Study On Isolation And Purification Of Active Components From Panax Ginseng,radix Gentianae By High-speed Counter-current Chromatography

High-speed counter-current chromatography (HSCCC), a support-free liquid-liquid partition chromatographic technique, has been widely applied for purication of functional components from traditional Chinese herbs and other natural products. It has asignificant advantage over the conventional liquid-solid column chromatography by eliminating the irreversible adsorption of the sample onto the solid support, What is more, HSCCC offers many other advantages such as choice of a wide range of the solvent systems, short separation time, high- purity of fractions, quantitative sample recovery and ease of scaling up and so on. The purpose of this study, was separated active components from Panax ginseng,the process ginseng and Gentiana scabra Bunge by HSCCC and different solvent system, the purity and chemical structures of target products was analyzed and identified by HPLC and MS.(1)Preparative isolation and purification of ginsenoside from Panax ginseng by HSCCCA preparative high-speed counter-current chromatography (HSCCC) method for isolation and purifcation of three compounds from Panax ginseng was successfully established by using ethyl acetate-butanol-water (4:1:3, v:v:v) as the two-phase solvent system in one-step run. The separated fractions are identified with standard samples by high-performance liquid chromatography (HPLC) and the electrospray ionization tandem mass spectrometry (ESI-MSn) data provided highly useful structural information for the saponins and the purities of these saponins were assessed to be over 95%.(2)Preparative isolation and purification of ginsenoside from processed ginseng by HSCCCHSCCC and D-101 macroporous resin column were applied to the separation and purification of one ginsenoside from processed ginseng by using a solvent system compose of n-hexane-ethyl acetate-methanol-water (1:3:1:2, v:v:v:v). The HPLC and ESI-MSn data was provided to determine the structural information for the separated fractions. The target compound was identified as Rg5 with the purity of 90.1%.(3)Preparative isolation and purification of secoiridoid from Gentiana scabra bunge by HSCCC Using HPD-100 macroporous resin chromatographic column and HSCCC, gentiopicroside and sweroside from Gentiana scabra bunge were separated. The two-phase solvent system compose of ethyl acetate-butanol-water (1:2:3, v:v:v) was applied to the HSCCC. The separation fraction was identified by ESI-MSn included Gentiopicrin and sweroside. The purity of gentiopicrin and sweroside were over 96.5% and 88.2% via HPLC, respectively.