|Journal of Liquid Chromatography & Related Technologies, 2014, 37(17):2546-2557. |
|ISOLATION AND PURIFICATION OF SEVEN COMPOUNDS FROM EXTRACT OF RHEUM PALMATUM L. BY HIGH SPEED COUNTER CURRENT CHROMATOGRAPHY AND RAPID PREPARATIVE CHROMATOGRAPHY[Reference: WebLink]|
|High speed counter current chromatography (HSCCC) combined with rapid preparative chromatography (RPC) was developed for the separation and purification of seven compounds from Rheum palmatum L. in this paper.
METHODS AND RESULTS:
Compounds dimmer-catechin, catechin, trans-3,5,4′-trihydroxystilbene-4′-O-β-D-(6′-O-gallayl)-glucoside, 1-O-Galloyl-6-O-cinnamoyl-glucose, 1-O-Galloyl-2-O-cinnamoyl-glucose(2-Cinnamoyl-1-galloylglucose), rhein and emodin were isolated and purified by HSCCC using an optimized two-phase solvent system composed of n-hexane–ethyl acetate–n-butanol–water (1:2:1:4, v/v/v/v) and RPC. The purities of compounds 1, 3, 4 and 6 from HSCCC separation were over 98.70%, 96.75%, 98.20% and 98.07%, and those of compounds 1, 2, 4, 5 and 7 from RPC separation were 95.33%, 89.40%, 93.7%, 92.89% and 94.76%, determined by HPLC. The HSCCC and RPC fractions were analyzed by HPLC and electrospray ion source mass spectroscopy (ESI-MSn) in negative ion mode. The identification of the seven compounds from the extract of R. palmatum L. was based on matching their retention time, the detection of the molecular ions and the fragment ions of the molecular ion obtained in the MSn experiments with those of the data reported in the literature.
The results demonstrated that HSCCC and RPC are feasible and efficient techniques for systematic isolation of bioactive components from R. palmatum L.