Preparative high-speed counter-current chromatography（HSCCC） and high performance liquid chromatography（HPLC） were employed firstly to separate chemical compounds in Vitex negundo L.. The procedure was performed in a two phase solvent system of chloroform-methanol-water （4∶3∶2,by volume） at a flow rate of 2.0 mL?min-1 by normal phase elution,in which the upper phase was the stationary phase and the lower phase was the mobile phase.The sample concentration was 20 g?L-1,while the rotation speed was 850 r?min-1 and the eluate was detected at 254 nm.Five compounds were obtained from the crude extract of the plant in a simultaneous process,which,according to their physical and chemical properties,were identified as:vitexin（Ⅰ）,casticin（Ⅱ）,3,5-di-O-caffeylquinic acid（Ⅵ）,luteolin（Ⅶ）,luteolin-7-O-β-D-glucoside（Ⅷ）.The purities of the five compounds were 98.0%,92.3%,96.8%,97.4%and 90.3%,respectively.