Study On HPLC Fingerprints And Simultaneous Determination Of Major Components Of The Hypencum Ascyron L.in Yanbian

Abstract:Objective:To establish the chemical composition HPLC fingerprints and HPLC method for simultaneous determination of major components in Hypericum ascyron L. which collected from different habitats in Yanbian.The chemical composition HPLC fingerprints were analyzed and the contents of active components were compared in10simples of Hypericum ascyron L.in yanbian.The methods will provide the reference for the quality control and identification of the Hypericum ascyron L.. Methods:The HPLC fingerprints and simultaneous determination methods were employed on Diamonsil C18column (250mmx4.6mm,5μm), the detecting wavelength was at360nm;eluted with gradient three mobile phase(methanol-0.3%phosophoric acid-acetonitrile) at the flow rate of1.0ml/min, the column temperature was35℃. Result:1.10samples of Hypericum ascyron L.were used to evaluate the whole HPLC spectrums and20common peaks of resolution was over1.0were identified and the common spectrogram was established. The data was processed on the similarity of fingerprint software, to gather statistics10samples’clustering results by SPSS analysis software.2.The HPLC method of simultaneous determination of major components showed a good linear relationship in the range of0.4～6.0μg/ml (r0.9995) for rutin,2.0～30.0μg/ml (r=0.9992) for hyperoside,0.8～12.0μg/ml (r=0.9991) for quereetin,1.2～18.0μg/ml (r=0.9990) for kaempferol; recoveries for the four components were all above90%.3.The average contents of rutin, hyperoside,quereetin,and kaempferol in Hypericum ascyron L. in Yanbian were0.0872-0.2673mg/g;0.4836-1.4354mg/g;0.1773-0.8327mg/g;0.1671-1.9681mg/g.Conclusion:1. The peaks of resolution and characteristic in the established HPLC fingerprints were very obviously. The method is stable, RSD of degree of precision and reproducible were less than5%.2.There were many difference about the contents of rutin, hyperoside, quereetin and kaempferol in Hypericum ascyron L.of yanbian.This method about simultaneous determination was accurate, sensitivite and reliable, which can be used for identification of the internal quality, and the results will provide the reference for the quality control of the Hypericum ascyron L. in Yanbian.To offer the science foundation for Hypericum ascyron L. about institution of quality standard.