Preparative isolation and purification of alkaloids from Corydalis yanhusuo W. T.!Wang by high speed counter-current chromatography

Four alkaloids including dehydrocorydaline, palmatine, coptisine, and columbamine were successfully isolated from the total alkaloids of Corydalis yanhusuo W. T. Wang using a stepwise elution by high speed counter-current chromatography (HSCCC). A pair of two-phase solvent systems consisting of carbon tetrachloride-trichloromethane-methanol-0.2 mol/L hydrochloric acid (1 : 7 : 3 : 4, v/v/v/v) and trichloromethane-methanol-0.2 mol/L hydrochloric acid (7 : 3 : 4, v/v/v) was successively used, which yielded four relatively pure alkaloids from 200 mg of the total alkaloids in a single run. The purities of dehydrocorydaline, palmatine, coptisine, and columbamine were 99.1, 97.1, 99.7, and 94.9%, respectively. The structures were identified by H-1 NMR and C-13 NMR.