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26Таблица 2
Результаты количественного определения содержания суммы веществ флавоноидного строения _в пересчете на рутин_
Количество полифенольных соединений в %, в сырье
соцветия клевера лугового цветки липы серд-целистной трава чабреца обыкновенного трава тысячелистника обыкновенного сбор
0,58±0, 01 1,61±0,02 1,26±0,01 1,16±0,02 0,45±0,03
Выводы.
1. Разработаны методики количественного и качественного определения полифенольных соединений в лекарственном растительном сырье и фи-токомпозиции.
2. Цветными реакциями определено наличие флавоноидов.
3. Методом ТСХ выявлено наличие биологически активных веществ, близких по строению к рутину и гиперозиду.
4. Предложено количественное содержание веществ полифенольного строения проводить спектрометрическим методом в пересчете на галловую кислоту, веществ флавоноидного строения в пересчете на рутин в лекарственном растительном сырье и фитокомпозиции.
5. Разработаные методики будут использованы для стандартизации биологически активных веществ гинекологического сбора и настоя.
СПИСОК ЛИТЕРАТУРЫ:
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2. Державна Фармакопея Украши. До-повнення 1 / ДП «Украшський науковий фармакопейний центр якосп лжарських засобiв». 2-е вид. Харшв: Державне шдприемство «Украшський науковий фармакопейний центр якосп лжарських за-^в», 2016. С. 187.
3. Коноваленко I. С., Половко Н. П. Склад та технология виготовлення в умовах аптек збору для корекци ммактеричних розлащв: №382-2018. К., 2018. 4 с.
4. Наукове обгрунтування складу композици лжарсько! рослинно! сировини для лжування ктмактеричного синдрому / 1.С. Коноваленко, Н. П. Половко// Зб, наук, праць сшвробггаишв НМАПО iM. П.Л. Шупика. - 2018. - 205-214 с.
5. Практикум по фармакогнозии: Учеб, пособие для студ. вузов / В. Н. Ковалёв, Н. В. Попова, В. С. Кисличенко и др.; Под общ. ред. В. Н. Ковалёва. - Х.: Изд-во НФаУ; Золотые страницы, 2003. - С. 129-130.
6. Рибак Л.М., Коновалова О.Ю., Колядич О.П. Порiвняльне дослщження кшьшсного вмюту полiфенолiв у рiзних видах гераш Geranium L. методом перманганатометричного титрування та спектрофотометричним методом // Фармацевтич-ний журнал. - 2010. - № 6. - С. 44-47.
7. Фармакогнозия: Учеб, пособ. Для студ. высш, учеб, завед. / В. Н. Ковалёв, В. С. Кисличенко, И. А. Журавель и др. - Х.: Изд-во НФаУ, 2007. - 272 с.
8. Evaluation of Aluminium Complexation Reaction for Flavonoid Content Assay / A. P^kal, K. Pyrzynska // Food Anal. Methods. - 2014. - № 7. - Р. 1776-1782.
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THE STUDY OF QUALITATIVE AND QUANTITATIVE CONTENT OF AMINO ACIDS IN CABBAGE LEAVES (BRASSICA OLERACEA L.)
Kuznetsova M.
Postgraduate student, Department of Chemistry of Natural Compounds and Nutritiology
National University of Pharmacy, Kharkiv, Ukraine
Zhuravel I.
Doctor of Pharmaceutical Science, professor, professor of Department of Chemistry of Natural Compounds and Nutritiology National University of Pharmacy, Kharkiv, Ukraine
Hutsol V.
Associate professor of Department of Pharmacy National Pirogov memorial medical university, Vinnytsya, Ukraine
Abstract
Amino acids from plants have an important influence on various systems and organs of the human body. They have broad pharmacotherapeutic properties, promote faster absorption of biologically active substances and potentiate their action.
Brassica oleracea L. (Brassicaceae family) - common vegetable plant that has long been used in folk medicine to treat many diseases. Therefore, it is important to determine the qualitative composition and quantitative content of amino acids in cabbage leaves.
