F a ) was estimated and predicted after oral dosing considering
Fa(human)=l-e " 3928 ° Peffi(rat) ( R 2= 0-91> p

P-248
CALLUS PRODUCTION IN CENTAUREA TCHIHATCHEFFII (YANARDONER)
H. Colgecen 1 . C. Yagci 2 , M.C. Toker 2 , G. Toker 3
'Zonguldak Karaelmas University, Faculty of Arts and Science, Department of Biology, 67100
Incivez, Zonguldak,Turkey,
2 Ankara University, Faculty of Science, Department of Biology 06100
Tandogan/Ankara, 3 Gazi University, Faculty of Pharmacy, Department of Pharmacognosy, Etiler -
Ankara 06330
Centaurea tchihatcheffii Fich. & Mey (Yanardoner, Peygamber 9i

P-249
SPECTROSCOPIC STUDIES ON THE INTERACTION OF HUMAN SERUM
(HSA) WITH SURFACTIN
ALBUMIN
1 2 3
G. Dehghan Noudeh , M. Housaindokht , B.S. Fazly Bazzaz
I Department of Pharmaceutics, School of Pharmacy, Kerman University of Medical Sciences,
2
Kerman, Iran , Department of Chemistry, School of Sciences, Ferdowsi University, Mashhad,
Iran, Department of Medicinal chemistry, School of Pharmacy, Mashhad University of Medical
Sciences, Mashhad, Iran
We were studied interaction of HSA with surfactin in solution as a function of surfactin
concentrations by using fluorescence and circular dichrosim spectroscopies. The different effects of
surfactin upon binding to HSA are interpreted based on considerations of the expected changes in
the vicinity of tryptophan residue W 2^ [i]. The results of fluorescence and CD showed that HSA
has partially unfolded at pH 3.0 and 10.0 in comparison with pH 7.0. At pH 7.0 and 10.0 surfactin
at low concentrations caused a red shift of maximum emission and at high concentrations a blue
shift of maximum emission, which should mean unfolding after partial folding. But, in compare
with other pH values (pH 7.0 and 10.0), in pH 3.0, surfactin does not have considerable effect.
Stability of HSA has been obtained at pH 7.0 and 10.0. DG(H20) was about lSkJmol" 1 at pH 7.0
and pH 10.0. The results led us to conclude that the degree of exposed groups is highly dependent
on pH in surfactin media and indicated the presence of a cooperative process in the surfactin
denaturation medium at pH 7.0 [1].
1. Gelamo, E.L, Tabak, M, 2000. Spectrochimica Acta Part A, 56, 2255-2271.
3

P-250
ALTERATION OF FLAVONOID COMPOSITION VIA BIOTECHNOLOGICAL
METHODS
U. Koca'. G.A. Moore 2
'Gazi University, Faculty of Pharmacy, Department of Pharmacognosy, Etiler, Ankara/Turkey,
horticultural Sciences Department, Plant Molecular and Cellular Biology Program, University of
Florida, Gainesville FL 32611. USA
Flavonoids are widely distributed diverse groups of plant secondary metabolites with important
functions as protective filters against UV-B light damage, as pigments to attract pollinators and
have key roles in pollen growth, and plant-microbe signaling. Moreover, they contribute numerous
dietary health benefits to human being due to their antioxidant, anticancer, anti-inflammatory,
immunostimulan, and antithrombogenic properties. Flavanone glycosides are the main group of
flavonoids in citrus, which accumulate in entire plants. The presence of some of these compounds
affects the taste of citrus fruits, rather than influencing their color. Flavanone rutinosides are
tasteless, whereas flavanone neohesperidosides, for instance naringin, give a bitter taste to fruit and
fruit juice products. The bitterness of naringin in some citrus species reduces the acceptability of
fresh fruit and juice products by taste-sensitive individuals. Therefore, those taste-sensitive people,
a high number of individuals in the population, can not get the health benefit of bitter tasted fruits.
The main objective of this research is to manipulate the citrus flavonoid biosynthetic pathway in
order to reduce bitter taste or increase flavonoids by altering the production of flavanone
neohesperidosides or rutinosides. That will enable plants to have higher level of all flavonoids or
only tasteless flavonoids which will effect the taste. Molecular genetic and transformation
techniques were applied to achieve this goal. The flavonoid pathway genes were utilized for the
genetic transformation of C. paradisi. Interestingly, a few transgenic plants showed changed
flavonoid composition compared to control plants, which will be discussed in our presentation. Our
initial studies are with the grapefruit; however, the approach could be applied to other citrus types
that have bitter tasting fruits.
3

P-251
INVESTIGATIONS ON CAP MICROSPHERES FOR ENCAPSULATION OF
PROTEINOUS DRUGS
N.O. Sahin 1 , K. Opelia 12 . G. Alexandra 1 ' 2
'Mersin University, Faculty of Pharmacy, Department of Pharmaceutics, Yenisehir Campus, Mersin
33169, Turkey, 2 Jagellonian University, Faculty of Pharmacy, Krakow, Poland
Developments in rDNA technology and genetic engineering have lead to development of potent
therapeutic proteins and peptides. Many biotechnology derived drugs have been rapidly entering
phase I and II of clinical studies. At present, the biggest obstacles to the widespread utilization of
these drugs are in vitro (during storage and manufacturing) and in vivo (e.g. enzymatic hydrolysis)
instability, which may lead to a loss of biological activity. This instability problem may be
overcome by encapsulation. From a formulation perspective, proteins are encapsulated to provide:
1) efficient protection for the active protein moiety against a physiological environment; 2)
targeting to the site of action allowing optimal therapeutic efficiency. Some proteins are influenced
by gastric medium. In order to prevent this instability, we suggest preparing cellulose acetate
phthalate (CAP) microspheres to encapsulate a model protein: BSA, employing solvent evaporation
method. Loading capacity was found 52%. Particle size distribution of enteric microspheres was
determined by sieving (525 ± 1.38 pm.). Electron micrographs were taken. In vitro release studies
were conducted using pedal method at 37 ± 0.5°C (pH 1.2 and 7.2, 50 rpm). Integrity of
encapsulated protein was determined by SDS-PAGE electrophoresis. FTIR studies and the results
of electrophoresis indicated that CAP microspheres can be utilized for delivering proteinous active
moiety to colon, maintaining its integrity throughout the manufacturing process, and protecting it
from the influence of gastric media. In vivo studies are still ongoing.
3

P-252
DNA TRANSFECTION INTO THE MAMMALIAN CELLS WITH HYDROLYSED
CHITOSAN FRACTIONS
K. Turan 1 . K. Nagata 2
'Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, MarmaraUniversity, Istanbul
Turkey; department of Infection Biology, Graduate School of Comprehensive Human Sciences
and Institute of Basic Medical Sciences, University of Tsukuba, Japan
Efficient DNA delivery into the target cells is very important not only to analyze the function of genes but also for
therapeutic treatment of hereditary diseases. There are two main types of gene-transfer approaches employed in
vitromd in vivo-, gene transfer w ith viral vectors and non-viral vectors. In general, less efficient DNA transaction
(especially in vivo) is achieved by non-viral gene delivery methods. However, the non-viral vectors have very
important advantages such as ease of synthesis, cell/tissue targeting, low immune response, and unrestricted
DNAsize. Natural and synthetic polymers facilitating the transfer DNA into the cells are usually being considered
as non-viral vehicles. Among the natural polymers, chitosan (Ch) as a non-viral carrier for gene transfer has
recently attracted much attention. Ch is positively charged and hydrophilic at acidic pHand enable to interact with
negatively charged macromolecules like DNA in an aqueous environment. In this study, to further understand the
potential of Ch as a gene carrier, we prepared Ch fractions hydrolyzed at different levels and investigated their
DNA binding capacity and DNA transfection efficiency into mammalian cells. Commercial Chswere dissolved
in 0.5 N hydrochloric acid at the concentration of 0.5 % (w/v) and heated at 50°C, 80°C, 100°C, or autoclaved at 1
atmat 121°C for 30 minutes. The viscosity average molecular weights of hydrolized Chswere calculated using the
classical Mark-Houwinkequation. Two types of plasmid vector were used for formation of Ch-DNA
nanoparticles; pSVp-Gal (Promega, USA) and pEGFP-Nl (Clontech, USA). For formation of Ch-DNA
nanoparticles, plasmid DNAsand Ch solutions diluted in DMEM (pH 7.0) at different concentrations were mixed
by vortexingfor 10 seconds and incubated at room temperature for 30 min for complex fonnation. The formation
of Ch-DNA complexes and unbound DNA was monitored by agarose gel electrophoresis. The size and
morphology of the Ch-DNA nanoparticles was analyzed by a scanning electron microscopy. The surface charge
densityof nanoparticlerwas measured by Zetasizer NanoZS (Malvern Instruments)Human embiyonic kidney
(HEK293), Maden-Darby canine kidney (MDCK), mouse fibroblastic Swiss3T3, and HeLa cell lines were used
for transient transfection experiments. The DNA-transfection efficiency was quantitatively detennined with (3-
galactosidase assay and/or qualitatively examined under a fluorescence microscope. The viscosity-average
molecular weight of Chswere gradually decreased from 450 kDato 14 kDaas a function of temperature. Gelretardation
assaysrevealed that Chs with the higher molecular weight havethe higherDNAbinding ability. The
average diameter of nanoparticles were found to be 200-220 nm. The surface charge of nanoparticles was varied
depending the Ch/DNA ratio.The cell lines differently respondto DNA transfection with Ch fractionsdepended
on molecular weight. HEK293 cells were efficiently transfected by nanoparticles prepared with Chshaving a wide
range of molecular weight (-14-195 kDa). However, Swiss3T3 cells were efficiently transfected by Ch polymers
with ~

ISOPS - 8
PARTICIPANT LIST

372

1 Surname Name Address. E-mail 1
Abacioglu Bengi Santa Farma lla< San. A.§. Istanbul, Turkey
babacioglu@santafarma.com.tr
Aboul-Enein Hassan Y. Pharmaceutical and Medicinal Chemistry Dept., The Pharmaceutical and Drug
Industries Research Division, National Research Center, Dokki, Cairo 12311,
Egypt
hyaboulenein@yahoo.com
Acarturk Fusun Gazi University Faculty of Pharmacy Dept. of Pharmaceutical Technology,
Ankara, Turkey
acarturk@gazi.edu.tr
Ada
Ahmet Oguz Ankara University, Faculty of Pharmacy, Dept. of Toxicology, 06100 Tandogan,
Ankara
ada@pharmacy.ankara.edu.tr
Adejare Adeboye Dept. of Pharmaceutical Sciences, Philadelphia College of Pharmacy, University
of the Sciences in Philadelphia, Philadelphia, Pennsylvania 19104, USA,
a.adejar@usip.edu
Agca Ash Ankara University, Faculty of Pharmacy, Dept. of Pharmacognosy, Tandogan,
06100, Ankara, Turkey
acan@pharmacy.ankara.edu.tr
Ahmed Jehad Ankara University, Faculty of Pharmacy, Dept. of Botany, Tandogan, 06100,
Ankara, Turkey
jehadahmed 4@hotmail.com
Akalin Gulsen Dept. of Biochemistry, Faculty of Pharmacy, Anadolu University, 26470 Eskisehir,
Turkey
gakalin@anadolu.edu.tr
Akdag Ozlem Pfizer llaglari Ltd. §ti. Turkey
Akin Ahmet Dept. of Pharmaceutical Microbiology, Faculty of Pharmacy, Ankara University,
06100 Ankara, Turkey
aakin@pharmacy.ankara.edu.tr
Akyon Yilmaz Yakut Hacettepe University, Faculty of Medicine, Dept. of Microbiology and Clinical
Microbiology, 06100, Sihhiye, Ankara, Turkey
yakyon@gmail.com
Algul Oztekin University of Mersin, Faculty of Pharmacy, Dept. of Pharmaceutical Chemistry,
33169, Mersin, Turkey
oztekinalgul@yahoo.com
Algin Evren Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Tandogan, 06100, Ankara, Turkey
evren9000@yahoo.com
AlKhatb Hatim S. Faculty of Pharmacy, University of Jordan, Queen Rania St., Amman, 11942,
Jordan
h.khatib@ju.edu.jo
Alp Mehmet Ankara University, Faculty of Pharmacy, Pharmaceutical Chemistry, 06100,
Tandogan, Ankara, Turkey
malp@pharmacy.ankara.edu.tr
Alparslan
Duygu
Fatma
Marmara University, Faculty of Pharmacy, Dept. of Pharmaceutical Botany,
Haydarpaa, Istanbul, Turkey
alparslanduygu@yahoo.com.tr
Alper-Hayta Sabiha Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Chemistry,
Tandogan, Ankara, Turkey
sabihaalper@yahoo.com
Altan V. Melih Ankara University, Faculty of Pharmacy, Dept. of Pharmacology, 06100,
Tandogan-Ankara, Turkey
maltan@pharmacy.ankara.edu.tr
Altinoz Sacide Dept. of Analytical Chemistry, Faculty of Pharmacy, Hacettepe University,
Sihhiye, Ankara, Turkey
saltinoz@hacettepe.edu.tr
Altinyay Qigdem Dept. of Pharmacognosy, Faculty of Pharmacy, Ankara University, 06100
Tandogan Ankara, Turkey
cigdemaltinyay@yahoo.com
Altiokka Goksel Dept. of Analytical Chemistry, Faculty of Pharmacy, Anadolu University, 26470,
Eski§ehir, Turkey
galtiokk@anadolu.edu.tr
Altun M. Levent Dept. of Pharmacognosy, Faculty of Pharmacy, Ankara University, 06100
Tandogan Ankara, Turkey
altun@pharmacy.ankara.edu.tr
3

