The medicinal plant Atractylodes macrocephala (Baizhu in Chinese) has been widely used in traditional Chinese medicine for energy and stomach complaints, treatment of dyspepsia and anorexia, anti-inflammation, anticancer and for increasing assimilation. A high-speed counter-current chromatography (HSCCC) method was developed for the preparative separation and purification of two main bioactive components,namely, atractylon and atractylenolide III from A. macrocephala by using light petroleum (60908C)ethyl acetateethanolwater (4 :1:4:1 v/v) as the two-phase solvent system in dual-mode elution. Compared with the separation using the normal-mode elution, the dual-mode HSCCC can be achieved with shorter elution time.Atractylenolide III (32.1 mg) at 99.0% purity and 319.6 mg atractylon at 97.8% purity could be obtained from 1000 mg crude sample in a single run. The recoveries of atractylenolide III and atractylon were 95.4 and 92.6%, respectively. 高速逆流色譜儀/GX逆流色譜儀 TBE-1000A制備色譜儀/萃取儀/快速分離制備色譜儀 TBE300B+AKTA高速逆流色譜儀/離心分配色譜/萃取儀/制備色譜儀 TBE-200V 高速逆流色譜儀