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Objective: To study the chemical constituents from plant of Artemisia frigida Willd. and its pharmacological activities. Methods: The compounds were isolated and purified by various chromatography methods. All compounds were identified by spectroscopic and chromatographic techniques, and were screening for their PPARγ activating activity and PTP1B inhibitory activity. Results: There were 26 compounds isolated from the plant of A. frigida and identified as pectolinarigenin (1), jaceosidin (2), chrysoeriol (3), tricin (4), 3-oxogermacra-1(10),11(13)-dien-6α,12-olide (5), achillin (6), 1,10β-epoxyachillin (7), scoparone (8), 4-hydroxyacetophenone (9), chrysosplentin (10), jaceidin (11), 11α,13-dihydroyomogin (12), chrysanthemin A (13), eupatilin (14), eupatrin (15), artemorin (16), 6-methoxytricin (17), hanphyllin (18), cirsimaritin (19), ridentin (20), desacetylmatricarin (21), subchrysine (22), luteolin (23), caffeic acid (24), agastachoside (25), and tilianin (26). Conclusion: Compounds 5, 7, 11, 18, 25, and 26 are firstly isolated from the Artemisia. Compounds 10, 12, 13, 16, 17, and 22 are firstly isolated from this plant. Compounds 2 and 15 exhibited weak activity of PPARγ. Compounds 1 and 3 had inhibitory effect on PTP1B.
Résumé
The pharmacognostic analysis of Artemisia frigida Willd. (from Buryatia, Zabaikailsky krai and Qinghai-Tibet plateau) was studied in this research. Macro- and microscopic characteristics of three samples were mostly the same. Purity tests and identification of main biologically active substances in A. frigida Willd were carried out using standard identity tests. The essential oils were obtained by hydrodistillation method, qualitative and quantitative composition was studied by GC-MS. The main components of the essential oils in three samples are camphor, 1,8 – cineol, endo-borneol and borneol, terpinen-4ol and others. Keywords: Artemisia frigida Willd., pharmacognostic analysis, essential oils, chemical composition, monoterpenes, sesquiterpenes. Artemisia frigida Willd. is a valuable medicinal plant which is widely spread on the territory of Republic of Buryatia. It is also used in a Tibetan medicine for the treatment of injuries, tuberculosis, deficiency disease and neurological disturbance [1]. Pharmacological activity is associated with the presence of different biologically active substances, especially, essential oils, which demonstrate anti-bacterial, antifungal and anthelmintic activities [2]. The aim of this work is to make a Pharmacognostic analysis of herba Artemisiae frigidae which was collected on the territory of Russia and China. To achieve this goal we should make the following tasks: 1) to do macro- and microscopic analysis; 2) to make purity tests of plant materials; 3) to identify the presence of main biologically active substances; 4) to make a qualitative composition and chemical assay of essential oils by gas chromatography mass spectrometry (GC-MS). For the research we used herba Artemisiae frigidae which was collected in 2014, in such regions like Ivolginsky region (Russia), Zabaikailsky krai (Russia) and Qinghai-Tibet plateau (China) in a flowering stage. Purity tests and the presence of main biologically active substances were identified by using standard identity tests [3,4].The essential oil from the whole aerial part of Artemisia frigida was obtained by hydrodistillation method and was analyzed by gas chromatography–mass spectrometry (GC– MS) to determine its chemical composition and assay [5,6]. Macroscopic analysis revealed that plant material is presented by numerous caules with a high around 8-40 sm, woody roots. Lower part of leaves are gray colored and on the top they are light green. Flower anthodes are large, spherical, droop and combined in a raceme and has characteristic aromatic odour. Microscopic analysis revealed that the main diagnostic features in 3 samples are the same: flexuous cells of the top epidermis, abaxial and adaxial stoma (anomocytic stomata), sword-like fuzzes and internal secretion glands on two sides of leaves [7]. Also, we made purity tests and identified such characteristics as moisture, total ash, 10% chlorohydric acid-insoluble ash, and we found out that 70% ethanol as a solvent is more suitable for the extraction. In this material we found out the presence of polysaccharides, essential oils, fatty acids, tannins and flavonoids using standard identity tests [8]. According to the quantitative analysis of essential oils in three samples of herba Artemisiae frigidae, made by gas chromatography mass spectrometry (GC-MS), it contains 1,00% of light green essential oil in the raw material from Zabaikailsky krai (Russia). The major components of this essential oil were presented by sesquiterpenes (50.33%), such as germacrene D, caryophyllene, α-zingiberene and bicyclogermacrene; among monoterpenes - camphor, 1,8-cineol, endo-borneol, terpinen-4ol, α-terpineol. Chemical assay