The International Conference
on Pharmaceutical Sciences and Pharmacy Practice
dedicated to 125th Birth Anniversary of
Prof. Benediktas Šiaulis
Achievements and Development
Perspectives of Pharmaceutical
chemistry and other sciences
The International Conference on Pharmaceutical Sciences and Pharmacy
practice is organized by Faculty of Pharmacy of Lithuanian University of
Health Sciences in collaboration with Lithuanian Pharmaceutical
Association and LSMU FF Alumni Association.
Conference organizers:
Vitalis BriedisEduardas Tarasevičius Vilma Petrikait÷ Antanas Tauras Mekas
Jerzy Lazowski (Warszaw, Poland) Ilze Barene (Riga, Latvia)
Main sponsor:
ISBN 978-9955-15-254-5
© LSMU, 2012 Language of abstracts was not corrected.
CONFERENCE PROGRAMME
V. Lašo hall, Lithuanian University of Health Sciences A. Mickevičiaus street 9, LT-44307 Kaunas
10.30-11.00 Registration
11.00-13.00 PLENARY SESSION, Chair: Prof. Eduardas Tarasevičius
11.00-11.10 Opening and wellcome speech Prof. Vitalis Briedis 11.10-11.40 Professor Benediktas Šiaulis: life and
activities: prof. Benediktas Šiaulis
Prof. Eduardas Tarasevičius,
Dr. Tauras Antanas Mekas 11.40-12.10 Needles sodium chlorophyllin – perspective
natural substance for health promotion
Sanita Siksna, Riga Stradins University 12.10-12.40 Underestimated problem in cancer treatment:
anorexia and cachexia
Dr. Jerzy Lazowski, Polish National Pharmaceutical Chamber
12.40-13.00 Diversity of phytochemical research of perspectives medicinal and spice (aromatic) plants in Vytautas Magnus University
Prof. Audrius Maruška, Vytautas Magnus University
13.00-14.00 Lunch
14.00-15.00 ORAL PRESENTATIONS (part I), Chair: Prof. Vitalis Briedis 14.00-14.15 Role of the pharmacist in contemporary society Jurgita Daukšien÷ 14.15-14.30 The association between mother consultation at
the pharmacy about primary prevention of diseases and pharmacy specialist – physician interaction
Švitrigail÷ Grincevičien÷
14.30-14.45 Structure – biological activity relationship of 4-thiazolidinone derivatives
Raimondas Radžiūnas 14.45-15.00 Modernization of traditional poly-combination
herbal medicinal product: development and validation of HPLC assay of the common markers quercetin, luteolin and emodin
Audronis Lukošius
15.00-15.30 Coffee break, EXHIBITION
15.30-16.30 ORAL PRESENTATIONS (part II), Chair: Prof. Vitalis Briedis 15.30-15.40 Rhodanine derivatives targeting
Mycobacterium InhA
Liudas Šlepikas 15.40-15.50 Optimization of extraction and HPLC analysis
of pharmacologically active compounds accumulated in Solanum tuberosum L. tubers
Vaiva Bražinskien÷
15.50-16.00 Evaluation of antioxidant activity of Green tea catechins using postcolumn HPLC method
Asta Špadien÷ 16.00-16.10 Hot water pressure assisted extraction and
chemical analysis of Roselle (Hibiscus
Sabdariffa L.)
Tomas Drevinskas
CONTENT
WELLCOME SPEACH 5
ORAL TALKS 6
Professor Benediktas Šiaulis: life and activities
E. Tarasevičius, T. Mekas, H. Rodovičius
6
Needles sodium chlorophyllin – perspective natural substance for health promotion
S. Siksna, I. Barene
8
Underestimated problem in cancer treatment: anorexia and cachexia
J. Łazowski
10
Diversity of phytochemical research of perspectives medicinal and spice (aromatic) plants in Vytautas Magnus University
A. Maruška, O. Ragažinskien÷
12
SHORT TALKS 14
Role of the pharmacist in contemporary society
J. Daukšien÷
14
The association between mother consultation at the pharmacy about primary prevention of diseases and pharmacy specialist – physician interaction
Š. Grincevičien÷, L. Kubilien÷, J. Grincevičius, A. Baranauskas
15
Structure – biological activity relationship of 4-thiazolidinone derivatives
F. Malinauskas, R. Pečiūra, V. Petrikait÷, R. Radžiūnas, L. Šlepikas, E. Tarasevičius
16
Modernization of traditional poly-combination herbal medicinal product: development and validation of HPLC assay of the common markers quercetin, luteolin and emodin
V. Jakštas, A. Lukošius, K. Vitkevičius, L. Ivanauskas
18
Rhodanine derivatives targeting Mycobacterium InhA
L. Slepikas, W.H. Bisson, R. Perozzo, E. Tarasevicius, L. Scapozza
20
Optimization of extraction and HPLC analysis of pharmacologically active compounds accumulated in Solanum tuberosum L. tubers
V. Bražinskien÷, A. Ražukas, L. Ivanauskas, V. Jakštas, V. Dirs÷
22
Evaluation of antioxidant activity of Green tea catechins using postcolumn HPLC method
A. Špadien÷, N. Savickien÷, L. Ivanauskas, V. Jakštas, H. Rodovičius
24
Hot water pressure assisted extraction and chemical analysis of Roselle (Hibiscus Sabdariffa L.)
T. Drevinskas, G. Sokait÷, Z. Kalv÷nien÷, A. Maruška
25
Phenolic acids release from experimental propolis semisolid formulations and penetration into the human skin in vitro
M. Žilius, K. Ramanauskien÷, V. Briedis
27
Active compounds of hawthorn released from natural basis
A. Stelmakien÷, K. Ramanauskien÷, V. Jakštas, V. Briedis
ABSTRACTS OF POSTERS 31
Iatrochemical books from the Library of Vilnius University‘s Pharmacy, 1773-1774
V. Gudien÷, A. Baranauskas
31
Quinoxaline derivatives – potential antimicrobial and anticancer compounds
A. Vegyt÷, J. Šarlauskas, V. Miliukien÷, V. Petrikait÷
33
Isolation and determination of Lectin enriched fractions of dry and fresh herb and dry extract of Urticadioica L., dry leaves and dry extract of Thymus vulgaris L. and dry leaves of Salvia officinalis L.
G. Balciunaite, V. Baksenaite, N. Savickiene, D. Baniulis
35
Research of emulsive features of polyacrylic acid polymer Pemulen TR-1
K. Pakalniškyt÷, J. Bernatonien÷
37
Spectrophotometrical determination of phenolic compounds in ethanolic extracts of lyophilised apples
M. Liaudanskas, J. Viškelis, D. Kviklys, V. Janulis
38
Augmented renal clearance and its association with vancomycin serum level
R. Minkut÷, V. Briedis, R. Steponavičiūt÷, A. Vitkauskien÷, R. Mačiulaitis
40
Pharmacy Education at Vilnius University in the first half of the 20th century
K. Ivanauskas, E. Tarasevičius
42
Solid phase extraction procedure for detection of eight antidepresants in human urine using high performance liquid chromatography
I. Halkevych, D. Kazlauskien÷, L. Ivanauskas
44
Evaluation of flavonoids in Baikal skullcap (Scutellaria baicalensis Georgi) extract and their solubility in ethanol
V. Čižinauskas, V. Briedis, K. Ramanauskiene, M. Žilius
45
Antioxidant activity of different extracts and semisolid preparation containing propolis
G. Kasparavičien÷, Z. Kalvenien÷, S. Velžien÷, A. Savickas
47
The assessment of pharmacists’ efforts on providing qualified pharmaceutical service
Rita Mikalauskien÷
48
New approach to the synthesis of 5-aminolevulinic acid pro-drugs for iontophoresis: addition of extra charge into the molecule
V. Armoškait÷, K. Ramanauskien÷, V. Briedis
50
Modulation of adenosine receptor expression by hypoxia in pulmonary endothelial cells and receptor agonist effects on cell proliferation
J. Salys, A. Kratzer, E. Tarasevičius, E.V. Gerasimovskaya, H. Rodovičius, 52
WELLCOME SPEACH
Dear participants of the conference,
It is becoming a tradition to meet at the annual conference of pharmaceutical sciences organized by Lithuanian university of health sciences, Alumni organization of pharmacy faculty, and Pharmacy society of Lithuania. It is already a tradition to dedicate the conference to outstanding personalities, whose achievements have become the milestones in history of pharmacy science and education in Lithuania. This conference is dedicated to prof. Benediktas Šiaulis founder and pioneer of pharmaceutical chemistry in Lithuania.
