EFFECTIVE COUNTER-CURRENT CHROMATOGRAPHIC METHOD FOR ONE-STEP PREPARATIVE ISOLATION AND PURIFICATION OF ANTHRAGLYCOSIDE B FROM Begonia fimbristipula USING ELUTION-EXTRUSION SEPARATION MODE

Abstract

Counter-current chromatography (CCC) is a chromatographic technique with a support-free liquid stationary phase. Elution-extrusion CCC (EECCC) takes full advantage of the liquid nature of the stationary phase and effectively extends the hydrophobicity window with any biphasic liquid system. In this study, a preparative EECCC method for isolation and purification of anthraglycoside B from the ethanol extract of Begonia fimbristipula was successfully established by using n-hexane–ethyl acetate–methanol–water as the two-phase solvent system. The upper phase of n-hexane–ethyl acetate–methanol–water (1:6:1:6, v/v) was used as the stationary phase of HSCCC. Under the optimum conditions, 26 mg of anthraglycoside B at 97.8% purity, as determined by UPLC, could be yielded from 500 mg of the crude extract in a single EECCC separation. The peak fraction of CCC was identified by 1H NMR and 13C NMR.