Faculty of Pharmacy - Mahidol University

Mahidol University Annual Research Abstracts, Vol. 36 

ATOMIC FORCE MICROSCOPY IMAGING OF 

NOVEL SELF-ASSEMBLING PECTIN-LIPOSOME 

NANOCOMPLEXES (NO. 772) 

Pornsak Sriamornsak 1,2 , Nathaya Wattanakorn 1,2 , Jurairat 

Nunthanid 1,2 , Satit Puttipipatkhachorn 3 , Jringjai Thongborisute 4 , 

Hirofumi Takeuchi 4 

1 Department of Pharmaceutical Technology and 2 Pharmaceutical 

Biopolymer Group (PBiG), Faculty of Pharmacy, Silpakorn 

University, Nakhon Pathom, 73000, Thailand; 3 Department of 

Manufacturing Pharmacy, Faculty of Pharmacy, Mahidol 

University, Bangkok, 10400, Thailand. E-mail-: pyspt@ 

mahidol.ac.th; 4 Laboratory of Pharmaceutical Engineering, Gifu 

Pharmaceutical University, 5-6-1 Mitahora-Higashi, Gifu, 502 

8585, Japan. 

Key words: Atomic force microscopy (AFM), Liposomes, Pectin 

Self-assembling pectin-liposome nanocomplexes (PLNs) 

were prepared by a simple mixing of cationic liposomes with pectin 

solution. Nanostructures of liposomes, pectin, and PLNs were 

observed by atomic force microscopy (AFM). The AFM images of 

pectin show a chain-like structure with a small number of branches 

while those of liposomes show a spherical form. The AFM images 

also provided a direct evidence for association of cationic liposomes 

on the pectin chain. This was confirmed by the FTIR analysis. 

(Published in Carbohydrate Polymers 2008;71(2):324-329. Funded 

by Commission of Higher Education, Thailand and the Thailand 

Research Fund.) 

ALGINATE-MAGNESIUM ALUMINUM SILICATE 

COMPOSITE FILMS : EFFECT OF FILM 

THICKNESS ON PHYSICAL CHARACTERISTICS 

AND PERMEABILITY (NO. 773) 

Thaned Pongjanyakul 1 , Satit Puttipipatkhachorn 2 

1 Faculty of Pharmaceutical Sciences, Khon Kaen University, 

Khon Kaen, 40002, Thailand; 2 Department of Manufacturing 

Pharmacy, Faculty of Pharmacy, Mahidol University, Bangkok, 

10400, Thailand. E-mail : pyspt@mahidol.ac.th 

Key words: Sodium alginate, Magnesium aluminum silicate, Film 

The different film thicknesses of the sodium alginatemagnesium 

aluminum silicate (SA-MAS) microcomposite films were 

prepared by varying volumes of the composite dispersion for casting. 

Effect of film thickness on thermal behavior, solid-state crystallinity, 

mechanical properties, water uptake and erosion, and water vapor 

and drug permeability of the microcomposite films were investigated. 

The film thickness caused a small change in thermal behavior of the 

films when tested using DSC and TGA. The crystallinity of the thin 

films seemed to increase when compared with the thick films. The 

thin films gave higher tensile strength than the thick films, whereas 

% elongation of the films was on the contrary resulted in the lower 

Young’s modulus of the films when the film thickness was increased. 

This was due to the weaker of the film bulk, suggesting that the 

microscopic matrix structure of the thick films was looser than that 

285 

of the thin films. Consequently, water uptake and erosion, water vapor 

permeation and drug diffusion coefficient of the thick films were 

higher than those of the thin films. The different types of drug on 

permeability of the films also showed that a positive charge and large 

molecule of drug, propranolol HCl, had higher lag time and lower 

diffusion coefficient that acetaminophen, a non-electrolyte and small 

molecule. This was because of a higher affinity of positive charge 

drug on MAS in the films. The findings suggest that the evaporation 

rate of solvent in different volumes of the composite dispersion used 

in the preparation method could affect crystallinity and strength of 

the film surface and film bulk of the microcomposite films. This led 

to a change in water vapor and drug permeability of the films. 

(Published in International Journal of Pharmaceutics 2008;346(1- 

2):1-9. Funded by Commission of Higher Education, Thailand and 

the Thailand Research Fund.) 

DEVELOPMENT OF TIME- pH-, AND ENZYME- 

CONTROLLED COLONIC DRUG DELIVERY USING 

SPRAY-DRIED CHITOSAN ACETATE AND HYDRO- 

XYPROPYL METHYLCELLULOSE (NO. 774) 

Jurairat Nunthanid 1,2 , Kampanart Huanbutta 1,2 , Manee 

Luangtana-anan 1,2 , Pornsak Sriamornsak 1,2 , Sontaya 

Limmatvapirat 1,2 , Satit Puttipipatkhachorn 3 





University, Bangkok, 10400, Thailand. E-mail-: 

pyspt@mahidol.ac.th 

Key words: Chitosan acetate, Colonic drug delivery, Compressioncoated 

tablets 

A colonic drug delivery with a new concept based on a 

combination of time-, pH-, and enzyme-controlled system was 

developed. Spray-dried chitosan acetate (CSA) prepared from low 

molecular weight chitosan was characterized. A combination of CSA 

and hydroxypropyl methylcellulose (HPMC) was used as new 

compression-coats for 5-aminosalicylic acid (5-ASA) tablets. Factors 

affecting in-vitro drug release, i.e. % weight ratio of coating polymers, 

enzyme activity, pH of media, and excipients in core tablets, were 

evaluated. The tablets compression-coated with HPMC:CSA at 60:40 

and 50:50% weight ratio providing lag times about 5–6 h were able 

to pass through the stomach (stage I, 0.1 N HCl) and small intestine 

(stage II, pH 6.8, Tris–HCl). The delayed release was time- and pHcontrolled 

owing to the swelling with gradual dissolving of CSA and 

HPMC in 0.1 N HCl and the less solubility of CSA at higher pH. 

After reaching the colon (stage III, pH 5.0, acetate buffer), the 

dissolution of CSA at low pH triggered the drug release over 90% 

within 14 h. Furthermore, the degradation of CSA by -glucosidase 

in the colonic fluid enhanced the drug release while adding the 

disintegrant or the osmotic agent in the core tablets would affect the 

drug release. 

(Published in European Journal of Pharmaceutics and 

Biopharmaceutics 2008;68: 253-259. Funded by the Thailand 

Research Fund.)

286 Faculty of Pharmacy 

CHITOSAN-MAGNESIUM ALUMINUM SILICATE 

COMPOSITE DISPERISONS : CHARACTERIZA- 

TION OF RHEOLOGY, FLOCCULATE SIZE AND 

ZETA POTENTIAL (NO. 775) 

Wanwisa Khunawattanakul 1 , Satit Puttipipatkhachorn 2 , Thomas 

Rades 3 , Thaned Pongjanyakul 1 

1 Faculty of Pharmaceutical Sciences, Khon Kaen University, 

Khon Kaen, 40002, Thailand; 2 Department of Manufacturing 


10400, Thailand. E-mail : pyspt@mahidol.ac.th; 3 School of 

Pharmacy, University of Otago, PO Box 913, Dunedin, New 

Zealand. 

Key words: Chitosan, Magnesium aluminum silicate, Composite 

dispersion 

Composite dispersions of chitosan (CS), a positively 

charged polymer, and magnesium aluminum silicate (MAS), a 

negatively charged clay, were prepared and rheology, flocculate size 

and zeta potential of the CS-MAS dispersions were investigated. High 

and low molecular weights of CS (HCS and LCS, respectively) were 

used in this study. Moreover, the effects of heat treatment at 60 C 

on the characteristics of the CS-MAS dispersions and the zeta 

potential of MAS upon addition of CS at different pHs were examined. 

Incorporation of MAS into CS dispersions caused an increase in 

viscosity and a shift of CS flow type from Newtonian to pseudoplastic 

flow with thixotropic properties. Heat treatment brought about a 

significant decrease in viscosity and hysteresis area of the composite 

dispersions. Microscopic studies showed that flocculation of MAS 

occurred after mixing with CS. The size and polydispersity index of 

the HCS-MAS flocculate were greater than those of the LCS-MAS 

flocculate. However, a narrower size distribution and the smaller size 

of the HCS-MAS flocculate were found after heating at 60 C. Zeta 

potentials of the CS-MAS flocculates were positive and slightly 

increased with increasing MAS content. In the zeta potential studies, 

the negative charge of the MAS could be neutralized by the addition 

of CS. Increasing the pH and molecular weight of CS resulted in 

higher CS concentrations required to neutralize the charge of MAS. 

These findings suggest that the electrostatic interaction between CS 

and MAS caused a change in flow behavior and flocculation of the 

composite dispersions, depending on the molecular weight of CS. 

Heat treatment affected the rheological properties and the flocculate 

size of the composite dispersions. Moreover, pH of medium and 

molecular weight of CS influence the zeta potential of MAS. 

(Published in International Journal of Pharmaceutics 2008;351(1- 

2):227-235. Funded by RGJ-Ph.D. Program, the Thailand Research 

Fund and Commission of Higher Education, Thailand) 

THE EFFECT OF CETYL PALMITATE CRYSTAL- 

LINITY ON PHYSICAL PROPERTIES OF GAMMA- 

ORYZANOL ENCAPSULATED IN SOLID LIPID 

NANOPARTICLES (NO. 776) 

Uracha Ruktanonchai 1 , Surachai Limpakdee 2 , Siwaporn Meejoo 2 , 

Usawadee Sakulkhu 1 , Nuntavan Bunyapraphatsara 3 , Varaporn 

Junyaprasert 4 , Satit Puttipipatkhachorn 5 

1 National Nanotechnology Center, National Science and 

Technology Development Agency, Pathumthani 12120,Thailand; 

2 Department of Chemistry, Faculty of Science, Mahidol 

University, Rama VI Road, Bangkok 10400, Thailand; 

3 Department of Pharmacognosy, 4 Department of Pharmacy, and 

5 Department of Manufacturing Pharmacy, Faculty of Pharmacy, 

Mahidol University, Sri-ayudhya Road, Bangkok 10400, 

Thailand. E-mail : pyspt@mahidol.ac.th 

Key words : Solid lipid nanoparticles, Gamma-oryzanol, Crystallinity 

This present study was aimed at investigating the effect 

of the crystallinity of cetyl palmitate based solid lipid nanoparticles 

(SLNs) on the physical properties of -oryzanol-loaded SLNs. SLNs 

consisting of varying ratios of cetyl palmitate and -oryzanol were 

prepared. Their hydrodynamic diameters were in the range 210–280 

nm and the zeta potentials were in the range “27 to “35 mV. The size 

of SLNs increased as the amount of cetyl palmitate decreased whereas 

no significant change of zeta potentials was found. Atomic force 

microscopy pictures indicated the presence of disc-like particles. The 

crystallinity of SLNs, determined by differential scanning calorimetry 

and powder x-ray diffraction, was directly dependent on the ratio of 

cetyl palmitate to -oryzanol and decreased with decreasing cetyl 

palmitate content in the lipid matrix. Varying this ratio in the lipid 

mix resulted in a shift in the melting temperature and enthalpy, 

although the SLN structure remained unchanged as an orthorhombic 

lamellar lattice. This has been attributed to a potential inhibition by 

-oryzanol during lipid crystal growth as well as a less ordered 

structure of the SLNs. The results revealed that the crystallinity of 

the SLNs was mainly dependent on the solid lipid, and that the 

crystallinity has an important impact on the physical characteristics 

of active-loaded SLNs. 

(Published in Nanotechnology 2008;19: 095701 (10pp). Funded by 

the National Nanotechnology Center (NANOTEC), Thailand) 

FORMATION, PHYSICAL STABILITY AND IN VITO 

ANTIMALARIAL ACTIVITY OF DIHYDROARTEMI- 

SININ NANOSUSPENSIONS OBTAINED BY CO- 

GRINDING METHOD (NO. 777) 

Jiraporn Chingunpitak 1 , Satit Puttipipatkhachorn 1 , Porntip 

Chavalitshewinkoon-Petmitr 2 , Yuichi Tozuka 3 , Kunikazu 

Moribe 4 , Keiji Yamamoto 4 


Mahidol University, Sri-ayudhya road, Bangkok 10400, Thailand. 

E-mail : pyspt@mahidol.ac.th; 2 Department of Protozoology, 

Faculty of Tropical Medicine, Mahidol University, Rajvithi road, 

Bangkok 10400, Thailand; 3 Graduate School of Pharmaceutical 

Sciences, Gifu Pharmaceutical University, 5-6-1 Mitahorahigashi, 

Gifu 502-8585, Japan; 4 Graduate School of Pharmaceutical 

Sciences, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba 263- 

8522, Japan. 

Key words : Dihydroartemisinin, Nanosuspension, Malaria 

The purpose of this study was to investigate the formation 

of drug nanoparticles from binary and ternary mixtures, consisting 

of dihydroartemisinin (DHA), a poorly water-soluble antimalarial 

drug, with water-soluble polymer and/or surfactant. Binary mixtures 

of drug/polyvinyl pyrrolidone K30 (PVP K30), binary mixtures of


drug/sodium deoxycholate (NaDC), and ternary mixtures of drug/ 

PVP K30/NaDC were prepared at different weight ratios and then 

ground by vibrating rod mill to obtain ground mixtures. 

Nanosuspension was successfully formed after dispersing ternary 

ground mixtures or DHA/NaDC ground mixtures in water. The ternary 

ground mixtures did not give superior nanosuspension in terms of 

particle size reduction and recovery of drug nanoparticles, but they 

provided more physically stable nanosuspensions than DHA/NaDC 

ground mixtures. The size of drug nanoparticles was decreased with 

increasing grinding time and lowering amount of PVP K30 and NaDC. 

About 95% of drug nanoparticles were found in the nanosuspension 

from ternary ground mixtures. Zeta potential measurement suggested 

that stable nanosuspension was attributable to adsorption of NaDC 

and PVP K30 onto surface of drug particles. Atomic force microscopy 

and transmission electron microscopy with selected area diffraction 

indicated that DHA in nanosuspension was existed as nanocrystals. 

The obtained nanosuspensions had higher in vitro antimalarial 

acitivity against Plasmodium falciparum than microsuspensions. The 

results suggest that co-grinding of DHA with PVP K30 and NaDC 

seems to be a promising method to prepare DHA nanosuspension. 

(Published in Drug Development and Industrial Pharmacy 

2008;34(3):314-322. Funded by the Thailand Research Fund 

through the RGJ-Ph.D. Program,) 

ALGINATE-BASED PELLETS PREPARED BY 

EXTRUSION/SPHERONIZATION : EFFECT OF THE 

AMOUNT AND TYPE OF SODIUM ALGINATE AND 

CALCIUM SALTS (NO. 778) 

Pornsak Sriamornsak 1,2 , Jurairat Nunthanid 1,2 , Manee 

Luangtana-anan 1,2 , Yossanun Weerapol 1 , Satit 

Puttipipatkhachorn 3 

1 Department of Pharmaceutical Technology and 

2 Pharmaceutical Biopolymer Group (PBiG), Faculty of 

Pharmacy, Silpakorn University, Nakhon Pathom 73000, 

Thailand; 3 Department of Manufacturing Pharmacy, Faculty of 

Pharmacy, Mahidol University, Bangkok 10400, Thailand. E-mail 

: pyspt@mahidol.ac.th. 

Key words : Alginate, Extrusion/Spheronization, Pellets 

Pellets containing microcrystalline cellulose (MCC), a 

model drug (theophylline) and a range of levels of sodium alginate 

(i.e., 10-50% w/w) were prepared by extrusion/spheronization. Two 

types of sodium alginate were evaluated with and without the addition 

of either calcium acetate or calcium carbonate (0, 0.3, 3 and 10% w/ 

w). The effects of amount and type of sodium alginate and calcium 

salts on pellet properties, e.g., size, shape, morphology and drug 

release behavior, were investigated. Most pellet formulations resulted 

in pellets of a sufficient quality with respect to size, size distribution 

and shape. The results showed that the amounts of sodium alginate 

and calcium salts influenced the size and shape of the obtained pellets. 

However, different types of sodium alginate and calcium salt 

responded to modifications to a different extent. A cavity was observed 

in the pellet structure, as seen in the scanning electron micrographs, 

resulting from the forces involved in the spheronization process. Most 

of pellet formulations released about 75-85% drug within 60 min. 

Incorporation of calcium salts in the pellet formulations altered the 

drug release, depending on the solubility of the calcium salts used. 

The drug release data showed a good fit into both Higuchi and 

Korsmeyer-Peppas equations. 


Biopharmaceutics 2008;69(1):274-284.) 

DESIGN AND EVALUATION OF FLOATING 

MULTI-LAYER COATED TABLETS BASED ON 

GAS FORMATION (NO. 779) 

Srisagul Sungthongjeen 1 , Pornsak Sriamornsak 2 , Satit 

Puttipipatkhachorn 3 

1 Department of Pharmaceutical Technology, Faculty of 

Pharmaceutical Sciences, Naresuan University, Phitsanulok, 

Thailand; 2 Department of Pharmaceutical Technology, 

Pharmaceutical Biopolymer Group (PBiG), Faculty of Pharmacy, 

Silpakorn University, Nakhon Pathom 73000, Thailand; 


Mahidol University, Bangkok 10400, Thailand. E-mail : 


Key words : Floating tablets, Drug delivery system; Sustained release 

Floating multi-layer coated tablets were designed based 

on gas formation. The system consists of a drug-containing core tablet 

coated with a protective layer (hydroxypropyl methylcellulose), a gas 

forming layer (sodium bicarbonate) and a gas-entrapped membrane, 

respectively. The mechanical properties of acrylic polymers 

(Eudragit RL 30D, RS 30D, NE 30D) and ethylcellulose were 

characterized by the puncture test in order to screen a suitable film 

for the system. Eudragit RL 30D was chosen as a gas-entrapped 

membrane due to its high flexibility and high water permeability. 

The obtained tablets enabled to float due to the CO2-gas formation 

and the gas entrapment by polymeric membrane. The effect of 

formulation variables on floating properties and drug release was 

investigated. The floating tablets using direct-compressed cores had 

shorter time to float and faster drug release than those using wetgranulated 

cores. The increased amount of a gas forming agent did 

not affect time to float but increased the drug release from the floating 

tablets while increasing coating level of gas-entrapped membrane 

increased time to float and slightly retarded drug release. Good 

floating properties and sustained drug release were achieved. These 

floating tablets seem to be a promising gastroretentive drug delivery 

system. 


Biopharmaceutics 2008;69(1):255-263. Funded by Commission of 

Higher Education, Thailand and the Thailand Research Fund.) 

MICRONIZATION OF DIHYDROARTEMISININ 

BY RAPID EXPANSION OF SUPERCRITICAL 

SOLUTIONS (NO. 780) 

287 

Jiraporn Chingunpitak 1 , Satit Puttipipatkhachorn 1 , Yuichi 

Tozuka 2 , Kunikazu Moribe 3 , Keiji Yamamoto 3 


Mahidol University, Sri-ayudhya road, Bangkok 10400, Thailand. 

E-mail : pyspt@mahidol.ac.th; 2 Graduate School of 

Pharmaceutical Sciences, Gifu Pharmaceutical University, 5-6-


1 Mitahorahigashi, Gifu 502-8585, Japan; 3 Graduate School of 

Pharmaceutical Sciences, Chiba University, 1-33 Yayoi-cho, 

Inage-ku, Chiba 263-8522, Japan. 

Key words: Dihydroartemisinin, Micronization, Malaria 

The purpose of this study was to prepare fine particles of 

antimalarial drug dihydroartemisinin (DHA) by rapid expansion of 

supercritical solutions (RESS) using carbon dioxide as supercritical 

fluid. The mechanical grinding by jet mill and additional vibration 

rod mill also was performed as a comparative method. In the RESS 

process, drug particles were prepared by varying processing 

conditions, including extraction condition, pre-expansion condition, 

nozzle diameter, nozzle temperature, and collecting distance. Particle 

size and morphology and physicochemical characteristics of the drug 

particles were investigated. The RESS process could produce the 

smaller drug particles (about 1-2 m) when compared to mechanical 

grinding method (about 7 m). All RESS processing parameters had 

an effect on size and morphology of drug particles. The particle size 

of drug was related to the solubility of drug in supercritical CO2 at 

each processing condition. The fine particles of DHA (about 1 m) 

with narrow size distribution could be obtained at extraction pressure 

of 18 MPa and extraction temperature of 32 C, which was closed to 

the critical temperature of supercritical CO2 whereas broad size 

distribution was obtained at extraction temperature of 60 C. Powder 

X-ray diffraction study indicated that the RESS-processed particles 

were in crystalline form. The results revealed that RESS process is 

applicable for micronization of DHA. 

