HighSpeed countercurrent chromatography is a recently developed separation method which has been remarkably improved in both partition efficiency and separation time. In the present study, this advanced countercurrent chromatographic method was applied to separation of sample mixture containing tetrandrine, fangchinoline, and cyclanoline originally extracted from Stephania tetrandra S. Moore. Separations were performed with a two-phase solvent system composed of n-hexane/ethyl acetate/methanol/water in two different elution modes. Sample mixture containing 3 mg of alkaloids was efficiently separated in 100 min. The peak fraction of each component was analyzed with a mass spectrometer for structure identification. © 1988, Taylor & Francis Group, LLC. All rights reserved.