Amino acids were identified in the cabbage leaves of different cultivars by ion-exchange liquid chromatography. The total number of amino acids was 18 and 7 of them are essential and 2 are conditionally essential amino acids. Glutamic acid and proline were dominant in Bilosnizhka cultivar (1.74% and 1.26%), Ya-roslavna cultivar (1.24% and 2.31%), Ukrainian autumn cultivar (2.49% and 1.55%) respectively.
Methionine and y-aminobutyric acid have been found in minorities in all samples of cabbage leaves: Bilosnizhka cultivar - 0.07% and 0.10%; Yaroslavna cultivar - 0.11% and 0.01%; Ukrainian autumn cultivar - 0.07% and 0.11% respectively.
The highest sum of amino acids was in the cabbage leaves of Yaroslavna cultivar (10.73 %).
Keywords: Brassica oleracea L., ion-exchange chromatography, amino acids.
Introduction. Brassica oleracea L. (Brassicaceae family) - is the one of the most common food plants that is grown in almost every area of the globe.
The plants have a diverse chemical composition, which is represented by steroid compounds, vitamins, polysaccharides, proteins, carotenoids, amino acids, minerals, according to the literature. These compounds cause the high nutritional and medicinal properties of the plants [3, 5, 6, 8].
Amino acids are vital organic substances. Amino acids are included in the biosynthesis of proteins, enzymes, nucleic acids, complex carbohydrates, fats, hormones, and other compounds required for living organisms. Amino acids are able to maintain normal function of organs and systems. Individual amino acids are important in the prevention and treatment of many pathological conditions. Thus, glutamic acid is involved in maintaining of brain cell respiration, stimulates the oxidation processes; alanine is an effective tool in the prevention of ischemic brain disorders; proline is a part of synthetic nootropic agents and is a major component of collagen; leucine, methionine, aspartic and glutamic acids improve cardiovascular function; aspartic and glutamic acids are used in arrhythmias, hypoxia, and diseases of the central nervous system. The main sources of amino acids for the human body are food products of animal and plant origin.
Plant amino acids affect the function of various systems and organs of the human body, exhibit broad pharmacotherapeutic properties, promote faster absorption of biologically active substances from plants and potentiate their action [2, 4-8].
The aim of our research was to study the amino acid composition of cabbage leaves of Bilosnizhka, Ya-roslavna, and Ukrainian autumn cultivars.
Materials and Methods. The samples for the study were dried and milled cabbage leaves of Bilos-nizhka, Yaroslavna, and Ukrainian autumn cultivars. The raw materials are grown on research areas of the Institute of Horticulture and Melon-growing of the National Academy of Agrarian Sciences of Ukraine (Merefa, Kharkiv Region, Ukraine). Raw materials were collected in 2017-2018.
The determination of the qualitative composition and quantitative content of amino acids in the raw material was carried out by ion-exchange chromatography using an automatic amino acid analyzer AAA T-339M (Czech Republic).
The study of amino acids was performed after pre-hydrolysis with hydrochloric acid. To this end, 6 N hydrochloric acid solution was added to the test tube with exactly measured portion of herb and after that the tube was cooled in a liquid nitrogen flow. When the test tube contents were frozen, the air was removed from it under vacuum to avoid amino acid oxidation. Then the test tube was sealed and preserved in thermostat at a constant temperature of 106°C within 24 hours. Then the test tube contents were cooled, totally transferred to a glass weighing box, and hydrochloric acid was evaporated on a water bath. 3-4 ml of deionized water was added to a dried sample, and the evaporation went on. The obtained sample was dissolved in 0.3 N lithium citrate buffer at pH 2.2 and applied on amino acid analyzer ion exchange column, whose cation exchanger was previously equalized with sodium citrate or lithium citrate buffer solution. Amino acids were separated by small-grain spherical shape cation exchangers (resins) of sty-rene-divinyl benzene copolymer with a functional sulfite group [1].
