| Objective:The herbs and seeds of Peganum harmala L. were traditionally used as medicinal substances by Uygur, Hazakh, and Mongolia for a long history. The fatty acids from Peganum harmala L. closely related to plant metabolism, which provide energy for the biosynthesis of its alkaloids and flavonoids. The contents of fatty acids from Peganum harmala L. are significantly higher, but there ars raely reports about fatty acids from Peganum harmala L. in xinjiang. In the present study, the determination methods and GC fingerprint of fatty acids was established, the variation of fatty acids from Peganum harmala L and the bioactivation of lipid extracts from Peganum harmala L. were studied preliminarily. Methods:1) The oil yield was selected as index, uniform design was used to optimize the parameters of the Soemmering’s extraction and ultrasound-microwaves synergistic extraction method for the fatty acids extraction, and the appropriate extraction method was selected by the comparison beween the two optimized methods.2) The overall score was selected as optimizing index, which include the contents of 5 kinds fatty acids, the numbers of methylated peak, and the percentage of peak area, the orthogonal test was used to optimize the conditions of 4 kinds of esterification process. The proper conditions of fatty acid methyl esterification were selected by comparison of 4 kinds of optimized esterification process.3) The type and relative percentage content of fatty acid from Peganum harmala L. were analysized by GC-MS.4) The GC method for the content determination of five fatty acids was established. And the contents of fatty acids were determined from different parts of Peganum harmala L. and in different seasons.5) The GC fingerprint of the fat soluble extracts from Peganum harmala L. was established.6) The actue toxicity about Peganum harmala L. was evaluated.7) The bioactivation of fat soluble extracts was studied.Results:1) Soemmering’s extraction method was the suitable extraction for the fatty oil from seedsof Peganum harmala L. The process conditions for the Soemmering’s extraction method were as following:light petroleum is selected as the extraction solvent and solvent multiple is 20, water bath under 80℃ for 11h, the oil yield is 30.1%.2) By optimizing 4 kinds of esterification process, 1%H2SO4-methanol circumfluence method was more suitable for fatty acid methyl esterification, and the optimum condition as follows:the ratio of methanol to oil is 4:1, the ratio of catalyst to oil is 2:1, reaction temperature is 80℃, circumfluence time is 30min. The esterification rate of four kinds of fatty acid was more than 97%.3) According to the results of GC-MS, the 18 chemical constituents, including 14 kinds of fatty acid were identified from above-ground part of Peganum harmala L. and the 19 chemical constituents, including 16 kinds of fatty acid were identified from root of Peganum harmala L.4) The method for determination of the contents of fatty acids in Peganum harmala L. meet the requirement of methodology. Different batches, picking seasons and positions of Peganum harmala L. contribute to the diversity in the contents of fatty acids, which can be reflected mostly in this aspect:the species of the fatty acids in the seed are basicly similar with the fatty acids in the above-ground parts, which includes unsaturated fatty acid such as linoleate acid, oleate acid. The fatty acids in root mainly includes saturated fatty acid such as tetracosanoic acid, docosanoic acid and so on.5) 26 common peaks were identified in GC fingerprint.6) The LD50 of extracts from rootof Peganum harmala L. is 4439mg/kg and 95%confidence interval is 2800-5000mg/kg, the LD50 of extracts from Peganum harmala L. seeds is 4439mg/kg and 95%confidence interval is 2800-5000mg/kg, and the toxicity of extracts from root of Peganum harmala L. is more obvious. The LD50 of extracts from above-ground parts of Peganum harmala L. picked in May is 9963.4mg/kg and 95%confidence interval is 9003.5～10637mg/kg, the LD50 of extracts from above-ground parts of Peganum harmala L. picked in June is 10262mg/kg and 95%confidence interval is 7809.7～11182rng/kg, the LD50 of extracts from above-ground parts of Peganum harmala L. picked in July is 10427mg/kg and 95%confidence interval is 9357.6～11554mg/kg, the LD50 of extracts from above-ground parts of Peganum harmala L. picked in August is 12031mg/kg and 95%confidence interval is 9357.6～11554mg/kg. The LD50 of fat extracts from Peganum harmala L. is 7618.2mg/kg and 95%confidence interval is 5839.9～10303mg/kg, and the LD50 of alkaloidsof Peganum harmala L.is 672.84mg/kg and 95%confidence interval is 529.9～882.16mg/kg.7) High dose group, middle dose group of fat soluble substances and high dose group of total alkali could reduce the metatarsal swelling of mice at different time, and there are obvious statistical significance between the blank group and test group (P<0.05). Compared with the model group, total alkali and high dose group, middle dose group of fat soluble substances can significantly reduce the writhing times of mice (P<0.05). Compared with the positive drug, high dose group of total alkaloids has obvious analgesic effect on mice.Conclusion:1) The optimized Soemmering’s extraction method was stable, reasonable, and feasible, which could be used to extract the fatty oils from Peganum harmala L. in lab.2) The optimized methyl esterification method has excellent stability and short reaction time, which laid a foundation for the fatty acids content determination of Peganum harmala L. by GC.3) The types and contents of fattyacids of Peganum harmala L. relate to the natural environment, and the change of contents of fatty acids relate to the picked reason. Compared with water soluble extracts from Peganum harmala L. and Peganum harmala L.total alkali, mice acute toxicity of fat soluble extracts from Peganum harmala L. is smaller, which has anti-inflammatory and analgesic activity. Fat soluble extracts from Peganum harmala L. is a potential plant resource. |
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