Altun Yuksel Gazi University, Gazi Faculty of Education, Dept. of Chemistry, 06500,
Teknikokullar, Ankara, Turkey
yukseioz@gazi.edu.tr
Amasya
Angin
Gulin
Ay la
Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Tandogan, 06100, Ankara, Turkey
gamasya@pharmacy.ankara.edu.tr
Abdi ibrahim ila San. A.§. istanbul, Turkey
ayla.angin@abdiibrahim.com.tr
Anuta V. University of Medicine & Pharmacy "Carol Davila", Faculty Of Pharmacy,
Bucharest, Romania
Apikogiu Rabu§ §ule Clinical Pharmacy Dept., Marmara University Faculty of Pharmacy, istanbul,
Turkey
suierabus@yahoo.com
Asil Eri§ Dept. of Pharmacy Management, Faculty of Pharmacy, Ankara University,
06100, Tandogan, Ankara, Turkey
asil@pharmacy.ankara.edu.tr
Asian Sinem Ankara University, Faculty of Pharmacy, Dept. of Pharmacognosy, Tandogan,
06100, Ankara, Turkey
sinemasianus@yahoo.com
Ai Ali Ataturk Universitesi, Faculty of Pharmacy, Dept. of Toxicology 25240, Erzurum,
Turkey
aliasci1980@yahoo.com
A$ik Murat Merck Sharpe Dohme ilaglari, Turkey
Atasayar Suna Hacettepe University Faculty of Pharmacy Dept. of Toxicology, Sihhiye, Ankara,
Turkey
suna@hacettepe.edu.tr
Ate§ llker Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Toxicology,
06100, Tandogan-Ankara, Turkey
iates@pharmacy.ankara.edu.tr
Ate§-Alagoz Zeynep Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Chemistry,
06100, Tandogan, Ankara, Turkey
zates@pharmacy.ankara.edu.tr
Atila Alptug Dept. of Analytical Chemistry, Faculty of Pharmacy, Ataturk University, 25240,
Erzurum, Turkey; alptugatila@yahoo.com
Atko§ar Zeki Dept. of Analytical Chemistry, Faculty of Pharmacy, Anadolu University, 26470,
Eski§ehir, Turkey
zatkosar@anadoiu.edu.tr
Avunduk Sibel Ege University, Faculty of Science, Dept. of Chemistry, Bornova, izmir, Turkey
sivunnat@yahoo.com
Aydin Sevtap Dept. of Toxicology, Faculty of Pharmacy, Hacettepe University, Ankara, Turkey
sevtapay@hacettepe.edu.tr
Aydogmu§ Zeynep Dept. of Analytical Chemistry, Faculty of Pharmacy, Istanbul University, 34116
Beyazit, istanbul, Turkey
zaydogmus@yahoo
Ayhan Fatma Mugla University Faculty of Science Dept. of Chemistry, Mugla, Turkey
fatmaayh@yahoo.com
Aypar Eda Dept. of Pharmacology, Faculty of Pharmacy, Gazi University, 06330 Etiler
Ankara Turkey
eczedaaypar@yahoo.com
Bakar Filiz Ankara University, Faculty of Pharmacy, Dept. of Biochemistry, 06100 Tandogan,
Ankara, Turkey
bakar@pharmacy.ankara.edu.tr
Barla Ash Dept. of General Chemistry, Faculty of Pharmacy, University of istanbul 34116,
istanbul, Turkey
asiibarla@gmail.com
Basan Hasan Gazi University, Faculty of Pharmacy, Dept. of Analytical Chemistry, 06330
Ankara, Turkey
basan@gazi.edu.tr
Ba§aran Ahmet Hacettepe University, Faculty of Pharmacy Dept. of Pharmacognosy, Sihhiye,
Ankara, Turkey
basaran@hacettepe.edu.tr
Ba§aran Berrin Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Tandogan, 06100, Ankara, Turkey
bbasaran@pharmacy.ankara.edu.tr
Ba§

Batmaz Gozde Merck Sharpe Dohme ilaclari, Turkey
Bauer Rudolf Institute of Pharmaceutical Sciences, Karl-Franzens-Universitat Graz,
Universitatsplatz 4, A-8010 Graz, Austria
rudolf.bauer@uni-graz.at
Bauer Viktor Inst. Exp. Pharmacol. SASc, Bratislava, Slovakia; Embassy of the Slovak
Republic, Ataturk Bulvari 245, Kavaklidere, Ankara, Turkey
exfabauv@excite.com
Baykara
Bazylak
Tamer
Grzegorz
Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Tandogan, 06100, Ankara, Turkey
baykara@pharmacy.ankara.edu.tr
Dept. of Pharmaco-Bromatology, Faculty of Pharmacy, Collegium Medicum,
Nicolaus Copernicus University, Jagiellonska 13, PL-85-067 Bydgoszcz, Poland
qbazylak@cm.umk.pl
Bayraktar Aygin Hacettepe University, Faculty of Pharmacy, Ankara, Turkey
Beba Leyla Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Tandogan, 06100, Ankara, Turkey
leylabeba@yahoo.com
Belboukhari Nasser Phytochemistry & Organic Synthesis Laboratory, University of Bechar, Algeria
Nasro14@maktoob.com
Bekmezci erife Bahriye Uok Cd. 17/17 Bahgelievler, Ankara, Turkey
serife.bekmezci@gmail.com
Benkli Kadriye Dept. of Pharmaceutical Chemistry, Faculty of Pharmacy, Anadolu University,
26470, Eski§ehir, Turkey
kbenkli@anadolu.edu.tr
Berkoz Mehmet Faculty of Pharmacy, Mersin University, Yeniehir Campus, 33169 Mersin,
Turkey; mehmet berkoz@yahoo.com
Beyoglu Diren Dept. of Pharmaceutical Toxicology, Faculty of Pharmacy, Marmara University,
Haydarpa§a 34668, istanbul, Turkey
d beyoglu@yahoo.com
Bilgener Emrah Dept. of Pharmacy Management, Faculty of Pharmacy, Ankara
University,06100,Tandogan, Ankara, Turkey
ebilgener@yahoo.com
Bingol Semra Mustafa Nevzat ila San. A.$, Istanbul, Turkey
semra bingol@mn.com.tr
Bolelli Kayhan Ankara University, Faculty of Pharmacy, Pharmaceutical Chemistry, 06100,
Tandogan, Ankara, Turkey
bolelli@pharmacy.ankara.edu.tr
Boonleang Jutima Dept. of Pharmaceutical Chemistry, Prince of Songkla University, Songkhla,
90112, Thailand
jutima@pharmacy.psu.ac.th
Bo§gelmez i. ipek Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Toxicology,
06100, Tandogan-Ankara, Turkey
gelmez@pharmacy.ankara.edu.tr
Bozdag-Dundar Oya Ankara University, Faculty of Pharmacy, Pharmaceutical Chemistry, 06100,
Tandogan, Ankara, Turkey
bozdag@pharmacy.ankara.edu.tr
Bozkaya S. Pinar Dept. of Pharmaceutical Chemistry, Faculty of Pharmacy, Ankara University,
06100, Tandogan, Ankara, Turkey
pbozkaya@pharmacy.ankara.edu.tr
Bozkir Asuman Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Tandogan, 06100, Ankara, Turkey
bozkir@pharmacy.ankara.edu.tr
Buharali Can istanbul Ekonomi Danifmaniik Ltd. §ti, istanbul, Turkey
cbuharaii@istanbul-ekonomi.com
Buyukbingol Zeliha Ankara University, Faculty of Pharmacy, Dept. of Biochemistry, 06100 Tandogan-
Ankara-Turkey
zeliha@pharmacy.ankara.edu.tr
Buyukbingol Erdem Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Chemistry,
06100, Tandogan, Ankara, Turkey
erdem@pharmacy.ankara.edu.tr
Cadikova Barbora Dept. of Social and Clinical Pharmacy, Faculty of Pharmacy, Charles University;
Hradec Kralove, Czech Republic
barbora.cadikova@faf.cuni.cz
Can Nafiz Oncu Dept. of Analytical Chemistry, Faculty of Pharmacy, Anadolu University, 26470,
Eski§ehir, Turkey
nafizoc@anadolu.edu.tr
3

Can
Ozgur
Devrim
Anadolu University, Faculty of Pharmacy, Dept. of Pharmacology, 26470
Eski§ehir, Turkey
ozgurdt@anadolu.edu.tr
Can Eke Benay Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Toxicology,
06100, Tandogan-Ankara / Turkey
eke@pharmacy.ankara.edu.tr
Cemiloglu Ulker Ozge Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Toxicology,
06100, Tandogan, Ankara, Turkey
oulker@pharmacy.ankara.edu.tr
Chirita
Codreanu
lleana
Cornelia
Marilena-
Viorica
Dept. of Pharmaceutical Chemistry, University of Medicine and Pharmacy "Carol
Davila", Str. Traian Vuia 6, Bucharest, 020956 Romania
ileana chirita@yahoo.com
Dept. of Pharmaceutical Botany Faculty of Pharmacy, "Carol Davila" University of
Medicine and Pharmacy, Bucharest, Romania
marilenaviorica@yahoo.com
Cofkun Erdem Gazi University, Faculty of Pharmacy, Dept. of Toxicology, Hipodrom, Ankara,
Turkey
erdemcos@gazi.edu.tr
Co§kun Maksut Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Botany, 06100,
Tandogan, Ankara, Turkey
coskun@pharmacy.ankara.edu.tr
Cristea
Aurelia
Nicoleta
Dept. of Pharmacology and Clinical pharmacy, Faculty of Pharmacy, University of
Medicine and Pharmacy "Carol Davila", Str. Traian Vuia 6, Bucharest, Romania
farmacolfarmbuc@k.ro
Cuez Tuba University of Mersin, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
33169, Mersin, Turkey
Caglar Sena Istanbul University, Faculty of Pharmacy, Dept. of Analytical Chemistry, 34452,
Beyazit, Istanbul, Turkey
senacaglar@yahoo.com
Caglayan Aydan Hacettepe University Faculty of Pharmacy Dept. Of Pharmaceutical Toxicology,
Sihhiye, Ankara, Turkey
aydancaglayan@yahoo.com
Qakmak Gonca Dept. of Toxicology, Faculty of Pharmacy, Gazi University,06330, Ankara Turkey
Demircigil
goncacd@gmail.com
Qalgan Zeynep Dept. of Pharmacy Management, Faculty of Pharmacy, Hacettepe University,
06100 Sihhiye, Ankara, Turkey
zcalgan@hacettepe.edu.tr
Qalik Durmaz Tuba Gazi University, Faculty of Pharmacy, Dept. of Toxicology, 06330, Hipodrom,
Ankara, Turkey
tusemecz@hotmail.com
Qali Sema Hacettepe University Fac. of Pharmacy, Dept. of Parmaceutical Technology,
Ankara, Turkey
calis@hacettepe.edu.tr
Cavdar Seda Dept..of Analytical Chemistry, Faculty of Pharmacy, Ege University, 35100
Bornova-izmir, Turkey
seda cavdar@yahoo.com
Celebi Nevin Gazi University Fac. of Pharmacy, Dept. of Pharmaceutical Technology, Ankara,
Turkey;nevin@tr.net
Celebier Mustafa Dept. of Analytical Chemistry, Faculty of Pharmacy, Hacettepe University, 06100
Ankara, Turkey
saltinoz@hacettepe.edu.tr
Citak Meryem Canakkale Onsekiz Mart University, Terzioglu Campus, Faculty of Arts and
Science, Canakkale, Turkey
meryem citak@hotmail.com
Qigek Engin Ankara University, Faculty of Pharmacy, Dept. of Analytical Chemistry 06100
Tandogan, Ankara,Turkey
engin cicek@yahoo.com
Colgegen Hatice Zonguldak Karaelmas University, Faculty of Arts and Science, Dept. of Biology,
67100 Incivez, Zonguldak,Turkey; haticecolgecen@hotmail.com
Dandekar Vikrant K.M.Kundanani College of Pharmacy, Mumbai, India
Dattatrav contactvicks@yahoo.co.in
Da Evcimen Net Ankara University, Faculty of Pharmacy, Dept. of Biochemistry, 06100 Tandogan-
Ankara-Turkey; evcimen@pharmacy.ankara.edu.tr
Degim Zelihagul Gazi University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
06330 Ankara, Turkey
zdegimqgazi.edu.tr
3

