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.

Keywords:

• anthraglycoside B
• Begonia fimbristipula
• counter-current chromatography
• elution-extrusion
• preparative chromatography
• traditional Chinese Medicine

ACKNOWLEDGMENT

Yanbin Lu thanks the Natural Science Foundation of China (No. 21005072), Department of Science and Technology of Zhejiang Province (No. 2010C12016), and Young Talented Scholar Foundation of Zhejiang Gongshang University (No. QZ11-6) for financial supports. Ruilan Wang thanks the financial support from Guangdong Provincial Natural Science Foundation (No. 9151052005000005).