Preparative Isolation and Purification of Four Flavonoids from Daphne Genkwa Sieb. Et Zucc. by High-Speed Countercurrent Chromatography

Xie, H.; Liang, Y.; Ito, Y.; Wang, X.; Chen, R.; He, J.; Li, H.; Zhang, T.

Journal of Liquid Chromatography and Related Technologies 34(19): 2360-2372


ISSN/ISBN: 1082-6076
PMID: 22379361
DOI: 10.1080/10826076.2011.589094
Accession: 055137857

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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.