Degim i. Tuncer Gazi University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
06330 Ankara, Turkey
tunc@tr.net
Dehghan-Noudeh
Demir
Gholamreza
Goke
Dept. of Pharmaceutics, School of Pharmacy, Kerman University of Medical
Sciences, Kerman, Iran
grdehghan@yahoo.com
Ankara University, Faculty of Pharmacy, Dept. of Medicinal Chemistry, 06100
Tandogan, Ankara-Turkey
qkcedemr@hotmail.com
Demircan §eyda Dept. of Analytical Chemistry, Faculty of Pharmacy, Hacettepe University, 06100,
Sihhiye, Ankara, Turkey
seydad@hacettepe.edu.tr
Demirci
Demirci
Betul
Fatih
Dept. of Pharmacognosy, Faculty of Pharmacy, Anadolu University, 26470-
Eskisehir, Turkey
bdemirca@anadolu.edu.tr
Anadolu University, Faculty of Pharmacy, Dept. of Pharmacognosy, 26470-
Eskisehir; Ministry of National Defence, Army Drug Factory, 06110-Diskapi,
Ankara, Turkey.
fdemirci@anadolu.edu.tr
Demirkan Kutay Hacettepe University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Sihhiye, Ankara
Demirkaya Fatma Dept. of Analytical Chemistry, Faculty of Pharmacy, Ataturk University, 25240,
Erzurum, Turkey
ironstone a@yahoo.com
Dessing Rudolf P. Ligusterweg 1, NL-2202-AD Noordwuk, The Netherlands
rdQq6ina@xs4all.nl
Devrim Burcu Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Tandogan, 06100, Ankara, Turkey
bdevrim@pharmacy.ankara.edu.tr
Dicinoski Greg Australian Centre for Research on Separation Science, School of Chemistry,
University of Tasmania, Private Bag 75, Hobart, Tasmania, Australia, 7001
Greg.Dicinoski@utas.edu.au
Dikmen Necati Dingsa ilag San. A.§., 1. Organize Sanayi Bolgesi, Sincan, Ankara, Turkey
Ding Erdal Dept. of Analytical Chemistry, Faculty of Pharmacy, Ankara University, 06100,
Tandogan, Ankara, Turkey
dinc@pharmacy.ankara.edu.tr
Dinipel Aysun Faculty of Medicine, Dept. of Pharmacology, Hacettepe University, Ankara,
Turkey
ayskara@hacettepe.edu.tr
Dogan Burcu Ankara University Faculty of Pharmacy, Dept. of Analytical Chemistry, Tandogan,
Ankara, Turkey
burcudogan80@yahoo.com
Doganay Tanver Gazi University Fac. of Pharmacy Dept. of Technology, Ankara, Turkey
tanver@gazi.edu.tr
Dogru Bilgehan Ankara University, Faculty of Pharmacy, Dept. of Biochemistry, 06100 Tandogan-
Ankara, Turkey
pekiner@pharmacy.ankara.edu.tr
Dogruer
Deniz
Songul
Gazi University, Faculty of Pharmacy, Dept. of Pharmaceutical Chemistry,
Ankara, Turkey
dogruer@gazi.edu.tr
Dogrukol-Ak Dilek Anadolu University Faculty of Pharmacy, Dept. of Analytical Chemistry, Eskifehir,
Turkey
dak@anadolu.edu.tr
Dumlu Melek U. Marmara University, Faculty of Pharmacy, Dept. of Pharmacognosy, 34668
Haydarpaa, Istanbul, Turkey
melekulusoylu@hotmail.com
Durmaz
Durmaz
Guliz
Emre
Dept. of Biochemistry, Faculty of Pharmacy, Ege University, Bornova 35100
Izmir, Turkey
guliz.durmaz@ege.edu.tr
Gazi University, Faculty of Pharmacy, Dept. of Toxicology, 06330, Hipodrom,
Ankara, Turkey
edurmaz@gazi.edu.tr
Duydu Yalgin Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Toxicology,
06100, Tandogan-Ankara / Turkey
duydu@pharmacy.ankara.edu.tr
3

Dundar Yasemin Dept. of Pharmaceutical Chemistry, Faculty of Pharmacy, Gazi University,
06330, Hipodrom-Ankara-Turkey
yasemina@gazi.edu.tr
Ehrhardt Carsten School of Pharmacy and Pharmaceutical Sciences, University of Dublin, Trinity
College, 00002, Dublin 2, Ireland
ehrhardtc@tcd.ie
Ekinci Goz S. Dept. of Basic Pharmaceutical Sciences, Faculty of Pharmacy, Marmara
University, Haydarpa§a 34668, istanbul, Turkey
nuryars@yahoo.com
Elmasta Mahfuz Dept. of Chemistry, Faculty of Arts and Science, University of Gaziosmanpasa,
60240, Tokat, Turkey
elmastas@gop.edu.tr
Emeje Martins 0. Dept. of Pharmaceutical Technology and Raw Materials Development
National Institute for Pharmaceutical Research and Development (NIPRD) Idu,
P.M.B.21 Garki-Abuja, Nigeria
olobayokunle@yahoo.co.uk
Em re Deniz Hacettepe University Faculty of Pharmacy Dept. of Analytical Chemistry Sihhiye
Ankara, Turkey
nozaltin@hacettepe.edu.tr
Emre Bulut Gizem Dept. of Pharmaceutical Botany, Faculty of Pharmacy, Marmara
University,34668, Haydarpa§a, istanbul, Turkey
emre gizem@yahoo.com
Erda Burcu Mersin University, Faculty of Pharmacy, Dept. of Pharmaceutics, Yenisehir
Campus, Mersin 33169, Turkey
Erdem Gursan
Kadriye
Arzum
Ege University, Faculty of Pharmacy, Analytical Chemistry Dept., Bornova, izmir,
Turkey
arzum.erdem@ege.edu.tr
Erdemgil F. Zerrin Plant & Drug & Scientific Research Center (AUBiBAM), Anadolu University,
Eskisehir 26470, Turkey
zerdemgi@anadolu.edu.tr
Erdemoglu Nurgun Gazi University, Faculty of Pharmacy, Dept. of Pharmacognosy, 06330 Ankara,
Turkey
nurgun@gazi.edu.tr
Erdogan
Ozlem
Nazan
Kocaeli University, College of Hereke Omer Ismet Uzunyol, Marshall Campus,
Korfez, Kocaeli 41800, Turkey
nazanerdogan@hotmail.com
Erdogan Cankat Mersin University, Faculty of Pharmacy, Dept. of Pharmaceutics, Yenisehir
Campus, Mersin 33169, Turkey
Ergun Bulent Dept. of Pharmaceutical Toxicology, Faculty of Pharmacy, Anadolu University,
26470, Eski§ehir, Turkey
berqun@anadolu.edu.tr
Erk Nevin Dept. of Analytical Chemistry, Faculty of Pharmacy, Ankara University, 06100
Tandogan,Ankara, Turkey
erk@pharmacy.ankara.edu.tr
Erkekoglu Pinar Ulfer Hacettepe University, Faculty of Pharmacy, Dept. of Toxicology, Ankara Turkey
Ersan Seyhan Gazi University Faculty of Pharmacy, Dept. of Pharmaceutical Chemistry,
Ankara, Turkey
seyhanersan@yahoo.com
Ertan Tugba Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Chemistry,
06100, Tandogan, Ankara, Turkey
tertan@pharmacy.ankara.edu.tr
Erta Nusret Gazi University, Faculty of Pharmacy, 06330 Etiler Ankara, Turkey
nertas@gazi.edu.tr
Erten-§ener Derya Dept. of Biochemistry, Faculty of Pharmacy, Gazi University, 06330, Etiler,
Ankara, Turkey; deerten@yahoo.com
Eryilmaz Mujde Dept. of Pharmaceutical Microbiology, Faculty of Pharmacy, Ankara University,
06100 Ankara,Turkey; muideyuce@yahoo.com
Estour Frangois Laboratoire de Pharmacochimie, Faculte de Medecine et de Pharmacie, de
Rouen 22 Boulevard Gambetta 76183 Cedexl, France
francois.estour@univ-rouen fr
Fompeydie Dominique Dept. of Analytical Chemistry, Faculty of Pharmacy, University Rene Descartes, 4
avenue de I'Observatoire 752070 Paris cedex 06, France
dominique.fompeydie@univ.paris.fr
Fontaine Jeanine Dept. of Physiology and Pharmacology, Institute of Pharmacy CP 205-7
University of Brussels (ULB) - 1050 Brussels, Belgium
jfontain@ulb.ac.be
38

Foth
Gen
Heidi
Lutfi
Institute of Environmental Toxicology, Martin Luther University, D-069097 Halle -
Saale, Germany
heidi.foth@medizin.uni-halle.de
Anadolu University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
26470, Eski§ehir, Turkey
lqenc@anadolu.edu.tr
Gibbons Simon Centre for Pharmacognosy and Phytotherapy, The School of Pharmacy,
University of London, 29-39 Brunswick Square, London WC1N 1AX, UK
simon.gibbons@pharmacy.ac.uk
Giray Belma Hacettepe University, Faculty of Pharmacy, Dept. of Toxicology, Ankara
Turkey;bgiray@hacettepe.edu.tr
Golenia Aleksandra Mersin University, Faculty of Pharmacy, Dept. of Pharmaceutics, Yenisehir
Campus, Mersin 33169, Turkey; Jagellonian University, Faculty of Pharmacy,
Krakow, Poland
Goger
Goker
Nilgun G.
Hakan
Gazi University, Faculty of Pharmacy, Dept. of Analytical Chemistry, 06330
Ankara, Turkey
ngoger@qazi.edu.tr
Ankara University, Faculty of Pharmacy, Pharmaceutical Chemistry, 06100,
Tandogan, Ankara, Turkey
goker@pharmacy.ankara.edu.tr
Golcu Ay§egul Kahramanmara Sutu imam University, Faculty of Science and Arts, Dept. of
Chemistry, 46100, Kahramanmara§, Turkey
ag518@ksu.edu.tr
Goneng Aymelek Dept. of Biochemistry, Faculty of Pharmacy, Gazi University, Hipodrom Ankara,
Turkey
aymelek@qazi.edu.tr
Gonul Bilge Dept. of Physiology Faculty of Medicine, Gazi University, 06500, Besevler,
Ankara, Turkey
bilgeg@gazi.edu.tr
GOnul Nur§in Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Tandogan, 06100, Ankara, Turkey
gonul@pharmacy.ankara.edu.tr
Goraler Sibel Vrije Universiteit Amsterdam, Amsterdam, The Netherlands
sgoraler@hotmail.com)
Gozpinar Burcu Fakulteler Mah., Oba Str. 3/5, Cebeci, Ankara, Turkey
burcu122@mynet.com
Grieb Pawel Fundacia Rozwoju Diagnostyki i Terapii, ul Sulupecka 11/12, 02309, Warsow,
Poland
pgrieb@cmdik.pan.pl
Guleli §ermin Ege University, Faculty of Pharmacy, Dept. of Biochemistry, Bornova, 35100
izmir, Turkey
sermin.guleli@ege.edu.tr
Guler
Volkan Faculty of Pharmacy, Mersin University, Yeni§ehir Campus, 33169 Mersin,
Gumu§
Gune§
Mustafa
Kemal
Turkey
Yildiz Technical University, Faculty of Science and Arts, Dept. of Chemistry,
Organic Chemistry, Davutpasa Campus, 34210, Merter-istanbul, Turkey
mustafakemalgumus@gmail.com
Gumu§ Fatma Gazi University Faculty of Pharmacy, Ankara, Turkey
fgumus@gazi.edu.tr
Guner §ahika Ankara University, Faculty of Pharmacy, Dept. of Pharmacology, 06100,
Tandogan-Ankara, Turkey; gunere@pharmacy.ankara.edu.tr
Gurbuz ilhan Dept. of Pharmacognosy, Faculty of Pharmacy, Gazi University, Hipodrom
6330,Ankara, Turkey; igurbuz@gazi.edu.tr
Gurkan A. Selen Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Chemistry,
06100, Tandogan, Ankara, Turkey
sgurkan@pharmacy.ankara.edu.tr
Gurkok Gokge Dept. of Pharmaceutical Chemistry, Ankara University, Faculty of Pharmacy,
06100, Tandogan, Ankara, Turkey
Gurson Oguzhan Dept. of Pharmacy Management, Faculty of Pharmacy, Ankara University,
06100, Tandogan, Ankara, Turkey
oguzhangurson@yahoo.com
Guveng Ay§egul Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Botany, 06100,
Tandogan, Ankara, Turkey
aguvenc@pharmacy.ankara.edu.tr
Guveng GQIin Institute of Health Sciences, Ankara University, 06500, Be§evler, Ankara, Turkey
guvenc@ankara.edu.tr
3

