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BRINGING<br />

PHARMACY<br />

KNOWLEDGE<br />

AND STUDENTS<br />

TOGETHER<br />

Volume 1 | Edition 1 | October 2013<br />

www.epsa-online.org


EPSA Students' Science Publication<br />

Dear friends,<br />

INTRODUCTION<br />

The purpose of the European Pharmaceutical Students' Association (EPSA)<br />

is to bring pharmacy, knowledge and students together. A way of bringing<br />

students' knowledge together, on a scientific level, is this new project which is<br />

a coalition between two departments: Education and Public Relations.<br />

In other words, Students + Science + Publications = EPSA Students' Science<br />

Publication (ESSP). We are proud to finally be able to collect abstracts of<br />

student research projects and compile them in a publication that students<br />

all over Europe can read and gain inspiration from. Additionally, authors get<br />

the opportunity to share their research, hypothesis, ways of tackling scientific<br />

obstacles, results and thoughts on the outcome of each scientific project. This<br />

publication will also contain some personal comments on the authors’ research<br />

experience alongside the abstracts.<br />

In order to ensure quality in the publication, we have been very lucky to be<br />

assisted by professional holds - with the help from Professor and Head of Drug<br />

Transport and Delivery Research Group, Natsa Skalko-Basnet, Department of<br />

Pharmacy (Dep. Phar) and Associate Professor at the Molecular Pharmacology<br />

Research Group (Dep. Phar), Yury Kiselev, both from the University of Tromsoe,<br />

Norway. We want, once again, to thank both of them for the energy put in this<br />

project and also thank all the students who have submitted their abstracts -<br />

because without them, this journal wouldn't exist!<br />

We hope you enjoy reading this publication, and we hereby invite and encourage<br />

you to send in your abstracts for the next editions!<br />

Yours in EPSA,<br />

Science Coordinator and Publications Coordinator 2013/2014<br />

Rebwar Saleh and Mara Lefter<br />

2


TABLE OF CONTENT<br />

Volume 1 | Edition 1 | October 2013<br />

Determination of nitrogen and organophosphorous pesticide<br />

residues in foods of plant origin – method validation, Srđan<br />

Radak......................................................................................... 4<br />

Figures and tables .......................................................................... 5<br />

Q&A of the authors ......................................................................... 6<br />

Determination of resveratrol in dietary supplements and<br />

assessment of their antioxidant activity, Karolina Podoga and<br />

Aleksandra Polak ...................................................................... 8<br />

Figures and tables .......................................................................... 9<br />

Q&A of the authors ....................................................................... 10<br />

Quantification of polyphenolic compounds with antioxidant<br />

action from various plant species, Carmen Sultan ................. 12<br />

Figures and tables ........................................................................ 13<br />

Q&A of the authors ....................................................................... 14<br />

Afterword ...................................................................................... 15<br />

3


EPSA Students' Science Publication<br />

DETERMINATION OF NITROGEN AND<br />

ORGANOPHOSPHOROUS PESTICIDE RESIDUES<br />

IN FOODS OF PLANT ORIGIN – METHOD<br />

VALIDATION<br />

Radak, Srđan. Medical Faculty, Department of Pharmacy, Novi Sad, Serbia<br />

Combination of QuEChERS method and gas or liquid chromatography with<br />

mass spectrometry has enabled pesticide residue analysis in a wide analytical<br />

range with high selectivity and sensitivity. These analytical tools have become<br />

a mainstay in most laboratories for pesticide monitoring in order to meet global<br />

standards.<br />

The aim of the study consisted in the validation of QuEChERS/GC-MS<br />

method for analysis of selected nitrogen and organophosphorous pesticide<br />

residues and evaluation of measurement uncertainty. The validation study<br />

was conducted on organically grown potato and carrot, fortified with standard<br />

mixture of nitrogen and organophosphorous pesticides, at two concentration<br />

levels, prepared by QuEChERS method and analysed by GC-MS.<br />

The results of the study showed that the limit of quantification for the analytical<br />

method of 10μg/kg reached 63% of target compounds in both examined<br />

matrices, and additional 21% compound in just one. Estimated limit of<br />

quantification of instrumental method was below or at the level of 10μg/kg<br />

for 89% of analytes in both matrices. Matrix-matched calibration curves were<br />

constructed in the range defined by the limit of quantification at lower level<br />

and 500μg/kg as the upper one. At 10μg/kg fortification level, 63% and 76%<br />

of analytes showed acceptable recovery values in potato and carrot matrix,<br />

respectively. At 100μg/kg fortification level, those values were 76% and 74%.<br />