T 339 automatic amino acid analyzer performs all analysis operations in a continuous eluent flow. At the column exit a micropump constantly mixed eluate with ninhydrine reagent (mixture of ninhydrine, buffer solution and stannic chloride in argon atmosphere). The obtained mixture was fed along a capillary pipe to reactor heated to 95°C - 98°C. Then it was fed to flow cell for photocolorimetric determination of obtained color intensity at 440 nm or 560 nm. The photocell signals were registered by potentiometric recorder in the form of chromatograms. The peak area in chromatograms was calculated and compared to amino acid peak areas from known chromatograms, and on this basis the absolute values of content of each amino acid in analyzed sample were calculated [1].
The content of each amino acid in sample in ^M (X1) in analyzed sample was calculated by formula: X1 = S1 / So,
Where Si - amino acid peak area in analyzed sample, So -peak area of this amino acid in solution of standard amino acid where in the content of each amino acid corresponds to 1 ^M. To express the content in mg the obtained amino acid amount in ^M was multiplied by corresponding molecular mass [1].
Results. The results of determination of the qualitative composition and quantitative content of amino acids in cabbage leaves of Bilosnizhka, Yaro-slavna, and Ukrainian autumn cultivars by the method of ion-exchange chromatography are presented in the table.
Table
The content of amino acids in cabbage leaf
№ Amino acid The content in cabbage leaves of different cultivars, %
Bilosnizhka Ukrainian autumn Yaroslavna
1 y-aminobutyric acid 0.10±0.01 0.11±0.01 0.01±0.001
2 Lysine * 0.36±0.01 0.28±0.01 0.63±0.02
3 Histidine ** 0.15±0.01 0.12±0.01 0.26±0.01
4 Arginine ** 0.88±0.04 0.56±0.02 1.04±0.05
5 Aspartic acid 0.90±0.04 0.83±0.04 1.19±0.05
6 Threonine * 0.23±0.01 0.19±0.01 0.32±0.01
7 Serine 0.29±0.01 0.24±0.01 0.44±0.02
8 Glutamic acid 1.74±0.08 1.24±0.05 2.49±0.11
9 Proline 1.26±0.05 2.31±0.09 1.55±0.06
10 Glycine 0.29±0.01 0.24±0.01 0.50±0.02
11 Alanine 0.42±0.01 0.33±0.01 0.51±0.02
12 Cystine 0.16±0.01 0.16±0.01 0.20±0.01
13 Valine* 0.24±0.01 0.22±0.01 0.25±0.01
14 Methionine* 0.07±0.01 0.07±0.01 0.11±0.01
15 Isoleucine* 0.20±0.01 0.20±0.01 0.26±0.01
16 Leucine* 0.31±0.01 0.28±0.01 0.43±0.02
17 Tyrosine 0.14±0.01 0.12±0.01 0.25±0.01
18 Phenylalanine* 0.20±0.01 0.18±0.01 0.29±0.01
E essential amino acids 2.64 2.10 3.59
E amino acids 7.94 7.68 10.73
Note. * - essential amino acid, ** - conditionally essential amino acids.
Discussion. The ion-exchange chromatography identified 18 amino acids in leaves of Bilosnizhka, Yaroslavna, and Ukrainian autumn cultivars. Amino acids included 7 essential amino acids and 2 conditionally essential amino acids.
The content of glutamic acid and proline were the greatest in cabbage leaves of Bilosnizhka cultivar -1.74% and 1.26%, in cabbage leaves of Yaroslavna cultivar - 1.24% and 2.31%, and Ukrainian autumn cultivar (2.49% and 1.55% respectively.
Methionine and y-aminobutyric acid were present in the least amount in all samples of cabbage leaves: Bilosnizhka cultivar - 0.07% and 0.10%; Yaroslavna cultivar - 0.11% and 0.01%; Ukrainian autumn cultivar - 0.07% and 0.11% respectively.
