运用高速逆流色谱分离纯化金莲花中黄酮苷成分

Abstract:Three flavonoid glycosides including orientin, vitexin, quercetin-3-O-neohesperidoside and one unknown compound were isolated and purified by high-speed counter-current chromatography (HSCCC) and semi-preparative HPLC from Trollius ledebouri Reichb.,a traditional Chinese medicine. Preparative HSCCC with a two-phase solvent system composed of ethyl acetate–n-butanol–water (2:1:3, v/v/v) was successfully performed by increasing the flow-rate of the mobile phase from 1.5 to 2.5 ml/min after 190min. Consequently, 95.8mg orientin,11.6mg vitexin, 9.3mg unknown compound with purities of over 97% and one partially purified peak fraction (contained quercetin-3-Oneohesperidoside at 85.1% purity)were obtained from 500mg of the crude extract. Then the partially purified fraction was further purified by reversed-phase semi-preparative high-performance liquid chromatography. The structure identification of all pure fractions was carried out by UV, MS, 1H NMR and 13C NMR.