Pesticides with good recovery values were also characterized with good<br />

precision. Evaluation of expanded measurement uncertainty showed that this<br />

parameter was below 50% for all pesticides meeting the validation criteria.<br />

Conclusion: Validation study and evaluation of measurement uncertainty<br />

confirmed the quality of multi residual QuEChERS/GC-MS method for the<br />

analysis of selected nitrogen and organophosphorous pesticides.<br />

4


FIGURES AND TABLES<br />

Volume 1 | Edition 1 | October 2013<br />

5


EPSA Students' Science Publication<br />

Q&A OF THE AUTHORS<br />

1. Why did you select this topic for your research / master thesis?<br />

Analytical chemistry is my favorite field of pharmacy. I have had the<br />

opportunity to use analytical instruments such as high performance liquid<br />

chromatography (HPLC) and gas chromatography (GC) since 2nd year of<br />

studies.<br />

2. How do you feel you have coped with the challenges that research brings?<br />

The topic is quite complex, but I have had a great opportunity to learn and<br />

work with Ass. Prof. Ljilja Torović. She is an expert in this field and one of<br />

the greatest professors in our faculty. I think I just scratched the surface of<br />

analytical chemistry and I am seeking for more knowledge.<br />

3. What personal skills have you developed as a researcher during your<br />

research period?<br />

I learned the importance of correct time management, patience and<br />

hardworking.<br />

4. If you had the opportunities, would you specialize more in the field?<br />

Yes, I have been interested in PhD and I hope to continue doing this type of<br />

research in the future.<br />

5. Any other comments you want to share with other students?<br />

Once again, I would like to thank Ass. Prof. Ljilja Torović for all the help she<br />

provided me during the research.<br />

More information:<br />

srdjanradak@yahoo.com<br />

6


Volume 1 | Edition 1 | October 2013<br />

DETERMINATION OF RESVERATROL IN DIETARY<br />

SUPPLEMENTS AND ASSESSMENT OF THEIR<br />

ANTIOXIDANT ACTIVITY<br />

Podoga, Karolina and Polak, Aleksandra from Nicolaus Copernicus University Collegium<br />

Medicum in Bydgoszcz, Poland<br />

Resveratrol (3,5,4'-trihydroxy-trans-stilbene) is a polyphenolic compound<br />

found in the skin of red grape skins, red wine and Japanese knotweed<br />

(Polygonum cuspidatum). During the last decade, resveratrol became a subject<br />

of many clinical trials, due to its influence on multiple biochemical pathways ,<br />

in particular as a strong antioxidant and chemopreventive agent. Resveratrol<br />

nutritional supplements are available over-the-counter on the pharmaceutical<br />

market and the sales are constantly increasing.<br />

The aim of this study was to optimize chromatographic method for the<br />

determination of resveratrol in food supplements, as well as evaluation of their<br />

antioxidant activity. Dietary supplements used in the project were available<br />

on the Polish pharmaceutical market: one containing only resveratrol (e.g.<br />

Trans Resveratrol) and the other multicomponent supplements (Cellasene,<br />

LongaVit, Natursteron, Nutrof). Determination of resveratrol was performed<br />

using isocratic reversed-phase high performance liquid chromatography (RP-<br />

HPLC) with spectrophotometric detection (306 nm). The antioxidant properties<br />

of tested supplements were evaluated in vitro by determining the efficiency of<br />

the inactivation of the synthetic stable radicals (DPPH •). For each sample the<br />

IC50 value was calculated, which determines the concentration of the sample<br />

causing 50% inhibition of radicals. In the final stage of the experiment, all tested<br />

supplements were compared in terms of the amount of active substance they<br />

contained, and also their antioxidant potential.<br />

Results show that the content of active substance in dietary supplements is<br />

not always the same as declared by the manufacturer. The differences in the<br />

concentration of active substance may have an influence over the expected<br />

effectiveness of dietary supplements. The study shows that preparations have<br />

various antioxidant activities. The strongest antioxidant is the multicomponent<br />

supplement – Nutrof, containing also, besides resveratrol, a number of other<br />

substances with antioxidant activity (selenium, zinc, vitamin C, vitamin E).<br />

