Studies On Separation And Purification Of Active Constituents From Ephedra Sinica, Diploclisia Glauces Cens, Gelsemium Elegans And Bufo Bufo Gargarizans By HSCCC

Objective:The present article is to develop new methods for separation and purification of active components from Ephedra sinica,Diploclisia glaucescens,Gelsemium elegans and Bufo bufo gargarizans by High-Speed Counter-Current Chromatography. The newly established methods will provide reference for separating and purifying active ingredients from traditional Chinese medicinal herbs. And they will also provide technical support for preparing standard substances and finding new drugs.Methods:Literature reviews relating to the progress of researchs on applying HSCCC for separating different kinds of active constituents from traditional Chinese medicine were summarized systematically. Use the combination of silica gel column chromatography and HSCCC to separate flavonoids from Ephedra sinica; apply HSCCC to isolate ecdysteroids from Diploclisia glaucescens; employ pH-zone-refining counter-current chromatography to prepare indole alkaloids from Gelsemium elegans; utilize HSCCC coupled with Preparative High Performance Liquid Chromatography to obtain bufadienolides from Bufo bufo gargarizans. The purities of the obtained compounds were determined by HPLC and their chemical structures were all identified by MS and NMR.Results:Two biflavonoids, mahuannin A and mahuannin D, together with two flavones, dihydroquercetin and catechin, were successfully obtained from Ephedra sinica and their purities were all over98%; six indole alkaloids including19-oxo-gelsenicine, gelsemine, koumine,11-methoxygelsemamide, gelsenicine and humantenine were prepared from Gelsemium elegans with high quantity and purity; eight bufadienolides including arenobufagin, bufotalin,19-oxo-cinobufagin, cinobufotalin, bufalin, cinobufagin, resibufogenin and hellebrigenin-3-hemisuberate were purified from Bufo bufo gargarizans.Conclusions:HSCCC could realize the object of both analytical separation and preparative purification with the quantity from several micrograms to several grams. Without solid supporter, HSCCC could avoid sample loss, inactivation and degeneration caused by irreversible adsorption, therefore, it is especially suitable for the isolation of bioactive compounds from natural products.