(Published in Drug Development and Industrial Pharmacy 

2008;34(6):609-617. Funded by the Thailand Research Fund 

through the RGJ-Ph.D. Program,) 

MODULATION OF DRUG RELEASE KINETICS OF 

SHELLAC-BASED MATRIX TABLETS BY IN-SITU 

POLYMERIZATION THROUGH ANNEALING 

PROCESS (NO. 781) 

Sontaya Limmatvapirat 1,2 , Chutima Limmatvapirat 3 , Satit 

Puttipipatkhachorn 4 , Jurairat Nunthanid 1,2 , Manee Luangtana- 

Anan 1,2 , Pornsak Sriamornsak 1,2 

1 Department of Pharmaceutical Technology; 2 Pharmaceutical 

Biopolymer Group (PBiG) and 3 Department of Pharmaceutical 

Chemistry, Faculty of Pharmacy, Silpakorn University, Nakhon 

Pathom 73000, Thailand; 4 Department of Manufacturing 

Pharmacy, Faculty of Pharmacy, Mahidol University, Bangkok 

10400, Thailand. E-mail : pyspt@mahidol.ac.th 

Key words : Shellac, Matrix tablets, Controlled release 

A new oral-controlled release matrix tablet based on shellac 

polymer was designed and developed, using metronidazole (MZ) as 

a model drug. The shellac-based matrix tablets were prepared by wet 

granulation using different amounts of shellac and lactose. The effect 

of annealing temperature and pH of medium on drug release from 

matrix tablets was investigated. The increased amount of shellac and 

increased annealing temperature significantly affected the physical 

properties (i.e., tablet hardness and tablet disintegration) and MZ 

release from the matrix tablets. The in-situ polymerization played a 

major role on the changes in shellac properties during annealing 

process. Though the shellac did not dissolve in acid medium, the 

MZ release in 0.1 N HCl was faster than in pH 7.3 buffer, resulting 

from a higher solubility of MZ in acid medium. The modulation of 

MZ release kinetics from shellac-based matrix tablets could be 

accomplished by varying the amount of shellac or annealing 

temperature. The release kinetics was shifted from relaxationcontrolled 

release to diffusion-controlled release when the amount 

of shellac or the annealing temperature was increased. 


Biopharmaceutics 2008;69(3):1004-1013. Funded by Commission 

of Higher Education, Thailand and the Thailand Research Fund.) 

FORMATION OF SHELLAC SUCCINATE HAVING 

IMPROVED ENTERIC FILM PROPERTIES 

THROUGH DRY MEDIA REACTION (NO. 782) 

Sontaya Limmatvapirat 1,2 , Danuch Panchapornpon 1,2 , Chutima 

Limmatvapirat 3 , Jurairat Nunthanid 1,2 , Manee Luangtana- 

Anan 1,2 , Satit Puttipipatkhachorn 4 

1 Department of Pharmaceutical Technology; 2 Pharmaceutical 

Biopolymer Group (PBiG) and 3 Department of Pharmaceutical 

Chemistry, Faculty of Pharmacy, Silpakorn University, Nakhon 

Pathom 73000, Thailand; 4 Department of Manufacturing 

Pharmacy, Faculty of Pharmacy, Mahidol University, Bangkok 

10400, Thailand. E-mail: pyspt@mahidol.ac.th 

Key words : Co-grinding, Enteric polymer, Shellac 

The aim of this study was to improve enteric properties of 

shellac by the formation of succinate derivative through dry media 

reaction. Shellac and succinic anhydride were mixed and then coground 

by planetary ball mill. The ground mixture was then activated 

by heating for various times and washed for removal of excess 

succinic anhydride. The ground mixtures and the heat-activated 

mixtures were characterized by physical and chemical tests, including 

acid value, FTIR spectroscopy, 1H NMR and 13C NMR spectroscopy, 

thermal analysis and film properties. The results demonstrated that 

acid values of heat-activated shellac mixtures increased with the 

increase of annealing time, suggesting the presence of carboxylic 

acid moieties of succinate at shellac molecules. The results were in 

good agreement with the DSC thermograms. The melting peak of 

shellac disappeared after heating, while melting peak of succinic 

anhydride gradually decreased, suggesting the utilization of succinic 

anhydride for the esterification. The shellac succinate formation was 

also confirmed by 1H NMR and 13C NMR spectroscopies. Film 

prepared from shellac succinate showed improved solubility, 

especially at the pH of small intestine (5.8-6.7), as compared to native 

shellac. The shellac succinate film also demonstrated better 

mechanical property, in terms of increased flexibility. In conclusion, 

solid-state formation of shellac succinate ester, which had improved 

enteric properties, was easily accomplished under the concept of 

“green approach”. 2008 Elsevier B.V. All rights reserved. 


Biopharmaceutics 2008;70(1):335-344. Funded by Commission of 

Higher Education, Thailand and the Thailand Research Fund.)


MUCOADHESION OF PECTIN AS EVIDENCE BY 

WETTABILITY AND CHAIN INTERPENETRA- 

TION (NO. 783) 

Pornsak Sriamornsak 1,2 , Nathaya Wattanakorn 1,2 , Jurairat 

Nunthanid 1,2 , Satit Puttipipatkhachorn 3 





University, Bangkok, 10400, Thailand. E-mail : 


Key words : ATR-FTIR, Mucoadhesion, Pectin 

Different types of pectin were characterized for 

gastrointestinal (GI) mucoadhesion. The mechanisms of 

mucoadhesion of these pectins were investigated by measurements 

of surface tension, contact angle, and FTIR studies. The surface 

tension of different GI fluids was relatively the same and found to 

decrease after addition of mucin. The contact angle of tested fluids 

on pectin surfaces behaved as time-dependent and the values at 30 s 

were used for comparison. The type of pectin, addition of mucin and 

pectin surfaces influenced the contact angle of GI fluids. The 

thermodynamic work of adhesion (Wad/therm) and spreading 

coefficient were calculated. The positive values of Wad/therm 

indicated that the pectin surfaces could be wet spontaneously by 

adhesional process. The pectin with higher molecular weight and 

higher degree of substitution showed a lower Wad/therm, resulting 

from the low wettability. The spreading coefficient of tested fluids 

containing mucin on pectin surface was negative, suggesting that no 

spreading could be occurred spontaneously. ATR-FTIR studies 

revealed the changes resulting from interpenetration of pectin-mucin 

chains at film interface and the formation of hydrogen bonds. The 

results obtained from this study demonstrated that the wetting 

behavior and work of adhesion could be used to explain the 

mechanism of mucoadhesion of various pectins in GI conditions, 

and that a chain interdiffusion occurred at the interface of pectin 

film and mucin solution which supported the diffusion theory of 

mucoadhesion. 

(Published in Carbohydrate Polymers 2008;74(3):458-467. Funded 

by Commission of Higher Education, Thailand and the Thailand 

Research Fund) 

EFFECT OF ACIDIC MEDIUM ON SWELLING 

AND RELEASE BEHAVIORS OF CHITOSAN- 

REINFORCED CALCIUM PECTINATE GEL BEADS 

(NO. 784) 

Pornsak Sriamornsak 1,2 , Kanokporn Burapapadh 1,2 , Satit 

Puttipipatkhachorn 3 , Jurairat Nunthanid 1,2 



University, Nakhon Pathom 73000, Thailand; 3 Department of 


University, Bangkok 10400, Thailand. E-mail : 


Key words : Chitosan, Pectin, Beads 

Chitosan-reinforced calcium pectinate (ChCP) gel beads 

were prepared by ionotropic gelation method. The swelling of ChCP 

gel beads and release behavior of indomethacin from the beads were 

investigated and compared with conventional calcium pectinate (CP) 

gel beads. The factors, such as molecular weight of chitosan, 

concentration of chitosan, and release medium, which can have a 

significant effect on the swelling and release behaviors from the beads, 

were discussed in this study. The mechanical test showed that the 

ChCP beads have slightly higher strength than that of CP beads. The 

swelling index of the ChCP beads in acidic medium was much lower 

than that in neutral medium. The release of indomethacin from ChCP 

beads under conditions mimicking intestinal transit were evaluated 

in pH 7.4 Tris buffer. The acid pretreatment caused a faster drug 

release from ChCP beads. The less swelling in acidic medium and 

faster drug release of acid-pretreated ChCP beads may be due to the 

dissolution of chitosan from the beads in acidic medium, as no 

fluorescence signal was seen at the shell of the beads. The results 

suggested that the acid, which essentially found in stomach, 

influenced the swelling and release behaviors of ChCP beads. 

(Published in Silpakorn University Science and Technology Journal 

2008;2: 37-44) 

ISOLATION OF ERGOSTEROL PEROXIDE FROM 

NOMURAEA RILEYI INFECTED LARVAE OF 

TOBACCO CUTWORM (NO. 785) 

289 

Pannipa Prompiboon 1 , Amaret Bhumiratana 2 , Somsak 

Ruchirawat 3 , Drion G. Boucias 4 , Chanpen Wiwat 1 

1 Department of Microbiology, Faculty of Pharmacy, Mahidol 

University, Bangkok 10400, Thailand; 2 Department of 

Biotechnology, Faculty of Science, Mahidol University, Bangkok 

10400, Thailand; 3 Chulabhorn Research Institute, Vipavadee- 

Rangsit Highway, Bangkok 10210, Thailand; 4 Department of 

Entomology and Nematology, Institute of Food and Agricultural 

Sciences, University of Florida, 110620, Gainesville, FL 32611, 

United States. 

Key words : Entomopathogenic fungus, Ergosterol peroxide, 

Nomuraea rileyi, Spodoptera litura 

In the search for potential cytotoxic substances produced 

by Nomuraea rileyi, an active compound was isolated from mycosed 

insects through an activity guided fractionation process. The 

compound, cytotoxic against the Sf9 insect cell line, was identified 

to be ergosterol peroxide (5 , 8 -epidioxy-24(R)-methylcholesta-6, 

22-dien-3 -ol) using nuclear magnetic resonance techniques, infrared 

spectrometry, and mass spectroscopy. Anticancer screens 

demonstrated that ergosterol peroxide at micromolar concentrations 

inhibited the growth of hormone-dependent breast cancer cell line 

(T47D), hormone-independent breast cancer cell line (MDA-MB- 

231), human epidermoid carcinoma in mouth cell line (KB), human 

cervical carcinoma cell line (HeLa), lung cancer cell line (H69AR) 

and human cholangiocarcinoma cell line (HuCCA-1). Ergosterol 

peroxide showed moderate effects against Spodoptera litura larvae; 

46.7% mortality via topical application after 7 day post-treatment 

whereas the insect’s death was not found in per os application. The 

amounts of ergosterol peroxide produced by N. rileyi cultures under 

in vitro and in vivo were determined. The physiological levels of


ergosterol peroxide detected in mycosed and mummified cadavers 

were very low (0.011 and 0.386 g/larva) less then levels that either 

inhibited insect cell proliferation or caused insecticidal activity. 

(World Journal of Microbiology and BiotechnologyVolume 24, Issue 

12, December 2008, Pages 2909-2917.) (This research was supported 

by the Royal Golden Jubilee Ph.D. Scholarship (PHD/0287/2545), 

the Thailand Research Fund and Mahidol University.) 

STUDY ON CYTOTOXICITY AND NUCLEOTIDE 

SEQUENCES OF ENTEROTOXIN FM OF 

BACILLUS CEREUS ISOLATED FROM VARIOUS 

FOOD SOURCES. (NO. 786) 

Boonchai, N. 1 , Asano, S.-I. 2 , Bando, H.2, Wiwat, C. 1,3 


University, Bangkok, Thailand; 2 Faculty of Agriculture, 

Hokkaido University, Sapporo, Hokkaido, Japan; 3 Department 

of Microbiology, Faculty of Pharmacy, Mahidol University, 447 

Sri-Ayudhya Rd, Rachatwewee, Bangkok 10400, Thailand. 

Key words : Bacillus cereus; Cytotoxicity; Enterotoxin FM; Food 

poisoning; Sequencing 

Objective: To determine the cytotoxicity of the crude 

culture broth containing enterotoxins of B. cereus and investigate 

the nucleotide sequences of ent FM among various isolates B. cereus. 

Material and Method: The 1.2 kb fragment of ent FM of B. cereus 

was amplified, cloned, sequenced, and compared with published 

sequences. The biological activity of crude culture broth containing 

enterotoxins with and without monoclonal antibodies against HBL, 

NHE, and EntFM enterotoxins was tested using Vero cells assay. 

Results: A percentage homology of nucleotide sequences and deduced 

amino acid sequences among test isolates and published strains were 

90-99% and 88-99%, respectively. An assays for cytotoxic activity 

revealed that seven and three of B. cereus isolates were positive for 

NHE enterotoxin and EntFM enterotoxin, respectively. In addition, 

the 4-repeating sequences “TCAAAC” of ent FM were found, which 

may or may not be probably correlated with cytotoxicity of B. cereus. 

Conclusion: The enterotoxin FM from B. cereus isolates is cytotoxic 

and the degree of cytotoxicity depends on the bacterial strain. 

(Journal of the Medical Association of Thailand Volume 91, Issue 9, 

September 2008, Pages 1425-1432.) 

ANTIBACTERIAL ACTIVITY OF THAI MEDICINAL 

PLANTS AGAINST METHICILLIN-RESISTANT 

STAPHYLOCOCCUS AUREUS (NO. 787) 

Mullika Traidej Chomnawang 1 , Suvimol Surassmo 1 , Karn 

Wongsariya 1 and Nuntavan Bunyapraphatsara 2 


University, Bangkok, Thailand; 2 Department of Pharmacognosy, 

Faculty of Pharmacy, Mahidol University, Bangkok, Thailand 

Key words: Methicillin-resistant S. aureus; Medicinal plants; 

Antibacterial activity; Bioautography; Garcinia mangostana; - 

mangostin 

Methicillin-resistant Staphylococcus aureus (MRSA) is a 

major nosocomial pathogen which causes severe morbidity and 

mortality worldwide. Seventeen Thai medicinal plants were 

investigated for their activity against MRSA. Garcinia mangostana 

was identified as the most potent plant, and its activity was traced to 

the prenylated xanthone, -mangostin (MIC and MBC values of 1.95 

and 3.91 g/ml, respectively). 

(Fitoterapia Article in Press ) (This work was supported by the 

Mahidol University and the Thailand Research Fund.) 

3D-QSAR STUDIES ON CHROMONE DERIVATIVES 

AS HIV-1 PROTEASE INHIBITORS : APPLICA- 

TION OF MOLECULAR FIELD ANALYSIS (NO. 788) 

Nunthanavanit, P. 1 , Anthony, N.G. 2 , Johnston, B.F. 2 , Mackay, S.P. 2 , 

Ungwitayatorn, J. 3,4 

1 Faculty of Pharmacy, Srinakharinwirot University, Nakhon 

Nayok, Thailand; 2 Strathclyde Institute for Pharmacy and 

Biomedical Sciences, University of Strathclyde, Glasgow, United 

Kingdom; 3 Faculty of Pharmacy, Mahidol University, Bangkok, 

Thailand; 4 Faculty of Pharmacy, Mahidol University, 447 Sri- 

Ayudhya Road, Bangkok 10400, Thailand. 

Key words : 3D-QSAR; Alignment; Chromone derivatives; HIV-1 

protease; Molecular field analysis (MFA) 

Three-dimensional quantitative structure-activity 

relationship (3D-QSAR) models were developed for chromone 

derivatives against HIV-1 protease using molecular field analysis 

(MFA) with genetic partial least square algorithms (G/PLS). Three 

different alignment methods: field fit, pharmacophore-based, and 

receptor-based were used to derive three MFA models. All models 

produced good predictive ability with high cross-validated r2 (r2cv), 

conventional r2, and predictive r2 (r 2pred) values. The receptorbased 

MFA showed the best statistical results with r2cv = 0.789, r2 = 

0.886, and r2pred = 0.995. The result obtained from the receptorbased 

model was compared with the docking simulation of the most 

active compound 21 in this chromone series to the binding pocket of 

HIV-1 protease (PDB entry 1AJX). It was shown that the MFA model 

related well with the binding structure of the complex and can provide 

guidelines to design more potent HIV-1 protease inhibitors. 2008 

Wiley-VCH Verlag GmbH & Co. KGaA. 

(Archiv der Pharmazie Volume 341, Issue 6, June 2008, Pages 357- 

364) (This study was granted by Faculty of Graduate Studies, 

Mahidol University.) 

3D-QSAR INVESTIGATION OF SYNTHETIC 

ANTIOXIDANT CHROMONE DERIVATIVES BY 

MOLECULAR FIELD ANALYSIS (NO. 789) 

Samee, W. 1 , Nunthanavanit, P. 2 , Ungwitayatorn, J. 2 

1 Department of Pharmaceutical Chemistry and Pharmacognosy, 

Faculty of Pharmacy, Srinakharinwirot University, Nakhon 

Nayok, 26120, Thailand; 2 Department of Pharmaceutical 

Chemistry, Faculty of Pharmacy, Mahidol University, Bangkok, 

10400, Thailand


Key words : 3D-QSAR; Antioxidants; Chromone; Genetic partial 

least squares (G/PLS) method; Molecular field analysis (MFA) 

A series of 7-hydroxy, 8-hydroxy and 7,8-dihydroxy 

synthetic chromone derivatives was evaluated for their DPPH free 

radical scavenging activities. A training set of 30 synthetic chromone 

derivatives was subject to three-dimensional quantitative structureactivity 

relationship (3D-QSAR) studies using molecular field 

analysis (MFA). The substitutional requirements for favorable 

antioxidant activity were investigated and a predictive model that 

could be used for the design of novel antioxidants was derived. 

Regression analysis was carried out using genetic partial least squares 

(G/PLS) method. A highly predictive and statistically significant 

model was generated. The predictive ability of the developed model 

was assessed using a test set of 5 compounds (r2pred = 0.924). The 

analyzed MFA model demonstrated a good fit, having r2 value of 

0.868 and cross-validated coefficient r2cv value of 0.771. 2008 

by MDPI. 

(International Journal of Molecular Sciences Volume 9, Issue 3, 

March 2008, Pages 235-246) 

SYNTHESIS AND HIV-1 REVERSE TRANSCRIP- 

TASE INHIBITORY ACTIVITY OF NON- 

NUCLEOSIDE PHTHALIMIDE DERIVATIVES 

(NO. 790) 

Ungwitayatorn, J. 1 , Wiwat, C. 2 , Matayatsuk, C. 1 , Pimthon, J. 1 , 

Piyaviriyakul, S. 1 

1 Department of Pharmaceutical Chemistry, Faculty of Pharmacy, 

Mahidol University, 447 Sri-Ayudhya, Rachatevee, Bangkok 

10400, Thailand; 2 Department of Microbiology, Faculty of 

Pharmacy, Mahidol University, 447 Sri-Ayudhya, Rachatevee, 

Bangkok 10400, Thailand. 

Key words : HIV-1 reverse transcriptase inhibitors; Non-nucleoside; 

Phthalimide derivatives 

A new type of non-nucleoside HIV-1 reverse transcriptase 

inhibitors in phthalimide series has been synthesized from either the 

reaction of N-carboethoxyphthalimide with amines or phthalimide 

with appropriate alkyl halides. The in vitro inhibitory activity of the 

synthesized compounds was studied by a radiometric assay at a 

concentration of 200 g/mL using poly(rA) oligo(dT) as a templateprimer 

and methyl-[3H]dTTP as a substrate. The three most potent 

compounds, N-(m,p-dihydroxybenzyl) phthalimide (11), N-[2-(?furyl)ethyl]phthalimide 

(29) and N-(5-methylpyrazin-2ylmethyl)phthalimide 

(25) exhibited IC50 values of 60.90, 98.10 and 

120.75 /mL, respectively, lower than IC50 of delavirdine (502.22 

g/mL, using poly(rA) oligo(dT) as a template-primer and [ 3H]dTTP 

as a substrate). 2008 SIOC, CAS, & Wiley-VCH Verlag GmbH & 

Co. KGaA. 