Guzel Sevda Dept. of Pharmacognosy, Faculty of Pharmacy, Mersin University, Yeniehir
Campus, 33169 Mersin, Turkey
Haci§evki Aysun Gazi University Fac. of Pharmacy Dept. of Biochemistry, Ankara, Turkey
abozkir@gazi.edu.tr
Hamburger Matthias Institute of Pharmaceutical Biology, Dept. of Pharmaceutical Sciences, University
of Basel, CH-4056 Basel, Switzerland
Matthias. Hamburger@unibas.ch
Hamed Saja H. Cosmetic Science Dept., College of Allied Health Sciences, The Hashemite
University, Zarqa, Jordan
saja_hamed@yahoo.com
Hartzema Abraham G. University of Florida, Florida, USA
hartzema@cop.ufl.edu
Harvey James S. GSK R&D, Park Road,Ware,Herts UK
james.s.harvey@gsk.com
Haspigek Canan Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Tandogan, 06100, Ankara, Turkey
cogan@pharmacy.ankara.edu.tr
Hassanzadeh Davoud Depatment of Pharmaceutics, Faculty of Pharmacy, Tabriz University of Medical
Sciences, Tabriz, Iran
davoudpharmatest@yahoo.com
Haznedaroglu M. Zeki Ege University, Faculty of Pharmacy, Dept. of Pharmaceutical Botany 35100
Bornova, Izmir, Turkey
zeki.haznedarogiu@ege.edu.tr
Hennink Wim E. Dept. of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences, Utrecht
University, PO Box 80082, 3508 TB Utrecht, The Netherlands
w.e.hennink@pharm.uu.nl
Hiltunen
Raimo V. K. Division of Pharmaceutical Biology, Faculty of Pharmacy, University of Helsinki,
P.O.Box 56 (Viikinkaari 5 E), FIN-00014 University of Helsinki, Finland
raimo.hiltunen@helsinki.fi
Hofer Michal Laboratory of Experimental Hematology, Institute of Biophysics, Academy of
Sciences of the Czech Republic, Kralovopolska 135, CZ-612 65 Brno, Czech
Republic
hofer@ibp.cz
Hosoi Shinzo Dept. of Pharmacognosy and Chemistry of Natural Products, School of
Pharmaceutical Sciences, Kyushu University of Health and Welfare, 1714-1
Yoshino-cho, Nobeoka 882-8508, Japan
shosoi@phoenix.ac.jp
Hudson Steve A. Dept. of Pharmaceutical Sciences, School of Pharmacy, University of
Strathclyde, Glasgow, Scotland UK
s.a.hudson@strath.ac.uk
l§ik
Murat Willi Brandt Cd. No.9, Kavaklidere, Ankara, Turkey
Muhibbi ecz.mmi@gmail.com
Ibif Sundus Monrol NQkleer Urunler A.§. Beytepe, Ankara, Turkey
sibis@monroi.com
ilbars Hilal Dept. of Pharmacy Management, Faculty of Pharmacy, Ankara University,
06100, Tandogan, Ankara, Turkey
hilbarsl 970@yahoo.com
ilbasmi Tamer Sibel Dept. of Pharmaceutical Technology, Faculty of Pharmacy, Gazi University,
Ankara, Turkey
ilbasmis@qazi.edu.tr
Inal Ozge Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Tandogan, 06100, Ankara, Turkey
inal@pharmacy.ankara.edu.tr
l§can Mumtaz Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Toxicology,
06100, Tandogan-Ankara, Turkey
iscan@pharmacy.ankara.edu.tr
Izzettin
Fikret Vehbi Marmara Universitesi Eczacilik Fakultesi Klinik Eczacilik B.D. Tibbiye Cad.
No:49 Haydarpaa istanbul, Turkey
fvizzettin@hotmaii.com
Javadzadeh Yousef Dept. of Pharmaceutics, Faculty of Pharmacy-Tabriz University of Medical
Sciences-Iran; Drug applied research center, Tabriz University of Medical
Sciences-Iran,
javadzadehy@yahoo.com
Jelen Frantisek Institute of Biophysics, Academy of Sciences of the Czech Republic,
Kralovopolska 135, 612 65 Brno Czech Republic
jelen@ibp.cz
38

Jelvehgari Mitra School of Pharmacy, Tabriz University of Medical Sciences, Iran
mitraJelvehgari@yahoo.com
Jenkins
Kabanova
Gareth
Tatiana
Swansea School of Medicine, Swansea Unviersity, Singleton Park Swansea
SA28PP, UK
q.j.jenkins@swansea.ac.uk
Dept. of Pharmaceutical Chemistry, State Medical University of Kazan, Butlerov
str., 49, 420012 Kazan, Tatarstan, Russian Federation
t-kabanova@mail.ru
Kadioglu
Kara Kadayifcilar
Yucel
Pinar
Ataturk University, Faculty of Pharmacy, Dept. of Analytical Chemistry, Erzurum,
Turkey
yucel@atauni.edu.tr
Ege University Faculty of Pharmacy Dept. of Analytical Chemistry, Bornova, izmir
Karaaslan Qigdem
35100, Turkey
pinar.kara@ege.edu.tr
Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Chemistry,
06100, Tandogan, Ankara, Turkey
cigdemka@gmail.com
Karabay
Arzu Zeynep Ankara University, Faculty of Pharmacy, Dept. of Biochemistry, 06100 Tandogan,
Ankara, Turkey
zeynepkarabay@yahoo.com
Karabey Yasemin Hacettepe University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Division of Biopharmaceutics and Pharmacokinetics 06100 Ankara, Turkey
yaseminkarabey@yahoo.com
Karadeniz Hakan Analytical Chemistry Dept., Faculty of Pharmacy, Ege University, 35100 Bornova,
izmir, Turkey
hakank@pharm.ege.edu.tr
Karakaya Selma Gazi University, Faculty of Pharmacy, Dept. of Analytical Chemistry, 06330,
Ankara, Turkey
selmakarakaya@hotmail.com
Karakaya Asuman Dept. of Toxicology, Faculty of Pharmacy, Ankara University, 06100, Tandogan,
Ankara, Turkey
karakaya@pharmacy.ankara.edu.tr
Karataf Ay§egul Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Tandogan, Ankara, Turkey
akaratas@pharmacy.ankara.edu.tr
Kartal Murat Ankara University, Faculty of Pharmacy, Dept. of Pharmacognosy, Tandogan,
06100, Ankara, Turkey
kartal@pharmacy.ankara.edu.tr
Kasiwong Srirat Dept. of Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences, Prince
of Songkla University, 90112, Thailand
kasiwons@yahoo.com
Kauffmann Jean Michel Universite Libre de Bruxelles, Institut de Pharmacie, Campus Plaine CP 205/6,
1050 Bruxelles, Belgium
jmkauf@ulb.ac.be
Kaya Ayla Anadolu University, Faculty of Pharmacy, Pharmaceutical Botany, 26470
Eskisehir Turkey
aykaya@anadolu.edu.tr
Kayser Oliver Dept. of Pharmaceutical Biology, GUIDE, University of Groningen, The
Netherlands
o.kayser@rug.nl
Kazemipour Maryam Dept. of Chemistry, School of Science, Azad University, Kerman Branch, Iran;
maryam kazemipour@yahoo.com
Kazimierczuk Zygmunt Polish Academy of Science Medical Research Center, 5 Pawinskiego St, 02-106
Warsaw, Poland
kazimierczuk@delta.sgqw.waw.pl
Kemprecos Jeff Merck Sharpe Dohme ilaglari, Turkey
Kendir Giilsen Dept. of Pharmaceutical Botany, Faculty of Pharmacy, Ankara University, 06100,
Tandogan, Ankara, Turkey
gulsenkendir@mynet.com
Kerimov llgar Dept. of Pharmaceutical Chemistry, Ankara University, Faculty of Pharmacy,
06100, Tandogan, Ankara, Turkey; ecz ilga@hotmail.com
Keski Donmez Menek§e Dept. of Toxicology, Faculty of Pharmacy, Gazi University, 06330, Ankara Turkey
meneksekeski@yahoo.com
Keskiner AN Unsal Turkey
Khazaeli Payam Dept. of Pharmaceutics, School of Pharmacy, Kerman Univ. of Med. Sci, Iran
pkhazaeli@yahoo.com
38

Kill Ceyda Sibel Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Botany, 06100,
Tandogan, Ankara, Turkey
erdurak@pharmacy.ankara.edu.tr
Kill Pelin Dept. of Pharmacology, Faculty of Pharmacy, Gazi University, 06330, Etiler,
Ankara, Turkey
Kiliparslan Muge Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Tandogan, 06100, Ankara, Turkey
kilicars@pharmacy.ankara.edu.tr
King George L. Harvard University, Medical School, Joslin Diabetes Center, Boston, USA
George.King@joslin.harvard.edu
Koblihova Helena Dept. of Social and Clinical Pharmacy, Faculty of Pharmacy, Heyrovskeho 1203,
500 05 Hradec Kralove, Czech Republic
helena.koblihova@faf.cuni.cz
Koca Ufuk Gazi University, Faculty of Pharmacy, Dept. of Pharmacognosy, Etiler, Ankara,
Turkey
ukoca@gazi.edu.tr
Kopyigit Mine Istanbul University, Faculty of Pharmacy, Dept. of Pharmaceutical Botany,
34452, Beyazit, istanbul, Turkey
minekocyigit@hotmail.com
Konuklugil Belma Dept. of Pharmacognosy, Faculty of Pharmacy, Ankara University, 06100
Tandogan, Ankara, Turkey
konuklug@pharmacy.ankara.edu.tr
Konyalioglu Sibel Ege University, Faculty of Pharmacy, Dept. of Biochemistry 35100 Bornova -
izmir, Turkey
sibelkonyalioglu@ege.edu.tr
Kopecky Jiri Institute of Experimental Biopharmaceutics, Joint Research Centre of
PRO.MED.CS Praha a.s. & Academy of Sciences of the Czech Republic,
Heyrovskeho 1207, CZ-50003 Hradec Kralove, Czech Republic
kopecky@uebf.cas.cz
Korkmaz Belma Dept. of Pharmacology, Faculty of Pharmacy, Mersin University, 33169, Mersin,
Turkey
belmakorkmaz@yahoo.com
Ko§ar Muberra Faculty of Pharmacy, Dept. of Pharmacognosy, Anadolu University, 26470
Eski§ehir, Turkey
mkosar@anadolu.edu.tr
Koyuncu Mehmet Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Botany, 06100,
Tandogan, Ankara, Turkey
Koyunoglu Semra Dept. of Basic of Pharmaceutical Science, Faculty of Pharmacy, Hacettepe
University, 06100, Sihhiye, Ankara, Turkey
eczsemra@hacettepe.edu.tr
Kokdil Gamze University of Mersin, Faculty of Pharmacy, Dept. of Pharmacognosy, 33169,
Mersin, Turkey
gkokdil@mersin.edu.tr
Kunle Olobayo 0. Dept. of Pharmaceutical Technology and Raw Materials Development
National Institute for Pharmaceutical Research and Development (NIPRD) Idu,
P.M B.21 Garki-Abuja, Nigeria
olobayokunle@yahoo.co.uk
Kurtoglu
Aslihan Hilal Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Tandogan, 06100, Ankara, Turkey
kurtoglu@pharmacy.ankara.edu.tr
Ku§ Canan Dept. of Pharmaceutical Chemistry, Faculty of Pharmacy, Ankara University,
06100-Tandogan, Ankara, Turkey
kus@pharmacy.ankara.edu.tr
Kugukoglu Kaan Ataturk University, Faculty of Pharmacy, Dept. of Pharmaceutical Chemistry,
Erzurum, Turkey
kucukogluk@hotmail.com
Kupeli Esra Dept. of Pharmacognosy, Faculty of Pharmacy, Gazi University, Hipodrom
6330,Ankara, Turkey
esrak@gazi.edu.tr
Lacaille-Dubois Marie-Aleth Laboratoire de Pharmacognosie, Unite de Molecules d'lnteret Biologique, UMIB,
UPRES-EA 3660, Faculte de Pharmacie, Universite de Bourgogne, 7, Bd Jeanne
d'Arc, 21079 Dijon Cedex, France
m-a.lacaille-dubois@u-bourqogne.fr
Lerkiatbundit Sanguan Dept. of Pharmacy Administration, Faculty of Pharmaceutical Sciences, Songkla,
90112, Thailand
lsanquan@makok.pharmacy.psu.ac.th
38

