[Preparative isolation and purification of the active components from Viticis Fructus by high-speed counter-current chromatography].

Vitex trifolia L.var.simplicifolia Cham.is widely distributed in Asia,and its fruits are used as a folk medicine for headaches,colds,migraine,eyepain,etc.In order to effectively separate high-purity active components from the seeds of Vitex trifolia L.var.simplicifolia Cham.,a high-speed counter-current chromatography(HSCCC)procedure was performed to separate four components from the crude extract of the fruits.A two-phase solvent system composed of light petroleum-ethyl acetate-methanol-water(3:6:3.6:3,v/v/v/v)was used.Within 230 min,23 mg of 4-hydroxybenzoic acid,15 mg of 3,6,7-trimethylquercetagetin,24 mg of casticin and 5 mg of artemetin were obtained from250 mg of the crude extract of Viticis Fructus in one-step elution under the conditions of a flow rate of 1.5 mL/min,800 r/min and the detection wavelength of 254 nm.The purities of the four fractions were 93.1%,97.3%,98.7%and 98.5%,respectively.The obtained fractions were analyzed by high performance liquid chromatography(HPLC),and identified by electrospray ionization mass spectrometry(ESI-MS),1 H-nuclear magnetic resonance(NMR)and 13C-NMR.The results indicate that HSCCC is a powerful technique for the purification of active components from Viticis Fructus.