The development of pharmaceuticals has become increasingly challenging and needs multilateral approach today. It introduces major challenges to research, development and manufacturing, to all aspects related to the safety and efficacy of products from quality control to regulatory affairs. Thus you are invited to present original approaches to existing unsolved issues in pharmacy theory and practice, to discuss the risks, opportunities and challenges. You are welcomed to enjoy the presentations and poster session on pharmacy history, pharmaceutical chemistry, analytical chemistry, clinical aspects of pharmacy, pharmacognosy, pharmaceutical technology, biopharmaceutics, and social pharmacy. We believe that established and young scientists will present their work, discuss new scientific findings and share their experience with colleagues on a broad range of topics related to pharmacy, and this will initiate fruitful discussions and collaborations.
Prof. Vitalis Briedis Dean of the Faculty of Pharmacy
ORAL TALKS
Professor Benediktas Šiaulis: life and activities
(dedicated to 125th Birth Anniversary)Eduardas Tarasevičius*, Tauras Antanas Mekas, Hiliaras Rodovičius
Lithuanian University of Health Sciences
*Corresponding author. E-mail address: [email protected]
Professor Benediktas Šiaulis was born 125 years ago (on July 21, 1887) in Kretinga County (now Klaipeda district) Juodupis village in 12 people peasant family. In his curriculum vitae B. Šiaulis wrote: “graduated from a local elementary school, I was taken to Petersburg, where graduated gymnasium in 1904”. He completed his higher pharmacy studies at Moscow University, where he graduated in 1913 receiving honorous degree. While in Moscow, in 1909-1919 B. Šiaulis worked in the famous Ferein pharmacy, in which at various times many Lithuanians worked, for example, P. Raudonikis, K. Grybauskas, A. Vienuolis Žukauskas. In 1921 he came back to Lithuania and one year later began working at the Lithuanian University as a teacher at the Department of Pharmacy and Pharmacognosy, beginning as a junior and then as a senior assistant.
In 1925 B. Šiaulis received travelling fellowship of the Rockefeller Foundation and he studied biology and pharmacology for 2 years at Harvard (Boston) and a Western Reserve (Cleveland) University. His scientific mentor was famous at that time U.S. pharmacologist professor T. Solman. Scientific data on the intestinal muscles particularities to chemical, physical and mechanical factors were published in three American scientific journals.
After returning to Lithuania in 1927 B. Šiaulis at Kaunas University was appointed senior assistant in the Department of Pharmacology at Kaunas University. Working as a teacher B. Šiaulis continued scientific research work. Those activities were the result of the thesis on the topic:”Pharmacological intestinal muscle features” which was defended on June 12, 1936. He was the first teacher at Kaunas University who received doctoral degree in pharmacy. One year later Council of Medical Faculty nominated B. Šiaulis as Associate Professor and appointed him as Head of the Department of Pharmacology. Associated professor B. Šiaulis focused on teaching methodological work: it was prepared in 1939 pharmacology course in two parts for students. During the same year it was released drug action in schemes for students. Consequently, at the Department of Pharmacology B. Šiaulis taught for 13 years. On June 1, 1941 he was appointed as Professor and Head of the Department of Pharmacy and Pharmacognosy.
Since 1942 professor B. Šiaulis was invited to give the pharmacology course at Vilnius University Faculty of Medicine and temporarily to mannage the Department of Pharmacy. During Nazi occupation B. Šiaulis has supported persecuted students. After the closure of the university B. Šiaulis took care to preserve the entire assets of the Department of Medicine. In 1947 Rector of Kaunas University has commit a task to B. Šiaulis to reorganize the pharmacy section into the Faculty of Pharmacy. Since January 1, 1948 he was appointed as a dean of the Faculty. On January 1951 it was made reorganization of Kaunas University into two independent institutes (Institute of Polytechnics and Institute of Medicine). At the same time pharmaceutical-dental faculty was established. The dean of this faculty was appointed professor B. Šiaulis. He headed Dean Office up to 1956, and at same time he was head of the pharmaceutical chemistry department. B. Šiaulis proposed the introduction of a weekly report on the progress of scientific work. Being Dean B. Šiaulis carefully examined the reports submitted by departments. While working in pharmaceutical chemistry prof. B. Šiaulis prepared the first Lithuanian language textbook for students "Basics of the Pharmaceutical chemistry”. He was in cooperation with the USSR Ministry of Health pharmacopoeial committee on the corrections and supplements of the State Pharmacopoeia (9th edition) and Pharmaceutical manual. Moreover, prof. B. Šiaulis wrote reviews of students' work, participated as an opponent by defending doctoral dissertations. In addition to research and teaching, professor B. Šiaulis characterized as an active lecturer. The most interesting and most popular professor’s lecture in the postwar period was "From Magic to modern drugs”. The lecture he gave in Kaunas to public, to pharmacists in conferences, adopting main material to the audience.
Professor B.Šiaulis has been an active advocate of a healthy lifestyle. About alcohol and homemade alcohol dangers he lectured residents of Kaunas while lecturing students in the pharmacology and pharmaceutical chemistry by scientifically explaining alcohol-related harm to the human body.
Professor B. Šiaulis personality captivated others by his modesty and simplicity, friendliness and intelligence, deep erudition.
For efficient scientific-pedagogical work and an active social activities prof. B. Šiaulis was awarded by the Presidium of the Supreme Soviet with Honor acknowledgment.
Sudden death on January 19, 1957 interrupted B. Šiaulis creative life. His Eternal resting place is Petrašiūnai Cemetery in Kaunas. Kaunas Institute of Medicine and teachers of his former students on November 4, 1987 celebrated professor’s B. Šiaulis centennial anniversary of his birth. On the occasion this event in Main building of the Faculty of Pharmacy unveiled a bronze bas-relief memorial. For many years, Faculty of Pharmacy, staff and students of prof. Benediktas Šiaulis at the eternal resting place, kindle memorial candles and placing bouquets of flowers as a symbol of thanks to Professor and Dean of the released for the lives of medical and pharmacy professional generations.
References:
1. Tarasevičius, E., Kaikaris, A. (1989). The centenary of professor Benediktas Šiaulis’s birth. Scientific works of the higher school of the Lithuanian SSR, Medicina, 29, 3-7 (in Russian).
2. Žukien÷, R., Kostiukevičius, A. (1998). Benediktas Šiaulis. Lietuvos farmacija, 1, 163-164 (in Lithuanian).
3. Rodovičius, H. (2010). Department of Drug Chemistry. The international Conference on Pharmaceutical Sciences and Pharmacy Practice, Abstract book, 22-26.
ORAL TALKS
Needles sodium chlorophyllin – perspective natural substance for health
promotion
Sanita Siksna*, Ilze Barene
Riga Stradins University, Latvia
*Corresponding author. E-mail address: [email protected]
In recent years there is a tendency in society to focus on ecological thinking, giving preference to more natural substances and choosing them for maintenance of environment as well as health. Overarching objective of public health policy in public health guidelines 2011-2017 (approved by the Cabinet of ministers of the Republic of Latvia) [1] is to extend healthy lived life years of citizens of Latvia and prevent premature death by maintaining, improving and restoring the health. As one of the main lines of action to achieve the target is decrease of non-infectious disease risk factors – chemical treatments have a huge role in these diseases, but certain compounds deficit in the body can be eliminated with herbal products.