(Chinese Journal of Chemistry,26,(2), February 2008: 379-387) 

(This study was granted by The Thailand Research Fund.) 

5-SUBSTITUTED PYRIDO[2,3-D]PYRIMIDINE, 

AN INHIBITOR AGAINST THREE RECEPTOR 

TYROSINE KINASES. (NO. 791) 

291 

Naparat Kammasud 1 , Chantana Boonyarat 2 , Kingkan 

Sanphanyaa 1 , Maleeruk Utsintonga 1 , Satoshi Tsunodac 3 , Hiroaki 

Sakuraic 3 , Ikuo Saikic 3 , Isabelle Andr 4 , David S. Griersond 4 and 

Opa Vajragupta 1 


Mahidol University, 447 Sri-Ayudhya Road, Bangkok 10400, 

Thailand; 2 Faculty of Pharmaceutical Sciences, Khon Kaen 

University, Khon Kaen 40002, Thailand; 3 Division of Pathogenic 

Biochemistry, Department of Bioscience, Institute of Natural 

Medicine, University of Toyama, Japan; 4 UMR 176 CNRS, 

Institut Curie, Section Recherche, Centre Universitaire, Bat.110- 

112, 91405 Orsay cedex, France 

Key words: FGFR-1; FGFR-1 inhibitor; SU6668; 5-Substituted 

indolin-2-one; Virtual screening; Docking; Binding mode; Antiproliferation; 

Anti-angiogenesis 

NP506, the 3-{2,4-dimethyl-5-[2-oxo-5-(N2 - 

phenylhydrazinocarbonyl)-1,2-dihydro-indol-3-ylidenemethyl]-1Hpyrrol-3-yl}-propionic 

acid, was designed as FGF receptor 1 inhibitor 

by computational study and found to be more active against 

endothelial proliferation of HUVEC after the rhFGF-2 stimulation 

than SU6668 with minimum effective dose of 10 M. NP506 

inhibited the tyrosine phosphorylation in FGF, VEGF, and PDGF 

receptors and the activation of extracellular signal-regulated kinase 

(ERK), c-Jun-N-terminal-kinase (JNK) and AKT after the rhFGF-2 

stimulation. The introduction of the phenyl hydrazide motif to the 

position 5 of the pyrido[2,3-d]pyrimidine scaffold led to the inhibitory 

effect in two signaling pathways: inhibition of AKT activation in the 

phosphatidyl inositol 32 -kinase (PI13K)/AKT signaling pathway and 

the inhibition of ERK and JNK activation in MAPK pathway. 

Graphical abstract 

The introduction of the phenyl hydrazide motif to the position 5 of 

the pyrido[2,3-d]pyrimidine scaffold in NP506 led to the inhibitory 

effect in two signaling pathway: inhibition of AKT activation in the 

phosphatidyl inositol 32 -kinase (PI13K)/AKT signaling pathway and 

the inhibition of ERK and JNK activation in MAPK pathway. 

(This work was funded by the Royal Golden Jubilee Project, Thailand 

Research Fund, The Commission of Higher Education, Ministry of 

Education, Thailand and the 21st Century COE project from the 

Ministry of Education, Culture, Sports, Science and Technology, 

Japan.) (Bioorganic & Medicinal Chemistry Letters, Article in Press)


VIRTUAL SCREENING AGAINST COBRA VENOM. 

(NO. 792) 

Maleeruk Utsintong 1 , Palmer W Taylor 2 , Arthur J Olson 3 , Opa 

Vajragupta 1 


Mahidol University 447 Sri-Ayudhya Road, Rajthevi, Bangkok 

10400, Thailand; 2 Skaggs School of Pharmacy and 

Pharmaceutical Sciences, University of California, San Diego 

9500 Gilman Dr., La Jolla, California 92093-0636, USA; 3 The 

Scripps Research Institute, Molecular Graphics Laboratory, 

Department of Molecular Biology 10550 North Torrey Pines 

Road, La Jolla, California 92037, USA 

α-Cobratoxin, the long chain neurotoxin from Naja 

kaouthia acts on the postsynaptic membrane of nicotinic acetylcholine 

receptors. It causes paralysis by preventing acetylcholine (ACh) 

binding to the nicotinic acetylcholine receptor.?? α-?Cobratoxin has 

been proposed as a potential target for anticobratoxin drug design. 

The


A STUDY ON ARTIFACTS FORMATION IN THE 

THAI TRADITIONAL MEDICINE PRASAPLAI 

(NO. 795) 

Prasan Tangyuenyongwatana and Wandee Gritsanapan* 

Department of Pharmacognosy, Faculty of Pharmacy, Mahidol 

University, Bangkok, Thailand. *Corresponding author, E-mail 

: pywgs@mahidol.ac.th 

Key words : Prasaplai, Zingiber cassumunar , Nigella sativa 

The artificial formation of three fatty acid esters, (E)-4- 

(3,4-dimethoxyphenyl)but-3-en-1-yl linoleate (1),(E)-4-(3,4dimethoxyphenyl)but-3-en-1-yl 

oleate (2) and (E)-4-(3,4dimethoxyphenyl)but-3-en-1-yl 

palmitate (3) originating during 

storage by the interaction of components in Prasaplai preparation 

was investigated. The artifacts were not formed when 0.1 mL of water 

or more was added to 1.0 g of the mixture (1:1) of Zingiber 

cassumunar and Nigella sativa even when stored for 20 days. This 

result showed that water was able to stop the esterification reaction. 

The formation of the artifacts by chemical reaction under water-free 

conditions was evaluated. (E)-4-(3,4-dimethoxyphenyl)but-3-en-1ol 

was mixed with linoleic acid in the presence of anhydrous Na 

(2)SO (4) and stored in a dessicator for 7 days. The artifact (1) was 

formed in 6.0 % yield. It was concluded that a water-free environment 

is necessary for the direct chemical formation of the artificial esters. 

(The manuscript was published in Planta Medica 74, (2008); 1403- 

1405.) 

ANALYSIS OF NAPHTHOQUINONE DERIVATIVES 

IN THE ASIAN MEDICINAL PLANT ELEUTHERINE 

AMERICANA BY RP-HPLC AND LC-MS (NO. 796) 

Sompol Paramapojn 1 , Markus Ganzera 2 , Wandee Gritsanapan 1 * 

and Hermann Stuppner 2 

1 Department of Pharmacognosy, Faculty of Pharmacy, Mahidol 

University,447 Sri-Ayudhaya Road, Ratchathewi, Bangkok 

10400, Thailand; 2 Institute of Pharmacy, Pharmacognosy, 

University of Innsbruck, Innrain 52, 6020 Innsbruck, Austria 

*Corresponding author, E-mail : pywgs@mahidol.ac.th 

Key words : Eleutherine Americana, Naphthoquinone, LC–MS 

The first analytical procedure for the determination of a 

new naphthopyrone, eleutherinoside A, together with the known 

bioactive compounds eleuthoside B, isoeleutherin, eleutherin and 

eleutherol in Eleutherine americana was established. Optimum HPLC 

separation of these naphthoquinone derivatives was possible on RP- 

12 column material, using water and acetonitrile as mobile phase. 

Flow-rate, detection wavelength and temperature were adjusted to 

1.0 mL/min, 254 nm and 40 C, respectively. Validation results 

indicated that the HPLC method is well suited for the determination 

of naphthoquinone derivatives in the bulbs of E. americana with a 

good linearity (r2 > 0.9996), precision (intra-day R.S.D.


Prasaplai is a drug preparation which is listed in the Thai 

traditional common household drug list for remedy of relieving 

dysmenorrhea and adjusting the cycle of menstruation. (E)-4-(3,4dimethoxyphenyl)but-3-en-1-yl 

oleate (compound 1) originated 

during storage by the interaction of component in Prasaplai formula 

was isolated by preparative HPLC together with (E)-4-(3,4dimethoxyphenyl)but-3-en-1-yl 

linoleate (compound 2) and (E)-4- 

(3,4-dimethoxyphenyl)but-3-en-1-yl palmitate (compound 3). After 

an investigation of the origin of the artifacts by a systematic 

preparation of two components mixture and subsequently HPLC 

analysis, we found that the artifacts come from the mixture of the 

rhizome of Zingiber cassumunar Roxb. and the seed of Nigella sativa 

L. These three compounds were undergone for antituberculosis test 

and all three compounds were active against Mycobacterium 

tuberculosis H37Ra for which compounds 1 and 3 had inhibitory 

concentration at 200 g/mL while compound 2 inhibited at 100 g/ 

mL. 

(The manuscript was published in Acta Horticulturae 786, ISHS 

(2008); 41-46.) 

VARIABILITY OF ANTIOXIDATIVE QUERCETIN 

CONTENT IN SIAMESES NEEM TREE LEAVES 

IN THAILAND BY TLC-DENSITOMETRY (NO. 799) 

Pongtip Sithisarn and Wandee Gritsanapan* 


University, Bangkok, Thailand. *Corresponding author, E-mail 


Key words : Siamese neem tree, quercetin, Azadirachta indica 

Siamese neem tree (Azadirachta indica A. Juss. 

var.siamensis Valeton) is a Thai medicinal plant of which young leaves 

and flowers are used as a bitter tonic. Siamese neem tree leaves 

exhibited antioxidant activity according to flavonoids including 

quercetin, rutin and rhynchosin-O-β-D-glucoside. Quercetin is 

reported to be the major antioxidative component. There has been 

no report about antioxidant content in Siamese neem tree leaves 

before. Therefore, the quantitative analysis of quercetin content in 

Siamese neem tree leaves collected from different locations in 

Thailand was performed. Leaf aqueous extracts of Siamese neem 

tree collected from 12 provinces in Thailand were quantitative analysis 

for quercetin content using a TLC-densitometric method. Quercetin 

content in the leaf aqueous extracts and in the dried leaf samples 

were in the limit of 6.19 0.16 to 48.11 1.50 mg% and 1.55 0.04 

to 19.30 0.60 mg%, respectively. The average high quercetin 

contents were found in samples from Prachinburi, Songkhla, and 

Petchabun provinces. Comparison between areas, it was found that 

samples from the East and the South of Thailand significantly 

contained high quercetin content while the samples from the Northeast 

and the West exhibited the low content. These results would be useful 

as a basic information for finding good sources of Siamese neem 

tree leaf raw material with high active content and also useful as a 

guidance for standardization of Siamese neem tree leaf extract used 

for medicinal and health supplement proposes. 

quercetin 

HO 

6 

7 

8 

5 

OH 

10 O 2 1' 

9 

O 

4 

(This study was granted by The Royal Golden Jubilee 

Ph.D.Program.) (The manuscript was published in Acta 

Horticulturae 786, ISHS(2008);161-168.) 

3 

QUANTITATIVE ANALYSIS OF CURCUMINOIDS 

IN CURCUMA ZEDOARIA RISOMES IN THAILAND 

BY HPLC METHOD (NO. 800) 

Sompol Paramapojn and Wandee Gritsanapan * 


University,447 Sri-Ayudhaya Road, Ratchathewi, Bangkok 

10400, Thailand.*Corresponding author, E-mail : 

pywgs@mahidol.ac.th 

Key words : Curcuma, curcumin, demethoxycurcumin 

A high performance liquid chromatographic (HPLC) 

method was developed and validated for quantitative determination 

of curcuminoids (curcumin, demethoxycurcumin and 

bisdemethoxycurcumin) contents in extracts of the rhizomes of 

Curcuma zedoaria collected from ten locations in Thailand. The 

analysis was carried out using a BDS Hypersil C18 column as a 

stationary phase, 0.1%glacial acetic acid aqueous solution and 

acetonitrile as mobile phases. The detection was done at 425 nm. 

The validation using curcumin, demethoxycurcumin and 

bisdemethoxycurcumin as standards demonstrated good linearity, 

precision and accuracy at the concentration range 2-6 μg/ml. The 

content of curcumin in all powdered samples was in the range of 

0.50 0.06 to 0.73 0.03%w/w (average 0.65 0.07%w/w) while the 

contents of demethoxycurcumin and bisdemethoxycurcumin were 

in the range of 0.23 0.02 to 1.43 0.55%w/w (average 0.91 0.35%w/ 

w) and 0.12 0.01 to 0.44 0.21%w/w (average 0.25 0.10%w/w), 

respectively. The highest average total curcuminoids content in the 

powdered samples was found to be 2.50 0.52%w/w while the lowest 

content was 1.06 0.31 %w/w. This information will be useful as a 

guidance for further standardization of C. zedoaria raw material of 

which the content has not been reported before. 

(This study was granted by The Royal Golden Jubilee 

Ph.D.Program.) (The manuscript was published in Acta 

Horticulturae 786, ISHS(2008); 169-174.) 

2' 

OH 

3' 

OH 

6' 

4' 

5' 

OH


VARIABILITY OF CURCUMINOIDS : ANTIOXI- 

DATIVE COMPONENTS IN ETHANOLIC 

TURMERIC EXTRACT DETERMINED BY UV 

AND HPLC METHODS (NO. 801) 

Werayut Pothitirat, and Wandee Gritsanapan* 


University, Bangkok 10400, Thailand*Corresponding author, Email 


Key words : curcuminoids, turmeric, Curcuma longa 

In Thailand, turmeric (Curcuma longa L.) is mainly used 

in forms of capsules/tablets of the powder for herbal medicine while 

its extract is also used in herbal cosmetic and functional food. So, 

the quality assessment of this plant is needed to control the limits of 

volatile oil and curcuminoids contents. This study was undertaken 

to evaluate total curcuminoid content in the ethanolic turmeric extract 

of C. longa rhizome collected from 10 locations from the North, North 

east, Central and South of Thailand by a UV spectrophotometry and 

HPLC. Each curcuminoid content (curcumin, demethoxycurcumin 

and bisdemethoxycurcumin) determined by HPLC were also reported. 

By a UV spectrophotometer, the total curcuminoids contents of all 

extracts found in the limits of 14.14 0.87 to 26.76 0.17 %w/w. 

By HPLC, the content of curcumin, demethoxycurcumin, 

bisdemethoxycurcumin and total curcuminoid were found in the limits 

of 8.55 0.42 to 15.88 0.46, 1.50 0.14 to 5.17 0.58, 5.54 

0.23 to 9.33 0.30, and 16.39 0.68 to 27.39 1.04 %w/w of the 

extract, respectively. This shows that the ethanol extract of C. longa 

should contain total curcuminoids not less than 16 %w/w when 

determined by HPLC and not less than 13 %w/w when determined 

using UV spectrophotometry. The samples from the South where 

raining is obtained all year were found to contain the highest amount 

of total and each curcuminoid. The results confirm that turmeric 

grown in the South of Thailand is the best source for high contents 

of individual curcuminoid and total curcuminoids. This information 

would be a useful database for medicinal plant of the country, to 

maximize the potential of local community, especially for C. longa 

which is a product champion of Thailand. The results are also useful 

as a guidance for further standardization of turmeric extract. 

(This study was granted by Mahidol University Research Fund and 

The National Research Council of Thailand.) (The manuscript was 

published in Acta Horticulturae 786, ISHS(2008); 175-184.) 

QUANTITATIVE ANALYSIS OF TOTAL 

MANGOSTINS IN GARCINIA MANGOSTANA 

FRUIT RIND (NO. 802) 

Werayut Pothitirat and Wandee Gritsanapan* 


University, Bangkok, Thailand, 10400 *Corresponding author, 

E-mail : pywgs@mahidol.ac.th 

Key words : Garcinia mangostana, mangostin, UVspectrophotometry 

The fruit of Garcinia mangostana Linn. (mangosteen) is 

very popular in Thailand. The fruit rind contains mangostins of which 

a major constituent is -mangostin. The fruit rind extract and 

mangostin have been known to possess antibacterial causing acne. 

In Thailand, the extract is popularly used in herbal cosmetics for 

anti-acne effect. Thus quality assessment of this plant needs to be 

controlled for the limit of mangostin content. This study was 

undertaken to evaluate the content of total mangostins in the dried 

powder and the ethanolic extract of the fruit rinds of G. mangostana 

collected from 13 locations from the East and the South of Thailand. 

The UV-spectrophotometric method was validated for linearity, 

precision, accuracy, limit of detection (LOD) and limit of quantitation 

(LOQ). The linearity was found over the range of 2-20 g/ml with 

regression coefficient (r2) 0.9999. Intra- and interday precisions 

showed relative standard deviation (%RSD) less than 2 %. Accuracy 

of the method was determined by a recovery study conducted at 3 

different levels, and the average recovery was found to be 100.68 %. 

The LOD and LOQ were 0.1622 and 0.4915 g/ml, respectively. The 

total mangostin contents in all dried powder samples were in the 

range of 8.51 0.05 to 11.50 0.02 % w/w while in the crude 

ethanolic extracts were 30.19 0.16 to 45.61 0.09 % w/w. The 

average content of total mangostins (10.39 1.04 % of the dried 

powder) was higher in samples from the South where it rains all 

year. The averages of total mangostin contents in all dried powder 

samples and in the ethanolic extracts were found to be 9.94 0.88 

and 36.25 4.66 %w/w, respectively. The proposed UVspectrophotometric 

method was found to be simple, rapid, and 

suitable for routine quality control of raw material of G. mangostana 

fruit rind and its extract. This information will be useful as a guidance 

for standardization of G. mangostana fruit rind and the extracts, and 

finding appropriate sources of high total mangostins content for good 

quality of G. mangostana raw materials in Thailand. 

(This project was granted by Mahidol University Research Fund.) 

(The manuscript was published in Thai Journal Health Research 

18(1), (2008) 

EXTRACTION METHOD FOR HIGH CONTENT 

OF ANTHRAQUINONES FROM CASSIA FISTULA 

PODS (NO. 803) 

Aurapa Sakulpanich and Wandee Gritsanapan* 

Department of Pharmacognoscy, Faculty of Pharmacy, Mahidol 

University, Bangkok, Thailand *Corresponding author, E-mail : 


Key words : Cassia fistula, anthraquinone, rhein 

295 

The ripe pod of Cassia fistula Linn. has long been used in 

traditional medicines as a laxative drug. The active principles are 

known to be anthraquinone glycosides of which rhein and aloeemodin 

are major components. The pulp from ripe pods of C. fistula 

was extracted with 70% ethanol by maceration, percolation, and 

soxhlet extraction, and by decoction with water according to Thai 

traditional uses. The contents of total anthraquinone glycosides and 

total anthraquinones in the crude extracts prepared by each of 

extraction method were determined using a UV-vis spectrophotometer 

and the contents were calculated as rhein and aloe-emodin. The extract 

prepared by decoction method contained the highest content of total 

anthraquinone glycosides which are the active laxative form in the 

range of 0.2383 0.0011 and 0.2194 0.0077 %w/w calculated as 

rhein and aloe-emodin, respectively. Maceration exhibited the extract 

containing the highest content of total anthraquinones at 0.3139


0.0129 % w/w calculated as rhein and 0.2194 0.0088 % w/w 

calculated as aloe-emodin. Comparing all extraction methods, 

decoction is simple, convenient, carried low cost in terms of solvent 

and instrumentation and found to be the appropiate extraction method 

for the pulp of C. fistula pods for a laxative drug. 

(This project was granted by The Thailand Research Fund (TRF) 

with Office of Small and Medium Enterprises Promotion (OSMEP). 

