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。

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

湖南农业大学大学生科技创新项目资助（K1205202-21）