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Preparative isolation and purification of flavonoids from the Chinese medicinal herb Belamcanda by high-speed countercurrent chromatography

机译：高速逆流色谱法从中草药贝拉菌根中制备黄酮类化合物的分离纯化

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High-speed countercurrent chromatography (HSCCC) was applied for separation and purification of flavonoids 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 1.2mL/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.
机译：高速逆流色谱法（HSCCC）用于从白lam提取物中分离和纯化黄酮类化合物。在正己烷-乙酸乙酯-甲醇-水（4：5：5：5，v / v）两相溶剂系统中，通过洗脱下层流动相，可实现HSCCC的高效分离。 1.2mL / min，转速为800 rpm。在HSCCC色谱图中获得了三个分离良好的峰，并通过HPLC-UV吸收光谱法测定了其纯度。这些峰用ESI-MS n 和NMR表征，数据与参考标准品进行了比较，其中三个峰被确定为异鼠李素，irigenin和组蛋白。每个峰的纯度分别为94％，95％和90％。在HSCCC实验中，分离出100 mg的粗提物，得到10 mg异鼠李素，8 mg irigenin和7 mg组织蛋白。因此，HSCCC为制备规模的HPLC提供了一种经济高效的替代方案，可用于半制备规模的分离和纯化来自Belamcanda的类黄酮。

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