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 1H NMR and 13C NMR.