The Study On The Chemical Constitents And Tyrosinase Activity Of Anacyclus Pyrethrum(L)DC

Objeceive:1. To analyse the chemical components of Anacyclus pyrethrum（L）DC, the pre-experiments were systemically made to determine.2. To study component and PTP1B inhibition of the fatty acid from Anacyclus pyrethrum（L）DC.3. To isolate and identify the chemical constituents of ethyl acetate extract in Anacyclus pyrethrum（L）DC, and to investigate the effect on tyrosinase activity. Methods:1. The pre-experiments were systemically made to determine the chemical constituents ofAnacyclus pyrethrum（L）DC by tube and filter paper method.2. Fats and oil wereextracted by soxhlet extraction from Anacyclus pyrethrum（L）DC and then fats and oils were dissolved in KOH of methonal solution with water-bath. Lastly, the fatty acids were extracted with n-Heptane, ether and chloroform, respectively. The chemical constituents of the fatty acids were identified and quantified by means of gas chromatography-mass spectrometry（GC-MS）, and determined their PTP1B inhibitory activity by microtest.3. To target on active fractions and constituents, the ethylacetate extract of Anacyclus pyrethrum（L）DC was isolated and purified by thin-layer chromatography, column chromatography and recrystallization, and tested theirtyrosinase activity by Dopa tyrosinase oxidation. Results:1. The results of pre-testshows that the chemical composition of Anacyclus pyrethrum（L）DC includes organic acids, tannins, alkaloids, quinones, flavonoids, cardiac glycosides, amino acids,sugars, and so on. This result agrees with “Uygur medicine”.2. The fatty acidswas extracted with n-hexane, included eight kinds of fatty acids: the linoleic acid（36.341%）,the hexadecanoic acid（34.268%）,the oleic acid（19.656%）,and so on; The fatty acids, with Petroleum ether, included10kinds of fatty acids: the linoleic acid（34.102%）,the oleic acid（20.000%）,the α-linolenic acid（5.329%）,andso on; The fatty acids, with chloroform, included five kinds of fatty acids: the linoleic acid（42.427%）, the hexadecanoic acid（40.952%）,the oleic acid（11.250%）,and so on; The fatty acids had the strong inhibiting effect on PTP1B,and its IC50value was5.27±0.40μg/mL,4.71±0.08μg/mL,7.01±1.98μg/mL respectively.3. Is olation and purification of the ethyl acetate extracts of Anacyclus pyrethrum（L）DC was carried out on the basis of1H-NMR,13C-NMR,DEPT,HMBC.The five compounds from the ethyl acetate extracts were as folloews: etradeca-2E,4E-diene-8,10-diynoic acid IBA（Ⅰ,5.9mg）,Dodeca-2E,4E-dienoic acid4-hydroxyphenylethylamide（Ⅱ,8.6mg）,6-methoxy-7-hydroxy-2-oxo-quinoline（Ⅲ,4.4mg）,Deca-2E,4E-dienoic acid4-hydroxyphenylethylamide（Ⅳ,7.5mg）,6,8-dimethoxy-7-hydroxy-2-oxo-quinoline（Ⅴ,3.9mg）.The results of tyrosinase activity showed the22fractions and five compounds of ethyl acetate had different levels on tyrosinase activation. Conclusion: Theextraction method is simple. Based on plant chemical theory, modern separation and analysis technology, the chemical components of Anacyclus pyrethrum（L）DC were systemically pre-test. On the basis, the fatty acid composition of Anacyclus pyrethrum（L）DC were analyzed and identified, and measured the inhibition of PTP1B.The chemical constituents of ethyl acetate extraction were isolated and identified,and filter tyrosinase activity. The results shows that the Anacyclus pyrethrum（L）DC had tyrosinase activity. The results provided the certain reference value and experimental basis for further study of Anacyclus pyrethrum（L）DC.