(The manuscript was published in Thai Journal Health Research 

18(1), (2008) 

VARIATION OF ANTRAQUINONE CONTENT IN 

THE LEAVES AND PODS OF CASSIA FISTULA 

(NO. 804) 

Wandee Gritsanapan*, Somsak Nualkaew 


University, 447, Sri-Ayudthaya Road, Ratchatewi, Bangkok, 

10400 Thailand.*Corresponding author, E-mail : 


Key words : anthraquinone, Cassia fistula, TLC-densitometry 

Cassia fistula Linn. is a medicinal plant of which the ripe 

pod has been traditionally used as a laxative drug in Thailand and 

many Asian countries. The pods contain anthraquinones in both 

aglycones and glycosides which are the active laxative form, while 

rhein is a major component [1, 2]. The leaves, which have not been 

used for laxative, were reported to contain sennosides, chrysophanic 

acid, phycion and a major constituent rhein [1]. The contents of 

anthraquinones in the pods and the leaves of C. fistula have not been 

compared. This study determined and compared the contents of total 

anthraquinone glycosides and a major anthraquinone rhein in the 

ripe pods collected from various parts of Thailand in summer (April) 

and in the leaves collected in summer, rainy season and winter. The 

contents of total anthraquinone glycosides in the ripe pods and in 

the leaves collected from the North, North-East, Central and South 

of Thailand were within a range of 0.21-0.67% (average 0.44%) and 

0.05-0.74% (average 0.32%) dry weight, respectively. The contents 

of rhein in the ripe pods and in the leaves of C. fistula determined by 

TLC-densitometric method were 0.05-0.14% (average 0.09%) and 

0.002-0.29% (average 0.12%) dry weight, respectively. The ripe pods 

containing higher contents of anthraquinone glycosides and of a major 

compound rhein, were found in the southern >northeastern>central>northern 

samples while the high anthraquinones 

content in the leaves was found in summer (March-June) samples 

from the North and the South where the weather is not warm as in 

the central and north-eastern parts. The results support a traditional 

way of collection of leaf drugs of Thai people and the recommendation 

that medicinal leaves should be collected before flowering period of 

plants. According to the Standard of ASEAN Herbal Medicine, the 

leaves of C. fistula collected from the North and the South of Thailand 

in Summer which contain > 0.5% of anthraquinone glycosides might 

be used as a source of anthraquinone laxative herbal drug as same as 

the ripe pods. 

(This work was presented at 7th Joint Meeting of AFERP, ASP, GA, 

PSE & SIF, Athens, Greece, August 3-8, 2008. and the Abstract was 

published in Planta Medica 74(9), (2008); 1085) 

ACUTE TOXICITY STUDY ON ARTIFACTS 

FROM PRASAPLAI PREPARATION, A THAI 

TRADITIONAL MEDICINE. (NO. 805) 

Prasarn Tangyuenyongwatana and Wandee Gritsanapan* 


University, Bangkok 10400, Thailand. *Corresponding author, 


Key words : acute toxicity, Prasaplai, traditional medicine 

Prasaplai is a drug preparation which is listed in the Thai 

traditional common household drug list for relieving dysmenorrhea 

and adjusting the cycle of menstruation [1,2]. Three fatty acid ester 

artifacts, (E)-4-(3,4-dimethoxyphenyl)but-3-en-1-yl linoleate (1), (E)- 

4-(3,4-dimethoxyphenyl)but-3-en-1-yl oleate (2) and (E)-4- 

(3,4dimethoxyphenyl)but-3-en-1-yl palmitate (3) originated during 

storage by the interaction of components in Prasaplai [3,4] were 

synthesized for toxicity testing. These three artificial esters were 

subjected to acute toxicity testing in mice and the results showed 

that all compounds had LD50 more than 300 mg/kg. In addition, 

the artifacts formation versus time was also studied by analysis with 

HPLC and we found that the amount of artifacts formation became 

saturation after 2 months of storage. The toxicity and the amount of 

artifacts formation information are crucial for safety usage of the 

Prasaplai preparation. 

(This work was presented at the 7th Joint Meeting of AFERP, ASP, 

GA, PSE & SIF, Athens, Greece, August 3-8, 2008. and the Abstract 

was published in Planta Medica 74(9), (2008); 1139.) 

ANTI-ACNE INDUCING BACTERIA ACTIVITY 

OF GARCINIA MANGOSTANA FRUIT RIND 

EXTRACTS FROM VARIOUS LOCATIONS OF 

THAILAND (NO. 806) 

Werayut Pothitirat 1 , Mullika (Traidej) Chomnawang 2 , Wandee 

Gritsanapan 1 * 

1 Department of Pharmacognosy, 2 Department of Microbiology, 

Faculty of Pharmacy, Mahidol University, 447 Sri-Ayuthaya Rd., 

Ratchathewi, Bangkok 10400, Thailand, *Corresponding author, 


Key words : anti-acne, Garcinia mangostana, Propionibacterium 

acnes 

Acne vulgeris is the most common cetaceous disorder 

found in both man and woman especially teenagers. The 

microorganisms involved Propionibacterium acnes and 

Staphylococcus epidermidis [1]. The fruit rind of Garcinia 

mangostana was reported a strong inhibitory effect on P. acnes and 

S. epidermidis [2]. The aim of this study was to compare the 

antimicrobial activity of the extracts of G. mangostana fruit rind 

collected from various locations in the South and the East of Thailand 

against these bacteria. Alpha-mangostin, a major constituent of the 

fruit rind extracts, was also tested for the activity. The MIC values 

were evaluated using the broth microdilution method [3]. Thirteen 

ethanolic extracts of G. mangostana fruit rinds were prepared by 

soxhlet extraction with 95 %ethanol. The MIC values of all crude


ethanolic extracts against P. acnes and S. epidermidis were in the 

range of 7.81-15.63 and 15.63-31.25 g/ml, respectively, while the 

MBC values were in the range of 15.63-31.25 and 62.50-125.00 g/ 

ml, respectively. The average MIC and MBC values indicate that 

samples from the South have the more potent antimicrobial effect 

than the samples from the East. Base on MIC and MBC values, alphamangostin 

showed a good inhibitory effect on P. acnes (MIC = MBC 

= 1.56 g/ml) and S. epidermidis (MIC = 1.56, MBC= 6.25 g/ml). 

Therefore, this compound is suitable for using as a marker for quality 

assurance of the extract and its product. The results can be used as a 

guidance for standardization of G. mangostana extract for anti-acne 

property and for finding sources of good quality of G. mangostana 

raw material. The anti-acne preparation from this plant extract will 

be further investigated. 

(This work was granted by Mahidol University.) (It was presented 

at the 7th Joint Meeting of AFERP, ASP, GA, PSE & SIF, Athens, 

Greece, August 3-8, 2008. and the Abstract was published in Planta 

Medica 74(9), (2008); 1127.) 

DETERMINATION OF FURANOCOUMARINS IN 

CITUS HYSTRIX BY HPLC PHOTODIODE ARRAY 

(NO. 807) 

Paramapojn S 1 , Gritsanapan W 1 *, Ganzera M 2 , Stuppner H 2 


University, 447 Sri-Ayudhaya Road, Ratchathewi, Bangkok 

10400, Thailand.; 2 Institut f r Pharmazie, Abteilung 

Pharmakognosie, Universit t Innsbruck, Innrain 52, 6020 

Innsbruck, sterreich *Corresponding author, E-mail : 


Key words : Citrus hystrix, oxypeucedanin, bergamottin 

A rapid and sensitive reversed-phase high-performance 

liquid chromatographic (RP-HPLC) method was used for 

determination of furanocoumarins in methanolic extracts of the peel 

of Citrus hystrix DC. HPLC separation was performed on a C-12 

Phenomenex column (Synergi Max-RP, 150 x 4.6 mm, 4 μm particle 

size) used as a stationary phase and acetonitrile–water gradient as a 

mobile phase. Flow-rate, detection wavelength and temperature were 

adjusted to 1.0 mL/min, 254 nm and 40 oC, respectively. Calibration 

plots were linear in a range of 31.74-241 μg/mL of oxypeucedanin 

hydrate and 32.79-249 μg/mL of 6’, 7’-dihydrobergamottin with a 

good linearity (r2 = 1), precision (intra-day R.S.D. < 0.77 %, interday 

R.S.D. < 3.09 %) and recovery rates of 96.83 to 101.23 %. Limit 

of detection (LOD) was found to be 0.31 μg/mL for both compounds. 

The analysis of C. hystrix peel samples from 10 different locations 

of Thailand revealed that 6’, 7’-dihydrobergamottin (0.38-0.88 %w/ 

w) was dominant in all specimens, followed by oxypeucedanin hydrate 

(0.32-0.54 %w/w). 

(This work was granted by The Royal Golden Jubilee Ph.D.Program.) 

(It was presented at the 7th Joint Meeting of AFERP, ASP, GA, PSE 

& SIF, Athens, Greece, August 3-8, 2008. and the Abstract was 

published in Planta Medica 74(9), (2008); 1089.) 

QUERCETIN AND RUTIN CONTENTS IN SIAMESE 

NEEM FLOWER EXTRACTS PREPARED BY 

DIFFERENT EXTRACTION METHODS (NO. 808) 

Worarat Chaisawangwong and Wandee Gritsanapan* 


University, 447 Sri-Ayudthaya Road, Ratchathevi, Bangkok 

10400, Thailand.*Corresponding author, E-mail : 


Key words : Siamese neem tree, Azadirachta indica, quercetin 

Siamese neem tree (Azadirachta indica A. Juss. var. 

siamensis Valeton) is a medicinal plant found in every part of 

Thailand. The young flowers of this plant are commonly consumed 

with sweet sauce as a bitter tonic vegetable for elemental tonic 

purpose [1]. The flower extract has been reported to exhibit in vitro 

free radical scavenging activity and inhibit lipid peroxidation of 

bronchogenic cancer cell line [2]. Active components in Siamese 

neem flowers are flavonoids such as quercetin and rutin. The content 

of these compounds in the crude extract is depended on the method 

of extraction. This work investigated the quantity of rutin and 

quercetin in the flower extracts of Siamese neem tree prepared by 

several extracting methods using high performance liquid 

chromatography (HPLC) to find the appropriate extraction method 

which gives the maximum contents of rutin and quercetin. The flowers 

were extracted using maceration, percolation, decoction, soxhlet 

extraction, ultrasonic extraction (UE) and microwave assisted 

extraction methods. The solvent used in maceration, percolation and 

soxhlet extraction was 50% ethanol, in decoction and MA was distilled 

water, while in UE was both 50% ethanol and distilled water. By 

HPLC, the results showed that soxhlet extraction method gave the 

extract containing the maximum contents of rutin and quercetin 

(10.21 and 0.12% w/w of the extract, respectively), and quercetin 

was detected only in the extract prepared using 50% ethanol as the 

solvent. Thus, soxhlet extraction should be the appropiate extraction 

method for Siamese neem flowers to yield the extract with high 

contents of active components, rutin and quercetin. 

(This work was granted by TRF-OSMEP-MAG Scholarship.) (It was 

presented at the 7th Joint Meeting of AFERP, ASP, GA, PSE & SIF, 

Athens, Greece, August 3-8, 2008. and the Abstract was published 

in Planta Medica 74(9), (2008); 1088.) 

DEVELOPMENT OF HERBEL MOUTHWASH 

(NO. 809) 

Warangkana Punjapratheep 1 , Usanee Pienputtarapong 1 , Wandee 

Gritsanapan 1 *, Chonticha Amornchat 2 


University; 2 Department of Microbiology, Faculty of Dentristry, 

Mahidol University *Corresponding author, E-mail : 


Key words : Herbal mouthwash, Tooth decay, Mangosteen 

297 

The Development of anti-tooth decay mouthwash 

formulation containing 3 herbal extracts which were mangosteen 

(Garcinia mangostana Linn.) fruit rind extract, Koi(Streblus asper


Lour.) leaf extract and greentea (Camellia sinensis (L.)Kuntze) extract, 

it was found that mangosteen fruit rind extract tested by Disc diffusion 

method could inhibit Streptococcus mutans, an important bacteria 

causing tooth decay, at a concentration more than 2.5 mg/mL (Clear 

zone diameter 7.16 mm). Koi and greentea extracts could not inhibited 

S. mutans. The mouthwash formulation containing the mangosteen 

extract which could inhibit S. mutans should contain not less than 4 

g of the extract in 100 mL of the preparation. However, the prepared 

preparation had a dark brown color. The active compound against S. 

mutans was found to be alpha mangostin, a major component in the 

mangosteen fruit rind extract. The inhibiting activity against S. 

mutans of the mangosteen mouthwash formulation was lower than a 

commercial mouthwash containing chlorhexidine as an active 

ingredient. When greentea extract and/or Koi leaf extract were added 

into the preparation, the dissolution of the formulation was decreased, 

but the dark color of the preparation was increased. Adding 

peppermint and spearmint oils into the formulation, could improve 

the flavor and taste of the preparation. However, the flavor and taste 

were diminished after 2 weeks. The formulation should be further 

developed. 

(This work was granted by IRPUS-The Thailand Research Fund.) 

(It was presented at IRPUS Exhibition 2008, Bangkok, March 28- 

30, 2008.) 

WHITENING HERBAL SKIN LOTION (NO. 810) 

Jiradchadapa Pluemlamai 1 , Suphamas Satniyom 1 , Wandee 

Gritsanapan 1 *, Wichet Leelamanitaya 2 


University; 2 Department of Biochemistry, Faculty of Pharmacy, 

Mahidol University *Corresponding author, E-mail : 


Key words : antityrosinase, melanin, Artocarpus lakoocha 

At present, Herbs are popularly used in cosmetics, 

especially to nourish and to whiten skin. Whitening substances have 

many mechanisms of which is the popular found in cosmetics is a 

tyrosinase inhibitor group. Tyrosinase is an enzyme involving in 

melanin synthesis that causes dark skin. In this project, three kinds 

of herbs were chosen for extraction and formulation of whitening 

skin lotion. Artocarpus lakoocha Roxb., Glycyrrhiza glabra Linn., 

Bombyx mori were extracted and their extracts were tested for 

antityrosinase activity using dopamine method. It was found that 

three herbal extracts at 1 mg/mL concentration exhibited tyrosinase 

inhibitory activity was 87.88 0.31%, 74.24 0.62% and 5.19 0.61% 

(IC50 = 50, 140 and >1000 g/mL), respectively, while the reference 

standard kojic acid showed the activity at 83.30 0.51% (IC50 = 130 

g/mL) at the same concentration. The herbal extracts were 

Artocarpus lakoocha Roxb., Glycyrrhiza glabra Linn. and Bombyx 

mori respectively in 100 mL of the preparation. The preparation 

exhibited antityrosinase activity at 98.60 045%, having a brown color 

with slightly alcoholic odor. In addition, the formula was evaluated 

for stability test within 4 weeks at 4 oC, room temperature (28-30 

o C) and 45 o C, and examined for tyrosinase inhibitory activity, active 

ingredients, color and odor at 0, 7, 14, 21, and 28 days of the 

preparation. It was found that only being kept at 4 o C, the 

antityrosinase activity was not changed during the period examined. 

Furthermore, active ingredients, color and odor of the preparation 

were not change during the kept period at all conditions. 

(This work was granted by IRPUS-The Thailand Research Fund.) 

(It was presented at IRPUS Exhibition 2008, Bangkok, March 28- 

30, 2008.) 

VARIABILITY OF MANGOSTINS: ANTI-ACNE 

COMPONENT IN MANGOSTEEN FRUIT RIND 

(NO. 811) 

Werayut Pothitirat and Wandee Gritsanapan* 

Department of Pharmaconosy, Faculty of Pharmacy, Mahidol 

University, Bangkok 10400, Thailand *Corresponding author, 

E-mail: pywgs@mahidol.ac.th 

Key words : mangostin, anti-acne, Garcinia mangostana 

In Thailand, the extract of fruit rind of mangosteen 

(Garcinia mangostana Linn.) is popularly used in herbal cosmetics 

for anti-acne effect. It contains mangostins of which a major 

constituent is -mangostin. The fruit rind extract and mangostin have 

been known to possess antibacterial causing acne. Thus quality 

assessment of this plant needs to be controlled for the limit of 

mangostin content. This study was undertaken to evaluate the content 

of total mangostins in the dried powder and the ethanolic extract of 

the fruit rinds of G. mangostana collected from 13 locations from the 

East and the South of Thailand determined by UV-spectrophotometric 

method. The total mangostin contents in all dried powder samples 

were in the range of 8.51 0.05 to 11.50 0.02 % w/w while in the 

crude ethanolic extracts were 30.19 0.16 to 45.61 0.09 % w/w. 

The average content of total mangostins (10.39 1.04 % of the dried 

powder) was higher in samples from the South where it rains all 

year. The averages of total mangostin contents in all dried powder 

samples and in the ethanolic extracts were found to be 9.94 0.88 

and 36.25 4.66 %w/w, respectively. This information will be useful 

as a guidance for standardization of G. mangostana fruit rind and the 

extracts, and finding appropriate sources of high total mangostins 

content for good quality of G. mangostana raw materials in Thailand. 

(This work was granted by Mahidol University Research Fund.) (It 

was presented at 25th Annual Research Conference in 

Pharmaceutical sciences, December 2, 2008,Bankok,Thailand.) 

VARIATION OF ANTIOXIDANT COMPONENTS 

ANDFREE RADICAL SCAVENGING ACTIVITY OF 

SIAMESE NEEM TREE FLOWERS (NO. 812) 

Worarat Chaisawangwong and Wandee Gritsanapan* 


University, Bangkok 10400, Thailand *Corresponding author, 

E-mail: pywgs@mahidol.ac.th 

Key words : Siamese neem tree ,antioxidant, total flavonoid 

Siamese neem tree (Azadirachta indica A. Juss. var. 

siamensis Valeton) is a medicinal plant traditionally consumed as a 

bitter tonic found in every part of Thailand. There is a report 

concerning antioxidant activity of Siamese neem tree leaves and 

flowers including free radical scavenging activity and inhibition of 

lipid peroxidation in cancer cell line. Moreover, it is found that the 

leaves and flowers of Siamese neem tree contain some antioxidant 

flavonoids including quercetin and rutin. Therefore, aqueous extracts 

of Siamese neem tree flowers collected from 12 provinces of Thailand 

were analyzed for the contents of total phenolic compounds and total 

flavonoids according to the Folin-Ciocalteu procedure, and aluminium 

chloride colorimetric method, respectively. The EC50 of free radical 

scavenging activity of the extracts was also evaluated using DPPH


scavenging assay. The total phenolic compounds and total flavonoid 

contents in all samples were found in the range of 3.92 to 10.44 

GAE mg/g dried powder and 4.18 to 8.44 RE mg/g dried powder, 

respectively. EC50 of the extracts was in the range of 9.37 to 46.97 

g/ml. The highest free radical scavenging activity, total phenolic 

compounds and total flavonoids content were found in the extract 

from Nakhonpathom province. The highest average content of total 

phenolic compounds (46.785 GAE mg/g dried powder) and total 

flavonoids (64.745 RE mg/g dried powder) were found in the central 

part of the country. It also promoted strong free radical scavenging 

activity (EC50=17.17 g/ml). This information can be useful as a 

guidance for standardization of Siamese neem tree flowers, and 

finding appropriate sources of natural antioxidants from this plant. 

(This work was granted by TRF-OSMEP-MAG Scholarship) (It was 

presented at 25th Annual Research Conference in Pharmaceutical 

sciences, December 2, 2008,Bankok,Thailand.) 

TYROSINASE INHIBITORY ACTIVITY OF POD 

PULP AND LEAF EXTRACT OF CASSIA FISTULA 

Linn. (NO. 813) 

Aurapa Sakulpanich 1 , Wandee Gritsanapan 1 * and Adelheid H. 

Brantner 2 


University, Bangkok 10400, Thailand; 2 Institute of 

Pharmaceutical Sciences, Pharmacognosy, University of Graz, 

Universitaetsplatz 4/I, 8010 Graz, Austria *Corresponding 

author, E-mail : pywgs@mahidol.ac.th 

Key words ; Tyrosinase , Cassia fistula, anthraquinone 

Tyrosinase is an enzyme involving in melanin synthesis 

causing dark color of skin. Tyrosinase inhibitor is used as a skinwhitenning 

agent. In this study, tyrosinase inhibitory activity of crude 

extracts of pod pulp and leaves of Cassia fistula Linn. consisting of 

anthraquinone compounds was investigated. The tyrosinase inhibition 

assay revealed that the leaf crude extract prepared by decoction, 

maceration, percolation and soxhlet extraction with 70% ethanol 

promoted IC50 at 11.52, 7.20, 2.50, and 0.71 mg/ml, respectively, 

whereas IC50 values of the pod pulp crude extracts prepared by those 

methods were 14.64, 13.34, 13.20 and 10.55 mg/ml, respectively. 

