Isolation and Separation of Flavones from Patrinia villosa by High-Speed Counter-Current Chromatography

OBJECTIVE To develop a preparative high-speed counter-current chromatography(HSCCC) for isolating the chemical constituents from Patrinia villosa Juss.METHODS A two-phase solvent system composed of n-hexane: ethyl acetate: methanol: water(1∶(0.6∶0.6)∶1) was used.at a flow rate of 2.0 mL·min~(-1),while the apparatus rotated at 800 r·min~(-1) and the effluent was detected at 254 nm.The obtained fractions were analyzed by HPLC and identified by MS,()~(1)H-NMR and()~(13)C-NMR.RESULTS 26 mg luteolin,15 mg 5-hydroxyl-7,4′-dimethoxy flavone and 21 mg 5-hydroxyl-7,3′,4′-trimethoxy flavone with purities of over 98% were obtained from 400 mg crude extract in one-step elution and less than 6 hours.CONCLUSION Three kinds of flavones are all discovered from the plant of Patrinia genius for the first time.The results indicate that HSCCC is a powerful technique of purification for natural products.