Abstract High-speed countercurrent chromatography (HSCCC) was applied for separation and purification of flavanoids from the extract of belamcanda. High efficiency of HSCCC separation was achieved on a two-phase solvent system of n-hexane–ethyl acetate–methanol–water (4:5:5:5, v/v) by eluting the lower mobile phase at a flow-rate of 1.2 mL/min and a revolution speed of 800 rpm. Three well-separated peaks were obtained in the HSCCC chromatogram and their purities were determined by HPLC-UV absorption spectrometry. These peaks were characterized by ESI-MSn and NMR, and the data compared with the reference standards where three peaks were identified as isorhamnetin, irigenin and hispidulin. The purities of each peak were 94, 95 and 90% respectively. In HSCCC experiment, 100 mg of the crude extract were separated yielding 10 mg of isorhamnetin, 8 mg of irigenin and 7 mg of hispidulin. HSCCC thus provides a cost-effective alternative to preparative scale HPLC for the semi-preparative scale separation and purification of flavonoids from Belamcanda.

中文翻译：

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高速逆流色谱法分离纯化中草药白兰地黄酮类化合物

摘要 应用高速逆流色谱(HSCCC) 分离纯化白兰地提取物中的黄酮类化合物。在正己烷-乙酸乙酯-甲醇-水 (4:5:5:5, v/v) 两相溶剂系统上通过以1.2 mL/min 和 800 rpm 的转速。在 HSCCC 色谱图中获得了三个分离良好的峰，它们的纯度通过 HPLC-UV 吸收光谱法测定。这些峰通过 ESI-MSn 和 NMR 进行表征，并将数据与参考标准进行比较，其中三个峰被确定为异鼠李素、鸢尾素和马皮苷。每个峰的纯度分别为 94、95 和 90%。在 HSCCC 实验中，分离出 100 mg 粗提物，得到 10 mg 异鼠李素，8 mg irigenin 和 7 mg hispidulin。因此，HSCCC 为半制备规模的 HPLC 分离和纯化 Belamcanda 中的黄酮类化合物提供了一种具有成本效益的替代方案。