Vitamin C was used as a reference standard which showed IC50 at 

31.4 g/ml. The IC50 of decoction extracts of both pod pulp and 

leaves showed the highest values while the IC50 of the soxhlet extract 

with 70% ethanol showed the lowest values. In the preliminary 

observation, it suggests the difference of extraction method and the 

effect of solvent on tyrosinase inhibitory activity. Comparison 

between the leaf crude extract and the pod pulp crude extract prepared 

by the same extraction method and same solvent showed that the 

leaf extract promoted higher tyrosinase inhibitory activity than the 

pod pulp extract. Therefore, the leaf extract of C. fistula might be 

used as a source of skin-whitenning agent. 

(This work was granted by TRF-OSMEP-MAG Scholarship) (It was 

presented at 25th Annual Research Conference in Pharmaceutical 

sciences, December 2, 2008, Bankok,Thailand.) 

PETROSAMINE, A POTENT ANTICHOLINE- 

STERASE PYRIDOACRIDINE ALKALOID 

FROM A THAI MARINE SPONGE PETROSIA 

N. SP. (NO. 814) 

Nukoolkarn, V.S. 1,4 , Saen-oon, S. 2 , Rungrotmongkol, T. 2 , 

Hannongbua, S. 2 , Ingkaninan, K. 3,4 , Suwanborirux, K. 4,5 



pyvnk@mahidol.ac.th; 2 Computational Chemistry Unit Cell, 

Department of Chemistry, Faculty of Science, Bangkok, 10330, 

Thailand; 3 Department of Pharmaceutical Chemistry and 

Pharmacognosy, Faculty of Pharmaceutical Sciences, Naresuan 

University, Phitsanulok, 65000, Thailand; 4 Center for Bioactive 

Natural Products from Marine Organisms and Endophytic Fungi 

(BNPME), Bangkok, 10330, Thailand; 5 Department of 

Pharmacognosy, Faculty of Pharmaceutical Sciences, 

Chulalongkorn University, Bangkok, 10330, Thailand. 

Key words: Anticholinesterase; Molecular docking; Petrosamine; 

Petrosamine-TcAChE interaction; Pyridoacridine 

Two pyridoacridine alkaloids, including a known 

petrosamine and a new 2-bromoamphimedine were isolated from a 

Thai marine sponge Petrosia n. sp. The alkaloids were characterized 

on the basis of 1D and 2D NMR, MS, and IR spectroscopy. Only 

petrosamine showed strong acetylcholinesterase inhibitory activity 

approximately six times higher than that of the reference 

galanthamine. A computational docking study of petrosamine with 

the enzyme from the electric eel Torpedo californica (TcAChE) 

showed the major contribution to the petrosamine-TcAChE 

interaction to be arising from the quaternary ammonium group of 

petrosamine. 

(Bioorganic and Medicinal Chemistry Volume 16, Issue 13, 1 July 

2008, Pages 6560-6567) (This work was supported by the Thailand 

Research Fund through the Royal Golden Jubilee Ph.D. program # 

PHD/00054/2541 and the New Researcher Grant # MRG4880041.) 

MOLECULAR DYNAMIC SIMULATIONS 

ANALYSIS OF RITONAVIR AND LOPINAVIR 

AS SARS-COV 3CL(PRO) INHIBITORS. (NO. 815) 

299 

Nukoolkarn, V. 1 , Lee, V.S. 2 , Malaisree, M. 3 , Aruksakulwong, O. 4 , 

Hannongbua, S. 3 



pyvnk@mahidol.ac.th; 2 Computational Simulation and Modeling 

Laboratory (CSML), Department of Chemistry, Faculty of 

Science, Chiang Mai, 50200, Thailand; 3 Computer Chemistry 

Unit Cell (CCUC) Department of Chemistry, Faculty of Science, 

Chulalongkorn University, Bangkok, 10330, Thailand; 

4Department of Chemistry, Faculty of Science, Rangsit 

University, Pathumtani, 12000, Thailand 

Key words: Lopinavir; MD simulations; Proteinase; Ritonavir; SARS 

Since the emergence of the severe acute respiratory 

syndrome (SARS) to date, neither an effective antiviral drug nor a


vaccine against SARS is available. However, it was found that a 

mixture of two HIV-1 proteinase inhibitors, lopinavir and ritonavir, 

exhibited some signs of effectiveness against the SARS virus. To 

understand the fine details of the molecular interactions between these 

proteinase inhibitors and the SARS virus via complexation, molecular 

dynamics simulations were carried out for the SARS-CoV 3CL(pro) 

free enzyme (free SARS) and its complexes with lopinavir (SARS- 

LPV) and ritonavir (SARS-RTV). The results show that flap closing 

was clearly observed when the inhibitors bind to the active site of 

SARS-CoV 3CL(pro). The binding affinities of LPV and RTV to 

SARS-CoV 3CL(pro) do not show any significant difference. In 

addition, six hydrogen bonds were detected in the SARS-LPV system, 

while seven hydrogen bonds were found in SARS-RTV complex. 

(Journal of Theoretical Biology Volume 254, Issue 4, 21 October 

2008, Pages 861-867) (This work was jointly supported by the 

Commission on Higher Education and the Thailand Research Fund 

(MRG4880041) 

FORENSIC DETECTION OF MARIJUANA TRACE 

(NO. 816) 

Thitika Kitpipit 1 , Nathinee Panvisavas 1,2 , Nuntavan 

Bunyapraphatsara 3 

1Forensic Science Graduate Programme, Faculty of Scinece, 

Mahidol Univerfsity, Bangkok 10400, Thailand; 2Department of 

Plant Science, Faculty of Science, Mahidol University, Thailand; 

3Deparment of Pharmacognosy, Faculty of Pharmacy, Mahidol 

University, Thailand. 

Key words : DNA, Forensic, Marijuana, TLC, Trace 

In this study, we compared the use of both chemical and 

biological tests for precise screening. Marijuana leaves had been 

treated in simulated conditions according to the way they are 

consumed; leaves materials were boiled in water for 5 min to 8 h. 

dried in hot-air oven, air-dried in shade and sunlight, and burned to 

black and white ashes. The THC band was detected in the TLC 

fingerprints of all samples, except the white ash extract. In contrast, 

the 197-bp mitochondrial trnL-F fragment was amplified in two 

samples, i.e., the DNA extracted from fresh marijuana leaves that 

were boiled for 5 min and some of the dried marijuana sample. The 

results suggested that TLC was a robust method for the detection of 

THC in marijuana. However, DNA analysis seems to be limited when 

DNA from heat-treated materials were analyzed 2008 Elsevier 

Ireland Ltd. All rights reserved. 

(Forensic Science International : Genetics Supplement Series; 1(1) 

August 2008: 600-602.) 

THE SMOOTHNESS OF BLOOD PRESSURE 

CONTROL OF RAMIPRIL IN ESSENTIAL 

HYPERTENSIVE THAI PATIENTS EVALUATION 

BY 24-HOUR AMBULATORY BLOOD PRESSURE 

MONITORING. (NO. 817) 

Uchaipichat, V. 1,4 , Koanantakul, B 2 , Suthisisang, C 3 

1 Department of Pharmacy Practice, Faculty of Pharmaceutical 

Sciences, Khon Kaen University, Khon Kaen, Thailand; 2 Division 

of Medicine, Bhumibol Adulyadej Hospital, Bangkok, Thailand; 

3 Department of Pharmacology, Faculty of Pharmacy, Mahidol 

University, Bangkok, Thailand; 4 Department of Pharmacy 

Practice, Faculty of Pharmaceutical Sciences, Khon Kaen 

University, Khon Kaen 40002, Thailand. 

Key words : Angiotensin converting enzyme inhibitor; Blood 

pressure variability; Hypertension 

Objective: Evaluate the efficacy of ramipril 2.5 and 5 mg 

once daily on the degree and homogeneity of 24-hour blood pressure 

reduction in essential hypertensive Thai patients. Material and 

Method: Nineteen male subjects, aged 30 to 60 years, with newly 

diagnosed essential hypertension were evaluated using the 24-hour 

ambulatory blood pressure (24-h ABP) measurement. Results: Twelve 

subjects responded and/or normalized with ramipril once daily, where 

the office and 24-h ABP were decreased significantly from baseline 

(p < 0.01). The percentage and magnitude of 24-h SBP/DBP loads 

after treatment were significantly decreased from 92 9.7/91 15.9 

to 67 23.8/65 27.6 (p < 0.01) and from 23 10.6/16 5.3 mmHg 

to 17 10.3/10 4.8 mmHg (p < 0.05). Trough to peak ratio for 

SBP/DBP was 0.59/0.52 (overall estimated) and 0.68 0.23/0.52 

0.22 (individual estimated), while the smoothness index was 0.89/ 

1.03. Conclusion: Ramipril 2.5 and 5 mg once daily exerted the 

smooth 24-hour blood pressure reduction in essential hypertensive 

Thai patients. 

(Journal of the Medical Association of Thailand Volume 91, Issue 9, 

September 2008, Pages 1468-1477.) 

EFFECT OF FENOFIBRATE THERAPY ON 

PARAOXONASE1 STATUS IN PATIENTS WITH 

LOW HDL-C LEVELS (NO. 818) 

Wimon Phuntuwate 1 , Chuthamanee Suthisisang 1 , Banhan 

Koanantakul .2 , Preecha Chaloeiphap 3 , Bharit Mackness 4 , Mike 

Mackness. 4 

1 Department of Pharmacology, Faculty of Pharmacy, Mahidol 

University, Sri Ayudhaya Road, Bangkok, 10400, Thailand; 

2 Cardiac and Preventive Center, Bhumibol Adulyadej Hospital, 

Bangkok, 10200, Thailand; 3 Division of Preventive Medicine, 

Royal Thai Air Force, Bangkok, 10200, Thailand; 4 University of 

Manchester, Department of Medicine, Manchester Royal 

Infirmary, Oxford Road, Manchester, M13 9WL, United 

Kingdom 

Key words : Fenofibrate; HDL-C; Oxidized LDL; Paraoxonase1 

Objectives: We evaluated the effect of micro-coated 

fenofibrate on lipid parameters, high sensitivity C-reactive protein 

and paraoxonase1 levels in dyslipidemic patients with low highdensity 

lipoproteins levels. In addition, the effects of the paraoxonase1 

polymorphisms on lipid and paraoxonase1 responses to fenofibrate 

therapy were examined. Methods: A total of 61 dyslipidemic patients 

with low high-density lipoproteins levels were recruited into this study 

to receive micro-coated fenofibrate (160 mg/day) for 12 weeks. Lipid 

parameters, C-reactive protein, paraoxonase1 concentration and 

activity were measured at baseline and after 6 and 12 weeks of 

fenofibrate treatment. Four polymorphisms in both the coding (L55M 

and Q192R) and regulatory regions (T-108C and G-909C) of human 

paraoxonase1 were also quantified. Results: Micro-coated fenofibrate


significantly decreased total cholesterol, triglycerides, non-highdensity 

lipoprotein cholesterol, oxidized low-density lipoprotein and 

apolipoprotein-B levels after 6 and 12 weeks (all p < 0.001). While 

high-density lipoprotein and apolipoprotein AI levels were 

significantly increased by 14.7% and 6.9%, respectively, after 6 weeks 

and by 17.3% and 7.2%, respectively, after 12 weeks (all p < 0.01). 

There were no significant differences in the mean of low-density 

lipoprotein and C-reactive protein after fenofibrate treatment. There 

were significant increases in paraoxonase1 concentration and activity 

by 7.7% and 5.7% after 6 weeks and by 14.6% and 10.1% after 12 

weeks, respectively (all p < 0.01). After micro-coated fenofibrate 

therapy, a significantly positive correlation between the change in 

high-density lipoprotein and the changes in paraoxonase1 

concentration and activity was observed (p = 0.001). On the other 

hand, the changes in paraoxonase1 activity were significantly and 

negatively correlated with the changes in triglycerides (p = 0.007). 

The therapeutic response of lipid parameters to micro-coated 

fenofibrate was independent of paraoxonase1 polymorphisms. 

However, paraoxonase1 Q192R and T-108C polymorphisms 

significantly affected the increase in paraoxonase1 activity (the 

highest increase in 192QQ and -108TT) and paraoxonase1 

concentration (the highest increase in -108TT). Conclusion: Lipidmodifying 

therapy with micro-coated fenofibrate in patients with low 

high-density lipoprotein levels not only reduced atherogenic lipids 

(total cholesterol, triglycerides, oxidized low-density lipoprotein and 

apolipoprotein-B) and increased atheroprotective lipids but also 

increased paraoxonase1 concentration and activity. Increasing 

paraoxonase1 levels by fenofibrate may play an important role in 

decreasing low-density lipoprotein oxidation. 

(Atherosclerosis Volume 196, Issue 1, January 2008, Pages 122- 

128 ) (This study was supported by grant from the Ministry of 

University Affairs, Thailand.) 

SYSTEMATIC REVIEW OF THE BENEFITS OF 

SELF-MONITORING OF BLOOD GLUCOSE ON 

GLYCEMIC CONTROL IN TYPE 2 DIABETES 

PATIENTS. (NO. 819) 

Nalinee Poolsup 1 , Naeti Suksomboon 2,3 , Warisara Jiamsathit 2 

1 Department of Pharmacy, Faculty of Pharmacy, Silpakorn 

University, Nakhon-Pathom, Thailand; 2 Department of 


Thailand E-mail: pynss@mahidol.ac.th ; 3 Department of 


10400, Thailand 

Key words: glucose, type 2 diabetes, self mornitoring 

Background: The benefit of self-monitoring of blood 

glucose (SMBG) in improving glycemic control in type 2 diabetes 

has been controversial. This systematic review evaluated the evidence 

of benefit of SMBG on glycemic control in non-insulin-treated type 

2 diabetes. Methods: Clinical trials of SMBG were identified through 

electronic searches (MEDLINE, EMBASE, and The Cochrane 

Library) up to and including September 2007. Studies were included 

if they met the following inclusion criteria: (1) randomized controlled 

trial comparing SMBG against non-SMBG in type 2 diabetes, (2) 

included non-insulin-dependent patients only, and (3) hemoglobin 

A1c (HbA1c) reported as an outcome measure. The efficacy was 

estimated with the mean difference in the changes of HbA1c from 

baseline to final assessment between the SMBG and the non-SMBG 

groups. Results: SMBG was effective in reducing HbA1c in noninsulin-treated 

type 2 diabetes (pooled mean difference -0.24%; 95% 

confidence interval [CI] -0.37% to -0.12%; P = 0.0002). HbA1c 

decreased significantly in the SMBG subgroup where the results of 

SMBG were used to modify therapeutic regimens (pooled mean 

difference -0.27%; 95% CI -0.41% to -0.14%; P < 0.0001). In contrast, 

there was no significant difference in effects between the SMBG 

subgroup without the use of its results to modify diabetes management 

and the non-SMBG group (pooled mean difference -0.12%; 95% CI 

-0.32% to 0.08%). Conclusions: The evidence suggests the beneficial 

effect of SMBG in improving glycemic control in non-insulin-treated 

type 2 diabetes as demonstrated by the reduction of HbA1c levels. 

Specifically, SMBG proved to be useful only when SMBG results 

were used to adjust therapeutic regimens. 

(Diabetes Technology and Therapeutics Volume 10, Issue SUPPL. 

1, 1 June 2008, Pages S51-S66). (This study was granted by Faculty 

of Graduate Studies, Mahidol University.) 

PREVALENCE AND CHARACTERISTICS OF 

ADVERSE DRUG EVENTS IN RHEUMATOID 

ARTHRITIS AND OSTEOARTHRITIS AMBULA- 

TORY PATIENTS AT A LARGE TEACHING 

HOSPITAL, THAILAND. (NO. 820) 

301 

Authors: Limsuwan T 1 , Tragulpiankit P 2 , Chulavatnatol S 2 , 

Janwityanujit S 1 , Sirikhedgon U 2 , Somjarit S 2 

1 Department of Medicine, Ramathibodi Hospital, Mahidol 

University, Bangkok, Thailand, 2 Faculty of Pharmacy, Mahidol 

Univwrsity, Bangkok, Thailand: E-mail : pyptg@mahidol.ac.th 

Key words: adverse drug event, rheumatoid arthritis, osteoarthritis 

Background: Adverse drug event (ADE) is an injury due 

to medication. Most studies of ADE were performed in hospitalized 

patients but few in ambulatory patients. In addition, ADE in 

rheumatoid arthritis (RA) and osteoarthritis (OA) patients have not 

been investigated in Thailand. Objective: To determine the prevalence 

and characteristics of ADE in RA and OA ambulatory patients at 

Ramathibodi Hospital, Thailand. Methods: The RA and OA patients 

at Rheumatology clinic were randomly selected and interviewed by 

research pharmacist to identify ADE occurring during 30th April and 

19th July, 2007. Causality assessment of all suspected ADEs was 

performed using Roussel Uclaf Causality Assessment Method 

(RUCAM) and validated by rheumatologists. ADE characteristics, 

including causative drug group, affected organ, severity, and patient 

outcomes were recorded. Preventability was determined using 

Schumock and Thornton’s criteria. Results: One hundred and forty 

three patients consisted of 129 RA and 14 OA were recruited. The 

patients’ mean age was 54.3 + 14.3 years old and 121 (84.6%) patients 

were female. The number of concomitant administered drugs was 

7.9 + 2.9 items. Total of 68 ADEs were detected in 51 patients. The 

prevalence and rate of ADE were 35.6% and 47.6 events per 100 

patients, respectively. Twenty-nine ADEs (42.6%) were preventable. 

Disease-modifying anti-rheumatic drugs (DMARDs) and nonsteroidal 

anti-inflammatory drugs (NSAIDs) resulted in ADEs by 41 

(59.4%) and 10 (14.5%) events, respectively. Common affected organ 

were skin, gastrointestinal tract and vision disorders; 20 (29.4%), 18 

(25.0%) and 8 (11.8%), respectively. Thirty-three ADEs (48.5%) were 

categorized in moderate severity, i.e., required treatment with


suspected drug be held, discontinued, or changed, but no antidote or 

other treatment required. Twenty-one ADEs (30.9%) outcome were 

not recovered during studied period, e.g., skin hyper-pigmentation 

and maculopathy. Conclusion: ADEs are common in RA and OA 

patients with prevalence of 35.6%. DMARDs and NSAIDs were main 

causative agents. High exposure to potentially harmful drugs might 

help explained higher rate of non-preventable ADEs in these patients. 

(3rd Drug Hypersensitivity Meeting, 11-13 April 2008, Paris, 

France.) (This study was granted by Faculty of Graduate Studies, 

Mahidol University.) 

COLLABORATION OF PHARMACOVIGILANCE 

IN THAILAND (NO. 821) 

Tangkeo W , Suwankesawong W , Trakulpiankit P 

Deputy Secretary, Food and Drug Administration, Ministry of 

Public Health, Thailand, Health Product Vigilance Center, Food 

and Drug Administration, Ministry of Public Health, Thailand, 

Faculty of Pharmacy, Mahidol University, Bangkok, Thailand: 

E-mail : pyptg@mahidol.ac.th 

Key words: Pharmacovigilance, Thailand 

Pharmacovigilance has been defined as the science of 

activities relating to the detection, assessment, understanding, and 

prevention of adverse effects. In Thailand, the first national activity 

of pharmacovigilance is likely to the spontaneous reporting system 

according to WHO international drug monitoring programme. 

Although so far the number of reports of adverse product reactions 

has been significantly increased and resulted in signal generation, 

the other activities of pharmacovigilance by regulatory agency i.e. 

Health Product Vigilance Center has been initiated by the concept of 

partnership for patient safety. Interestingly, the impact of clinical 

pharmacy service has been established for individual patient care 

according to pharmacy practice sector. The academic institutional 

sector also has been stimulated according to pharmacovigilance/ 

clinical pharmacy global trend and public awareness. As a result, the 

collaboration of pharmacovigilance should be developed in order to 

be more powerful all aspects of activities and scientific research still 

need to support and encourage. 