Surprisingly, a low IC50 value (high activity) was obtained for Longavit containing<br />

only 390 mg of standardized extract from the black grape skins. This may<br />

be a result of the presence of additional natural polyphenolic compounds in<br />

grape skin extract . Dietary supplement – TransResveratrol – according to the<br />

manufacturer‘s assertion, containing 200 mg pure trans-resveratrol in capsule,<br />

has the lowest antioxidant activity. The real content of trans-resveratrol in 1<br />

capsule is 4-fold less than declared.<br />

7


EPSA Students' Science Publication<br />

FIGURES AND TABLES<br />

Graph 1: Determination of resveratrol antioxidant activity.<br />

Graph 2: Relation/Nexus between radical activity inhibition (%) and the<br />

concentration of dietary supplement - Nutrof.<br />

8


Q&A OF THE AUTHORS<br />

Volume 1 | Edition 1 | October 2013<br />

1. Why did you select this topic for your research / master thesis?<br />

The reason seems to be simple. It is due to the fact that the regulations for<br />

dietary supplements are different and less strict than those for prescription<br />

or over-the-counter drugs. Then, these studies in our opinion become<br />

extremely valuable. Obviously, dietary supplement companies affirm their<br />

products’ efficacy and good quality. The aim of our research was to check<br />

how far the companies’ declarations are from the research results.<br />

2. How do you feel you have coped with the challenges that research brings?<br />

We’ve manage to test some of the dietary supplements which were<br />

available in the Polish Pharmaceutical market. The goal for the future is to<br />

test another part of these products and compare them among each other.<br />

This would allow to choose the best product, containing resveratrol, in the<br />

Pharmaceutical market.<br />

3. What personal skills have you developed as a researcher during your<br />

research period?<br />

The first one would be assiduity, especially when at the beginning of the<br />

research some of our results weren’t reproducible. We have learnt that no<br />

matter how many difficulties will occur on your way, you should follow and<br />

fulfill your passion. The satisfaction at the end is the best prize you could<br />

imagine. The other personal skills that we have managed to obtain during<br />

the whole process of our research were definitely: being more accurate and<br />

focused.<br />

4. If you had the opportunities, would you specialize more in the field?<br />

Yes, of course. If we were given such a chance, we would definitely like to<br />

specialize ourselves in this field of science.<br />

5. Any other comments you want to share with other students?<br />

To all students - If there is something that you are really interested in, do<br />

not waste your time and go for it!<br />

Even if it seems to be impossible<br />

to achieve, don’t give up! Together,<br />

passion and joy for what you are<br />

doing connected with hard work,<br />

are the formula which make dreams<br />

come true!<br />

More information:<br />

karolina.pogoda@gmail.com<br />

9


EPSA Students' Science Publication<br />

QUANTIFICATION OF POLYPHENOLIC<br />

COMPOUNDS WITH ANTIOXIDANT ACTION<br />

FROM VARIOUS PLANT SPECIES<br />

Sultan, Carmen., The university of Carol Davila, Bucharest, Romania<br />

Nowadays many diseases are influenced and some are generated by oxidative<br />

stress which results from free radicals and reactive oxygen species. This is why<br />

research is oriented towards finding a natural solution: vegetable antioxidants.<br />

Poly-phenolic compounds (flavonoids, caffeic acid, chlorogenic acid) from<br />

plants are known to have antioxidant properties so that they may be used in<br />

therapy.<br />

Crataegi fructus, Daucus carota tuber, Aurantii pericarpium and Myrtilli fructus<br />

extract were subject to pharmacognostic analysis and HPLC analysis to<br />

determine the free radical scavenging and antioxidant properties.<br />

In Crataegi and Myrtilli fructus, there were identified poly-phenolic derivatives<br />

(flavonoids, anthocyanins, proanthocyanidins, catechol). In Daucus carota<br />

tuber, there were determined tetraterpenoid compounds such as carotenoids .<br />

Regarding Aurantii pericarpium there were found flavonoids, proanthocyanidins<br />

and catechol. HPLC analysis separated caffeic acid in Crataegi fructus and<br />

Aurantii pericarpium, ferulic acid in Aurantii pericarpium and Myrtilli fructus,<br />

quercetol in Daucus carota tuber and Crataegi fructus.<br />

The analysed plants proved to be an important source of compounds with antiradical<br />

activity thus justifying their use associated with treatment for different<br />

types of ailments (disorders caused by capillary fragility, angiopathy and<br />

diabetic retinopathy, hemeralopia, dermal diseases, cardiovascular disorders).<br />

