Sorbifolia Chinese Bunge Saponins E Content By Hplc And The Reference Standard Preparation

Xanthoceras sorbifolia Bunge(Sapindaceae)is a plant widely distributed in Liaoning, Hebei and Shanxi Provinces of China.It is used to treat rheumatism and enuresis of children as a folk medicine in China.It has been reported that Xanthoceras sorbifolia Bunge showed good biological activities of anti-inflammation,anti-tumor,inhibition of HIV protease and improving memory,etc.The total saponins is determined to be the effective composition of Xanthoceras sorbifolia Bunge,and bunkanka saponin E is main effective component of the total saponins.It is the first time to establish the RP-HPLC method for quantitative determination of bunkanka saponin E content in Xanthoceras sorbifolia Bunge.The Chromsphere HypersilODS₂(250 mm×4.6 mm,5μm)column was used to determine the content of bunkanka saponin E.The mobile phase was methanol-water-phosphoric acid(70:30:0.02). The flow rate is 1.0 mL·min^-1.Column temperature is 34℃and the detection wavelength is 210 nm.The retention time of bunkanka saponin E was about 15 minutes under its chromatograms conditions,and the R-value between the main peak and the nearby peak was bigger than 1.5,which means reaching the standard of chromatograms separation.The linear range of bunkanka saponin E was from 0.0098 g·L^-1to 0.1960 g·L^-1.The rang of average recoveries in husk,carpophore,tunic and kernel was 99.2%to 100.4%.The results indicated: the content of bunkanka saponin E in the carpophore,husk and tunic were 0.45%,0.20%and 0.15%respectively.The bunkanka saponin E was not determinated in kernel.The method is simple,accurate,sensitive and stable used for the quality control for Xanthoceras sorbifolia Bunge.The results provided scientific basis for comprehensive development and utilization of different parts from Xanthoceras sorbifolia Bunge.From the 70%ethanol extract of the carpophore of Xanthoceras sorbifolia Bunge,the reference substance--bunkanka saponin E was isolated by means of solvent extraction, repeated column chromatography on silica gel and recrystallization.Its content determined by HPLC was 98.6%which meets the requirement of reference substance.During this course,12 compounds in total were isolated.On the basis of physico-chemical properties and spectroscopic analysis,their structures were identified as follows.Tritepenes:R₁-barrigenol (1),21.22-di-O-angeloyl-R₁-barragenol(2),3-O-(3-O-α-L-arabinofuranosyl-2-O-β-D-galactopy ranosyl)-β-D-glucuronopyranosyl-21,22-di-O-angeloyl-R₁-barrigenol(3),21-O-angeloyl-21 -angeloyl-R₁-barrigenol(4),3-oxotirucalla-7,24-dien-21-oic(5);Sterols:β-sitosterol(6), daucosterol(7),α-spinasterol(8),α-spinasterol-3-O-β-D-glucopyranoside(9);Others: n-hentriacontanic acid(10),n-heptacosanol(11),n-butyl-fructopyranoside(12).