[关键词]
[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。
[Key word]
[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
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[基金项目]
湖南农业大学大学生科技创新项目资助(K1205202-21)