10


FIGURES AND TABLES<br />

Volume 1 | Edition 1 | October 2013<br />

11


EPSA Students' Science Publication<br />

Q&A OF THE AUTHORS<br />

1. Why did you select this topic for your research/master thesis?<br />

At first I chose the department in which I wanted to do the research and then<br />

decided to work on something that could answer a current issue. With the<br />

help of my coordinating teacher I settled on antioxidants, more specifically<br />

the quantification of antioxidant compounds from 4 different plant species.<br />

2. How do you feel you have coped with the challenges that research brings?<br />

I was very enthusiastic about the research in the beginning and had a lot of<br />

ideas, then got a little less enthusiastic when I realised that not all of them<br />

could be applied. In order to get some results and after a great amount of<br />

work has been done, I had to change my initial plan because the results<br />

didn’t match with the hypothesis. I was really happy in the end when I<br />

realised the achievement.<br />

3. What personal skills have you developed as a researcher during your<br />

research period?<br />

Patience, perseverance, hard work, desire and knowledge.<br />

4. If you had the opportunities, would you specialize more in the field?<br />

Lab work is interesting and rather addictive: after finishing a research you<br />

want to continue with a different aspect of the same topic in order to have a<br />

complete view over the discussed issue. But working in this field only suits<br />

some of us - personally, talking with patients seems more challenging and<br />

fills me with the needed satisfaction.<br />

5. Any other comments you want to share with other students?<br />

Don't hesitate to get involved in research papers during university! It is an<br />

experience that shall contribute to your professional development even<br />

if you decide to take a different career path after graduation. There's no<br />

bigger accomplishment than sending the message across to others that<br />

find your work interesting and want to know more about it.<br />

More information:<br />

marocana14@yahoo.com<br />

12


AFTERWORD<br />

Volume 1 | Edition 1 | October 2013<br />

The idea of this publication was born at the first Team Meeting in Novi Sad,<br />

Serbia where members of the Education Board discussed the possibility of<br />

creating a platform where students can share knowledge. Also, together with<br />

the Vice President of Public Relations (VPPR) Stefan Rack, the idea got more<br />

air under its wings. Later on in the process, the publication coordinator, Mara<br />

Lefter, got involved in the projects in those areas where the project touched<br />

her portfolio.<br />

For this first publication, participants of the 5th EPSA Science day were<br />

called and asked for their interest in sharing their abstracts with the rest of<br />

the members of EPSA that did not have the opportunity to experience their<br />

awesome presentation in Catania, Italy. Luckily, we got enough responses to<br />

launch the project.<br />

They sent in their abstracts and answered those questions that were prepared<br />

beforehand, and our two professional researchers, Prof. Skalko-Basnet and<br />

Ass. Prof. Kiselev, went through all the abstracts in order to ensure its quality.<br />

The authors got the recommendations from the researchers, edited some parts<br />

and it was all was compiled into what you have seen today.<br />

Personally, I would like to grab my opportunity to once again direct a huge<br />

“Thank you” to all bodies involved. From our researchers, to our main involved<br />

VPs, Stefan Rack and Jorge Batista for their support. I would also thank<br />

specifically our Publication and Design coordinators, Mara Lefter and Svenja<br />

Laarhuis, for guiding through the, in my opinion, magical world of “how to<br />

make things look good”. Without their energy and effort, this would look like a<br />

regular word document (Black on white) with lots of text and without that real<br />

spice that they have managed to add.<br />

Now it is time for me to invite all our members students to share their knowledge<br />

and research outcomes. I would like to encourage everyone to send in abstracts<br />

and take part in this project. This is your opportunity to practice your skills in<br />

writing summaries of your research that you may need in your future career.<br />

Also, this is your chance to inspire other students for their future research!<br />

If you are interested, please send an e-mail to science@epsa-online.org and<br />

you will receive a list of requirements for documents that are needed for this<br />

publication!<br />

Yours in EPSA,<br />

Rebwar Saleh<br />

EPSA Science Coordinator 2013/2014<br />

13

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