Lewis Norman G. Institute of Biological Chemistry, Washington State University, Pullman, WA
99164-6340, USA
lewisn@wsu.edu
Limban Carmen Dept. of Pharmaceutical Chemistry, University of Medicine and Pharmacy "Carol
Davila", Bucharest, Traian Vuia 6, sect. 2, 020956, Romania
carmen limban(a>yahoo.com
Lingeman Henk Vrije Universiteit Amsterdam, Amsterdam, The Netherlands
lingeman@chem.vu.nl
Lotfipour Farzaneh Faculty of Pharmacy and Drug Applied Research Center, Tabriz University of
Medical Sciences, Tabriz, Iran.
lotfipoor@tbzmed.ac.ir
Maghsoodi Maryam School of Pharmacy, Dept. of Pharmaceutics, Tabriz University of Medical
Sciences, 51664, Tabriz, Iran
maghsoodim@tbzmed.ac.ir
McNeill John H. Division of Pharmacology and Toxicology, Faculty of Pharmaceutical Sciences,
2146 East Mall, The University of British Columbia, Vancouver, Canada, V6T
1Z3.
jmcneill@interchanqe.ubc.ca
Mente^e Arzu Dept. of Pharmaceutical Chemistry, Ankara University, Faculty of Pharmacy,
06100, Tandogan, Ankara, Turkey
arzumentese@yahoo.com
Men Asiye Anadolu Univ. Faculty of Pharmacy, Dept. of Pharmaceutical Chemistry, 26470,
Eskisehir, Turkey
americ@anadoiu.edu.tr
Merig Buket Ege University, Faculty of Pharmacy, Dept. of Analytical Chemistry, Bornova,
Izmir 35100, Turkey; mericbuket@yahoo.com
Mircioiu Constantin University of Medicine & Pharmacy "Carol Davila", Bucharest, Romania
cmirc@gg.unibuc.ro
Mitrea Niculina University of Medicine and Pharmacy, Faculty of Pharmacy, Dept. of
Biochemistry, Bucharest, Romania; biochimiebucuresti@yahoo.com
Mohammadi Ali Dept. of Drug and Food Quality Control, Faculty of Pharmacy, Medical Sciences
University of Tehran, Tehran, Iran
alimohammadi@sina.tums.ac.ir
Monajemzadeh Farnaz Dept. of Pharmaceutics, Faculty of Pharmacy, Tabriz University of Medical
Sciences, Tabriz, Iran
farnaz m76@yahoo.com
Moru§ciag Laurentiu Pharmaceutical Chemistry Dept, Faculty of Pharmacy, University of Medicine and
Pharmacy "Carol Davila", Bucharest, Traian Vuia 6, 020957, Romania
georgemihai2002@yahoo.com
Moshafi
Mohammad
Hassan
Dept. of Pharmaceutics, School of Pharmacy, Kerman University of Medical
Science, Kerman, Iran
Moshafi14@yahoo.com
Mumcj Arisan 0. Hacettepe University, Faculy of Pharmacy, Dept. of Pharmaceutical Botany,
06100, Sihhiye, Ankara, Turkey; omumcu@hacettepe.edu.tr
Mut Kamer Mersin University, Faculty of Pharmacy, Dept. of Pharmaceutics, Yeniehir
Campus, Mersin 33169, Turkey
Nebioglu Serpil Ankara University, Faculty of Pharmacy, Dept. of Biochemistry, 06100 Tandogan-
Ankara, Turkey
nebioglu@pharmacy.ankara.edu.tr
Nebioglu Dogu Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Chemistry,
06100 Tandogan- Ankara, Turkey
dnebiogl@pharmacy.ankara.edu.tr
Numanoglu F. Ulya Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Tandogan, 06100, Ankara, Turkey
unuman@pharmacy.ankara.edu.tr
Okada Yoshihito Dept. of Natural Medicine & Phytochemistry, Meiji Pharmaceutical University, 2-
522-1, Noshio, Kiyose, Tokyo 204-8588, Japan
y-okada@my-pharm.ac.jp
Omri Abdel Dept. of Chemistry and Biochemistry, Laurentian University, Sudbury, Ontario,
Canada;aomri@laurentian.ca
Onay Be§iki Arzu Ankara University, Faculty of Pharmacy, Dept. of Pharmacology, 06100,
Tandogan-Ankara, Turkey
onay@pharmacy.ankara.edu.tr
Onur Feyyaz Ankara University, Faculty of Pharmacy, Dept. of Analytical Chemistry, 06100
Tandogan, Ankara, Turkey
onur@pharmacy.ankara.edu.tr
38

Opelia Katarzyna Mersin University, Faculty of Pharmacy, Dept. of Pharmaceutics, Yenisehir
Campus, Mersin 33169, Turkey
Jagellonian University, Faculty of Pharmacy, Krakow, Poland
Orhan ilkay Dept. of Pharmacognosy, Faculty of Pharmacy, Gazi University, 06330, Ankara,
Turkey
iorhan@gazi.edu.tr
Orubu Samuel E.
F.
Faculty of Pharmacy College of Health Science Niger Delta University Bayelsa
State Nigeria
samuelorubu@mailcity.com
Olgen Sureyya Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Chemistry,
06100, Tandogan, Ankara, Turkey
olgen@pharmacy.ankara.edu.tr
Onkol
Ozalp
Tijen
Yildiz
Gazi University, Faculty of Pharmacy, Dept. of Pharmaceutical Chemistry,
Ankara, Turkey
tijen@gazi.edu.tr
Eczaciba§i Saglik Urunleri San. Tic. A§, Luleburgaz
yildizo@eczacibasi. com .tr
Ozaltin Nuran Hacettepe Universitesi Faculty of Pharmacy, Dept. of Analytical Chemistry,
Ankara, Turkey
nozaltin@hacettepe.edu.tr
Ozbilgin Ba§ak Dept. of Pharmacognosy, Faculty of Pharmacy, Mersin University, Yeniehir
Campus, 33169 Mersin, Turkey
basakozbilgin@yahoo.com.tr
Ozgagli Eren Gazi University, Faculty of Pharmacy, Dept. of Toxicology, Etiler, 06330, Ankara,
Turkey
eren@gazi.edu.tr
Ozgelikay Gulbin Dept. of Pharmacy Management, Faculty of Pharmacy, Ankara University,
06100, Tandogan, Ankara, Turkey
Gulbin.Ozcelikay@pharmacy.ankara.edu.tr
Ozgelikay A. Tanju Ankara University, Faculty of Pharmacy, Dept. of Pharmacology, 06100,
Tandogan-Ankara, Turkey
ozcelika@pharmacy.ankara.edu.tr
Ozgetin Aybike Hacettepe University Faculty of Science, Dept. of Chemistry, Beytepe, Ankara,
Turkey; aybikeozcetin@yahoo.com
Ozdemir M. T. Dept. of Pharmaceutical Microbiology, Faculty of Pharmacy, Ankara University,
06100, Ankara, Turkey
Ozden Segkin Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Chemistry,
06100, Tandogan, Ankara, Turkey
ozden@pharmacy.ankara.edu.tr
Ozer Qigdem Dept. of Physiology, Faculty of Medicine, Gazi University, 06500, Be§evler,
Ankara, Turkey
ozercigdem@yahoo.co.uk
Ozer A. Yekta Hacettepe University Faculty of Pharmacy Dept. of Pharmaceutical Technology,
Sihhiye, Ankara, Turkey
ayozer@yahoo.com
Ozer Unal Duri§ehvar Dept. of Molecular Biology and Genetics, Bogazigi University, Bebek, 34342,
istanbul, Turkey
unalduri@boun.edu.tr
Ozgen Ufuk Dept. of Pharmacognosy, Faculty of Pharmacy, Ataturk University, 25240,
Erzurum, Turkey
uozgen@atauni.edu.tr
Ozhatay Neriman istanbul University Faculty of Pharmacy Dept. of Pharmaceutical Botany,
Beyazit, istanbul, Turkey
nozhatay@lstanbul.edu.tr
Ozkan Semiha Gazi University, Faculty of Pharmacy, Dept. of Pharmaceutical Microbiology,
Ankara, Turkey; fatmaayh@yahoo.com
Ozkan Yalgin GATA Center of Pharmaceutical Sciences, Etlik, Ankara, Turkey
yozkan@gata.edu.tr
Ozkan Sibel A. Ankara University, Faculty of Pharmacy, Dept. of Analytical Chemistry, 06100
Tandogan, Ankara, Turkey
Sibel.Ozkan@pharmacy.ankara.edu.tr
Ozkan Ay§e Mine Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Botany, 06100,
Tandogan, Ankara, Turkey
gender 65@yahoo.com
Ozkan Ariksoysal Dil§ad Dept. of Analytical Chemistry, Faculty of Pharmacy, Ege University, 35100
Bornova-lzmir, Turkey; ozkand@pharm.eqe.edu.tr
38

Ozkay
Ozturk
Yusuf
Mehmet
Anadolu University, Faculty of Pharmacy, Dept. of Pharmaceutical Chemistry,
26470 Eski§ehir, Turkey
yozkay@anadolu.edu.tr
Bilkent University, Faculty of Science, Dept. of Molecular Biology and Genetics,
Ozturk Mehmet
06800 Ankara, Turkey
ozturk@fen.bilkent.edu.tr
Dept. of Analytical Chemistry, Faculty of Pharmacy, istanbul University 34116,
Istanbul, Turkey; Dept. of Chemistry, Faculty of Science and Arts, Mugla
University Mugla 48000, Turkey
mehmetsadettin@hotmail.com
Ozsoz
Ozturk
Mehmet
Murat
Dept. of Analytical Chemistry, Faculty of Pharmacy, Ege University, 35100
Bornova-izmir, Turkey
mehmet.ozsoz@ege.edu.tr
Dept. of Analytical Chemistry, Faculty of Pharmacy, Ataturk University, 25240,
Erzurum, Turkey
m ozturk78@yahoo.com
Ozturk Nilgun Dept. of Pharmacognosy, Faculty of Pharmacy, Anadolu University, 26470
Eski§ehir, Turkey
nozturk@anadolu.edu.tr
Pajeva llza Center of Biomedical Engineering, Bulgarian Academy of Sciences, 1113 Sofia,
Bulgaria
pajeva@bio.bas.bg
Palabiyik
Ismail Murat Deparment of Analytical Chemistry, Faculty of Pharmacy, Ankara University,
06100, Tandogan, Ankara, Turkey
mpala@pharmacy.ankara.edu.tr
Palecek Emil Institute of Biophysics, Academy of Sciences of the CR, 61265 Brno, Czech
Republic; palecek@ibp.cz
Pardakhty Abbas Dept. of Pharmaceutics, Kerman University of Medical Sciences, PO Box 76175-
493, Iran
abpardakhty@hotmail.com
Pattharachayakul Sutthiporn Faculty of Pharmaceutical Sciences, Prince of Songkla University, Songkhla,
90112Thailand
sutthiporn. p@psu.ac.th
Pellicciari Roberto Dipartimento di Chimica e Tecnologia del Farmaco, Universita di Perugia, via del
Liceo, 1, 06123 Perugia, Italy
rp@unipg.it
Pongwecharak Juraporn Dept. of Clinical Pharmacy, Faculty of Pharmaceutical Science, Prince of
Songkla University, Hatyai, Songkhla, 90112Thailand
pjurapor@pharmacy.psu.ac.th
Pourchaire Sebastien BlueLinea, Paris, France
sebastien. pourchaire@bluelinea.eu
Reanmongkol Wantana Dept. of Clinical Pharmacy Faculty of Pharmaceutical Sciences, Prince of
Songkla University Hat Yai, Songkhla, 90112 Thailand
wantana.r@psu.ac.th
Rezaeifar Mehdi Dept. of Pharmaceutics, School of Pharmacy, Kerman University of Medical
Science, Kerman, Iran
mrezaeifar@yahoo.com
Rodriguez Jaime Departamento de Ciencias Basicas Biomedicas, Facultad de Ciencias de la
Salud, Universidad de Talca, Casilla 747, Talca, Chile
jrodrig@utalca.cl
Rombaut Bart School of Pharmacy, Vrije Universiteit Brussel, Laarbeeklaan 103, B-1090
Brussels, Belgium, brombaut@vub.ac.be
Sagmanligil
Ozdemir
Hulya Dept. of Pharmacology, Faculty of Medicine, University of Yuzuncu Yil, 65200,
Van, Turkey; hulyaozdemir@yyu.edu.tr
Sakalli Ozgul Faculty of Pharmacy, Dept. of Toxicology, Tandogan, 06100, Ankara, Turkey
ozgulsakalli@yahoo.com
Salem Hesham Analytical Chemistry Dept., Faculty of Pharmacy, Minia University, Minia, Egypt
h_salem eg@yahoo.com
Saltan Qitoglu Gulgin Dept. of Pharmacognosy, Faculty of Pharmacy, Ankara University, 06100
Tandogan Ankara, Turkey
saltan@pharmacy.ankara.edu.tr
Sancar Mesut Dept. of Clinical Pharmacy, Faculty of Pharmacy, University of Marmara, Tibbiye
Cad. No:49 Haydarpasa, 34817, istanbul, Turkey
sancarmesut@yahoo.com
Sarikaya Mutlu Ankara Universty, Faculty of Pharmacy, Dept. of Biochemistry, 06100 Tandogan,
Ankara, Turkey; leonid.mutlu@qmail.com
38