The plant chlorophyll positive characteristics are known – especially its derivates quickly normalize blood haemoglobin content and prevents problems with the lack of blood haemoglobin. Many studies have shown that chlorophyll derivates have been successfully used in treatment of anaemia, chemical and radio therapy consequences. In addition, they have deodorant, body-rejuvenating, anti-inflammatory, antioxidant, antimutagenic and anticarcinogenic properties [2, 3], as well as body clearance supportive characteristics. Used topically, chlorophyll preparations regenerate tissues, heal wounds, burns and bedsores, prevent inflammation and neutralize toxins [4].
One of the practically used chlorophyll derivatives is needles sodium chlorophyllin, which is obtained from the crushed coniferous greenery in alkaline hydrolysis reaction, where the methyl and phytyl ester groups are replaced by potassium or sodium ions. During the process the central magnesium ion is often replaced with copper, iron, cobalt, chromium, etc. ions. Sodium chlorophyllin is considered to be protective for people who are at risk of contamination by aflatoxins since it prevents the bioavailability of carcinogens and avoid the potential development of liver cancer [5-9]. Clinical studies confirm the effectiveness of sodium copper chlorophyllin in leukopenia treatment [10]. Chlorophyllins can also slow down the formation of the calcium oxalate dihydrate which is considered to be the primary phase of calcium oxalate stones or kidney stones development [11, 12]. Sodium chlorophyllin is widely used in practice in many countries as natural vegetable dye (E141), which DHS is 15 mg per kg of body weight [13]. Assessment of chlorophylls and its salts properties has shown that they are non-toxic (LD50 > 0.5 g/kg, in some cases it is not even possible to determine) and does not accumulate in the body, causing chronic toxicity [14].
Chlorophyllins can be isolated from different plants, but particularly large amount of chlorophyll is in Norway spruce needles. With development of the forest biochemistry, coniferous greenery mass processing technologies to obtain valuable natural substances are designed in Latvia. It should be noted that forests are Latvian wealth, because it takes about half of the territory of the country, and the dominant tree species are coniferous trees, occupying 54% of all groves areas [15]. Every year 0.8 million tons of greenery remain in Latvian woodcutters – cheap, high-quality and ecologically clean raw material with high content of biologically active substances, which improved practical use should be
cost-with specific needles scent [17, 18]. There is a lack of literature data directly on the needle sodium chlorophyllin properties, therefore this issue requires additional research.
Distribution of individual active substances for therapeutic purposes is only one step in preparation development, they may be incorporated in the various oral and topical forms. The main aim of dosage form development is to achieve the active substance into the body in a predictable amount of the composition, that can be produced industrially and with reproducible product quality – it must be stable and safe in use. Each of dosage forms have different properties – they can ensure different absorption places in the body for the active substance, thus increasing the bioavailability and reducing side effects.
Currently in Latvia needles sodium chlorophyllin aqueous solution has been registered as a food supplement [19], however this natural substances wide range of positive activities is not fully used. Combining knowledge of the active substance and its properties as well as acquirement of the dosage form technological development opportunities, a preparation which has not only health-promoting, but also resources efficiency improving value can be created.
1. The Cabinet of Ministers 05.10.2011. order Nr. 504 Public health guidelines 2011 –2017 //
http://phoebe.vm.gov.lv/misc_db/web.nsf/626e6035eadbb4cd85256499006b15a6/ba89d22083b17edac2 2575a6002bb060/$FILE/SVP_2011_2017.pdf
2. Subramoniam A., Asha V.V., Nair S.A., Sasidharan S.P., Sureshkumar P.K., Rajendran K.N.,
Karunagaran D., Ramalingam K. Chlorophyll Revisited: Anti-inflammatory Activities of Chlorophyll a and Inhibition of Expression of TNF-į Gene by the Same //
http://www.ncbi.nlm.nih.gov/pubmed/22038065 (accessed 04.11.2012.)
3. Ferruzzi M. G., Blakeslee J. Digestion, absorption, and cancer preventative activity of dietary chlorophyll derivatives // Nutrition Research, 2007; 27: 1–12.
4. Bespalov V. G. Clinical use of conifer green needle complex: A review of medical applications. – St. Petersburg, 2006; Pp. 6.
5. 5.Egner P.A., Muńoz A., Kensler T.W. Chemoprevention with chlorophyllin in individuals exposed to dietary aflatoxin // The Cochrane Central Register of Controlled Trials (CENTRAL), 2011 Issue 4, mutation research, 2003; 523-524: 209–16.
6. Identification and characterization of chlorin e(4) ethyl ester in sera of individuals participating in the chlorophyllin chemoprevention trial // The Cochrane Central Register of Controlled Trials
(CENTRAL), 2011 Issue 4, chemical research in toxicology, 2000; 13–9: 900–6.
7. Egner P. A., Wang J. B., Zhu Y. R., et al. Chlorophyllin intervention reduces aflatoxin–DNA adducts in individuals at high risk for liver cancer // PNAS, 2001; 98 (25): 14601–14606.
8. Breinholt V., Hendricks J., Pereira C., et al. Dietary chlorophyllin is a potent inhibitor of aflatoxin B, hepatocarcinogenesis in rainbow trout // Cancer Research, 1995; 55 (1): 57–62.
9. Simonich M. T., Egner P. A., Roebuck B. D., et al. Natural chlorophyll inhibits aflatoxin B1-induced multiorgan carcinogenesis in the rat // Carcinogenesis, 2007; 28 (6): 1294–1302.
10. Gao F, Hu XF. Analysis of the therapeutic effect of sodium copper chlorophyllin tablet in treating 60 cases of leukopenia // Chinese journal of integrative medicine, 2005; 11–4: 279–82.
11. Tawashi R., Cousineau M., Denis G. Cristallisation of calcium oxalate dihydrate in noraml urine in presence of sodium copper chlorpohyllin // Urological Research, 1982; 10: 173-176.
12. Desjardins A., Tawashi R. Growth retardation of calcium oxalate by sodium copper chlorophyllin // European Urology, 1978; 4: 294.
13. Wood R., Foster L, Damant A., et al. Analytical Methods for Food Additives // CRC Press, New York, USA, 2004; Pp. 24.
14. Daugavietis M. Informācija par hlorofilīnu (Ho – fi) // A/S „Biolat” informācija, 08.05.2006. 15. Valsts meža dienesta informācija par Latvijas mežu sugu sastāvu //
http://www.vmd.gov.lv/doc_upl/sugas_2012_04_20.pdf (accessed 14.08.2012.)
16. York P. The design of dosage forms // Pharmaceutics: The science of Dosage Form Design / Ed. by Aulton M. E. – 2nd ed. – Edinburgh: Churchill & Livingston, 2002; Pp. 1–12.
17. Levent Inanc A. Chlorophyll: Structural Properties, Health Benefits and Its Occurence In Virgin Olive Oils // Akademik Gida, 2011; 9 (2): 26-32.
18. Xiu-Rong S., Shou-Hua J., Shao-Fang S., et al. Study on Preparation and Physico-chemical Properties of Sodium Chromium Chlorophyllin from Spinach // Food Science, 2007; 28 (6): 142-145.
19. Food and Veterinary Service, Food supplements database //
ORAL TALKS
Underestimated problem in cancer treatment: anorexia and cachexia
Jerzy ŁazowskiPolish National Pharmaceutical Chamber, Warsaw, Poland E-mail address: [email protected]
In recent years our cumulative understanding of a number of scientific breakthroughs in the field of tumor biology and genomics translated into novel antineoplastic therapeutic approaches.
Tumor growth is associated within profound metabolic and neurochemical alterations, which can lead to the onset of anorexia-cachexia syndrome. The word “anorexia” comes from the Greek word orexis (meaning “appetite”), literally translating to “lack of appetite”. The word “cachexia” is derived from the Greek words kakos (meaning “bad”) and hexis (meaning “condition”).
