高速逆流色谱法分离玫瑰茄中的花色苷

Isolation of Anthocyanin from Hibiscus sabdariffa by High Speed Counter-current Chromatography

摘要:

实验以玫瑰茄花萼为原料，建立高速逆流色谱法制备分离玫瑰茄花色苷的方法。玫瑰茄花萼经40%乙醇浸提，通过D101大孔吸附树脂吸附初步纯化，然后水-正丁醇-甲基叔丁基醚-乙腈-三氟乙酸（6:3:1:1：0.001，V/V）为两相溶剂系统进行HSCCC分离纯化，上相为固定相，下相为流动相,流速2.0 mL/min，上样量120 mg，经一次分离得到花色苷1粉末21.8 mg，HPLC面积归一法计算纯度为97.8%，回收率为95.3%；花色苷2纯度为84.5%，经相同溶剂体系二次分离，得到5.3 mg纯度为96.2%的花色苷2，回收率为92.3%。通过质谱、核磁等技术鉴定所分离得到的两个花色苷类化合物分别为飞燕草素-3-O-桑布双糖苷以及矢车菊素-3-O-桑布双糖苷。该方法操作简便，重现性好，适于玫瑰茄中高纯度花色苷大量制备，为花色苷进一步药理研究及质量控制提供物质基础。

Abstract:

The calyx of Hibiscus sabdariffa was extracted with 40% ethanol at room temperature, and poured through D101 resin. A solvent system of water-n-butanol-methyl tert- acetonitrile-trifluoroacetic acid (6:3:1:1:0.001, V/V) was used for isolation and purification by high speed counter-current chromatography (HSCCC) using upper phase as stationary phase and lower phase as mobile phase ,with the flow rate of 2 mL/min and injection volume of 120 mg. 21.8 mg anthocyanin 1 power was obtained with the purity and the recovery rate being of 97.8% and 95.3%, respectively. Anthocyanin 2 was isolated twice with purity of 96.2% (5.3 mg) and recovery rate of 92.3%. The chemical structures of the two anthocyanins were identified as delphindin-3-O-sambubioside for anthocyanin 1 and cyanidin-3-O-sambubioside for anthocyanin 2 by ESI-MS and NMR. The method is simple, reproducible, which is suitable for preparation of anthocyanins from Hibiscus sabdariffa, and also provides basis for further pharmacological studies and quality control of anthocyanins