用正交试验法优选白花蛇舌草中三萜类有机酸的提取工艺.DOC

白花蛇舌草中总三萜酸的提取纯化工艺 蒙瑞波1，汤庆发1，曾永长2，郭阳1，傅钧庭1，范昊宁1，罗佳波*  （1. 南方医科大学中药新药实验室 广东省中药制剂重点实验室）,利用正交实验法考察最佳提取工艺；以静态吸附和解析实验筛选树脂型号，考察动态吸附与解析最佳条件.结果:最佳提取工艺为: 70%乙醇10倍量回流提取1.5 h。AB-8型大孔树脂最佳工艺为上样流速2 BVh-1，上样液pH值，上样浓度 mg·mL-1，洗脱剂pH值，乙醇浓度0％洗脱速度BV·h-1。结论：该工艺操作简单、稳定可行，重复性好，适合总纯化。 关键词白花蛇舌草；三萜酸；正交设计；提取纯化 The extraction and purification process of total triterpene acids in Hedyotis Diffusa WilldMENG Rui-bo1,TANG Qing-fa1,ZENG Yong-chang2 ,GUO Yang1,FU Jun-ting1, FAN Hao-ning1,LUO Jia-bo*（1. Southen university key Lab of TCM and new drug ；Guangdong provincial key Lab of TCM， Guangzhou Yangtze River Pharmaceutical Group Guangzhou Harris pharmaceutical CO.LTD，Guangzhou 510633) [Abstract] Objective: To optimize of extraction and purification process of triterpene acids in Hedyotis Diffusa Willd.Methods: To investigate the optimum extraction process by Triterpenoid acid extraction rate ; Screening resin model by static adsorption and parsing experiments,To investigate the optimal elution conditions by dynamic adsorption and resolution experiments.Results: The optimum extraction process was reflux extraction with 10- fold 70 % alcohol for 1.5h. The optimum purification process of AB-8 macroporous resin was sampling flow rate 2BV·h-1,pH=5, concentration 0.25 mg.mL-1,and the concentration of elution was 80％ alcohol,eluting velocity2 BV·h-1，with pH =7.Conclusion: The technology is simple,stable with good reproduciblity,which is suitable for extraction and purification of total HD triterpene acid.  [ Keywords] Hedyotis Diffusa Willd; Triterpenes acid; Orthogonal experiment；Extraction and purification  UV-HP8453紫外-可见光分度仪（美国惠普），CP2250微量电子天平；回流提取装置；移液管；水浴锅(上海精宏实验仪器有限公司，型号DK-S26)；白花蛇舌草（HD）（购自广东省药材公司，批号，经南方医科大学中药鉴定室张宏伟教授鉴定为茜草科耳草属植物白花蛇舌草(Hedyotis diffusa Willd)；齐墩果酸对照品（购自中国药品生物制品检定所，批号：110709-200505）；香草醛（上海易利生物科技有限公司，批；大孔树脂HPD-862、AB-8、D101均由广州东巨试剂公司提供；乙醇、氢氧化钠、盐酸、石油醚、乙酸乙酯、冰醋酸、高氯酸等均为分析纯。2 方法和结果 2.1 总三萜酸含量测定方