Occurring in approximately 80% of patients with cancer, the prevalence and severity of anorexia and cachexia varies with tumor type. Reduced energy intake of calories is seen almost in all patients with cachexia. The causes of reduced intake are:
• Dysphagia
Head and neck pain in upper gastrointestinal cancers can inhibit the ability to swallow. • Disease treatment
In patients receiving chemotherapy or radiation therapy, nausea, vomiting, taste changes and stomatitis can all contribute to reduced caloric intake and malabsorption. • Gastrointestinal obstruction
May be due to cancer or intestinal surgery. • Pain
Is common in severe disease and can inhibit the desire to eat. • Psychological distress
Anxiety and depression are common in patients with cancer and other severe diseases as result of uncertainties about disease, its diagnosis, treatment and prognosis – these too often reduce food intake.
• Hypercalcemia
Cancer associated hypercalcemia is a fairly common medical emergency and leads to nausea, vomiting and loss of appetite.
Thus, an integrated multidisciplinary therapeutic approach should be devised to treat cancer anorexia-cachexia syndrome with a combination of non-pharmacological and pharmacological approaches which should include nutritional counseling, psychological and educational support and pharmacologic approach.
Due to its high prevalence and negative impact on quality of life, it is important for pharmacists to recognize the hallmark signs of loss of appetite and unintentional weight loss as well as the coexisting symptoms of the illness, which contribute to the patient discomfort and distress.
As members of interdisciplinary palliative care they can be involved at various levels of participation in patient care, developing treatment plans, managing medication therapy, educating patietnts and staff.
• answering the quality of life of both patient and family.
Attempts of drug therapy for cachexia with a variety of agents have been met with limited success.
Megestrol acetate has been the most thoroughly studied pharmacologic agent for the treatment of anorexia and cachexia. Appetite stimulating properties of corticosteroids (dexamethasone, methylprednisolone and prednisolone) are well known, but their beneficial effects are limited to two to four weeks.
Increased appetite is also an effect of synthetic cannabionoids (dronabinol and nabilone
Other agents, sometimes recommended for the treatment of anorexia and cachexia, such as metoclopramide, hydralazine sulphate, cyproheptadine and pentoxifylline has limited evidence to support a beneficial effect on appetite and weight gain. Metoclopramide is useful in patients with symptoms of delayed gastric emptying that may contribute to early satiety and nausea.
Other medications, such as mirtazapine and olanzapine are known to cause significant weight gain in some populations and may be useful, but controlled trials are still needed to decide about optimal dosing regimen and potential benefits of these agents.
New researches are looking beyond traditional agents to improve appetite and weight. Promising results have been obtained with melatonin, which appears to influence cytokine activity during tumor growth by reducing circulating levels of TFN and interleukin-6. It is well tolerated. In one study of 100 patients with untreatable metastatic solid tumors, lower weight loss was reported by those taking melatonin than by those who has best supportive care. Melatonin has also been shown to benefit patients on chemotherapy in terms of reduced toxicity and increased survival time, as well as performance status in patients with advanced cancer. It is also generally well tolerated, with the main side-effect being drowsiness.
As showed recent limited trials, supplementation with omega-3 fatty acids, especially eicosapentanoic acid, which reduces the production of inflammatory cytokines, decreased weight loss, promoted weight gain and improved survival I patients with cancer cachexia. But existing evidences are not sufficient to establish whether this way is better than placebo. Conclusions: Cancer related anorexia and cachexia is a myriad of symptoms which significantly affect the course of the disease. It is highly prevalent among cancer patients and causes reduction of their quality of life and negatively predicts treatment outcomes, as well as seriously increases their morbidity and mortality.
Oncology pharmacist can help identify, assess and manage cancer patients with anorexia-cachexia syndrome, playing a key role in holistic care of these patients.
ORAL TALKS
Diversity
of phytochemical
research
of perspectives
medicinal and spice
(aromatic) plants in Vytautas Magnus University
Audrius Maruska1, Ona Ragazinskiene2 1
Department of Biochemistry and Biotechnologies, Faculty of Natural Sciences, Vytautas Magnus University, [email protected]
2
Kaunas Botanical Garden of Vytautas Magnus University, [email protected]
The biodiversity of medicinal, spices (aromatic) plants species and varieties is important from the scientific and practical point of view. Plants are the renewable resources of biologically active compounds. The biogenesis of biologically active compounds depends on several issues: origin of the plant, geographical zone, where plant is growing, plant genotype, phenotype and chemotype and annual climatic fluctuations. Most of natural compounds possess several biological activities: antioxidant, antimicrobial, anti-inflammatory and play a certain physiological role in human organism (4). Therefore, it is very important to look for the new natural compounds and study less investigated plants as a possible source for such compounds. The aim of this study is to investigate an introduction of medicinal plants from various geographical regions, to analyse a local flora, perform phytochemical analyses, and to select most perspective species and varieties, help to protect genetic resources and biodiversity. Phytochemical analysis of medicinal plants extracts were performed using following methods: gas chromatography, high performance liquid chromatography, capillary zone electrophoresis, spectrophotometry (1, 2, 3).
The research is supported by the Council of Lithuania:
Application of solid-state fermentation for development of higher value and safety food products (GRANT No. SVE-09/2011 BIOFITAS).
Separation of anticancer fractions from willow-herb and their molecular and biological analysis. (GRANT No. MIP-084/2012 ONKOFITAS).
References:
1. Kaškonien÷, Vilma; Kaškonas, Paulius; Maruška, Audrius; Ragažinskien÷, Ona. Chemical composition and chemometric analysis of essential oils variation of Bidens tripartita L. during vegetation stages // Acta Physiologiae Plantarum Heidelberg: Springer. ISSN 0137-5881. 2011, Vol. 33, iss. 6, p. 2377-2385. [Duomenų baz÷s: Science Citation Index Expanded (Web of Science); SpringerLINK].
2. Maruška, Audrius [Marušk, Audrius]; Proscevičius, Juozas; Bimbirait÷-Survilien÷, Kristina; Kornyšova, Olga; Ragažinskien÷, Ona; Ratautait÷, Vilma. Comparison of phytochemical composition of medicinal plants by means of chromatographic and related techniques // Procedia Chemistry [elektroninis išteklius]. Amsterdam: Elsevier B.V. ISSN 1876-6196. Vol. 2, iss. 1 (2010), p. 83-91. Prieiga per internetą: <http://dx.doi.org/10.1016/j.proche.2009.12.014>. [Duomenų baz÷s: Science Citation Index Expanded (Web of Science); Science Direct].
3. Maruška A., Kornyšova O. (2006): Application of monolithic (continuos bed) chromatographic columnus for phytochemical analysis // Journal of Chromatography A. 1112: 319-330.
4. Raila, Algirdas; Lugauskas, Albinas; Kemzūrait÷, Aurelija; Zvicevičius, Egidijus; Ragažinskien÷, Ona; Railien÷, Marija [Railien÷, Marija]. Different drying technologies and alternation of mycobiots in the raw material of Hyssopus Officinalis L // Annals of Agricultural and Environmental Medicine. ISSN 1232-1966. Vol. 16, No. 1 (2009), p. 93-101. Prieiga per internetą: <http://www.aaem.pl/pdf/AAEM.htm>. [Duomenų baz÷s: Science Citation Index Expanded (Web of Science); MEDLINE; CAB Abstracts].