of the essential oil of A.frigida collected in Buryatia (Russia) is around 0,97%. There were detected about 63 components in this essential oil and, mostly, they are monoterpenes (87.67%), such as camphor, 1,8 – cineol, endo-borneol, myrtenol, α-terpineol acetate. What concerns the raw material from Qinghai-Tibet plateau (China), it contains 0,67% of essential oils. The major components were presented by monoterpenes like in Buryatian sample (93.85%): camphor, 1,8 – cineol, borneol, terpinen-4-ol and bornyl acetate. Thus, the most chemical assay of the essential oil of A.frigida was from the raw material collected in Zabaikailsky krai (Russia) – 1,00%. But the macro components of the essential oils in three samples are presented by monoterpenes, like camphor, 1,8 – cineol, endo-borneol and borneol, terpinen-4-ol and others. Thus, pharmacognostic analysis of three samples of herba Artemisiae frigidae revealed that macro- and microscopic characteristics are mostly the same. And purity tests and identification of main biologically active substances in A. frigida Willd has been carried out using standard identity tests. The quantitative composition of essential oils is different depending on growing conditions, but the chemical composition of the main components is constant and presented, mostly, by monoterpenes. So this research shows that Artemisia frigida Willd. can be used as a valuable raw material for the medicine. References: 1. Batorova S.M., Ubasheev I.O. Medical herbs of Zabaikalye, using in Tibetan medicine for wound healing // Resources of Zabaikalyan plant formation and its using. - Ulan-Ude, 1991. P. 169-182. 2. Mikhailova T.N. Antimicrobial activity of essential oils of some species of Artemisia in Siberian flora // some aspects of pharmacognosy and cultivated plants of Siberia. – Tomsk, 1969. P.32-39. 3. State Pharmacopoeia of the USSR. Ed. XI (part 2). Common methods of analysis. Medical plant material / USSR Ministry of Health. - 11th ed., Ext. - Moscow: Medicine, 1990. - 400 p. 4. Grinkevich N.I., Safronich L.N. Chemical analysis of medical plants. -Moscow: Higher school, 1983.-176p. 5. Chemical composition of essential oils of Artemisia frigid Willd. from Buryatian flora / S.V. Zhigzhitzhapova and others // Vestnik of Buryat State University. – 2013. - №3. P.71-74. 6. Chemical composition of essential oils of Artemisia frigid Willd., collected in Zabaikalye / N.V. Bodoev and others // Chemistry of plant materials. – 2000. – №3. P.41-44. 7. Macroscopic, microscopic and phytochemical analysis of medicinal plants: teaching aids / S.V. Zhigzhitzhapova, T.E. Randalova, L.D. Radnaeva - Ulan -Ude: Publishing Buryat Scientific Center. - 2014. 93 p. 8. Plant resources of USSR. Flowering plants, its chemical composition, using. Genus Asteraceae. St.Petersburg: Nauka, 1993. 352 P.
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Objective: Determining the content of five flavonoids in Artemisia frigida by high performance capillary electrophoresis (HPCE) method to provide reference data for the evaluation of processing products of A. frigida. Methods: Treated by different processing technologies, the content of five flavonoids compounds can be determined and compared by HPCE. Results: Regression equations were 5,7,3′-triterhydroxy-4′-methoxyflavone Y = 0.114 3 X + 0.032 7, r = 0.999 8; 5,3′-dihydroxy-6,7,4′-tritermethoxyflavone Y = 0.100 9 X + 0.048 5, r = 0.999 6; quercetin Y = 0.055 5 X + 0.026 8, r = 0.999 6; 5,7,3′-triterhydroxy-6,4′-dimethoxyflavone Y= 0.113 2X-0.016 1, r = 0.999 3; luteolin Y = 0.097 9X-0.029 5, r = 0.999 4. Five kinds of flavonoids were good linear relationship at 1.00-40.00, 10.00-200.00, 5.00-100.00, 1.00-40.00, 1.00-40.00 μg/mL. The average recoveries were 98.44%, 97.75%, 97.73%, 97.98%, and 98.07%. And RSD were 1.60%, 1.03%, 1.57%, 0.94%, and 1.20%. Conclusion: The results reveal that the different processing technologies have different effects on the content determination of five flavonoids compounds in A. frigida. Using the five flavonoids as testing indexes, the best processing technology of A. frigida is heating and drying to constant weight at 60 °C.
Résumé
Objective To study the chemical constituents of the CHCl3 and EtOAc extracts from Artemisia frigida.Methods The chemical constituents in A.frigida.were isolated with silica gel and LH-20 chromatography and their structures were identified by means of spectra,in same cases by direct comparison with authentic samples.Results Thirteen compounds were obtained and identified as quercetin(Ⅰ),luteolin(Ⅱ),5,7,3'-triterhydroxy-4'-methoxy flavone(Ⅲ),5,7,3'-triterhydroxy-6,4'-dimethoxy flavone(Ⅳ),5,3'-dihydroxy-6,7,4'-tritermethoxy flavone(Ⅴ),5,3'-dihydroxy-3,6,7,4'-tetramethoxy flavone(Ⅵ),methyl phenol(Ⅶ),7-hydroxy coumarin(Ⅷ),7-methoxy coumarin(Ⅸ),caffeic acid(Ⅹ),?-sitosterol(Ⅺ),6,7-dihydroxy coumarin(ⅩⅡ),and 7-hydroxy-5,6-dimethoxy coumarin(ⅩⅢ).Conclusion All these compounds are isolated from this plant for the first time.