Ligusticum chuanxiong Hort has been used widely in traditional medicines for the treatment of various kinds of disorders such as cardiovascular and cerebrovascular diseases. High-speed counter-current chromatography was applied to the isolation and purification of the bioactive component chuanxiongzine from L. chuanxiong Hort. The crude chuanxiongzine was obtained by extraction with ethanol from the dried roots of L. chuanxiong Hort under sonication. Preparative high-speed counter-current chromatography with a two-phase solvent system composed of n-hexane-ethyl acetate-ethanol-water (5:5:3:7, v/v) was successfully performed yielding 11.5 mg chuanxiongzine at 96.8% purity from 300 mg of the crude extract (4.2% chuanxiongzine) with the recovery of 91% in a one-step separation.