Role of the pharmacist in contemporary society
Jurgita Daukšien÷Lithuanian University of Health Sciences, Medical Academy, Faculty of Pharmacy, Department of Drug Technology and Social Pharmacy
E-mail address: jurgita.dauksiene@gmail
The profession of pharmacy has experienced significant growth and development over the past hundred years. Pharmacy entered twentieth century performing the social role of apothecary – preparing and selling medicinal drugs. The traditional role of the pharmacist as a compounder and supplier of pharmaceutical products began to wane as the preparation of pharmaceuticals was gradually taken over by the pharmaceutical industry. Compounding has been largely replaced by the commercial manufacture of nearly all formulations. There are also a number of new ways of medication supply -Medicines can be bought in supermarkets, in drug stores or at markets. They can also be obtained by mail order or over the Internet, they are sold by medical practitioners and dispensed by computerized dispensing machines. And as the choice of therapeutic agents passed to the physicians, the pharmacist professional role was narrowly constrained. At the same time – growing numbers in not rational drug use and drug related morbidity and mortality did show some serious problems for the society and did show the new opportunities for the pharmacist. Pharmacists get ready to change its original focus on medicine supply towards a more inclusive focus on patient care. Pharmaceutical care is a new concept of pharmacy practice which emerged in the mid-1970s. It states that all practitioners should assume responsibility for the outcomes of drug therapy in their patients. The scope of pharmacy practice now includes patient-centred care with all the cognitive functions of counselling, providing drug information and monitoring drug therapy, as well as technical aspects of pharmaceutical services, including medicines supply management. In patient-centred health care, the first challenges are to identify and meet the changing needs of patients. Pharmacists need to ensure that people can access medicines or pharmaceutical advice easily and, as far as possible, in a way and at a time and place of their own choosing. This calls for huge change in pharmacy student’s education. The knowledge about drug forms and preparation is not enough for future specialists. The pharmacy curriculum should be adapted, in order for the pharmacist to be able to acquire new knowledge and skills. This is the problem that is discussed world widely. Pharmacy students and practitioners must be educated to assume the responsibility for managing drug therapy, so that they can maintain and expand their position in the health care system and are compensated for their role in providing pharmaceutical care. Pharmaceutical care does not exist in isolation from other health care services. It must be provided in collaboration with patients, physicians, nurses and other health care providers. To be effective health care team members, pharmacists need skills and attitudes enabling them to assume many different functions. The concept of the “seven-star pharmacist” was introduced by WHO and taken up by FIP in 2000 in its policy statement on Good Pharmacy Education Practice to cover these roles: caregiver, decision-maker, communicator, manager, life-long learner, teacher and leader.
These are times of enormous change in health care and in the pharmacy profession. At no time in its recent history has the profession been faced with such challenges and opportunities.
1. Heppler Ch. D., Strand L.M., Oportunities and responsibilities in phamaceutical care, AJHP 1990, 47 p. 533-542. 2. Foppe van Mil J.W., Schulz M., Tromp (Dick) Th. F.J. Pharmaceutical care, European developments in concepts, implementation, teaching and research: A review, Pharm World Sci 2004; 26:303-311.
3. FIP Statement of Policy on Good Pharmacy Education Practice. Hague, 2000.
4. Walker R. Pharmaceutical public health: the end of pharmaceutical care? Pharm J 2000; 264:340–341. 5. Hepler CD. Clinical pharmacy, pharmaceutical care, and the quality of drug therapy. Pharmacotherapy. 2004 Nov; 24(11):1491–98.
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The association between mother consultation at the pharmacy about
primary prevention of diseases and pharmacy specialist - physician
interaction
Š. Grincevičien÷*, L. Kubilien÷, J. Grincevičius, A. Baranauskas
Lithuanian University of Health Sciences
*Corresponding author. E-mail address: [email protected]
Background. Integration and differentiation conceptual model for community pharmacists and general physician1, based on qualitative data, showed that there is relationship between physician-pharmacist interaction and additional services provided at the pharmacies. The research, based on quantitative methodology is necessary to support this correlation.
Methods. 409 Lithuanian pharmacy specialists were questioned in 2012 March-October. The questionnaire was developed based on International Pharmacy Federation reference paper on the effective utilization of pharmacists in improving maternal, newborn and child health. Data was analyzed using SPSS package. Chi-square was used to calculate where appropriate. Spearman coefficient was calculated for correlation.
Results. 52.1 % of respondents were contacting with physician. More than 30 % of respondents were contacting with physician to clarify dose, to inform about new products, discounts, services or when they can‘t read the prescription. They were contacted by physician about drug information. 23.7 % of respondents were contacting with family physicians, 24.7 % - with obstetrics /gynecologist and 0.7 % - with pediatricians. Pharmacy specialists, who were contacting with physician, were consulting mothers about primary prevention of disease during pregnancy and breastfeeding. Pharmacy specialist, who were contacting with family physicians, more likely were consulting mother about vaccination against flu during pregnancy. Pharmacy specialist, who were contacting with obstetricians/gynecologists, more likely were consulting mother about vaginal infection during pregnancy. Respondents who were more often contacted with physicians, more likely were consulting pregnant (Spearman’s correlation coefficient 0.275, with p < 0.001) and breastfeeding mothers (Spearman’s correlation coefficient 0.291, with p < 0.001) about primary prevention of diseases.
Conclusions. The research has confirmed the hypothesis, that advanced service at the pharmacy, such as consultation about primary prevention of diseases during pregnancy and breastfeeding was associated with pharmacy specialist‘s interaction with physicians. The topic for consultation at the pharmacy was associated with the specialization of physician to whom pharmacy specialist was contacting.
References
1. Bradley, F., Ashcroft, D., Noyce, P (2012). Integration and differentiation: a conceptual model of general practitioner and community pharmacist collaboration. Research in Social Administrative
Structure – biological activity relationship of thiazolidinone derivatives
Faustas Malinauskas1, Rimantas Pečiūra2*, Vilma Petrikait÷1,3, Raimondas Radžiūnas2, Liudas Šlepikas1, Eduardas Tarasevičius11
Department of Drug chemistry, Faculty of Pharmacy, Medical Academy of Lithuanian University of Health Sciences
2
Department of Drug technology and Social Pharmacy, Faculty of Pharmacy, Medical Academy of Lithuanian University of Health Sciences
3
Laboratory of Biothermodynamics and Drug Design, Institute of Biotechnology, Vilnius University
*Corresponding author. E-mail address: [email protected]
1. First synthesized compounds biological activity scale
The aim – search for the new biologically active substances in group of 2,4-thiazolidinones. It has been synthesized 426 new compounds (Table 1). Their structure was confirmed by methods of chemical and instrumental analysis, and biological activity was tested using microbiological and pharmacological activity analysis techniques.
Table 1. Biological activity of new thiazolidinone derivatives
Group No.
The new compound structural basis Number of synthesized compounds Number of biologically active compounds Percentage of biologically active compounds I 2-thio(2-imino)-4-thiazolidinone 105 82 78 II 2,4-thiazolidinedione 92 70 76 III 2,4-thiazolidinedione-2-aminoguanidine 90 39 43 IV 2,4-thiazolidinedione-β-hydrazide 88 32 36 V 2,4-thiazolidinedione-N-carboxylic acid 40 15 37 VI 2,4-thiazolidinedione-N-benzyl 11 7 63 TOTAL 426 245 58
2. Pharmacophore structure and biological activity relationship
Structures of new compounds were provided with sulfonamides, benzoic and salicylic acid, morpholine, 5-nitrobenzothiazole, aminoguanidine, guanylhydrazine and β-hydrazide fragments, predicting their pharmacophoric features (Table 2).
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Table 2. Pharmacophoric groups and biological activity of thiazolidinone derivatives
Pharmacophore structure and biological activity relationship
Number of synthesized compounds Number of biologically active compounds Percentage of biologically active compounds Sulfonamide 67 67 100 Benzoic acid 16 9 56 Salicylic acid 30 15 50 Aminoguanidine 28 9 32 Sulfamoylphenylaminoguanidine 24 11 46 Guanilhydrazone 38 19 50 β-hydrazide group 88 32 36 Triptophan 12 10 83 Morpholine 18 18 100 5-nitrobenzothiazole 9 9 100 TOTAL 330 199 60
3. The biological activity scale of new compounds
The new compounds were tested evaluating their antimicrobial (antibacterial, antifungal, antiviral, tuberculostatic), diuretic, neurotropic, anticonvulsive, pesticidal and ascaricidal activity. Majority of thiazolidinone derivatives were proved to be antimicrobial, diuretic ant neurotropic substances.
