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Резюме УДК 615. 07:582.949.27:547.466 Національний фармацевтичний університет (Харків) Национальный фармацевтический университет (Харьков) 61002, г. Харьков, ул. Пушкинская, 53 National University of Pharmacy (Kharkov) Ukraine, 61002, Kharkov, 53, str. Pushkinska pharmsy1@rambler.ru INTRODUCTION. Melissa officinalis L. (balm mint or lemon balm) – one of the most popular medicinal herb. Like many plants from family Lamiaceae, it is known as an aromatic and medicinal plant in traditional medicine of many countries of Europe and the Newly Independent States (NIS), it is widely used in the gastronomy. They began to cultivate melissa in ancient Greece and Rome as medicinal and honey plant [1, 2]. Chemical composition of the essential oil and some phenolic compounds of melissa herb and leaves were studied in detail [1, 2, 3, 4, 7, 8, 9, 10]. It was analyzed hydroxycinnamic acid derivatives composition (rosemarinic, caffeic and chlorogenic acids) and it was established their content in raw materials and extracts of Melissa officinalis [4, 5]. Flavonoids and components of essential oil of lemon balm are responsible not only the sedative and antiherpes activity, but also for anti-thyroid and regulatory activity. It was found that an extract of lemon balm is effective in the treatment of Graves' disease. The active ingredients of Melissa blocks antibodies and action of thyroid-stimulating hormone, preventing the enlargement of the thyroid gland [1, 2, 7, 8, 9, 10]. Research of composition and dynamics of accumulation of carboxylic acids is necessary for understanding the metabolism (glycolysis, Krebs cycle, the formation and decomposition of lipids, etc.). Some acids are important biologically active compounds (unsaturated fatty acids), as energy source, they are main structural components of cell membranes and so on. These heterofunctional compounds are common in nature and found in the leaves of plants. Therefore, an integrated approach to the study of biologically active compounds balm is a key issue pharmacy. The goal was to invastigate the different classes of organic acids in lemon balm herb. METHODS AND MATERIALS OF RESEARCH Melissa herb was harvested during full blossom period in 2012 at the experimental place of Botanical garden of National Pharmaceutical University. Studies of organic acids were carried out by chromatography-mass spectrometry [6]. It was added 50 mg tridecane in hexane (internal standard) and 1.0 ml of 14% BCl3 in methanol, Supelco 3-3033 (agent for methylation) to 50 mg of dried plant material in 2 ml vial. The mixture was kept in a vial about 8 hours at 65oC. During this peroid biologically active substances were extracted, hydrolized and methylation of organic acids were carried out. The reaction mixture was decanted from the sediment and plant material was diluted with 1 ml of distilled water. Methyl esters of fatty acids were extracted with 0.2 mL of methylene chloride, stirred several times an hour, and then extract obtained methyl esters of organic acids were analyzed by chromatography. The study was performed on chromatograph Agilent Technologies 6890 with mass spectrometric detector 5973. Chromatographic column - INNOWAX capillary with an inner diameter of 0.25 mm and a length of 30 m. Injection of samples (2 ml) in a chromatography column was performed using splitless, i.e. without flow separation, allowing the sample to enter without loss distribution and significantly (10 - 20-fold) increase sensitivity of chromatography analysis. The speed of the sample was 1.2 ml / min for 0.2 min. Flow rate of carrier gas (helium) 1.2 ml / min. Temperature of the heater input samples - 250 oC. Temperature programmable thermostat from was 50 to 250° C at a rate 4° / min. It was used library mass spectra NIST05 WILEY 2007 with a total of more than 470,000 spectra, combined with programs of identification AMDIS and NIST. RESULTS AND DISCUSSION In the melissa herb were identified and determined the content of 31 organic acids, which are belong to 3 chemical groups: mono-di-tri-carboxylic acids, phenolic and fatty acids, as shown in Figure 1 and Table 1. Among the mono-di-carboxylic acids (table 1) three were identified 9 compounds, by which concentration they may be represented as follow (from high to low): hepta-2 ,4-dienic> oxalic> citric> malic> succinic> malonic> azelaic>2-hexanic> fumaric (950–14mg / kg). Phenolic acids were identified in low concentrations (from 34 to 12 mg / kg). In previous articles were shown the results of the determination of other phenolic acids: rosemarinic, caffeic and chlorogenic which are characterized by a higher concentration (0.2 to 3%) [4, 5]. Fatty acid composition (table 1) includes 13 saturated and 4 unsaturated compounds. The dominant fatty acids are palmitic (1999 mg / kg) and linolenic (931 mg / kg). Figure.1. Chromatogram of organic acids of lemon balm herb. Table 1 Organic acids of lemon balm herb
CONCLUSIONS 1. There were identified 31 organic acids in the melissa herb using chromatography-mass spectrometry. 2. The presence of 9 mono-di-tri-carboxylic acids was determined. Dominant acid is hepta-2.4-diene acid (more than 900 mg / kg). The concentration of phenolic acids is low and is about 35 mg / kg. 3. The highest concentration among the 17 fatty acids belongs to palmitic (about 2000 mg / kg) and linolenic acid (930 mg / kg). Література
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