(31st Annual Meeting of the WHO Programme for International Drug 

Monitoring, 20-23 October 2008, Uppsala, Sweden.) 

AN INTRODUCTION OF ADR’S COMMUNITY OF 

PHARMACY PRACTICE IN THAILAND. (NO. 822) 

Tragulpiankit P 1 , AdCoPT working team 2 

1 Faculty of Pharmacy, Mahidol University, Bangkok, Thailand, 

2 The Association of Hospital Pharmacy (Thailand), Bangkok, 

Thailand: E-mail : pyptg@mahidol.ac.th 

Key words: pharmacy practice, adverse drug reaction, Thailand 

Objective: To introduce the development, aims, and major 

activities of Adverse drug reaction’s Community of pharmacy Practice 

in Thailand (AdCoPT). Methods: Eleven voluntary Thai hospital 

pharmacists who have an experience in ADR monitoring and reporting 

established the working team. Results: The influence of clinical 

pharmacy activity is growing up for patient safety. The Association 

of Hospital Pharmacy (Thailand) [Thai HP] is a professional 

organization which has a role to play in hospital pharmacy 

improvement in terms of academic and professional activities. As a 

result, AdCoPT was initiated in 2007 under the auspices of Thai HP 

and patient safety is a goal of this community of practice. The aims 

consist of experience sharing, learning and supporting among the 

AdCoPT’s members. So far, the number of member is 470 whereas 

Thai HP’s member is 6,650. The AdCoPT’s activities included 1) 

training courses for pharmacists focused on improvement of ADR 

monitoring and reporting 2) writing practical textbooks about ADR 

assessment and 3) creating website for communication among the 

members. In the near future, the next activities will be conducted as 

following 1) survey for pharmacist’s improvement and satisfaction 

according to AdCoPT’s training course 2) creating AdCoPT’s 

newsletter and 3) conducting multi-center research. Conclusions: 

Although the national network of ADR’s community practice was 

established and takes part in improvement of ADR monitoring and 

reporting, the proactive of membership for more participation should 

be encouraged. In addition, the impact of AdCoPT activities resulting 

in patient safety should be evaluated. 

(22nd Federation of Asian Pharmaceutical Associations Congress 

(FAPA2008), 7- 10 November 2008, Singapore.) (This study was 

granted by The Association of Hospital Pharmacy (Thailand). 

TRAINING COURSE FOR IMPROVEMENT OF 

THAI ADR MONITORING AND REPORTING 

(NO. 823) 

Tragulpiankit P 1 , AdCoPT working team 2 

1 Faculty of Pharmacy, Mahidol University, Bangkok, Thailand, 

2 The Association of Hospital Pharmacy (Thailand), Bangkok, 


Key words : pharmacy practice, adverse drug reaction, Thailand 

Objectives: To report adverse drug reaction (ADR) training 

courses by Adverse drug reaction’s Community of pharmacy Practice 

in Thailand (AdCoPT) under the auspices of the Association of 

Hospital Pharmacy (Thailand) and find characteristics of pharmacists 

who attended the course. Methods: AdCoPT working team provided 

the three-days training courses for pharmacist’s competency 

improvement. It included principle of ADR’s causality assessment, 

monitoring, preventing and reporting. The case-base workshop also 

was integrated. The demographic data of attending pharmacists were 

analyzed by descriptive analysis. Results: In Thailand, although the 

national ADR spontaneous reporting scheme has a major role to play 

in ADR monitoring and reporting, the under-reporting and quality of 

ADR reporting are still low probably because of relative insufficient 

training course for ADR in academic institutions according to a new 

trend of global pharmacovigilance. Therefore, 1,100 pharmacists 

attended seven training courses which were provided between 

September, 2006 and January, 2008. Most of attending pharmacists 

were female (87%). The pharmacists who work in the central 

(including Bangkok) and south region of Thailand accounted by 34% 

and 20%, respectively. The pharmacists from primary hospital 

attended by 46% whereas the pharmacists who work at private 

hospital attended by 17%. The training courses seem to be contributed 

nationwide hospital pharmacists. Conclusions: AdCoPT takes part


in pharmaceutical education for improvement of ADR monitoring 

and reporting. Although many pharmacists attended, the improvement 

of ADR monitoring and reporting is not evaluated. The survey of 

pharmacists’ achievement should be further studied. 

(22nd Federation of Asian Pharmaceutical Associations Congress 

(FAPA2008), 7- 10 November 2008, Singapore.) (This study was 

granted by The Association of Hospital Pharmacy (Thailand). 

EVALUATION ON REPORTING OF SEVERE SKIN 

DRUG REACTION IN THAI SPONTANEOUS 

REPORTING SYSTEM : COMPLETENESS, 

TIMELINESS, AND QUALITY OF REPORTS. 

(NO. 824) 

Prachachalerm W 1 , Suwankesawong W 1 , Kaewkungwal J 2 , 

Singhasivanon P 2 , Tragulpiankit P 2 

1 Food and Drug Administration, Thailand, 2 Mahidol University, 


Background: A spontaneous reporting system (SRS) for 

adverse drug reaction (ADR) is a principle of post-marketing 

surveillance. Several international researches have shown that severe 

skin reactions (SSR) related to drugs, namely Stevens-Johnson 

syndrome and toxic epidermal necrolysis, have been seriously underreported. 

No definitive research into SRS evaluation is currently 

being undertaken in Thailand. Objective: This assess the Thai SRS 

by determining rates of completeness, under-reporting, timeliness, 

and quality of SSR reporting. Method: Fourteen selected hospitals in 

Thailand participated in this research. Those in-patient medicalrecords, 

selected by ICD-10 codes of suspected SSR in 2005 were 

validated and compared to SSR reports in the Thai Food and Drug 

Administration (FDA) database. Data analysis used descriptive 

statistics and defined parameters. Completeness rate of reporting was 

the numbers of validated SSR events, which were matched in both 

databases divided by all the numbers in the hospital database. Quality 

of report was adequacy of information of FDA reports according to 

WHO documentation grading. Results: The completeness rates of 

reporting and under-reporting rate were 55.49% (101/182), and 

44.51% (81/182), respectively. The timeliness rate for labeled SSR 

(within 2 months) was 18.60% (16/86). The quality of SSR reports 

could be rated as grade 1 and 2 for 51.49% (52/101) and 41.58% 

(42/101) respectively. Conclusion: Although SSR can be detected 

visually, this research reveals the extent to which SSR reporting 

problems existing in the Thai SRS. It is recommended that the Thai 

FDA authorities encourage healthcare professionals to report ADRs 

with a high level of completeness, timeliness, and quality. 

(The 3rd Asian Conference on Pharmacoepidemiology-Korea 2008, 

3-5 November 2008, Seoul, Korea) (This study was granted by 

Faculty of Graduate Studies, Mahidol University.) 

PHARMACIST PERCEPTIONS OF NEW 

COMPETENCY STANDARDS (NO. 825) 

Maitreemit, P. 1 , Pongcharoensuk, P. 2 , Kapol, N. 3 , Armstrong, E.P. 4 

1 Faculty of Pharmacy, Silpakorn University, Nakhon Pathhom, 

Thailand; 2 Faculty of Pharmacy, Mahidol University, Bangkok, 

Thailand; 3 Faculty of Pharmacy, Silpakon University, Nakhon 

Pathom, Thailand; 4 College of Pharmacy, University of Arizona, 

Tucson, AZ, United States 

Key words : Education; Pharmacists; Pharmacy; Professional 

Competence; Thailand 

Objectives: To suggest revisions to the Thai pharmacy 

competency standards and determine the perceptions of Thai 

pharmacy practitioners and faculty about the proposed pharmacy 

competency standards. Methods: The current competency standards 

were revised by brainstorming session with nine Thai pharmacy 

experts according to their perceptions of society’s pharmacy needs. 

The revised standards were tested for reliability and validity by 50 

pharmacy practitioners and faculty members by using a written 

questionnaire. The respondents were classified based on their practice 

setting. Results: The revision of pharmacy competency standard 

proposed the integration and addition to current competencies. Of 

830 distributed questionnaires, 574 completed questionnaires were 

received (69.2% response rate). The proposed new competency 

standards contained 7 domains and 46 competencies. The majority 

of the respondents were supportive of all 46 proposed competencies. 

The highest ranked was Domain 1 (Practice Pharmacy within Laws, 

Professional Standards, and Ethics). The second and third highest 

expectations of pharmacy graduates were Domain 4 (Provide 

pharmaceutical care) and Domain 3 (Communicate and disseminate 

knowledge effectively). Conclusion: The expectation for pharmacy 

graduates’ competencies were high and respondents encouraged 

additional growth in multidisciplinary efforts to improve patient care. 

(Pharmacy Practice Volume 6, Issue 7, July 2008, Pages 113-120) 

(This one is partially supported by Faculty of Pharmacy, Silpakorn 

University) 

COST-EFFECTIVENESS ANALYSIS OF THIAZO- 

LIDINEDIONES IN UNCONTROLLED TYPE 2 

DIABETIC PATIENTS RECEIVING SULFONY- 

LUREAS AND METFORMIN IN THAILAND 

(NO. 826) 

303 

Chirakup, S. 1,2 , Chaiyakunapruk, N 1,3,4,9 , Chaikledkeaw, U. 5,6 , 

Pongcharoensuk, P. 5 , Ongphiphadhanakul, B. 7 , Roze, S.h, 

Valentine, W.J. 8 , Palmer, A.J. 8 

1 Department of Pharmacy Practice, School of Pharmacy, 

Naresuan University, Phitsanulok, Thailand; 2 Faculty of 

Pharmacy, Payup University, Chiangmai, Thailand; 3 Ministry 

of Public Health, Bangkok, Thailand; 4 School of Population 

Health, University of Queensland, Brisbane, QLD, Australia; 

5 Department of Pharmacy, Faculty of Pharmacy, Mahidol 

University, Bangkok, Thailand; 6 Health Intervention and 

Technology Assessment Program, Ministry of Public Health, 

Bangkok, Thailand; 7 Faculty of Medicine, Ramathibodi Hospital, 

Mahidol University, Bangkok, Thailand; 8 CORE-Center for 

Outcomes Research, Binningen/Basel, Switzerland; 9 Department 

of Pharmacy Practice, School of Pharmacy, Naresuan University, 

Phitsanulok, 65000, Thailand. 

Key words : Cost-effectiveness analysis; Pioglitazone; Rosiglitazone; 

Thiazolidinediones 

Objective: The national essential drug committee in 

Thailand suggested that only one of thiazolidinediones be included 

in hospital formulary but little was know about their cost-effectiveness 

values. This study aims to determine an incremental cost-effectiveness 

ratio of pioglitazone 45 mg compared with rosiglitazone 8 mg in 

uncontrolled type 2 diabetic patients receiving sulfonylureas and


metformin in Thailand. Methods: A Markov diabetes model (Center 

for Outcome Research model) was used in this study. Baseline 

characteristics of patients were based on Thai diabetes registry project. 

Costs of diabetes were calculated mainly from Buddhachinaraj 

hospital. Nonspecific mortality rate and transition probabilities of 

death from renal replacement therapy were obtained from Thai 

sources. Clinical effectiveness of thiazolidinediones was retrieved 

from a meta-analysis. All analyses were based on the government 

hospital policymaker perspective. Both cost and outcomes were 

discounted with the rate of 3%. Base-case analyses were analyzed as 

incremental cost per quality-adjusted life year (QALY) gained. A 

series of sensitive analyses were performed. Results: In base-case 

analysis, the pioglitazone group had a better clinical outcomes and 

higher lifetime costs. The incremental cost per QALY gained was 

186,246 baht (US$ 5389). The acceptability curves showed that the 

probability of pioglitazone being cost-effective was 29% at the 

willingness to pay of one time of Thai gross domestic product per 

capita (GDP per capita). The effect of pioglitazone on %HbA1c 

decrease was the most sensitive to the final outcomes. Conclusions: 

Our findings showed that in type 2 diabetic patients who cannot 

control their blood glucose under the combination of sulfonylurea 

and metformin, the use of pioglitazone 45 mg fell in the cost-effective 

range recommended by World Health Organization (one to three times 

of GDP per capita) on average, compared to rosiglitazone 8 mg. 

Nevertheless, based on sensitivity analysis, its probability of being 

cost-effective was quite low. Hospital policymakers may consider 

our findings as part of information for the decision-making process. 

2008, International Society for Pharmacoeconomics and Outcomes 

Research (ISPOR). 

(Value in Health Volume 11, Issue SUPPL. 1, March 2008, Pages 

S43-S51) (This one is supported by the Thailand Research Fund) 

EVALUATION OF CURRICULA CONTENT BASED 

ON THAI PHARMACY COMPETENCY 

STANDARDS (NO. 827) 

Kapol, N. 1,4 , Maitreemit, P. 1 , Pongcharoensuk, P. 2 , Armstrong, 

E.P. 3 

1 Faculty of Pharmacy, Silpakorn University, Nakhon Pathom, 

Thailand; 2 Faculty of Pharmacy, Mahidol University, Bangkok, 

Thailand; 3 College of Pharmacy, University of Arizona; 4 Faculty 

of Pharmacy, Silpakorn University, Nakhon Pathom, 73000, 

Thailand. 

Key words : Competency; Curriculum; Evaluation; Pharmacy 

education; Thailand 

Objective: To evaluate the curricula content of Thai 

pharmacy schools based on the Thai pharmacy competency standards. 

Methods: Course syllabi were collected from 11 pharmacy schools. 

A questionnaire was developed based on the Thai pharmacy 

competency standards. Course coordinators completed the 

questionnaire assessing the curricula content. Results: The curricula 

for both the Bachelor of Science in pharmacy degree (BS Pharm) 

and Doctor of Pharmacy (PharmD) degree programs included the 

minimum content required by the 8 competency domains. The 

dominant content area in BS Pharm degree programs was productoriented 

material. The content ratio of patient to product to social 

and administrative pharmacy in the BS Pharm degree programs was 

2:3:1, respectively. However, the content ratio suggested by the Thai 

Pharmacy Council was 3:2:1, respectively. For the PharmD programs, 

the largest content area was patient-oriented material, which was in 

agreement with the framework suggested by the Thai Pharmacy 

Council. Conclusions: The curricula of all Thai pharmacy schools 

met the competency standards; however, some patient-oriented 

material should be expanded and some product-oriented content 

deleted in order to meet the recommended content ratio. 

(American Journal of Pharmaceutical Education Volume 72, Issue 

1, 2008, Article number 9) (This one is partially supported by 

Faculty of Graduate Studies, Silpakorn University) 

PHYSICOCHEMICAL PROPERTIES AND 

BIOCOMPATIBILITY OF N-TRIMETHYL 

CHITOSAN: EFFECT OF QUATERNIZATION 

AND DIMETHYLATION. (NO. 828) 

Anchalee Jintapattanakit 1,2 , Shirui Mao 2,3 , Thomas Kisse l,2 , 

Varaporn Buraphacheep Junyaprasert 1 

1Department of Pharmacy, Faculty of Pharmacy, Mahidol 

University, Bangkok, Thailand; 2Department of Pharmaceutics 

and Biopharmacy, Philipps-Universitat, Marburg, Germany; 

3School of Pharmacy, Shenyang Pharmaceutical University, 

Shenyang, China. 

Key words: Cytotoxicity; Degree of dimethylation; Degree of 

quaternization; Mucoadhesive properties; N-trimethyl chitosan; 

Solubility 

The aim of this research was to investigate the effect of 

degrees of quaternization (DQ) and dimethylation (DD) on 

physicochemical properties and cytotoxicity of N-trimethyl chitosan 

(TMC). TMC was synthesized by reductive methylation of chitosan 

in the presence of a strong base at elevated temperature and polymer 

characteristics were investigated. The number of methylation process 

and duration of reaction were demonstrated to affect the DQ and 

DD. An increased number of reaction steps increased DQ and 

decreased DD, while an extended duration of reaction increased both 

DQ and DD. The molecular weight of TMC was in the range of 60- 

550 kDa. From the Mark-Houwink equation, it was found that TMC 

in 2% acetic acid/0.2 M sodium acetate behaved as a spherical 

structure, approximating a random coil. The highest solubility was 

found with TMC of an intermediate DQ (40%) regardless of DD and 

molecular weight. The effect of DD on the physicochemical properties 

and cytotoxicity was obviously observed when proportion of DD to 

DQ was higher than 1. TMC with relatively high DD showed 

reduction in both solubility and mucoadhesion and hence decreased 

cytotoxicity. However, the influence of DD was insignificant when 

DQ of TMC was higher than 40% at which physicochemical 

properties and cytotoxicity were mainly dependent upon DQ. 

(European Journal of Pharmaceutics and Biopharmaceutics Volume 

70, Issue 2, October 2008, Pages 563-571.) (Acknowledgements: 

The authors are grateful for financial support from 1The Thailand 

Research Fund (TRF) through the Royal Golden Jubilee Ph.D. 

program (Grant No. PHD/0226/2545) and the German Academic 

Exchange Service (Deutsche Akademische Austauschdienst, DAAD). 

We are very pleased to acknowledge the National Metal and 

Materials Technology Center (MTEC, Pathumthani, Thailand) for 

GPC experiment.)


EFFECT OF CHARGED AND NON-IONIC 

MEMBRANE ADDITIVES ON PHYSICOCHEMICAL 

PROPERTIES AND STABILITY OF NIOSOMES. 

(NO. 829) 

Varaporn Buraphacheep Junyaprasert, Veerawat Teeranachaideekul, 

Tasaneeya Supaperm. 

Department of Pharmacy, Faculty of Pharmacy, Mahidol 

University, 447, Sri-Ayutthaya Rd., Rajathavee, Bangkok, 10400, 

Thailand. 

Key words : entrapment efficiency; membrane additives; niosomes; 

salicylic acid; transmission electron microscopy 

The aim of this study was to investigate an influence of 

different types of membrane additives including negative charge 

(dicetylphosphate, DCP), positive charge (stearylamine, STR) and 

non-ionic molecule (cholesteryl poly-24-oxyethylene ether, SC24) 

on the physicochemical properties of drug-free and drug-loaded 

niosomes. Salicylic acid having different proportions of ionized and 

unionized species at different pH was selected as a model drug. The 

niosomes were composed of 1:1 mole ratio of Span 60: cholesterol 

as vesicle forming agents. The results show that incorporation of 

salicylic acid to the niosomes did not affect zeta potential values; 

however, addition of the membrane additives changed the zeta 

potential depending on the type of the additives. Transmission electron 

microscopy revealed that niosomes had unilamellar structure. The 

particle sizes of all developed niosomes were between 217 to 360 nm. 

The entrapment efficiency (% E.E.) of all salicylic acid niosomes at 

pH 3 was higher than that of niosomes at pH 5, indicating that 

salicylic acid in unionized form was preferably incorporated in 

niosomes. Furthermore, the positively charged niosomes showed the 

highest % E.E. of salicylic acid owing to electrostatic attraction 

between STR and salicylic acid. After 3 months of storage at 4 C, 

the particle size of the niosomes remained in the nanosize range except 

for DCP salicylic acid niosomes at pH 3 whose size increased due to 

an instability of DCP at low pH. In addition, all niosomes showed no 

leakage of the salicylic acid after 3 months of storage indicating the 

good stability. 

(AAPS PharmSciTech 9 (3), (2008) pp. 851-859.) 

