Systematic Study On The Lignans Compounds Of Herpetospermum Caudigerum

Herpetospermum seed,a common medication used by Tibetan medieinal, is the dried ripe seed of Herpetspermum Caudigerum Wall.. It is cold in nature and bitter in taste. Its efficacy include purging the fire of liver and gall and detoxifying. It produced in Sichuan, Yunnan, Tibet and India etc.,at an altitude of2300-3500m.In recent years, the study of the pharmacological effect of Herpetospermum seed showed, its effective lignanoids part and partial monomer have a good effect in anti-liver injury and anti-hepatitis. Through the systematic study of lignans from the seeds of Herpetspermum Caudigerum., the study provide the basis for establishing quality standard of the seeds of Herpetspermum Caudigerum. and its further development and utilization.The seeds of Herpetspermum Caudigerum. were extracted by refluxing method with80%alcohol. Portions of petroleum ether, ethyl acetate, n-butanol and water were obtained by extraction. In this paper, the part of ethyl acetate,which weighs146g, was isolated and purified. Fifteen compounds were isolated by the method of ordinary pressure normal phase silica gel column chromatography, reversed phase silica gel column chromatography, Sephadex LH-20, recrystallization etc.. The structures of eleven compounds were elucidated by means of TLC,2H-NMR,13C-NMR, DEPT, ESI-MS, two-dimensional NMR spectroscopy. Detailed Results As Follows, Herpetetrone(1), Herpetal(2), Herpetin(3), Herpetfluorenone(4), BL-6(6), Herpetrione(7), Herpetotriol(8),(-)-tanegool-71-methyl ether(10), Herpetetrol(11), BL-13(13), Herpepentol(15). All the compounds are Lignans compounds. Compound6and13are new compounds, Compound10was isolated from this plants for the first time.The separated compounds were analyzed by HPLC. The contents of Herpetfluorenone, Herpetotriol, Herpetin and Herpetrione were determined. The chromatographic analysis of Herpetfluorenone was carried out on a Kromasil C18(4.6mm×150mm,5μm) column with acetonitrile-water(24:76) as the mobile phase. The detection wavelength was set at260nm,the column temperature was40°, and the flow rate was about1.0mL/min. The regression equation was y=5966.0x-2492.80(r=0.9999). The linear was0.0294-0.147ug. The chromatographic analysis of Herpetotriol was carried out on a Kromasil C18(4.6mm× 150mm,5μm) column with acetonitrile-water(24:76) as the mobile phase. The detection wavelength was set at277nm,the column temperature was40°, and the flow rate was about1.0mL/min. The regression equation was y=3395.4x-13353(r=0.9998). The linear was0.62-3.1ug.The chromatographic analysis of Herpetin and Herpetrione was carried out on a Kromasil C18(4.6mm×250mm,5μm) column with acetonitrile-water(23:77) as the mobile phase. The detection wavelength was set at230nm,the column temperature was30°, and the flow rate was about1.0mL/min. The regression equation of Herpetin was y=10309x-24381(r=0.9998). The linear was0.4708-4.5540ug. The regression equation of Herpetrione was y=11433x-305603(r=0.9997). The linear was1.664-8.32ug. Three batcher of Herpetspermum Caudigerum. were determined. The average content of Herpetfluorenone was about0.135mg/g, The average content of Herpetotriol was about2.66mg/g. The average content of Herpetin was about1.73mg/g.The average content of Herpetrione was about5.0mg/g.The HPLC methods were simple, precision, repeatable and be used to provide a reference for the quality control of Herpetspermum Caudigerum..