Conclusion. Obtained experimental results proved our primary hypothesis to use 4-thiazolidinone scaffold with different substituenst for obtaining new biologically and pharmacologically active compounds.
References
1. Peciura, Rimantas. Synthesis of 2 aminotiazolindione-4 and medicines belonging to aminecompounds and their study: Abstract of Dissertation Pharmacy Ph.D degree / Lviv: Lviv State Medical Institute (Ukraine) in 1982, 23 pages.
2. Malinauskas, Faustas. Synthesis of 2 aminotiazolindione-4 oksalilecompounds and study of biological activity: Abstract of Dissertation Pharmacy Ph.D degree / Kharkov State Pharmaceutical Institute (Ukraine), 1987, 22 pages
3. Radžiūnas, Raimondas. Synthesis of 2 aminotiazolindione-4 aminoguanidinecompounds and study of biological activity: Abstract of Dissertation Pharmacy Ph.D degree / Kharkov State Pharmaceutical Institute (Ukraine), 1988, 21 pages.
4. Petrikaite, Vilma. Synthesis of 2,5-substituted thiazolidinone derivatives and evaluation of their antifungal and antibacterial activity: Summary of doctoral dissertation / Kaunas University of Medicine, 2007, 39 pages.
Modernization of traditional poly-combination herbal medicinal product:
development and validation of HPLC assay of the common markers
quercetin, luteolin and emodin
Valdas Jakštasa*, Audronis Lukošiusa, Konradas Vitkevičiusb, Liudas Ivanauskasb a
LUHS Department of Pharmocognosy, bLUHS Department of Analytical and toxicological chemistry
*Corresponding author. E-mail address: [email protected]
Trejos devynerios is a traditional herbal medicinal product with its origins going
back to the times of the Both Nations Republic (1). This preparation was mentioned in the article Wiadomosci farmaceutycsne by prof. J. Muszynski in 1927 (2). In the 20th century, during the Soviet period, Trejos devynerios also known as ,trojanka’ was produced in Kaunas Pharmacy of Medicinal Herbs, in Poland company Herbapol produced ,Trojanka litewska’, in Chicago Lithuanian pharmacists working in J&J pharmacy, also labelled the similar poly-ingredient mixture as ,Trejos devynerios’ (3). Formula of this traditional phytopreparation includes 27 herbal drugs: poplar buds, capsicum, clove, oak bark, calendula flower, laurel leaf, peppermint leaf, dill fruit, hawthorn berries, wormwood, fragrant pepper, lovage root, valerian root, hop strobilus, bitter-orange epicarp and mesocarp, frangula bark, willow bark, caraway fruit, birch buds, meadowsweet, St. John's wort, hyssop, yarrow, thyme, tormentil rhizome, rowan berries, dog rose hip. Long-time use of this poly-ingredient preparation substantiates its phytotherapeutic properties; however, the substantial increase of quality requirements for herbal medicinal products brings new challenges to pharmaceutical science. The aim of the work: to select quality indicating common markers for assaying of product and to validate HPLC analytical method.
Complexity of combination herbal medicinal product makes it difficult and, sometimes, impossible to give analytical identification and quantitative evaluation of the medicinal substances in the product (4). In these situations, with respect to scientific knowledge, application of combined markers, such as aglyconic fragments of various biologically active glycosidic forms is expedient. Hydrolysis of flavonoids and anthracene glycosides to aglycones is included in analytical method of the complicated product that contains 27 medicinal substances of plant origin. To optimise the analytical methodics, the experimental research involves optimization of the following characteristics of analytical column: length, sorbent, suitability of the system and selection of the test sample preparation conditions. Validation of the optimized methodology includes evaluation of the parameters recommended by the International Commission of Harmonization (ICH). Specificity of HPLC methodology is approved by evaluation of matched chromatograms of the test sample, reference compounds and „placebo“ solutions, which compare the presence of analytes peaks, duration of analytes retention, chromatographic picture and peak spectrum data read by diode-array (PDA) detector. Cohesion of the methodology results is determined by several series of measurements. Considering the recommendations of ICH and CPMP scientific guidelines, 6 analyses are conducted within time limits of the same analytical sample series investigation (24 h), using real test samples and conducting the analysis on different days. The suitability criterion is a variation coefficient that does not exceed 2.6 per cent during one day repetition test and 5.1 per cent during intermediate precision test. The linearity is evaluated by the calibration curve method. The suitability criterion – correlation coefficient is ≥ 0.9999. Data of calibration solutions chromatograms is used to substantiate application range. Within the application limits the response deviation from the applied linear calibration equation should not exceed 5 per cent, therefore, the application limits based on quercetin and luteolin analytes make at least 0.005 to 0.100 mg/ml (in test sample solution), those based on emodin analytes – respectively at least 0.002-0.100 mg/ml. As the test solution contains 0.0127 mg/ml of quercetin, 0.0075 mg/ml of luteolin and 0.0211 mg/ml emodin on average (intermediate
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precision data), thus the substantiated application limits cover 40-787 per cent for quercetin analyte, 66-1333 per cent for luteolin analyte and 10-473 per cent for emodin analyte, which ensures recommendations of scientific guidelines for quantitative method with respect to variation of test samples and reference compounds.
Conclusion. Determination of common analytical markers – flavonoids quercetin,
luteolin and anthracene emodin content in combination herbal medicinal product by developed HPLC method is scientifically substantiated and meets the requirements of scientific guidelines for quantitative method.
References:
1. Mekas, T. Senoji vaistininkyst÷. Kopa; 2005, p. 100.
2. Muszynski, J. Wilenskie ziola ludowe. Wiadomosci Farmaceutyczne, Warszawa; 1927:21-22, p. 475 3. Mekas, T. Senoji vaistininkyst÷. Kopa; 2005, p. 101.
4. EMEA. Quality of Combination Herbal Medicinal Products / Traditional Herbal Medicinal Products. 2008.
Rhodanine derivatives targeting Mycobacterium InhA
L. Slepikas1,2*, W.H. Bisson1, R. Perozzo1, E. Tarasevicius2, L. Scapozza1 1
School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, 1211 Geneva, Switzerland
2
Lithuanian University of Health Sciences, Faculty of Pharmacy, LT 44307 Kaunas, Lithuania
*Corresponding author. E-mail address: [email protected]
Many compounds bearing the 4-thiazolidinone chemical scaffold have been reported to have antimicrobial activity. Two aims have been defined for this project. The first one aim is to develop new 5-substituted rhodanine-N-amino acids 4-thiazolidinone derivatives with improved activity and selectivity against Mycobacterium and eventually other pathogens such as Klebsiella pneumoniae, Pseudomonas aeruginosa, Legionella pneumophil, Staphylococcus
aureus establishing the relationship between structure and activity ((Q)SAR). The second one
is to identify potential cellular targets for the 4-thiazolidinone class of compounds.
The new compounds from the series of 5-substituted-4-thiazolidinone were prepared in a three steps synthesis, as following. In the first step the conversion of amino acid into amino acid dithiocarbamate with carbon disulfide and potassium hydroxide was realized at room temperature overnight. In the second phase the amino acid dithiocarbamate was cyclized to 4-thiazolidinone ring with the amino acid at position N-3 by boiling them in presence of hydrochloric acid for few hours. Then the condensation of 4-thiazolidinone with aldehydes in boiling acetic acid with ammonium acetate lasting few hours was performed resulting in the formation of 5-aryliden-4-thiazolidinone-N-amino acid derivatives. The yield of the final product depended on the nature of the amino acid substitution and was varying between 30% and 90%. The identity (chemical structure) and quality/purity of the synthesized compounds were assessed by means of IR, high resolution MS, 1H and 13C NMR studies.
