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PREPARATIVE ISOLATION AND PURIFICATION OF FOUR FLAVONOIDS FROM DAPHNE GENKWA SIEB. ET ZUCC. BY HIGH-SPEED COUNTERCURRENT CHROMATOGRAPHY. 2011

Huichun Xie, and Yong Liang, and Yoichiro Ito, and Xiaohong Wang, and Renshan Chen, and Juanting He, and Hang Li, and Tianyou Zhang
School of Chemistry and Environment, South China Normal University, Guangzhou 510631, China.

Four flavonoids including luteolin, apigenin, 3'-hydroxygenkwanin and genkwanin were isolated and purified from Daphne genkwa Sieb. et Zucc. by high-speed countercurrent chromatography (HSCCC). Preparative HSCCC with a two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (5:7:5:5, v/v) was successfully performed by increasing the flow rate of the mobile phase from 1.2 to 2.0 mL/min after 260 min. In a one-step operation, 150 mg of the extracts of D. genkwa was separated to yield 8 mg of luteolin, 25.8 mg of apigenin, 23.6 mg of 3'-hydroxygenkwanin and 35.3 mg genkwanin with the purities of 91.2, 97.4, 94.3 and 95.8%, respectively, analyzed by HPLC using area normalization method. The chemical structures of the four compounds were identified by HPLC, ESI-MS, and (1)H NMR.

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