The highest sum of amino acids was in the cabbage leaves of Yaroslavna cultivar (10.73 %).
The sum of amino acids in the cabbage leaves of Bilosnizhka cultivar and Ukrainian autumn cultivar was almost the same - 7.94 % and 7.68 % respectively. These tendency was observed for the amount of essential amino acids. The highest content of the sum essential amino acids was in cabbage leaves of Yaro-slavna cultivar.
Conclusions. The analysis of the qualitative composition and quantitative content of amino acids of cabbage leaves of Bilosnizhka, Yaroslavna, and Ukrainian autumn cultivars was carried out.
The presence of 18 amino acids was found. Amino acids included 7 essential amino acids and 2 conditionally essential amino acids.
The content of glutamic acid and proline were the greatest in cabbage leaves of all cultivars.
The results can be used to produce complex drugs and functional food supplements and also to create methods of quality control of raw material of Brassica oleracea L..
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ANALYSIS OF ALBENDAZOLE PROFILE AS THE MOST POPULAR ANTHELMINTIC AGENT
Semchenko K.
PhD of Pharmacy, Associate Professor of Department of Pharmaceutical Technology of Drugs, NUPh
Vyshnevska L.
Doctor of Pharmacy, Professor, Head of Department of Pharmaceutical Technology of Drugs, NUPh
Polovko N.
Doctor of Pharmacy, Professor of Department of Pharmaceutical Technology of Drugs, NUPh
Romas K.
PhD of Pharmacy, Senior lecturer of Department of Pharmaceutical Technology of Drugs, NUPh
Abstract
The problem of helminthiases is particularly relevant in the recent years as this group of diseases is met in all countries of the world. With the purpose of the further creation of new complex anthelminthic drug for the treatment of helminthiases of digestive system, the profile of albendazole was studied. On the basis of the findings it was set that it is rational to develop complex anthelminthic drugs, which must include albendazole, as well as an additional anthelmintic agent to provide a broad spectrum of action.
Keywords: albendazole, helminthiasis, PASS analysis, drug profile
Helminthiases is the worldwide healthcare problem
Introduction. The development of dosage forms involves careful selection of active and auxiliary substances. Particular attention should be paid to the study of the profile of active substances.
Parasitic disease is one of the most pressing problems of the present. The existing on the pharmaceutical market range of medicines for the treatment of helminthiasis currently includes 19 drugs based on praziquantel, mebendazole, albendazole, piperazine, piranthel, levamisole. It has been established that among the an-thelmintic agents presented, foreign medicines dominate (63.2%), while Ukrainian producers offer fewer drugs (36.8%) [10].
In view of the above, it is promising to develop new domestic medicines with anthelmintic activity, which will ensure the needs of the population of the country in quality and affordable domestic drugs.
The national program for the import substitution of drugs provides the development of new drugs of domestic production, which can successfully compete with similar drugs of foreign production not only in terms of price, but also in efficacy and safety.
In view of the above, it is promising to develop new domestic medicines with anthelmintic activity,
which will ensure the needs of the population of the country in quality and affordable domestic drugs.
Therefore, in order to create a new anthelmintic drug, we aimed to study the profile of albendazole as an active ingredient, since this active pharmacological ingredient is included in the overwhelming majority of protocols for the treatment of helminthiasis of the digestive system [1, 4, 8].
Materials and methods. As the material of the research the proper open access literature sources about albendazole were used.
The methods of bibliosemantic analysis of the literature data and PASS-analysis of albendazole structural formula was used.
Data processing was performed using the methods of economic and statistical analysis and the results processing - with the help of computer programs. Prediction of activity of the main biologically active substances was performed with PASS-analysis (http://www.way2drug.com).
Results.
Physical and chemical properties
Albendazole is a carbamate of benzimidazole based on a bicyclic ring structure in which the benzene ring is fused to the 4- and 5- position of the imidazole ring. Gross formula: C12H15N3O2S; molar mass 265.34 g/mol (Fig. 1).