In silico results indicates that newly synthesized rhodanine derivatives could inhibit
enoyl acyl carrier protein reductase (InhA) from Mycobacterium tuberculosis1, which is involved in the type II fatty acid biosynthesis pathway, as the compounds show inhibitory effect to the homologous FabI from Plasmodium falciparum2 and Staphylococcus aureus. According to the results from molecular docking, several possible interactions have been identified between the newly synthesized compounds and M. tuberculosis InhA. H-bond network interactions between, (i) the sulfur atom of thiocarbonyl group, (ii) the carbonyl group at the position 4 of the rhodanine ring, (iii) the carbonyl group of -COOH from the tryptophan moiety as acceptors, and the hydroxyl group of tyrosine 158 in InhA and the hydroxyl group of NAD+ as donors, respectively. The binding modes are displayed in the figure 1.
Figure 1. The possibles binding modes:
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iii)
i) The binding mode of ligand 159 in InhA showing the interaction between the thiocarbonyl group of the rhodanine-tryptophan derivative with the hydroxyl group of tyrosine 158 and hydroxyl group of NAD+.
ii) The binding mode of ligand 159 in InhA showing the interaction between the carbonyl group in thiazolidinedione ring of the rhodanine-tryptophan derivative with the hydroxyl group of tyrosine 158 and hydroxyl group of NAD+.
iii) The binding mode of ligand 146 in InhA showing the interaction between the carboxylate group of the tryptophan moiety, the carbonyl group of the rhodanine ring with the hydroxyl group of tyrosine 158 and hydroxyl group of NAD+.
Ligands are color coded by atom type with the white carbons and displayed in balls and sticks, residue tyrosine-158 and NAD+ are colored by atom type (carbon atoms in orange) and displayed as capped sticks. The secondary structure of the InhA is displayed as ribbons. Left upper two dimensional figures showing hydrogen bonding between i) the sulfur atom of the thiocarbonyl group ii) the carbonyl group iii) the carboxylate group of the tryptophan moiety, the carbonyl group of the rhodanine ring and the tyrosine-158 hydroxyl group of InhA and NAD+. Left lower images representing open surface of binding pocket. The secondary structure of the InhA is displayed as ribbons and the ligands displayed in balls and sticks.
Based on experimental data from inhibition test against InhA many of newly synthesized derivatives showed inhibitory activity. The IC50 values of the most active compounds were determined and vary from 3µM to ~28µM. The most active derivative 167 ( IC50=2.7µM) have tryptophan moiety at the position 3, anthralyden residue at the position 5 of the rhodanine ring and is twice more potent than as a standard used triclosan against InhA. Inhibitory test against MRSA Staphylococcus aureus FabI show that 9 derivatives were more potent or had the same activity as triclosan at 10 µM concentration. The most active derivative 167, has IC50= 2µM against saFabI and was more active than as standard used triclosan. SAR studies show that increased lipophilicity play an important role as derivatives bearing lipophilic substituents have greater inhibitory activity. The higher activity of anthralyden vs naphthyliden vs benzyliden residue are due to hydrophobic interaction with hydrophobic groups from side chains in the InhA and saFabI binding pocket.
The newly synthsized 4-thiazolidinone derivatives were tested against M. marinum, K.
pneumoniae, P. aeruginosa and L. pneumophila in a host pathogen setting using eukaryotic
cells Dictyostelium and Acanthamoeba as a host. The test results showed, that some of the compounds have a biological activity in particular against M. marinum and L. pneumophila. The most potent derivatives have the rather hydrophobic and bulky tryptophan and norleucine residues as substitute in position 3. The test results also show that newly synthesized compounds have a broad range activity (MIC from 250 to 3.9 µg/ml) against S. aureus.
The type II fatty acid biosynthesis pathway has been characterized in silico as a putative target of newly synthesized rhodanine derivatives in the different pathogens. Further work has been started in optimizing the chemical structure in order to go with the design of new derivatives with increased antibacterial and antimycobacterial activity3.
Optimization of extraction and HPLC analysis of pharmacologically active
compounds accumulated in Solanum tuberosum L. tubers
Vaiva Bražinskien÷a*, Almantas Ražukasa, Liudas Ivanauskasb, Valdas Jakštasb, Vidmantas Dirs÷b
a
Voke branch of Lithuanian Research Centre for Agriculture and Forestry, Zalioji a. 2, Traku Voke, LT-02232 Vilnius, Lithuania
b
Department of Analytical and Toxicological Chemistry, Lithuanian University of Health Sciences, A. Mickeviciaus g. 9, LT-44307, Kaunas, Lithuania
*Corresponding author. E-mail address: [email protected]
Potatoes cause interest not solely as nutritive crop; their health improving properties are being studied widely. A wide variety of compounds is recorded in potato tubers: anthocyanins, carotenoids, glycoalcaloids, phenolic compounds, amino acids, etc. Potatoes also are important as a source of amino acids. Dealing with such complexity of compounds accumulated in Solanum tuberosum the importance and necessity for selective analysis methods is crucial. Thus, the goal of this work was to develop high-performance liquid chromatography method for qualitative and quantitative determination of pharmacologically active compounds accumulated in potatoes grown in Lithuania and to optimize their extraction conditions.
2. Materials
2.1. Chemicals
Standards of fifteen materials were used in the chromatographic analysis: chlorogenic acid, (-)-epicatechin, DL-catechin, gallic acid, eriodictyol, caffeic acid, rutin trihydrate, vanillic acid, naringenin, trans-cinnamic acid, trans-p-coumaric acid, trans-ferulic acid, L-tryptophan, L-tyrosine
2.2. Plant material
Tubers of twelve Lithuanian-grown potato varieties (Sante, Fresco, Courage, Saturna,
VB Liepa, VB Venta, Red Lady, VB Aista, Lady Rosetta, VB Rasa, Goda, Acapella) of the
2011 yield from potato variety collection of Vok÷ Branch of Lithuanian Research Centre for Agriculture and Forestry were used for analysis.
3. Results and discussion
3.1. Optimization and validation of the HPLC method.
When high performance liquid chromatography is used in search for separation systems suitable for determination of active compounds accumulated in potato tubers, gradient elution, with acidulated water and methanol or acetonitrile as mobile phase is most commonly used (Lewis et al., 1998; Mattila & Hellström, 2007; Soloft, Nielsen, Laursen, Husted, Halekoh & Knuthsen, 2010). In this case, the best results were achieved when 0.5% acetic acid solution in water (A) and methanol (B) were used for gradient elution: 0 min – 95% A and 5% B, 40 min – 40% A and 60% B, 41 min – 10% A and 90% B, 55 min – 10% A and 90% B, 56 min – 95% A and 5% B. The best resolution has formed using 4.6×250 mm, 5 µm ACE C18 column. In total, fifteen compounds were separated with the following retention times about: 6.2 min – tyrosine, 7.9 min – gallic acid, 13.0 min – 3,4-dihydroxybenzoic acid, 15.9 min – tryptophan, 17.3 min – catechin, 18.8 min – chlorogenic acid, 21.1 min – vanillic acid, 21.7 min – caffeic acid, 22.5 min – epicatechin, 27.2 min – coumaric acid, 28.4 min – ferulic acid, 32.8 min – rutin, 37.0 min – eriodictyol, 40.2 min – cinnamic acid, 41.1 min – naringenin. All peaks were completely separated (resolution > 1.5). The separated active compounds were analyzed at a wavelength ensuring their maximum absorption. The method specificity was checked according to two parameters: retention time of the tested compound peak and PDA spectrum matching the standard. Method validation parameters were chosen according to The International Conference on Harmonisation Guidelines.
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In order to optimize the extraction method, the best way of extraction was searched for comparing shaking with sonication and its duration; comparing ethanol with methanol as extraction solvents, as well as different acetic acid concentrations. Aiming to get most accurate results on the effect of these factors, all tested samples were prepared within-day; the closest extraction conditions were chosen.
