Separation and Identification of Flavonoids from Smilacina Japonica by High Speed Counter-Current Chromatography

Five flavoniods including 5,7,3',4'-tetrahydroxy-3-methoxy-8-methylflavone (1), 8-methylluteolin (2), 3-methoxyluteolin (3), luteolin (4) and quercetin (5) were isolated and purified by high speed counter-current chromatography(HSCCC) and other chromatography from the rhizomes of Smilacina japonica. All these compounds were isolated for the first time from the species of the Smilacina genus. Preparative HSCCC with a two phase solvent system composed of chloroform-methanol-water(4:3:2, V/V), the upper phase as the stationary phase and the lower phase as the mobile phase was successfully performed to isolated extract of S. japonica rhizomes. Consequently, three major pure fractions composed of compound(1), (4), (5) corresponded were obtained, the purity was 98.3%, 96.7%, 95.3%, respectively by HPLC analysis. And one partially purified peak fraction containing compound (2) and (3) was obtained in same time, then the partially purified fraction was further separated by Sephadex LH-20 column chromatography with methanol-water (1:1, V/V) solvent system, recovered pure compound (2) and (3). The structures of five flavones were identified by physical and chemical properties and UV, IR, MS, NMR spectroscopy.