Sayar Esin Hacettepe University, Faculty of Pharmacy, 06100-Ankara, Turkey; Ministry of
Defence, Army Drug Factory, 06310-Ankara, Turkey
esinsayar@yahoo.com
Schaefer Marion Charite-Universitatsmedizin Institut fur Klinische Pharmakologie Invalidenstrasse
115 10115 Berlin, Germany
schaefer@zeg-berlin.de
Schmeda-
Hirschmanri
Guillermo
Laboratorio de Quimica de Productos Naturales, Instituto de Quimica de
Recursos Naturales, Universidad de Talca, Casilla 747, Talca, Chile
schmeda@utalca.cl
Schmidt Thomas J. Institut fur Pharmazeutische Biologie und Phytochemie der Westfalischen
Wilhelms-Universitat Munster, HittorfstralJe 56, D-48149 Munster, Germany
thomschm@uni-muenster.de
Sedef Turgay Turkey
Sever Yilmaz Betul Ankara University, Faculty of Pharmacy, Dept. of Pharmacognosy, Tandogan,
06100, Ankara, Turkey
sever@pharmacy.ankara.edu.tr
Sevim Serpil Dinpsa llap San. Ve Tic. A.§., 1. Organize Sanayi BOIgesi Sincan, Ankara,
Turkey
serpil@dincsa.com
Sezgin Zerrin Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Tandogan, 06100, Ankara, Turkey
zsezgin@pharmacy.ankara.edu.tr
Sharififar Fariba Faculty of Pharmacy, Kerman university of Medical Science, Kerman, Islamic
Republic of Iran
fsharififar@kmu.ac.ir
Sonakin Ozlem Dept. of Pharmaceutical Technology, Faculty of Pharmacy, Ankara, University
06100, Tandogan, Ankara, Turkey
osonakn@yahoo.com
Stawny Maciej Dept. of Pharmaceutical Chemistry, Karol Marcinkowski University of Medical
Sciences, 6 Grunwaldzka, 60-780 Poznari, Poland
bmarcin@amp.edu.pl
Sunada Hisakazu Faculty of Pharmacy, Meijo University, 150 Yagotoyama, Tempak-ku, Nagoya
468-8503, Japan
sunada@ccmfs.meijo-u.ac.jp
Sunthornpit Arunsri Dept. of Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, Prince
of Songkla University, Hat Yai, Songkhla, 90112, Thailand
arunsri.s@psu.ac.th
Suleymanoglu Erhan Dept. of Pharmaceutical Chemistry, Gazi University, Faculty of Pharmacy, 06330,
Ankara, Turkey
esuleymanoglu@gazi.edu.tr
Sur-Altiner Dehen Marmara University, Faculty of Pharmacy, Dept. of Biochemistry, Haydarpasa,
istanbul, Turkey
dehensur@ttnet.net.tr
Suslu incilay Dept. of Analytical Chemistry, Faculty of Pharmacy, Hacettepe University, 06100,
Sihhiye, Ankara, Turkey
isuslu@hacettepe.edu.tr
Suzen Sinan Ankara University, Faculty of Pharmacy, Dept. of Toxicology, 06100
Tandogan/Ankara, Turkey
suzen@pharmacy.ankara.edu.tr
Suzen Sibel Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Chemistry,
06100, Tandogan, Ankara, Turkey
sibel@pharmacy.ankara.edu.tr
§ahin F. Pinar Dept. of Pharmaceutical Botany, Faculty of Pharmacy, Hacettepe University,
06100, Sihhiye, Ankara, Turkey
psahin@hacettepe.edu.tr
§ahin
Nefise Ozlen University of Mersin, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
33169, Mersin, Turkey
scitech@superonline.com
§ar Sevgi Dept. of Pharmacy Management, Faculty of Pharmacy, Ankara University,
06100, Tandogan, Ankara, Turkey
sar@pharmacy.ankara.edu.tr
§arer Engin Ankara University, Faculty of Pharmacy, Dept. of Pharmacognosy, Tandogan,
06100, Ankara, Turkey
sarer@pharmacy.ankara.edu.tr
§atana Eda Gazi University, Faculty of Pharmacy, Dept. of Analytical Chemistry 06330
Ankara, Turkey; eda@qazi.edu.tr
38

§encan
Muzeyyen
Nazli
Yeditepe University Faculty of Pharmacy 26 Agustos Yerleimi, Kayi§dag,
istanbul, Turkey
nsencan@yeditepe.edu.tr
§enel Sevda Hacettepe University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
06100, Ankara, Turkey
sensel@hacettepe.edu.tr
§engel-TQrk
Ceyda Tuba Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Tandogan, 06100, Ankara, Turkey
ctsengel@pharmacy.ankara.edu.tr
§enturk Zuhre Yuzuncu Yil University, Faculty of Science and Letters, Dept. of Analytical
Chemistry, 65080 Van, Turkey
zuhre@yyu.edu.tr
§ohretoglu Didem Dept. of Pharmacognosy, Faculty of Pharmacy, Hacettepe University, 06100,
Sihhiye, Ankara, Turkey
didems@hacettepe.edu.tr
§umnu Murat Hacettepe University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
06100, Ankara, Turkey
msumnu@hacettepe.edu.tr
Tamer Ugur Dept. of Analytical Chemistry, Faculty of Pharmacy, Gazi University, 06330,
Etiler, Ankara, Turkey
utamer33@yahoo.com
Tarimci Nilufer Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Tandogan, 06100, Ankara, Turkey
ntarimci@pharmacy.ankara.edu.tr
Ta§demir Deniz Centre for Pharmacognosy and Phytotherapy, School of Pharmacy, University of
London 23/39 Brunswick Square WC1N 1AX, London, UK
deniz.tasdemir@pharmacy.ac.uk
Tafoz Ayla Vrije Universiteit Amsterdam, Amsterdam, The Netherlands
aylatasoz@hotmail.com
Tatar Ulu Sevgi Dept. of Analytical Chemistry, Faculty of Pharmacy, Istanbul University, Beyazit,
istanbul, Turkey
sevgitatar@yahoo.com
Tatli Qankaya 1. irem Hacettepe University, Faculty of Pharmacy, Dept. of Pharmaceutical Botany,
Sihhiye, 06100 Ankara, Turkey
itatli@hacettepe.edu.tr
Tay Aydin Ankara University, Faculty of Pharmacy, Dept. of Pharmacology, 06100,
Tandogan-Ankara, Turkey
tay@pharmacy.ankara.edu.tr
Tekiner-Gulba Betul Ankara University, Faculty of Pharmacy, Pharmaceutical Chemistry, 06100,
Tandogan, Ankara, Turkey
btekiner@pharmacy.ankara.edu.tr
Theoduloz Cristina Departamento de Ciencias Basicas Biomedicas, Facultad de Ciencias de la
Salud, Universidad de Talca, Casilla 747, Talca, Chile
ctheodul@utalca.cl
Tokag Mahmut Ministry of Health, General Directorate of Pharmaceuticals and Pharmacy,
Ankara, Turkey
Tozan Ayfer Marmara University, Faculty of Pharmacy, Dept. of Toxicology, 34668
Haydarpaa, istanbul, Turkey; ayfertozan@yahoo.com
Tuncay Ozdemir Meltem Dept. of Pharmaceutical Microbiology, Faculty of Pharmacy, Ankara University,
06100, Ankara, Turkey
meltemtuncay@gmail.com
Tungbilek Meral Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Chemistry,
06100, Tandogan, Ankara, Turkey
tuncbile@pharmacy.ankara.edu.tr
Tungel Segil Ankara University, Faculty of Pharmacy, Dept. of Analytical Chemistry, 06100
Tandogan, Ankara, Turkey
seciltuncel@hotmail.com
Turan Kadir Dept. of Pharmaceutical Biotechnology, Faculty of Pharmacy, Marmara
University, Istanbul Turkey
kadirturan@marmara.edu.tr
Turung Sinem Ezgi Dept. of Biochemistry, Faculty of Pharmacy, Ege University Bornova 35100 izmir,
Turkey; ezgiturunc@hotmail.com
Turker Gulen Canakkale Onsekiz Mart University, Terzioglu Campus, Faculty of Science and
Arts, Canakkale, Turkey
Turkmen Berna Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Tandogan, 06100, Ankara, Turkey; bturkmen@pharmacy.ankara.edu.tr
38

Ugakturk Ebru Dept. of Basic Pharmaceutical Sciences, Faculty of Pharmacy, Hacettepe
University, 06100, Sihhiye, Ankara, Turkey
ebruu@hacettepe.edu.tr
Ugurlu Gamze Dept. of Analytical Chemistry, Faculty of Pharmacy, Hacettepe University,
Sihhiye, Ankara, Turkey;
gugurlu@hacettepe.edu.tr
Ulusoy Zeyriep Nobel lla San. ve Tic. A.§, istanbul, Turkey
zeynep.ulusoy@nobel.com.tr
Ulutan Gizem Bayar Cd. Bolen Str. 2/5 Kozyatagi, Istanbul, Turkey
ulugiz@gmail.com
Uluta§
Onur Kenan Dept. of Toxicology, Faculty of Pharmacy, Gazi University, 06330 Hipodrom,
Ankara, Turkey
onurkenan@gazi.edu.tr
Uslu Cagla Yeditepe University, Faculty of Pharmacy, 26 Agustos Yerleimi, Kayi§dag,
istanbul, Turkey
Uslu Bengi Ankara University, Faculty of Pharmacy, Dept. of Analytical Chemistry, 06100
Tandogan, Ankara, Turkey
buslu@pharmacy.ankara.edu.tr
Utku Semra Dept. of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Mersin,
33169, Mersin, Turkey
utkusemra@mersin.edu.tr
Uyar Banu Dept. of Analytical Chemistry, Faculty of Pharmacy, Hacettepe University,
Sihhiye, Ankara, Turkey
banuuyar@hacettepe.edu.tr
Ulgen Mert University of Marmara, Faculty of Pharmacy, Dept. of Pharmaceutical Chemistry,
81010, Haydarpaa, Istanbul, Turkey
oztekinalgul@yahoo.com
Ustel Ismail Halit Ziya Sk. No:22/3, 06540 Cankaya/Ankara/Turkey
usteli 1953@yahoo.com
Ustundag Ozgur Ankara University, Faculty of Pharmacy, Dept. of Analytical Chemistry, 06100
Tandogan, Ankara, Turkey
ustundag@pharmacy.ankara.edu.tr
Ustundag Aylin Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Toxicology,
06100, Tandogan, Ankara, Turkey
dur@pharmacy.ankara.edu.tr
Valizadeh Hadi Dept. of Pharmaceutics, Faculty of Pharmacy, Tabriz University of Medical
Sciences, Tabriz, Iran
valizadeh@tbzmed. ac. ir
Velescu Bruno University of Medicine and Pharmacy "Carol Davila" Bucharest, Fac. of
Pharmacy, Dept. of Inorganic Chemistry, 6-th Traian Vuia Str., Bucharest-
Romania; bruno velescu@yahoo.co.uk
Yagmur Sultan Canakkale Onsekiz Mart University, Terzioglu Campus, Faculty of Science and
Arts, Canakkale, Turkey
sultanyagmur@hotmail.com
Yalin Gorkem Dept. of Analytical Chemistry, Faculty of Pharmacy, Ege University, 35100
Bornova-lzmir, Turkey
qorkem.yalcin@ege.edu.tr
Yalpin Guler Marmara University Faculty of Pharmacy, Analytical Chemistry Dept. 34668
Haydarpa§a istanbul, Turkey
quyalcin@yahoo.com
Yalin Serap Mersin University Faculty of Pharmacy Biochemistry Dept., Mersin, Turkey
syalinOI @yahoo.com
Yanev Stanislav Dept. Pharmacology and Toxicology, Faculty of Pharmacy, Medical University -
Sofia, Bulgaria
tox@bio.bas.bg
Yardim Yavuz Yuzuncu Yil University, Faculty of Science and Letters, Dept. of Analytical
Chemistry, 65080 Van, Turkey
yavuzyardim2002@yahoo. com
Yardimci Ceren Dept. of Analytical Chemistry, Faculty of Pharmacy, Hacettepe University, 06100,
Sihhiye, Ankara, Turkey; cereny@hacettepe.edu.tr
Yarimkaya Sezen Dept. of Analytical Chemistry, Faculty of Pharmacy, Gazi University, 06330,
Etiler, Ankara, Turkey;sezen yarimkaya@yahoo.com
Yars Ozarslan Nur Dept. of Basic Pharmaceutical Sciences, Faculty of Pharmacy, Marmara
University, Haydarpa§a 34668, istanbul, Turkey nuryars@yahoo.com
Yazan Yasemin Anadolu University, Faculty of Pharmacy, Dept. of Pharmaceutical and Cosmetic
Technology, Eskiehir, Turkey;yyazan@anadolu.edu.tr
38

Yeniceli Duygu Dept. of Analytical Chemistry, Faculty of Pharmacy, Anadolu University,26470,
Eskisehir, Turkey
dyeniceli@anadolu.edu.tr
Yerdelen
Kadir Ozden Dept. of Pharmaceutical Chemistry, Faculty of Pharmacy, Ataturk University,
25240, Erzurum, Turkey; dadasozden@gmail.com
Yetimoglu Fusun Santa Farma ilag San. A.$., istanbul, Turkey
Yildirim Ozlem Ankara University, Faculty of Science, Dept. of Biology, Tandogan, Ankara,
Turkey
ozlemesn@hotmail.com
Yildiz Sulhiye Dept. of Pharmaceutical Microbiology, Faculty of Pharmacy, Ankara University,
06100 Ankara-Turkey
yildiz@pharmacy.ankara.edu.tr
Yilmaz Bilal Dept. of Analytical Chemistry, Faculty of Pharmacy, Ataturk University, 25240,
Erzurum, Turkey; bilalylmaz@yahoo.com
Yilmaz Selahattin Canakkale Onsekiz Mart University, Terzioglu Campus, Faculty of Science and
Arts, Canakkale, Turkey; seletyilmaz@hotmail.com
Yilmazer Semra Sakarya University, Faculty of Arts and Sciences, Chemistry Dept., 54187,
Sakarya, Turkey
syilmazer@sakarya.edu.tr
Yurdasiper Aysu Dept. of Pharmaceutical Technology, Faculty of Pharmacy, Ege University,
35100, Bornova, izmir, Turkey; aysu.yurdasiper@ege.edu.tr
Yuksel Gunseli Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Tandogan, 06100, Ankara, Turkey
gyuksel@pharmacy.ankara.edu.tr
Yuksel Altan University of Mersin, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
33169, Mersin, Turkey; altany@anet.net.tr
Yuksel Nilufer Ankara University, Faculty of Pharmacy, Dept. of Pharmaceutical Technology,
Tandogan, 06100, Ankara, Turkey
Nilufer.Yuksel@pharmacy.ankara.edu.tr
Yurekoglu Ipek Pfizer ilalari Ltd. Sti. Turkey
Zaghloul Abdel-Azim
A.
Faculty of Pharmacy, Kuwait University, PO BOX 24923, Safat 13110, Kuwait
abdei@hsc.edu.kw
Zakeri-Milani Parvin Dept. of Pharmaceutics, Faculty of Pharmacy, Tabriz University of Medical
Sciences, Tabriz, Iran; pzakeri@tbzmed.ac.ir
Zhdanova Elena Dept. of Pharmaceutical Chemistry, State Medical University of Kazan, Butlerov
str., 49, 420012 Kazan, Tatarstan, Russian Federation
ekonom2@sia.tatmail.ru
Zoulova Jana Institute of Experimental Biopharmaceutics, Joint Research Centre of
PRO.MED.CS Praha a.s. & Academy of Sciences of the Czech Republic, Hradec
Kralove, Czech Republic
zoulova@uebf. cas.cz
38

