高速逆流色谱法快速分离制备雷帕霉素工艺研究  

Separation of rapamycin from fermentation broth using high-speed counter-current chromatography

作　　者：李进[1,2] 邹祥[1] 

机构地区：[1]西南大学药学院,重庆400715 [2]泸州职业技术学院,泸州646005

出　　处：《中国抗生素杂志》2012年第10期752-756,共5页Chinese Journal of Antibiotics

基　　金：国家新药创制重大专项项目(2010ZX09401-306-4-4;2010ZX09401-306-1-4);重庆市创新能力建设项目(CSTC;2009CB1010);西南大学博士基金项目(SWU109034)

摘　　要：目的研究从雷帕霉素发酵液中快速分离制备雷帕霉素工艺方法,提高雷帕霉素提取效率。方法用高速逆流色谱法,考察不同两相溶剂系统对雷帕霉素粗提物分配效果。结果菌丝体经丙酮、乙酸乙酯粗提后,以正己烷-乙酸乙酯-乙醇-水(7:5:5:5,V/V)为两相溶剂系统,上相为固定相,下相为流动相,在流速2 mL/min,转速800 r/min,检测波长为280 nm条件下,从菌丝体初提物中一步纯化得到雷帕霉素,纯度为89.97%,纯化回收率达90%以上,同时初步分析其杂质主要为其同分异构体,含量为8.79%。结论本研究结果为从雷帕霉素发酵液中快速分离制备雷帕霉素提供新技术路线。Objective The quick separation method of the rapamycin from fermentation broth was investigated for improving the extraction recovery ratio and efficiency. Methods High-speed counter-current chromatography （HSCCC） technology was employed, and different solvent system was analyzed for the distribution efficiency. Results The crude extract of rapamycin from mycelium was separated by HSCCC. A mixture of hexane, ethyl acetate, ethanol and water （7：5：5：5, V/V） was selected as solvent system, the upper phase and the lower phase separated from which provided mobile phase and stationary phase, respectively. The HSCCC separation of rapamycin was carried out under the following conditions： flow rate 2 ml/min, rotation speed 800 r/min and detection wavelength 260 nm. The recovery ratio was over 90%. The purity of rapamycin from the ethyl acetate extract from mycelium was 89.97%, and the inpurity was confirmed as its isomeride by ESI/MS analysis. Conclusion The study provides a novel route for quick separation and purification of rapamycin from fermentation broth in large-scale.

关 键 词：高速逆流色谱 纯化:雷帕霉素 菌丝体 纯度 

分 类 号：R979.5[医药卫生—药品]