Abstract: To establish a method for separating high purity arctigenin from Arctii Fructusby combining acid hydrolysis with high-speed counter-current chromatography (HSCCC).The crude extract of Arctii Fructus was prepared by alcohol extraction and acid hydrolysis,followed by chloroform extraction.The separation conditions were as follows:the solvent system was petroleum ether-ethyl acetate-methyl-water (2:5:3:4,v/v) where the upper phase was used as the stationary phase and the lower phase as the mobile phase;the flow rate was 10.0 mL/min,the rotary speed was set at 850 rpm and the detection wavelength was 280 nm.As a result,318 mg of arctigenin with the purity of 99.12 % was separated from 1200 mg of crude sample within 80 minutes and the productive rate reached to 26.5%.The developed method was simple,rapid and efficient.It was suitable for the preparative separation of arctigenin and provided a reference for the development and utilization of Arctii Fructus.

Key words: acid hydrolysis, high-speed counter-current chromatography, Arctii Fructus, arctigenin