ISOPS - 8
AUTHOR LIST

I Surname Name Presentation I
Abacioglu N. P-91, P-113, P-119
Abbasoglu U. P-197, P-239
Aboul-Enein H.Y. PL-1
Abu-Asaker M. P-98
Ada A.O. 0-1
Adak S. P-195
Adejare A. PL-2
Adiguzel N. P-89
Adrangi M. P-63
Agca A.C. P-75
Ahmadi S. P-63
Ahmed J. P-34
Aiedeh K.M. P-220. P-222
Akalin G. P-1, P-191
Akay C. P-38, P-39
Akbiyik F. P-15
Akin A. P-32
Akinci M. P-4
Aki-§ener E. P-189
Akmansu M. P-15
Akyon Yilmaz Y. PL-3, P-33
Aldea V. P-216
Alemdar Y. 0-1
Alexandra G. P-251
AlgOl 0. P-215
Algin E. P-221
Alipour M. P-238
AlKhatib H.S. P-220, P-222
Alparslan D.F. P-36
Alper-Hayta S. P-189
Alsancak G. P-86
Altanlar N. P-203, P-204, P-207
Altinoz S. P-127, P-139, P-142. P-171. P-172. P-180
Altinyay C P-76, P-77. P-79
Altiokka G. P-60, P-128. P-137
Altun Y. P-129
Altun M.L. P-76, P-77, P-78, P-79, P-109, P-110
Amini M. P-208
Angin A. IP-3
Ansari M. P-63
Anuta V. P-130
Anzenbacher P. P-120
Apikoglu Rabu§ • 0-2. P-16, P-17
Areche C. P-125
Asian S. P-5
Asian S. P-103
Asnaashari S. P-101, P-226, P-227, P-228
Astudillo L. P-123
Atalay M. P-218
Ate§-Alagoz Z. P-190

Atila H. P-146
Atila A. P-131
Atko§ar Z. P-132
Atli 0. P-32
Avci G. P-95
Av§ar T. P-151
Awang A. P-21
Aydin S. P-80
Aydin A. P-34, P-45, P-73, P-93
Aydinuraz B. P-57, P-64
Aydogmu§ Z. P-133
Aydogmu§ Ozturk F. P-46
Ayhan F. P-239
Ayhan-Kilcigil G. P-203, P-204
Aypar E. P-113. P-119
Azarmi Y. P-188, P-247
Bagi§ S. P-14
Bailleul F. P-124
Baleanu D. 0-4
Bamdad S. P-101
Banoglu E. P-10
Barla A. P-35
Barzegar S. P-247
Barzegar Jalali M. P-188, P-247
Ba§aran A.A. P-80
Ba§aran N. P-80
Ba§i N.E. 0-3, P-146, P-178
Ba§er K.H.C. P-83, P-94
Ba§tug A.S. P-148
Bauer R. PL-4
Bauer V. PL-5
Bauer F. PL-5
Baykara T. P-221.P-242
Bayram i. P-79
Bazylak G. PL-6, P-134, P-135
Beihaghy A. P-235
Beker H.K. P-198, P-199, P-200
Belboukhari N. P-81
Benkli K. P-191
Berkoz M. P-14
Beynek N. P-191
Beyoglu D. P-52
Bilgener E. P-24
Bilir N. P-25
Bingol S. I P-2
Blankert B. PL-25
Boga M. P-35
Boissy R.E. 0-5
Bolelli K. P-197
Boonleang J. P-136
Boonlon T. P-20
Borm P.J.A. P-56
34

Bosgelmez 1. 1. P-53
Bozdag-Dundar 0. P-207
Bozkaya S.P. P-192, P-193
Bozkurt A. P-145, P-146
Bozkurt G. P-15
Bozkurt E. P-16
Buharali C. IP-5
Buharalioglu K. P-121
Burgaz S. 0-1, P-55, P-56
Bustanji Y. P-222
Butoescu V. P-115
Buttajeen P. P-20
Buyukbingol Z. P-8
Buyukbingol E. P-190
Cadikova B. P-18
Cahova K. PL-37
Can N.O. P-137
Can O.D. P-114, P-212
Can-Eke B. P-203
Carlson R.G. P-136
Cedillo-Rivera R. P-202
Cemiloglu Olker 0. P-54
Cengiz N. P-79
Cevheroglu S. P-38, P-39, P-84, P-168
Chalimoniuk M. P-202
Chamani G. P-232
Chayakul P. P-20
Cheriti A. P-81
Chirita i.e. PL-7, P-115. P-206. P-210
Chladek J. P-120, P-126
Choudhary M.I. 0-10, P-88
Cimrin A. P-56
Codreanu M.V. P-82
Contuk G. P-68, P-70
Cokun E. P-55, P-56
Co§kun M. P-105
Comert E. P-62
Cristea A. N. PL-8, P-115, P-116
Cui F.D. 0-11
Qaglar S. P-138
Qaglayan F. P-145
Qaglayan A. P-61
Qakir B. P-211
Qakir K. P-159
Qakmak Demircigil G. P-55, P-56
Qalgan Z. P-25
Qalik Durmaz T. P-57. P-59
Qali§ 1. P-44, P-160
Qali§ U. P-205, P-219
Qali§ S. PL-43
Qali§kan-Ergun B. P-10
Qavdar S. PL-34, P-163, P-182

Qelebier M. P-139. P-180
Qetindag F. P-55
Qetinel • P-68, P-70
Qevikba§ U. P-22, P-23
Qirakli Z. P-22
Qitak M. P-140
Qigek E. P-141. P-151
Qizmecioglu M. P-182, P-205
Qoban T. P-42, P-190, P-193, P-201
Qok 1. P-57, P-59, P-64, P-71
Qomoglu S. P-15
Qolgegen H. P-248
Qomelekoglu U. P-14
Dadandi M.Y. P-83
Dagci T. P-7
Dandekar V. D. P-194
Daneshvar H. P-2 36
Da§-Evcimen N. P-10
Davin L.B. PL-29
De Luca M. 0-4
Degim T. P-225
Dehghan-Noudeh G.R. P-241. P-249
Delazar A. P-101
Delialioglu N. P-104
Demir U. P-114, P-212
Demir H. P-33
Demirbag A.E. 0-1
Demirba§ P. P-62
Demircan • P-142, P-160. P-172
Demirci B. P-83
Demirci F. P-83. P-84
Demirkaya F. P-143, P-144
Demirta§ 1. P-3
Dessing R.P. CP-1
Dettlaff K. P-169
Dhanasekaran M. P-225
Dicinoski G. PL-9
Dicle D. P-108
Dilgin Y. P-140
Din E. 0-4. P-142, P-181
Dinpel A. P-145. P-146
Dinger A. 0-2
Dinu M. P-82
Dociu N. P-82
Dogan B. P-129. P-147, P-192
Dogruer D.S. P-195
Dogrukol-Ak D. P-186
Dorcak V. PL-37
Dorman H.J.D. PL-19
Donmez F. 0-2
Draghici C. P-206
Duman H. P-49, P-50
3

Durnlu M. U. P-58, P-85
Durmaz G. P-2, P-13
Durmaz E. P-59
Duru M. E. P-46, P-47, P-48
Du/du Y. P-66, P-72, P-73
Dundar Y. P-196
Dundar K. P-73
Edinsel H. P-55
Efe S. 0-1
Ehrhardt C. PL-10
Ejaz A. 0-10
Eken A. P-34, P-45, P-73, P-93
Ekinci 0. P-117
Ekinci Goz S. P-148
Ekiz A. 0-2
Elmasta§ M. P-3
Emeje M. 0. P-223. P-233
Emekda§ G. P-104
Em re D. P-149
Emre Bulut G. P-36
Erba§ Y. P-56
Erciyas E. P-205
Er«Sa§ B. P-26
Erdem A. PL-34, 0-7, P-150, P-158, P-161
Erciemgil F.Z. P-86
Erdemoglu N. P-87, P-88, P-89, P-90, P-91, P-97
Erdogan 0. N. P-27, P-28
Erdogan C. P-237
Erdugan H. P-140
Ergun B. P-60
Erk N. P-141. P-151
Erkekoglu P. P-61
Eroglu P. P-14
Ertan M. P-27
Ertan R. P-207
Ertan T. P-197
Erta§ N. P-152, P-155, P-159, P-173, P-174
Erten-§ener D. P-4
Eryavuz A. P-95
Eryilmaz M. P-32
Estour F. PL-11
Ezer N. P-49, P-50, P-51, P-124 .
Faber E. 0-8
Faroongsarng D. P-231
Fazly Bazzaz B.S. P-249
Fojta M. PL-37
Fompeydie D. P-153
Fontaine J. PL-12
Foth H. PL-13
Franck R. PL-16
Gedik N. P-67, P-68, P-70
Geffken D. P-167

Gen L. P-154
Genaslan G. P-110
Gibbons S. PL-14
Giray B. P-61
Glahn F. PL-13
Goger F. P-94
Goger N.G. P-152. P-155, P-159, P-173, P-174
G6ke G. P-37
Goke M. P-196
Golcu A. P-156
Gonen A. P-4, P^, P^
Gonul B. P-117, P-122
Gramond J.P. P-153
Gutu E. P-130
Gul H.I. P-205, P-217, P-218, P-219
Guleli S. P-7
Guler V.G. P-14
Guler 0. 0-2
Gumu§ M.K. P-198, P-199, P-200
Gumu§ F. P-214
Guncu G.N. P-145
Gur S. P-214
Gurbuz i. P-92
Gurkan E. P-85
Gurkok G. P-201
Gurlek A. P-184
Giirson 0. P-29
Guven K. P-83, P-150
Guven G. P-62
Guven A. P-34, P-41, P-44, P-45, P-77
Guvendik G. P-53
Guzel S. P-104
Haci§evki A. P-6
Haddad P.R. PL-9
Hamburger M. PL-15
Hamed S.H. 0-5
Hamedani M.P. P-208
Hanninen 0. P-218
Hantash T. P-22
Harmanci A. P-184
Harmandar M. P-48
Hartzema A.G. PL-16
Harvey J.S. PL-17
Hason S. P-157
Hassanzadeh D. P-224, P-234, P-235
Hatungil R. P-14
Havran L. PL-37
Haznedaroglu M.Z. 0-6, P-37. P-38, P-39
Heidari H. P-208
Henkelmann B. P-64
Hennink W.E. PL-18
Hincal F. P-61
38

Hincal A.A. P-168, P-230
Hiltunen R. PL-19
Hofer M. P-118
Hola J. P-118
Hosoi S. PL-20
Hosseini S.M. P-111, P-240
Hosseininajad F. P-111
Housaindokht M. P-249
Hudson S. PL-21. CP-2
Hur§itoglu M. P-23
loele G. 0-4
lonica G. P-130
Irth H. PL-30
Islambulchilar Z. P-188, P-247
Istudor V. P-82
l§ikdag i. P-1, P-212
l§ildak M. P-184
Ito s.
P-92, P-211
Ito Y. PL-5
Izquierdo R. P-123
ilbars H. P-30
ilbasmi§ Tamer S. P-225
Inal 0. P-221
incesu Z. P-1
l§can M. 0-1, P-203
lzde§ S. P-65
Izzettin F.V. PL-22. 0-2, CP-3, P-16, P-17, P-22, P-23
Jafari Navimipour B. P-228
Javadzadeh Y. P-226, P-227. P-228
Jelen F. PL-37, 0-7. P-157
Jelvehgari M. P-229
Jenkins G.J.S. PL-23
Jintapakorn W. P-231
Jourdes M. PL-29
Julsing M.K. PL-26
Kaban S. P-198, P-199, P-200
Kabanova T.V. P-246
Kabuku C. P-57
Kadioglu Y. P-131, P-143, P-144, P-164, P-187
Kadioglu E. P-65
Kan Y. 0-10, P-103
Kanbak 0. P-65
Kara Kadayifgilar P. PL-34, P-163, P-182
Karabay A.Z. P-8
Karabey Y. P-230
Karadeniz H. PL-34. 0-7. P-150. P-158. P-161
Karakaya S. P-159
Karakaya 0. P-17
Karakaya A. PL-24. P-54
Karata§ A. P-242
Kargin R. P-17
Kartal M. 0-10, P-98, P-103

