Separation and purification of rutin and acaciin from the chinese medicinal herb Herba Cirsii by combination of macroporous absorption resin and high-speed counter-current chromatography.

A preparative high-speed counter-current chromatography (HSCCC) method for isolation and purification of rutin and acaciin from the Chinese medicinal herb Herba Cirsii was successfully established. The crude extracts obtained from Herba Cirsii by water under reflux were subjected to a macroporous resin column and eluted with 10% and 60% ethanol, respectively. The fraction of 60% ethanol was used as the sample for HSCCC separation of rutin and acaciin. The two-phase solvent system used for the separation was ethyl acetate-n-butanol-water (5:1.5:5, v/v) and the upper phase was used as the stationary phase. Rutin (25.2 mg) and acaciin (21.8 mg) with a purity of 99.2% and 99.6%, respectively, were purified successfully from 60 mg of sample. The chemical structures of rutin and acaciin were identified by 1H-NMR and 13C-NMR.