High-speed countercurrent chromatography isolation of flavans from Ixeris chinensis and their identification by NMR spectroscopy.

High-speed countercurrent chromatography, combined with macroporous resin chromatography were applied to the separation and purification of flavans from Ixeris chinensis. Four flavans, namely, 5-methoxy-7,4'-dihydroxyflavan-3-ol (1), 5,7-dimethoxy-4'-hydroxyflavan-3-ol (2), 5,7-dimethoxy-4'-hydroxyflavan (3), and 5,7-dimethoxy-8-methyl-4'-hydroxyflavan (4), were obtained from I. chinensis for the first time. Their chemical structural identification was carried out by spectroscopic methods, including 1D and 2D NMR spectroscopy. Amounts of 13.2 mg of compound 1, 6.4 mg of compound 2, 5.8 mg of compound 3, and 14.5 mg of compound 4 were separated from 120 mg 75% ethanol fraction. The purities of 1-4 were 99.1, 99.2, 97.3, and 98.6 %, respectively.