The most effective method of extraction and its duration were determined by extraction of lyophilized potatoes with 0.5% methanolic solution of acetic acid. The extraction solvent was chosen basing on the references: methanol is better extraction solvent than ethanol (Mohdaly, Sarhan, Smetanska & Mahmoud, 2010), and phenolic acids are unstable at high pH (Friedman & Jurgens, 2000). The results show that sonication is much more effective than shaking: after 10 min of sonication the amount of extracted active compounds was higher than after one hour of shaking. The best result was obtained after application of 20 min sonication.
Extraction solvent should be acidulated due to the above-mentioned instability of phenolic acids at high pH (Friedman & Jurgens, 2000); therefore it was decided to test the impact of acetic acid concentration in extraction solvent on the total quantity of extracted compounds. The test was conducted using pure methanol as extraction solvent and acidulating it with acetic acid. Sonication of 30 min was used. The obtained results show that acetic acid improved the extraction of active compounds from potato lyophilisate. The study of the effect of 5 different concentrations of acetic acid (0.5%, 1%, 1.5%, 2%, 2.5%) on the extraction of active compounds revealed best results in case of methanol and 2% acetic acid solution as extraction solvent.
Searching for the most suitable extraction solvent for potato lyophilisate, total amounts of active compounds of potato tubers extracted with different concentrations (50%, 60%, 70%, 80%, 90%) of methanol and ethanol were compared. Extraction solvents were not acidulated; 30 min sonication was used. The results show that in all compared concentrations, methanol extracts more active substances than ethanol. Thus, the obtained results are in compliance with conclusions of Mohdaly, Sarhan, Smetanska & Mahmoud, 2010. The best extraction results were obtained using 60% methanol.
The results of all experiments conducted in order to optimize the extraction conditions suggest that acetic acid, methanol and water (5:147:98) mixture together with 20 min sonication is the most suitable for extraction of lyophilized potatoes.
4. Conclusions
The results of all experiments conducted in order to optimize the extraction conditions suggest that acetic acid, methanol and water (5:147:98) mixture together with 20 min sonication is the most suitable for extraction of lyophilized potatoes.
High-performance liquid chromatography method allowing qualitative and quantitative determination of fifteen different active compounds has been designed. The conducted analysis of potato tubers showed that tyrosine, tryptophan, chlorogenic and caffeic acids were mostly accumulated by all tested varieties.
References
Friedman, M., & Jurgens, H. (2000). Effect of pH on the stability of plant phenolic compounds. Journal of agricultural and food chemistry, 40, 2101-2110
Lewis,C.E., Walker, J.R.L., Lancaster,J.E., & Sutton, K.E. (1998). Determination of anthocyanins, flavonoids and phenolic acids in potatoes. I: Coloured cultivars of Solanum tuberosum L. Journal of the science of food and agriculture, 77, 45-57
Evaluation of antioxidant activity of Green tea catechins using postcolumn
HPLC method
Asta Spadiene1*, Nijole Savickiene2, Liudas Ivanauskas3, Valdas Jakstas2, Hiliaras Rodovicius1
1
Department of Drug Chemistry, Faculty of Farmacy, Lithuanian University of Health Sciences
2
Department of Pharmacognosy, Faculty of Farmacy, Lithuanian University of Health Sciences
3
Department of Analytical and Toxicological Chemistry, Faculty of Farmacy, Lithuanian University of Health Sciences
*Corresponding author. E-mail address: [email protected]
Evaluation of antioxidant activity of Green tea (Camellia sinensis L.) catechins is a part of international "Eureka“ project „Creation of the methodology for effects of natural antioxidants on the development of the Diabetes mellitus complications“. Extract of Green tea leaves has antioxidative, anti-inflammatory properties and may be used for the prevention and suppression of diabetes complications.
The main active constituents of Green tea leaves are 75% polyphenols: catechins (catechin, epicatechin, epigallocatechin, epicatechin gallate and epigallocatechin gallate) and proanthocyanins (prodelphinidin). There are also caffeine, teophylline, polysaccharides, essential amino acids (leucine, phenylalanine, valine), vitamins (B2, E and C), minerals (potassium, magnesium, calcium, manganese, natrium, zinc, fluoride, phosphorus etc.). Catechins are responsible for antioxidant effects [1].
The aim of this investigation was to evaluate the antioxidant activity of catechins in capsules of Green tea dry extract (JSC ”Sanitas”) using postcolumn HPLC method as described by Raudonis et al. [2]. The mobile phase was composed of water/acetonitrile/methanol/ethyl acetate/glacial acetic acid (89:6:1:3:1 v/v/v/v/v). Flow rate gradient system was optimized to assay the trolox and mixed solution of standards. Trolox content equivalent and antiradical activity equivalent were used to evaluate antioxidant activity of catechins in the capsules of Green tea dry extract.
Total antiradical activity equivalent of catechins 9018.75 µg/g. According to antioxidant activity catechins can be arranged as follows: epigallocatechin gallate > epigallocatechin > epicatechin gallate > epicatechin (antiradical activity equivalent 5537.89 µg/g, 1576.25 µ g/g, 1386.89 µg/g and 518.59 µg/g respectively). It was determine linear correlation between the antioxidant activity of each catechin and the amound of this compound.
Considering that other compounds of Green tea extract have no significant influence on total Green tea antioxidant activity it can be stated that one gram of dry Green tea extract has the same antioxidant activity as well as 9 mg of standard antioxidant trolox.
References
1. Chacko SM, Thambi PT, Kuttan R and Nishigaki I. Beneficial effects of green tea: A literature review. Chin. 2010, 5:13
2. Raudonis R, Jakstas V, Burdulis D, Benetis R, Janulis V. Investigation of contribution of individual constituents to antioxidant activity in herbal drugs using postcolumn HPLC method. Medicina. 2009, 45(5):382-394
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Hot water pressure assisted extraction and chemical analysis of roselle
(Hibiscus Sabdariffa L.)
Tomas Drevinskas1*, Gintar÷ Sokait÷2, Zenona Kalv÷nien÷2, Audrius Maruška1 1
Vytautas Magnus University, Faculty of Natural Sciences, Dept. of Biochemistry and Biotechnology, Vileikos 8, LT-44404 Kaunas.
2
Lithuanian University of Health Sciences, Faculty of Pharmacy, Dept. of Drug Technology and Social Pharmacy, Mickevičiaus 9, LT-44307 Kaunas
*Corresponding author. E-mail address: [email protected]
Roselle (Hibiscus Sabdariffa L.) is a valuable herbal material due to its natural colorants (mainly Delphinidin), phenolic compounds (chlorogenic acid, protocatechuic acid etc.), vitamins and other substances that provide pharmacological effects [1]. Roselle is known to regulate lipid metabolism [2], reduce hypertension [3] and take role in apoptosis [4]. Different procedures and different solvents have been implemented in extraction of roselle material [5]. Less polar solvents are known to extract less polar compounds (such as polyphenols, phenolic acids, flavonoids, anthocianins etc.) with greater recovery. Some disadvantages when preparing end product as additional steps of extract preparation (vaporization, preparative cleaning etc.) are brought in by such solvents. Water is one of the very few green solvents. It provides different dissolvation abilities at different temperatures. Such effect is caused by change of dielectric constant of water when temperature changes [6]. Classical extraction procedure is limited to only 100oC (boiling temperature of water), but boiling temperature can be increased by increasing pressure in extraction reservoir. Hot water pressure assisted extraction provides a possibility to increase temperature up to 374 oC (supercritical temperature of water) if the pressure is sufficient.
Hot water pressure assisted extraction was performed at 4 bar pressure that allowed reaching of 144 oC temperature. Milled raw material (fraction of 0.02-1.2 mm) of roselle was used during procedure. Total amount of phenolic compounds of obtained extracts was assessed by Folin-Ciocalteu method and colorants were determined by capillary electrophoresis (Fig. 1). 0.08 0.10 0.12 0.14 A U Electroosmotic Flow Colorants of Roselle