INFLUENCE OF OIL CONTENT ON PHYSICO- 

CHEMICAL PROPERTIES AND SKIN DISTRI- 

BUTION OF NILE RED-LOADED NLC. (NO. 830) 

Veerawat Teeranachaideekul 1,2 , Prapaporn Boonme 3 , Eliana 

Barbosa Souto 2,4 , Rainer Helmut M ller 2 , Varaporn 

Buraphacheep Junyaprasert 1 


University, Thailand; 2 Department of Pharmaceutics, 

Biopharmaceutics and Quality Management, Free University of 

Berlin, Germany; 3 Department of Pharmaceutical Technology, 

Faculty of Pharmaceutical Sciences, Prince of Songkla University, 

Thailand; 4 Department of Pharmaceutical Technology, Faculty 

of Health Sciences, Fernando Pessoa University, Portugal 

Key words : Confocal laser scanning microscopy; Nanostructured 

lipid carriers; Nile red; NLC; Occlusion factor; Skin distribution 

The aims of this study were to investigate the effect of the 

oil content on the physicochemical properties of NLC and to elucidate 

the potential of NLC for skin targeting. The obtained results showed 

that an increase in the oil content did not affect the mean particle 

size of NLC but impacted on the zeta potential. The inner structure 

of NLC was influenced by the increasing proportion of oil towards 

the less ordered structure as confirmed by differential scanning 

calorimetry (DSC) and X-ray diffraction (XRD), particularly for the 

higher medium chain triglycerides (MCT) loading. The data from 

proton nuclear magnetic resonance (1H NMR) revealed that cetyl 

palmitate nanoparticles did not completely recrystallize after cooling 

down to room temperature. 1H NMR and DSC results indicate that 

MCT molecules were restricted in the NLC as compared to the 

nanoemulsions (NE). Nile red distribution and penetration into skin 

from NLC were pronounced as compared to NE and dependent on 

the MCT loading. The deep penetration and high amount of Nile red 

were related to the occlusion factor. Moreover, the epidermal targeting 

was achieved by NLC applications, particularly those containing 5% 

MCT (NLC-5) depending on the amount of MCT loading. 

(Journal of Controlled Release Volume 128, Issue 2, 4 June 2008, 

Pages 134-141.) 

(Acknowledgements: 2Financial support from the Thailand Research 

Fund (TRF) through the Royal Golden Jubilee Ph.D. Program (Grant 

No. PHD/0160/2546), the TRF-Master Research Grants (MRG- 

OSMEP505S176) and the German Academic Exchange Service 

(DAAD) is gratefully acknowledged. Yanhee General Hospital 

Bangkok, Thailand is thanked for providing the skin for the in vitro 

skin distribution studies.) 

AEROSOL OT MICROEMULSIONS AS CARRIERS 

FOR TRANSDERMAL DELIVERY OF HYDRO- 

PHOBIC AND HYDROPHILIC LOCAL ANESTHE- 

TICS. (NO. 831) 

305 

Varaporn Buraphacheep Junyaprasert 1,5 , Prapaporn Boonme 2 , 

Dale Eric Wurster 3 , Thomas Rades 4 


University, Bangkok, Thailand; 2 Department of Pharmaceutical 

Technology, Faculty of Pharmaceutical Sciences, Prince of 

Songkhla University, Songkhla, Thailand; 3 College of Pharmacy, 

University of Iowa, Iowa City, IA, United States; 4 School of 

Pharmacy, University of Otago, Dunedin, New Zealand; 


University, Bangkok 10400, Thailand 

Key words : Aerosol OT microemulsions; Local anesthetics; Skin 

permeation 

The skin permeation enhancement of many kinds of drugs 

and cosmetic substances by microemulsions has been widely known; 

however, the correlations between microemulsion microstructures and 

the efficiency of skin permeation are not fully elucidated. Therefore, 

the aim of our study was to investigate the influence of microemulsion 

types on in vitro skin permeation of model hydrophobic drugs and 

their hydrophilic salts. The microemulsion systems were composed 

of isopropyl palmitate (IPP), water, a 2:1 w/w mixture of Aerosol OT 

(AOT) and 1-butanol, and a model drug. The concentrations of 

surfactant mixture and model drug were maintained at 45% and 1% 

w/w, respectively. The concentrations of IPP and water were 15%


and 39% w/w, respectively, for oil-in-water (o/w) type and vice versa 

for water-in-oil (w/o) type. The samples were prepared by simple 

mixing and characterized by visual appearance, pH, refractive index, 

electrical conductivity, viscosity, and determination of the state of 

water and IPP in the formulations using differential scanning 

calorimetry. Transdermal flux of lidocaine, tetracaine, dibucaine, and 

their respective hydrochloride salts from the drug-loaded AOT-based 

microemulsions through heat-separated human epidermis was 

investigated in vitro using modified Franz diffusion cells. The o/w 

microemulsions resulted in the highest fluxes of the model drugs in 

base form as compared with the other formulations within the same 

group of drugs. Moreover, the skin permeation of drug from 

microemulsions depended on drug molecular structure and interaction 

between drug and surfactant. 

(Drug Delivery Volume 15, Issue 5, June 2008, Pages 323-330.) 

(The research was supported by The Thailand Research Fund (TRF) 

through the Royal Golden Jubilee Ph.D. program.) 

FLOATING PROPERTIES AND RELEASE 

CHARACTERISTICS OF HOLLOW MICRO- 

SPHERES OF ACYCLOVIR. (NO. 832) 

Varaporn Buraphacheep Junyaprasert 1,2 , Supaporn Pornsuwannapha 

1 


University, Bangkok, Thailand; 2 Department of Pharmacy, 

Faculty of Pharmacy, Mahidol University, Bangkok 10400, 

Thailand. 

Key words: Acyclovir; Controlled release; Eudragit S100; Floating 

microspheres; Hollow microspheres; Solvent evaporation diffusion 

Acyclovir, a selective antiherpes virus agent, was loaded 

in the hollow microspheres to improve bioavailability and patient 

compliance by prolonging the residence time in the gastrointestinal 

tract. The hollow microspheres of acyclovir were prepared by solvent 

evaporation diffusion method using Eudragit S 100 as a controlled 

polymer. We found that the process conditions that provided the high 

% yield of the hollow microspheres were the use of 5:8:2 of 

dichloromethane: ethanol: isopropanol as a solvent system and stirring 

at 300 rpm for 60 min. The size of the microspheres prepared from 

different ratios of acyclovir and Eudragit S 100 was 159-218 m. 

When the drug-to-polymer ratio was increased, the size and percent 

drug content increased. The highest percent drug entrapment was 

obtained at the ratio of 600 mg acyclovir: 1 g Eudragit S 100. The 

hollow microspheres tended to float over 0.1 M hydrochloric acid 

containing 0.02% Tween 20 solution for 24 hr. The rate of acyclovir 

released from the microspheres was generally low in simulated gastric 

fluid without enzyme due to the low permeability of the polymer. 

However, in phosphate buffer pH 6.8, the drug release increased as 

the drug load increased due to the swelling property of the polymer. 

In simulated intestinal fluids without enzymes, the polymer 

completely dissolved resulting in instant release of the drug in this 

medium. 

(Drug Delivery Volume 15, Issue 5, June 2008, Pages 331-341.) 

(The research was supported by Mahidol University Research 

Funding.) 

DEVELOPMENT OF ASCORBYL PALMITATE 

NANOCRYSTALS APPLYING THE NANOSUS- 

PENSION TECHNOLOGY. (NO. 833) 

Veerawat Teeranachaideekul 1,2 , Varaporn B. Junyaprasert 1 , 

Eliana B. Souto 3 , Rainer H. M ller 2 


University, 447 Sri-Ayutthaya Road, Rajathavee, Bangkok 10400, 

Thailand; 2 Department of Pharmacy, Pharmaceutical 

Technology, Biopharmaceutics and NutriCosmetics, Kelchstr. 31, 

D-12169 Berlin, Germany; 3 Department of Pharmaceutical 

Technology, Faculty of Health Sciences, Fernando Pessoa 

University, Rua Carlos da Maia 296, 4200-150 Porto, Portugal 

Key words: Ascorbyl palmitate; Atomic force microscopy; 

Lyophilization; Nanosuspensions; Scanning electron microscopy; 

Ascorbyl palmitate (AP) is an antioxidant used in both 

cosmetics and food industry. Owing to its poor solubility and 

instability caused by oxidation having been observed in several 

colloidal systems, the aim of this study was to investigate the 

feasibility of applying the nanosuspension technology by highpressure 

homogenization (HPH) (DissoCubes technology) to 

enhance the chemical stability of AP, followed by lyophilization. 

Sodium dodecyl sulfate (SDS) and Tween 80 were chosen as 

emulsifying agents to stabilize the developed AP nanosuspensions. 

After 3 months of storage at three different temperatures (4 C, 25 

C and 40 C), the photon correlation spectroscopy (PCS) analysis 

of AP nanosuspensions revealed that the mean particle size of those 

stabilized with SDS significantly increased compared to those 

stabilized with Tween 80. The results observed from both atomic 

force microscopy (AFM) and scanning electron microscopy (SEM) 

revealed AP nanocrystals of cubic-like shape. The percentage of AP 

remaining in nanosuspensions stabilized with Tween 80 was higher 

than 90% after 3 months storage at 4 C, 25 C and 40 C. To increase 

the chemical stability of AP nanosuspensions, a drug powder was 

prepared by lyophilization. The effect of the presence of 

cryoprotectant trehalose on the physical stability was evaluated at 

different concentrations. After redispersing the lyophilized product, 

the mean size of AP nanosuspensions without trehalose was 

significantly higher compared with the system with trehalose. After 

3 months of storage at 25 C the mean size of lyophilized AP 

nanosuspensions remained constant. X-ray diffraction revealed the 

crystalline character of AP nanocrystals after HPH and lyophilization. 

(International Journal of Pharmaceutics Volume 354, Issue 1-2, 16 

April 2008, Pages 227-234.) (Acknowledgements: Financial 

support from 3The Thailand Research Fund (TRF) through the Royal 

Golden Jubilee Ph.D. Program (Grant no. PHD/0160/2546) and from 

the German Academic Exchange Service (DAAD) is gratefully 

acknowledged. The AFM machine is supported by the Nanoscience 

and Nanotechnology, Faculty of Science, Mahidol University, 

Thailand.)


RELEASE PROFILE COMPARISON AND 

STABILITY OF DILTIAZEM-RESIN MICROCAP- 

SULES IN SUSTAINED RELEASE SUSPENSIONS. 

(NO. 834) 

Varaporn Buraphacheep Junyaprasert 1 , Greepol Manwiwattanakul 

2 

1Department of Pharmacy, Faculty of Pharmacy, Mahidol 

University, 447 Sri-Ayutthaya Road, Bangkok, 10400, Thailand; 

2Faculty of Pharmacy, Mahasarakham University, Mahasarakham, 

Thailand. 

Key words : Diltiazem; Emulsion-solvent evaporation; Ion-exchange 

resin; Sustained release suspension 

A sustained release suspension of diltiazem, a short halflife 

calcium channel blocker, was developed to reduce frequency of 

drug administration, ease of dose adjustment and improve patient 

compliance. In this study, the sustained release of diltiazem was 

obtained by complexing the drug with Dowex 50W 4 and Dowex 

50W 8, strong cationic exchange resins with 4% and 8% degree of 

cross-linking, respectively. The diltiazem-Dowex 50W 4 complexes 

provided the highest drug release and were subsequently used to 

prepare the microcapsules by emulsion-solvent evaporation method, 

using 0.75-5.00% cellulose acetate butyrate (CAB) in methylene 

chloride as a coating solution. As the concentration of CAB increased, 

the size of microcapsule increased and the drug release from the 

microcapsule was retarded. From release profile comparison using 

f1 and f2 factors, it was found that the microcapsules coated with 

1.75% CAB provided a release profile equivalent to the commercial 

product of diltiazem sustained release capsule, Herbesser 90SR. 

Furthermore, sustained release suspensions of the diltiazem 

microcapsules were formulated with the use of 0.8% sodium 

carboxymethylcellulose or 0.4% xanthan gum as a suspending agent. 

The suspension of 0.4% xanthan gum showed superior in physical 

appearance after 120-day storage at 30 and 45 C. In addition, all 

sustained release suspensions possessed good stability with low drug 

leaching and their release profiles were unchanged when compared 

with the dried microcapsules for 120 days at 30 and 45 C. 

(Acknowledgement: 4The authors are grateful for financial support 

received from Mahidol University Research Funding.) (International 

Journal of Pharmaceutics Volume 352, Issue 1-2, 20 March 2008, 

Pages 81-91.) 

PHYSICOCHEMICAL CHARACTERIZATION AND 

IN VITRO RELEASE STUDIES OF ASCORBYL 

PALMITATE-LOADED SEMI-SOLID NANOSTRUC- 

TURED LIPID CARRIERS (NLC GELS). (NO. 835) 

Veerawat Teeranachaideekul 1,2 , Eliana B. Souto 2,3 , Rainer H. 

Mller 2 , Varaporn B. Junyaprasert 1 

1 Faculty of Pharmacy, Department of Pharmacy, Mahidol 

University, Bangkok, Thailand; 2 Department of Pharmacy, 

Department of Pharmaceutical Technology, Freie Universitat 

Berlin, Berlin, Germany; 3 Faculty of Pharmacy, Department of 

Pharmacy, Mahidol University, Bangkok, Thailand. 

Key words : Ascorbyl palmitate; Atomic force microscopy (AFM); 

Nanostructured lipid carriers (NLC); Viscoelastic properties 

The aim of this study was to characterize the 

physicochemical properties and to study in vitro release of ascorbyl 

palmitate from semi-solid lipid nanoparticles based on nanostructured 

lipid carriers (NLC gels) systems with the desired viscosity for dermal 

delivery. NLC gels were obtained by a one-step production procedure 

employing a high pressure homogenization technique using different 

solid lipid matrices. Ascorbyl palmitate (AP) was selected as a 

lipophilic active ingredient due to its range of cosmetic applications. 

After the production, particles within the size range 170-250 nm 

having polydispersity index lower than 0.3 were obtained from all 

formulations. After the AP incorporation into the NLC gels, the zeta 

potential increased to values higher than 30 mV . Almost 100% 

encapsulation efficiency was observed. The obtained SEM and AFM 

data revealed non-spherical shaped nanoparticles. From DSC and Xray 

diffraction studies, it was shown that the lipid recrystallized in 

the solid state possessing a less ordered structure as compared to the 

bulk material. The release study of active-loaded NLC gel 

formulations using Franz diffusion cells revealed that the type of 

lipid matrix affects both the rate and the release pattern. The 

viscoelastic measurements revealed a more elastic than viscous 

behaviour of NLC formulations indicating a typical gel-like structure. 

(Journal of Microencapsulation Volume 25, Issue 2, March 2008, 

Pages 111-120.) 

THE EFFECT OF CETYL PALMITATE CRYSTAL- 

LINITY ON PHYSICAL PROPERTIES OF GAMMA- 

ORYZANOL ENCAPSULATED IN SOLID LIPID 

NANOPARTICLES. (NO. 836) 

307 

Uracha Ruktanonchai 1 , Surachai Limpakdee 2 , Siwaporn Meejoo 2 , 

Usawadee Sakulkhu 1 , Nuntavan Bunyapraphatsara 3 , Varaporn 

Junyaprasert 4 , Satit Puttipipatkhachorn 5 

1 National Nanotechnology Center, National Science and 

Technology Development Agency, 111 Thailand Science Park, 

Paholyothin Road, Klong 1, Klong Luang, Pathumthani 12120, 

Thailand.; 2 Department of Chemistry, Faculty of Science, 

Mahidol University, Rama VI Road, Bangkok 10400, Thailand.; 


University, Sri-ayudhya Road, Bangkok 10400, Thailand.; 


University, Sri-ayudhya Road, Bangkok 10400, Thailand; 


Mahidol University, Sri-ayudhya Road, Bangkok 10400, 

Thailand. 

This present study was aimed at investigating the effect 

of the crystallinity of cetyl palmitate based solid lipid nanoparticles 

(SLNs) on the physical properties of -oryzanol-loaded SLNs. SLNs 

consisting of varying ratios of cetyl palmitate and -oryzanol were 

prepared. Their hydrodynamic diameters were in the range 210-280 

nm and the zeta potentials were in the range -27 to -35 mV. The size 

of SLNs increased as the amount of cetyl palmitate decreased whereas 

no significant change of zeta potentials was found. Atomic force 

microscopy pictures indicated the presence of disc-like particles. The 

crystallinity of SLNs, determined by differential scanning calorimetry 

and powder x-ray diffraction, was directly dependent on the ratio of 

cetyl palmitate to -oryzanol and decreased with decreasing cetyl


palmitate content in the lipid matrix. Varying this ratio in the lipid 

mix resulted in a shift in the melting temperature and enthalpy, 

although the SLN structure remained unchanged as an orthorhombic 

lamellar lattice. This has been attributed to a potential inhibition by 

-oryzanol during lipid crystal growth as well as a less ordered 

structure of the SLNs. The results revealed that the crystallinity of 

the SLNs was mainly dependent on the solid lipid, and that the 

crystallinity has an important impact on the physical characteristics 

of active-loaded SLNs. 

(Nanotechnology 19(9); 5 February 2008: Article number 095701.) 

(The research was supported by The National Nanotechnology 

Center (NANOTEC), Thailand (Research grant number B22 

CR0102). 

CHARACTERIZATION OF “YAA CHUD”MEDICINE 

ON THE THAILAND-MYANMAR BORDER : 

SELECTING FOR DRUG-RESISTANT MALARIA 

AND THREATENING PUBLIC HEALTH. (NO. 837) 

Newton, P.N. 1,2,4 , Hampton, C.Y. 2 , Alter-Hall, K. 2 , Teerwarakulpana, 

T. 5 , Prakongpan, S. 6 , Ruangveerayuth, R. 7 , White, N.J. 

4,8 , Day, N.P.J. 4,8 , Tudino, M.B. 9 , Mancuso, N. 9 , Ferna ndez, 

F.M. 2 

1 Wellcome Trust-Mahosot Hospital-Oxford Tropical Medicine 

Research Collaboration, Microbiology Laboratory, Mahosot 

Hospital, Vientiane, Laos; 2 School of Chemistry and 

Biochemistry, Georgia Institute of Technology, Atlanta, GA 

30332, United States; 3 Wellcome Trust-Mahosot Hospital-Oxford 

Tropical Medicine Research Collaboration, Mahosot Hospital, 

Vientiane, Laos; 4 Centre for Clinical Vaccinology and Tropical 

Medicine, University of Oxford, Churchill Hospital, Oxford, OX3 

7LJ, United Kingdom; 5 Faculty of Pharmacy, Mahidol University, 

Bangkok, Thailand; 6 Mae Sot Hospital, Mae Sot, Tak Province, 

Thailand; 7 Faculty of Tropical Medicine, Mahidol University, 

420/6 Rajvithi Road, Bangkok 10400, Thailand; 8 Departamento 

de Qui mica Inorga nica, Anali tica Y Qui mica Fi sica, Instituto 

de Quimica de Los Materiales, Medio Ambiente Y Energia, 

Ciudad Universitaria, 1428, Buenos Aires, Argentina 

Multidrug-resistant Plasmodium falciparum malaria is a 

severe public health problem on the Thailand-Myanmar border. Many 

villagers buy packets of 4-5 mixed medicines (“yaa chud”) from shops 

without medical assessment as their first-line malaria treatment. In 

2000-2001 a local researcher purchased 50 yaa chud from 44 shops 

around Mae Sot, Thailand and Myawaddy, Myanmar (Burma), for 

his wife who was said to be pregnant with fever and drowsiness. The 

tablets/capsules were provisionally identified by appearance and 

active ingredients determined in a subset by using mass and atomic 

spectrometry. The most frequently detected active ingredients were 

acetaminophen (22%), chlorpheniramine (13.4%), chloroquine 

(12.6%), tetracycline/doxycycline (11.4%), and quinine (5.1%). Only 

seven bags contained potentially curative medicine for malaria. A 

total of 82% of the bags contained medicines contraindicated in 

pregnancy. Inappropriate, ineffective antimalarial drugs on the 

Thailand-Myanmar border are likely to increase malaria morbidity, 

mortality and health costs and engender the emergence and spread 

of antimalarial drug resistance. 

(Acknowledgments: We thank Yongyuth Losuppakarn (Mae Sot 

Hospital) and Kamolrat Silamut for assistance; the students of the 

Faculty of Pharmacy, Mahidol University (Bangkok) for help in 

provisionally identifying the yaa chud ingredients; Catherine 

Goodman, Elizabeth Ashley, Rose McGready, Shunmay Yeung, and 

Francois Nosten for helpful comments on the manuscript; and Amin 

Abdinasir, Karen Barnes, and Mayfong Mayxay for advice) 

(Financial support: 5The collection of samples and part of the 

chemical analysis were supported by the Wellcome Trust of Great 

Britain as part of the Wellcome Trust–Mahidol University–Oxford 

Tropical Medicine Research Programme. Facundo M. Fern ndez 

was supported by a National Science Foundation CAREER grant 

for DART-MS analysis. Christina Y. Hampton was supported by a 

Molecular Biophysics Training Program from the Georgia Institute 

of Technology.) 

