PREPARATIVE SEPARATION AND PURIFICATION OF FOUR PHENYLPROPANOID GLYCOSIDES FROM RHODIOLA ROSEA BY HIGH-SPEED COUNTER-CURRENT CHROMATOGRAPHY

Abstract

Phenylpropanoid glycosides are a class of the major bioactive components from Rhodiola rosea. In this study, an efficient method for isolation and purification of four phenylpropanoid glycosides (PPGS) from Rhodiola rosea was established by High-speed Counter-current Chromatography (HSCCC). Rhodiola rosea extract was pre-separated by HPD-200 macro porous resin and polyamide column adsorption chromatography for the enrichment of PPGS and the removal of flavonoid impurities. The enriched PPGS were prepared for subsequent HSCCC. The efficient preparative HSCCC separation and purification of PPGS was developed at a flow rate of 1.5 mL, using a 2-phase solvent system composed of ethyl acetate–butanol–water (1:0.35:1.35, v/v/v) which was selected according to the measurement of partition coefficient (K). By normal HSCCC separation model, 150 mg of the enriched sample was separated, yielding 27.8 mg of compound A, 34.3 mg of compound B, 3.6 mg of compound C, 54.8 mg of compound D, at a purity of 97.4%, 96.8%, 95.2%, and 98.5%, respectively. The purities of target compounds were tested by HPLC and the structure was identified by ESI-MS, 1H NMR, and 13C. The results demonstrate that HSCCC is a very efficient method for the simultaneous preparative separation of phenylpropanoid glycosides from Rhodiola rosea.