Kasiwong S. P-231
Kauffmann J.M. PL-25
Kaya A. P-40
Kaya S. P-28
Kaynarsoy B. P-245
Kayser 0. PL-26
Kazaz C. P-105
Kazemipour M. P-63
Kazimierczuk Z. P-2 02
Kendir G. P-41
Kerimov 1. P-203
Keski Donmez M. P-64
Khazaeli P. P-232. P-241
Kill C.S. P-42
Kill P. P-119
Kiliarslan M. P-242
Kir S. P-160, P-172
Kircali K. P-60
Kivrak i. P-47, P-48
King G.L. PL-27
Kirpi E. P-198, P-199, P-200
Koblihova H. 0-8
Koca U. P-103, P-250
Kocaba§ N.A. P-55
Koyigit M. P-43
Konuklugil B. P-93
Konyalioglu S. P-7
Koohzad M. P-236
Kopecky J. P-120. P-126
Korkmaz B. P-121
Ko§ar M. P-94
Kotalik J. P-64
Koulman A. PL-26
Kourilova A. P-157
Kovoh F. P-124
Koyunoglu S. P-160
Kozak M. P-170
Kozana E. P-96
Kozubik A. P-118
Kokdil G. P-104
Krstic D. PL-37
Kul A. P-108
Kummee S. P-243
Kunle 0.0. P-223, P-233
Ku§ C. P-204
Kuukkurt 1. P-95
Kuukoglu K. P-205
Kultur S. P-35
Kupeli E. P-44. P-87. P-91. P-95. P-96. P-97. P-196
Kusmenoglu §• P-103
Kvetina J. P-120, P-126
Lacaille-Dubois M.A. PL-28, P-98, P-99. P-100
4

Lafont 0. PL-11
L.audy A.E. P-202
Lehr C.M. PL-10
Lerkiatbundit S. P-19
L.event A. P-175
Levillain P. P-153
Lewis N.G. PL-29
Limban C. P-206, P-210
Limsuwannachot S. P-21
Lingeman H. PL-30
Lotfipour F. P-101
Luten J. PL-18
Macek K. P-120, P-126
Maghsoodi M. P-234
Malak A. P-134, P-135
Manavba§i Y. P-213
Maneenuan D. P-243
Marciniec B. P-169, P-170
Margina D. P-9
Marineci C. D. P-115
Martin M. P-124
Masarik M. PL-37
Matyas S. PL-5
McNeill J.H. PL-31
Mehrabani M. P-232
Memi§ L. P-117
Mente§e A. P-207
Meri B. PL-34, P-163, P-182
Merip A. P-161
Mikelova R. P-157
Mircioiu C. 0-9. P-130
Mircioiu 1. 0-9
Miron D. 0-9
Missir A.V. PL-7, P-206, P-210
Mitaine-Offer A.C. P-98
Mitrea N. P-9
Miyamoto T. P-99, P-100
Mohammadi A. P-208
Mohammadi N. P-232
Mohesin S. P-222
Momekov G. 0-14
Monajemzadeh F. P-224, P-235
Moo-Puc R. P-202
Moore G.A. P-250
Moru§ciag L. P-209. P-210
Moshafi M.H. P-236
Moustafine R.I. P-246
Mumcu 0. P-124
Mumcu Arisan 0. P-45
Musaalrezaee L. P-227
Mut K. P-237
Nagata K. P-252

Nawaz S. A. P-88
Nayci A. P-14
Naz Q. 0-10
Nazemiyeh H. P-101
Nebioglu D. P-192, P-193
Nemutlu E. P-139
Nitulescu G.M. P-209
Nokhodchi A. P-226, P-227, P-228, P-234, P-244
Novak J. P-18
Noyanalpan N. P-196
Nuta D. P-209
Ogrodowczyk M. P-169
Okada Y. PL-32
Okuyan B. P-22
Olgena S. P-193
Omri A. P-238
Omurtag G.Z. P-52, P-58, P-67, P-68, P-69, P-70
Onur F. P-165
Opelia K. P-251
Orhan i. 0-10, P-102, P-103
Orman M.N. P-6
Ostatna V. PL-37
Oussoren C. PL-18
Okstiz S. P-35
Onal A. P-138
Onkol T. P-195. P-211
Ozaltin N. P-149, P-179, P-184
Ozbek H. P-79, P-109
Ozbilgin B. P-104
Ozgagli E. P-65
Ozgelik B. P-102, P-103, P-211
Ozgelikay G. P-24, P-29, P-30
Ozdemir M.T. P-33
Ozer

Palabiyik i.M. P-165
Palecek E. PL-37. 0-7. P-157
Parameshwaran K. P-225
Pardakhty A. P-236, P-240
Pastera J. P-120, P-126
Pattharachayakul S. P-20
Paululat T. P-98
Pechan Z. PL-37
Pellicciari R. PL-38
Phadoongsombut N. P-231
Pinar P.T. P-176
Pi§kin E. P-150
Pongwecharak J. P-21
Popelkova V. 0-8
Posplsil M. P-118
Potoglu Erkara 1. P-106
Pourchaire S. IP-1
Pucovsky V. PL-5
Quax W.J. PL-26
Radulescu F. 0-9
Raemisch A. PL-13
Ragno G. 0-4
Rahmani V. P-229
Rashidi MR. P-229
Ratanajamit C. P-231
Razmilic 1. P-125
Reanmongkol W. P-107
Reiazi B. 0-2
Rezaeifar M. P-241
Rodriguez J.A. PL-41, P-123. P-125
Rombaut B. PL-39
Saglik Q. P-93
Sagmanligil Ozdemir H. P-108
Sakalli 0. P-66
Sakar M.K. P-112
Salem H. P-166. P-167
Saltan-Qitoglu G. P-79. P-109. P-110
Sancar M. P-17, P-22, P-23
Sardaf S. P-65
Sarikaya M. P-10
Satil F. P-40
Sautour M. P-99
Sava§er A. P-168
Sayal A. P-93
Sayar E. P-168
Sayar F. P-150
Schaefer M. PL-40
Schins R.P.F. P-56
Schmeda-
Hirschmann
G. PL-41, P-123, P-125
Schmidt T.J. PL-42
Schramm K.W. P-64
Scurti V. 0-8

Seen H. P-105
Segal R. PL-16
Seller Z. P-191
Sen A. P-71
Senyer N. P-113
Sever Yilmaz B. P-78, P-79, P-109, P-110
Sevgi F. P-245
Seyhan S. P-183
Sezgin A. P-124
Shannon C. 0-12
Sharififar F. P-111. P-241
Sharma V. PL-31
Siahi M.R. P-226
Snedecor S. PL-16
Snel C.J. PL-18
Sonakin 0. P-242
Soyagir A. P-6
Sogut 0. P-182
Soylemezoglu T. P-53
Sriwiriyanont P. 0-5
Starosciak B. J. P-202
Stawny M. P-169. P-170
Stehfest E. PL-13
Stobaugh J.F. P-136
Storm G. PL-18
Streitova D. P-118
Subhadhirasakul S. P-107
Suksanan P. P-20
Sunada H. 0-11
Sunter 0. P-55
Sunthornpit A. P-243
Supplramaniam V. P-225
Suleymanoglu E. P-213
Sur-Altiner D. P-11. P-12
Surucu S. P-15
Suslu i. P-142. P-171. P-172
Suzen H.S. 0-1, P-62, P-66,
Suzen S. P-10, P-192, P-201
Svoboda Z. P-126
§ahbaz s. P-124
§ahin S. P-230, P-168
§ahin N. 0. P-26, P-237, P-251
§ahin F. P-217
§ahin F.P. P-49. P-50, P-51
§ahin M.F. P-195, P-196, P-211
§amdancioglu S. PL-43
§anli N. P-86, P-154
§arer E. P-75
§atana E. P-152. P-159. P-173. P-174
§atiroglu M.H. P-57, P-64
ehirli 0. P-67, P-68, P-70
§encan N. P-31
4

§ener G. P-67, P-68, P-70
§ener B. 0-10, P-88, P-91
§enturk Z. P-175. P-176
§im§ek B. P-5, P-6
§ohretoglu D. P-112
§umnu M. PL-43. P-27
§ukuroglu M. P-10
Taha M. 0. P-220
Tahir E. P-25
Tajerzadeh H. P-188, P-247
Talebpour A.H. P-101
Tamer U. 0-12
Tanvejsilp P. P-20
Tanyildiz M. P-25
Tapondjou A. L. P-100
Ta§ Q- P-38, P-39
Ta§demir D. 0-13
Tatar Ulu S. P-177
Tatli Qankaya i.i. P-124
Taylor M. PL-16
Tekin 1.0. P-54
Tekiner-Gulba§ B. P-197
Temiz-Arpaci 0. P-189, P-197
Temizer A. P-131
Theoduloz C. PL-41, P-123, P-125
Thongroichang P. P-21
Toker G. P-102, P-248
Toker M.C. P-248
Topalov M. 0-14
Topu G. P-46
Toper S. P-105
Torky A. PL-13
Torun M. P-4, P-5, P-6
Tosun F. P-89, P-90
Tozan A. P-67, P-68, P-69. P-70
Trefulka M. PL-37
Trnkova L. P-157
Tungbilek M. P-204
Tunnel M. P-106, P-186
Tungtan B. P-121
Turan K. P-252
Turan N.N. P-91, P-119
Turan G. P-191
Turanli M. P-62
Turculet L.I. P-116
Turung S.E. P-2, P-13
Tuzlaci E. P-36, P-85
Turkben C. P-86
Turker G. P-140
Turkoglu A. P-47
Tutuncu B. P-71
Ugakturk E. P-178

Ugurlu G. P-179
Uivarosi V. P-216
Ulubayram K. P-178
Ulugam G. P-191
Ulusoy Z. IP-4
Uluta§ O.K. P-71
Urairat S. P-19
Uras F. P-22
Uslu Q. P-31
Uslu B. P-129, P-147
Uthaythas S. P-225
Utku S. P-214
Uyar B. P-139, P-180
Uzun L. P-195
Uzun G. P-73
Ulgen M. P-215
Ustel i. PL-44
Ustundag A. P-72, P-73
Ustundag 0. P-181
Vacek A. P-118
Valizadeh H. P-188, P-244, P-247
Varshosaz J. P-240
Vasilev N.P. PL-26
Velescu B. P-216
Velingkar V.S. P-194
Velioglu-Ovung A. P-67
Vidinli N. P-56
Vlcek J. 0-8, P-18
Vuorela H. PL-19
Wagner H. P-98
Wang L. 0-11
Watanabe H. P-107
Weiterova L. P-118
Wlckett R. 0-5
Wiese M. PL-36
Wijayawardhane N. P-225
Winterstein A.G. PL-16
Wolf H.K. PL-18
Yagci C. P-248
Yagmur S. P-140
Yakut C. P-17
Yalgin A. P-2, P-13
Yalgin G. PL-34. P-163, P-182
Yalgin G. P-183
Yalgin i. P-189
Yalin S. P-14
Yanev S. 0-14
Yanez T. P-123, P-125
Yardimci C. P-184
Yardimci S. P-15
Yarimkaya S. P-185
Yars Ozarslan N. P-148
4

Yavuz P. P-156
Yazan Y. 0-15
Yegenoglu S. P-25
Yenice B. P-11, P-12
Yeniceli D. P-186
Yerdelen K.O. P-217. P-218. P-219
Ye§ilada A. P-178
Ye§ilada E. P-87, P-92, P-95, P-96, P-97
Yildirim 0. P-15
Yildirim K. P-74
Yildiz S. P-33
Yildiz i. P-189, P-197
Yilmaz B. P-187
Yilmaz M. P-56
Yilmaz S. P-140
Yilmaz G. 0-10
Yilmazer S. P-74
Yilmazer M. 0-1
Yuenyongsawad S. P-243
Yurdasiper A. P-245
Yuce A. P-33
Yiicesoy B. P-54
Zakeri-Milani P. P-188, P-244, P-247
Zbarcea C. P-116
Zeybek U. 0-6
Zhdanova E.R. P-246
Znojil V. P-118
Zoulova J. P-120, P-126

Kogak Farma;
Ila^ ve ilag hammaddesi iiretim tesisleri ile tibbm hizmetinde...
Serving health with medicine and API manufaciuring facilities
AVRUPA BiRLUSl*
GMP UYGUNLUK
SERTiFiKASI
LAAKELAITOS
LAKEMEDELSVERTKET
NATIONAL AGENCY
FOR MEDICINES
Ceritificate N
FI/205H/2005
EUROPEAN UNION
CERTIFICATE OF GMP
COMPLIANCE OF
S

Secretariat:
Prof. Dr. Sibel A. OZKAN
Ankara University, Faculty of Pharmacy,
Tandogan, 061 00 / Ankara-TURKEY
, Participants from ISOPS - 7
Phone +90 312 223 82 43
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E-mail ankpharm@pharmacy.ankara.edu.tr
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