(American Journal of Tropical Medicine and Hygiene Volume 79, 

Issue 5, November 2008, Pages 662-669.) 

COMBINATION CHEMOTHERAPY AND PHOTO- 

DYNAMIC THERAPY WITH FAB2 FRAGMENT 

TARGETED HPMA COPOLYMER CONJUGATES 

IN HUMAN OVARIAN CARCINOMA CELLS. 

(NO. 838) 

Jarunee Hongrapipat, Pavla Kope kova , Jihua Liu, Sompol 

Prakongpan and Jindri h Kope ek 

Department of Pharmaceutics and Pharmaceutical Chemistry 

and Department of Bioengineering, University of Utah, Salt Lake 

City, Utah 84112, and Department of Pharmacy, Mahidol 

University, Bangkok 10400, Thailand. 

The biological activities of sequential combinations of 

anticancer drugs, SOS thiophene (SOS) and mesochlorin e6 

monoethylenediamine (Mce6), in the form of free drugs, nontargeted 

N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer”drug 

conjugates, P-GFLG-Mce6 and P-GFLG-SOS (P is the HPMA 

copolymer backbone and GFLG is the glycylphenylalanylleucylglycine 

spacer), and Fab2 -targeted HPMA copolymer”drug 

conjugates, P-(GFLG-Mce6)-Fab2 and P-(GFLG-SOS)-Fab2 (Fab2 

from OV-TL16 antibodies complementary to CD47), were evaluated 

against human ovarian carcinoma OVCAR-3 cells. Mce6, SOS, P- 

GFLG-Mce6, P-GFLG-SOS, P-(GFLG-Mce6)-Fab2 , and P-(GFLG- 

SOS)-Fab2 , when used as single agents or in binary combination, 

exhibited cytotoxic activities against OVCAR-3 cells, as determined 

using a modified MTT assay. The binding and internalization of P- 

(GFLG-Mce6)-Fab2 and P-(GFLG-SOS)-Fab2 by OVCAR-3 cells 

were visualized by confocal microscopy and flow cytometry. The 

results confirmed an enhanced biorecognition by OVCAR-3 cells of 

Fab2 -targeted HPMA copolymer conjugates over nontargeted 

conjugates. The median-effect analysis and the determination of the 

combination index (CI) were used to describe the drug interaction 

and quantify the synergism, antagonism, or additivity in anticancer 

effects. The sequential combinations of SOS+Mce6 and P-GFLG- 

SOS+P-GFLG-Mce6 displayed very strong synergism to synergism 

in the entire range of cell inhibition levels (fa = 0.5 “ 0.95). The P- 

(GFLG-SOS)-Fab2 +P-(GFLG-Mce6)-Fab2 exhibited a strong 

synergism for fa values up to about 0.85, but showed synergistic 

effect and nearly additive effect at fa = 0.9 and 0.95, respectively. 

These observations support the continuation of in vivo investigations 

of these conjugates for the treatment of ovarian cancer. 

(Mol. Pharmaceutics, 2008, 5 (5), pp 696–709)


ENHANCED ANTITUMOR ACTIVITY OF COMBI- 

NATIONS OF FREE AND HPMA COPOLYMER- 

BOUND DRUGS (NO. 839) 

J. Hongrapipat 1,2 , P. Kopeckova 1 , S. Prakongpan 2 , J. Kopecek 1,3 

1 Department of Pharmaceutics and Pharmaceutical Chemistry, 

University of Utah, Salt Lake City, UT 84112, USA; 2 Department 

of Pharmacy, Mahidol University, Bangkok 10400, Thailand; 

3 Department of Bioengineering, University of Utah, USA. 

Key words: N-(2-Hydroxypropyl) methacrylamide (HPMA) 

copolymer; 2,5-bis(6-hydroxymethyl-2-thienyl)furan; Doxorubicin; 

Mesochlorin e6 monoethylenediamine;Combination index; Renal 

cancer 

The synergism in anticancer effect toward human renal 

carcinoma A498 cells by binary combinations of free and N-(2hydroxypropyl)methacrylamide 

(HPMA) copolymer-bound 

anticancer drugs, SOS thiophene (SOS), doxorubicin (DOX), and 

mesochlorin e6 monoethylenediamine (Mce6), was evaluated. The 

combination index (CI) analysis was used to quantify the synergism, 

antagonism, and additive effects. Both free drugs 

andHPMAcopolymer conjugates, when used as single agents or in 

combination, exhibited cytotoxic activities against A498 cells, as 

determined using a modifiedMTTassay.As single agents, SOS and 

P-GFLG- SOS(HPMAcopolymer conjugates containing SOS bound 

via glycylphenylalanylleucylglycine [GFLG] spacer) were 

significantly more effective than the other agents evaluated. The 

synergistic effects ranked in the order SOS +DOX> P-GFLG-DOX+ 

P-GFLG-Mce6 H”DOX+ Mce6 > P-GFLG-SOS + P-GFLG- 

DOXH”SOS + Mce6 > P-GFLG-SOS + PGFLG- Mce6. The 

combination of SOS +DOX proved to be synergistic over all cell 

growth inhibition levels. All other combinations exhibited synergism 

in a wide range of drug effect levels. The SOS + Mce6 and P-GFLG- 

SOS + P-GFLG-Mce6 combinations displayed synergism up to drug 

affected fraction (fa) values of about 0.8 and reached slight 

antagonism and nearly additivity at fa = 0.95, respectively. However, 

all other combinations were synergistic up to fa < 0.9 and were 

additive at higher fa values. The observations that most combinations 

produced synergistic effects will be important for clinical translation. 

(International Journal of Pharmaceutics. 351 (2008) 259–270.) (The 

research was supported in part by the NIH grant CA51578 from the 

National Cancer Institute and by the Royal Golden Jubilee Ph.D. 

Program (PHD/0176/2545) from Thailand.) 

SYSTEM DYNAMIC MODELING : AN ALTER- 

NATIVE METHOD FOR BUDGETING. (NO. 840) 

Witsanuchai Srijariya 1,2,3 , Arthorn Riewpaiboon 1 , Usa 

Chaikledkaew. 1 


University, Bangkok, Thailand; 2 Ministry of Public Health, 

Bangkok, Thailand; 3 Department of Pharmacy, Faculty of 

Pharmacy, Mahidol University, 447 Sri-Ayutthaya, Ratchathevi 


Key words: Budgeting; Emergency services; Financial model; 

System dynamic model 

Objectives: To construct, validate, and simulate a system 

dynamic financial model and compare it against the conventional 

method. Methods: The study was a cross-sectional analysis of 

secondary data retrieved from the National Health Security Office 

(NHSO) in the fiscal year 2004. The sample consisted of all emergency 

patients who received emergency services outside their registered 

hospital-catchments area. The dependent variable used was the 

amount of reimbursed money. Two types of model were constructed, 

namely, the system dynamic model using the STELLA software and 

the multiple linear regression model. The outputs of both methods 

were compared. Results: The study covered 284,716 patients from 

various levels of providers. The system dynamic model had the 

capability of producing various types of outputs, for example, 

financial and graphical analyses. For the regression analysis, 

statistically significant predictors were composed of service types 

(outpatient or inpatient), operating procedures, length of stay, illness 

types (accident or not), hospital characteristics, age, and hospital 

location (adjusted R2 = 0.74). The total budget arrived at from using 

the system dynamic model and regression model was 

US$12,159,614.38 and US$7,301,217.18, respectively, whereas the 

actual NHSO reimbursement cost was US$12,840,805.69. 

Conclusions: The study illustrated that the system dynamic model is 

a useful financial management tool, although it is not easy to 

construct. The model is not only more accurate in prediction but is 

also more capable of analyzing large and complex real-world 

situations than the conventional method. 2008, International 

Society for Pharmacoeconomics and Outcomes Research (ISPOR). 


S115-S123. 8Contract grant sponsor: The National Health Security 

Office (NHSO), Thailand.) 

A COST FUNCTION ANALYSIS OF SHIGELLOSIS 

IN THAILAND. (NO. 841) 

Arthorn Riewpaiboon 1,7 , Sitaporn Youngkong 2 , Nutta 

Sreshthaputra, 3 , John F. Stewart 4 , Seksun Samosornsuk 5 , Wanpen 

Chaicumpa 5 , Lorenz Von Seidlein 6 , John D. Clemens. 6 

1 Faculty of Pharmacy, Mahidol University, Bangkok, Thailand; 

2 Faculty of Pharmacy, Srinakarinwirot University, Nakhonnayok, 

Thailand; 3 Faculty of Economics, Chulalongkorn University, 

Bangkok, Thailand; 4 Department of Economics, University of 

North Carolina-Chapel Hill, Chapel Hill, NC, United States.; 

5 Faculty of Allied Health Sciences, Thammasart University, 

Patumthani, Thailand.; 6 International Vaccine Institute, Seoul, 

South Korea.; 7 Department of Pharmacy, Faculty of Pharmacy, 

Mahidol University, 447 Sri Ayutthaya Road, Ratchathevi, 


Key words : Cost function; Public treatment cost; Shigellosis 

309 

Objective: The purpose of this study was to develop a cost 

function model to estimate the public treatment cost of shigellosis 

patients in Thailand. Methods: This study is an incidence-based costof-illness 

analysis from a provider’s perspective. The sample cases 

in this study were shigellosis patients residing in Kaengkhoi District, 

Saraburi Province, Thailand. All diarrhea patients who came to the 

health-care centers in Kaengkhoi District, Kaengkhoi District Hospital 

and Saraburi Regional Hospital during the period covering May 2002 

to April 2003 were tested for Shigella spp. The sample for our study 

included all patients with culture that confirmed the presence of


shigellosis. Public treatment cost was defined as the costs incurred 

by the health-care service facilities arising from individual cases. 

The cost was calculated based on the number of services that were 

utilized (clinic visits, hospitalization, pharmaceuticals, and laboratory 

investigations), as well as the unit cost of the services (material, labor 

and capital costs). The data were summarized using descriptive 

statistics. Furthermore, the stepwise multiple regressions were 

employed to create a cost function, and the uncertainty was tested by 

a one-way sensitivity analysis of varying discount rate, cost category, 

and drug prices. Results: Cost estimates were based from 137 episodes 

of 130 patients. Ninety-four percent of them received treatment as 

outpatients. One-fifth of the episodes were children aged less than 5 

years old. The average public treatment cost was US$8.65 per episode 

based on 2006 prices (95% CI, 4.79, and 12.51) (approximately US$1 

= 38.084 Thai baht). The majority of the treatment cost (59.3%) was 

consumed by the hospitalized patients, though they only accounted 

for 5.8% of all episodes. The sensitivity analysis on the component 

of costs and drug prices showed a variation in the public treatment 

cost ranging from US$8.29 to US$9.38 (-4.20% and 8.43% of the 

base-case, respectively). The public treatment cost model has an 

adjusted R 2 of 0.788. The positive predictor variables were types of 

services (inpatient and outpatient), types of health-care facilities 

(health center, district hospital, regional hospital), and insurance 

schemes (civil servants medical benefit scheme, social security 

scheme and universal health coverage scheme). Treatment cost was 

estimated for various scenarios based on the fitted cost model. 

Conclusion: The average public treatment cost of shigellosis in 

Thailand was estimated in this study. Service types, health-care 

facilities, and insurance schemes were the predictors used to predict 

nearly 80% of the cost. The estimated cost based on the fitted model 

can be employed for hospital management and health-care planning. 

2008, International Society for Pharmacoeconomics and Outcomes 

Research (ISPOR). 


S75-S83. ) (Source of financial support: International Vaccine 

Institute, Seoul, Korea. This study was also supported by the Diseases 

of the Most Impoverished (DOMI) Program, funded by the Bill and 

Melinda Gates Foundation and coordinated by the International 

Vaccine Institute.) 

ECONOMIC BURDEN OF ROAD TRAFFIC 

INJURIES : A MICRO-COSTING APPROACH 

(NO. 842) 

Riewpaiboon, A. 1,2 , Piyauthakit, P. 1 , Chaikledkaew, U. 1 

1 Department of Pharmacy, Division of Social and Administrative 

Pharmacy, Mahidol University, Bangkok, Thailand; 2 Faculty of 

Pharmacy, Mahidol University, 447 Sri Ayutthaya Road, 

Ratchathevi, Bangkok 10400, Thailand. 

This study aimed to determine the economic burden 

incurred from road traffic injuries in Thailand. It was designed as a 

prevalence-based cost-of-illness analysis from a societal perspective, 

employing a micro-costing bottom-up approach. It covered direct 

medical cost, direct non-medical cost, and indirect cost or productivity 

loss. Productivity loss. covers the costs of work absence or death due 

to road traffic injuries suffered by persons of working age. We 

collected data on road traffic injuries and resource utilization which 

occurred in the fiscal year 2004. A simple random sampling was used 

to select 200 patients for analysis. The average cost of road traffic 

injuries per patient was USD 2,596 at 2004 prices. This can be divided 

into direct cost (USD 102, or 4%) and indirect cost (USD 2,494, or 

96%). From these results, we can see that the indirect cost far 

outweighed the direct cost. To base decisions regarding road safety 

campaigns on savings of direct costs, particularly direct medical costs, 

is inadequate. Therefore, data on the complete cost of illness should 

be taken into account in the planning and creation of a road safety 

policy. 

(Southeast Asian Journal of Tropical Medicine and Public Health 

Volume 39, Issue 6, November 2008, Pages 1139-1149.) 

A DRUG COST MODEL FOR INJURIES DUE TO 

ROAD TRAFFIC ACCIDENTS. (NO. 843) 

Riewpaiboon A, Piyauthakit P, Srijariya W, Chaikledkaew U. 

Division of Social and Administrative Pharmacy, Department of 

Pharmacy, Mahidol University, Bangkok, Thailand. 

Key words : Accidents, Drug Costs, Thailand, Traffic 

Objective: This study aimed to develop a drug cost model 

for injuries due to road traffic accidents for patients receiving 

treatment at a regional hospital in Thailand. Methods: The study was 

designed as a retrospective, descriptive analysis. The cases were all 

from road iiaiic accidents receiving treatment at a public regional 

hospital in the fiscal year 2004. Results: Three thousand seven 

hundred and twenty-three road accident patients were included in 

the study. The mean drug cost per case was USD 18.20 (SD)=73.49, 

median=2.36). The fitted drug cost model had an adjusted R2 of 

0.449. The positive significant predictor variables of drug costs were 

prolonged length of stay, age over 30 years old, male, Universal Health 

Coverage Scheme, time of accident during 18:00-24:00 o’clock, and 

motorcycle comparing to bus. To forecast the drug budget for 2006, 

there were two approaches identified, the mean drug cost and the 

predicted average drug cost. The predicted average drug cost was 

calculated based on the forecasted values of statistically significant 

(p


4 Setting Priority Using Cost-effectiveness Analysis, Ministry of 

Public Health, Nonthaburi, Thailand; 5 School of Population 

Health, University of Queensland, Brisbane, QLD, Australia; 

6 Faculty of Medicine, Ramathibodi Hospital, Mahidol University, 

Bangkok, Thailand; 7 Department of Pharmacy, Faculty of 

Pharmacy, Mahidol University, Bangkok 10400, Thailand. 

Key words: Diabetes; Health-care costs; Hospitalizations; Risk 

factors 

Objective: The study investigated the factors affecting 

health-care costs and hospitalizations among diabetic patients in Thai 

public hospitals. Methods: A retrospective study was conducted by 

using administrative claims data obtained from diabetic patients 

during October 1, 2002 and September 30, 2003. Dependent variables 

were total health-care costs and the occurrence of hospitalizations. 

Independent variables included demographic factors, health-care 

utilizations, complications, comorbidities, and payment methods. 

Multivariate statistical analyses were applied. Results: The results of 

this study suggested that demographic factors of patients (i.e., age 

and male sex), payment methods (i.e., capitation, fee-for-service, and 

out-of-pocket) were significantly associated with higher health-care 

costs and probability of hospitalization. Patients receiving treatment 

from teaching hospitals significantly consumed higher health-care 

costs. In addition, the more health-care utilizations (i.e., occurrence 

of hospitalization, number of outpatient visit, and insulin utilization), 

the higher health-care costs the patients significantly had. Diabetic 

patients taking insulin had significantly higher health-care costs and 

risk of hospitalization. Furthermore, comorbidities (e.g., hypertension 

and cancer) and diabetes-related complications (e.g., nephropathy, 

neuropathy, retinopathy, coronary artery disease, cardiovascular 

disease, and peripheral vascular disease) were significantly associated 

with an increase in health-care costs and hospitalization. Conclusion: 

Factors affecting health-care costs and hospitalizations may help 

health-care providers intervene to improve patient management and 

possibly reduce health-care costs in the future. 2008, International 

Society for Pharmacoeconomics and Outcomes Research (ISPOR). 


S69-S74.) (This study is supported by a grant from the Thailand 

Research Fund.) 

TYPE 1 DIABETES AND HYPERCHOLESTERO- 

LAEMIA REVEAL THE CONTRIBUTION OF 

ENDOTHELIUM-DERIVED HYPERPOLARIZING 

FACTOR TO ENDOTHELIUM-DEPENDENT 

RELAXATION OF THE RAT AORTA. (NO. 845) 

Wachirawadee Malakul 1,2 , Suwan Thirawarapan 1 , Wisuda 

Suvitayavat 1 , Owen L Woodman 2,3,4 

1 Department of Physiology, Faculty of Pharmacy, Mahidol 

University, Thailand; 2 Department of Pharmacology, University 

of Melbourne, VIC, Australia; 3 School of Medical Sciences, RMIT 

University, PO Box 71, Bundoora, VIC 3083, Australia; 4 School 

of Medical Sciences, RMIT University, VIC, Australia. 

Key words: Diabetes; Endothelium-dependent relaxation; 

Endothelium-derived hyperpolarising factor; Hypercholesterolaemia; 

Superoxide anion 

311 

1.The present study evaluated the effect of diabetes, 

hypercholesterolaemia and their combination on the contribution of 

nitric oxide (NO) and endothelium-derived hyperpolarizing factor 

(EDHF) to relaxation of rat isolated aortic rings and the potential 

contribution of oxidant stress to the disturbance of endothelial 

function. 2. Thoracic aortic rings from control, diabetic, 

hypercholesterolaemic and diabetic plus hypercholesterolaemic rats 

were suspended in organ baths for tension recording. Generation of 

superoxide by the aorta was measured using lucigenin-enhanced 

chemiluminescence. 3. The maximal response to acetylcholine (ACh) 

was significantly reduced in diabetic or hypercholesterolaemic rats 

compared with control rats. In rats with diabetes plus 

hypercholesterolaemia, both the sensitivity and maximal response to 

ACh was impaired. In control rats, the response to ACh was abolished 

by the NO synthase inhibitor NG-nitro-l-arginine (l-NNA) or 

inhibition of soluble guanylate cyclase with 1H- 

[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ). In contrast, in rats 

with diabetes, hypercholesterolaemia or both, relaxation to ACh was 

resistant to inhibition by l-NNA or ODQ, but abolished by additional 

inhibition of KCa channels with charybdotoxin plus apamin. 4. The 

generation of superoxide was not significantly enhanced in aortic 

rings from either diabetic or hypercholesterolaemic rats, but was 

significantly increased in aortic rings from rats with diabetes plus 

hypercholesterolaemia. 5. These results suggest that when diabetes 

and hypercholesterolaemia impair endothelium-dependent relaxation, 

due to a diminished contribution from NO, a compensatory 

contribution of EDHF to endothelium-dependent relaxation of the 

aorta is revealed. The attenuation of NO-mediated relaxation, at least 

in the presence of both diabetes and hypercholesterolaemia, is 

associated with enhanced superoxide generation. 

(Clinical and Experimental Pharmacology and Physiology Volume 

35, Issue 2, February 2008, Pages 192-200.) (The authors thank 

The Commission of Higher Education, Ministry of Education, 

Thailand, for financial support of WM and this project.)

Faculty of Pharmacy - Mahidol University

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