制备液相色谱分离纯化埃博霉素B  

Study on the Separation and Purification of Epothilone B by Preparation Liquid Chromatography

作　　者：郑玲辉 陈伟 ZHENG Linghui;CHEN Wei(Hangzhou Zhongmeihuadong Pharmaceutical Co.Ltd,Zhejiang Hangzhou 310000)

机构地区：[1]杭州中美华东制药有限公司,浙江杭州310000

出　　处：《工业微生物》2023年第2期98-101,共4页Industrial Microbiology

摘　　要：基于制备液相色谱法开发与优化埃博霉素B的分离纯化工艺,制备得高纯度埃博霉素B样品。实验首先对埃博霉素B粗品进行了液相色谱(HPLC)分析,定位目标峰和杂质情况。其次,对两款正相色谱填料进行筛选和模拟制备,考查不同填料对埃博霉素B相关杂质的去除效果和回收率。结果表明,上样量为0.2%时,埃博霉素B在NPLC-2分析柱(250 mm×4.6 mm,10μm)上保留得较好,可与杂质有效分离。最后,将优化好的纯化方法在制备水平上进行放大,以正庚烷和乙酸丁酯为洗脱剂,使用NPLC-2制备柱(260 mm×50 mm,10μm)对埃博霉素B粗品进行分离纯化,样品的色谱纯度可达99.63%,回收率为90%,各项有关杂质均符合限量规定。该方法对埃博霉素B的相关杂质去除效果好,纯化效率和回收率均很高,为埃博霉素B分离纯化生产工艺的开发提供了新方法。Based on the preparative chromatography,the separation and purification of Epothilone B was developed and optimized,and Epothilone B with high purity was prepared.Firstly,the crude product of Epothilone B was analyzed by HPLC,and the target peak and impurities were located.Then,two kinds of stationary phases were selected,and the separation and yield of related impurities and on different stationary phases were investigated.The results showed that when the sample loading was 0.2%,the retention of Epothilone B on NPLC-2 column(250 mm×4.6 mm,10μm)was better,and it could be effectively separated from impurities.Finally,the optimized purification method was scaled up to the preparative level,and the crude product was separated and purified using n-heptane and butyl acetate as eluents on a NPLC-2 column(260 mm×50 mm,10μm);the HPLC purity of the sample was 99.63%;the yield was 90%,and all the impurities met the limit requirements.This method exhibited good removal of impurities and high efficiency and yield which was of help for the development of separation and purification of Epothilone B.

关 键 词：制备液相色谱 分离纯化 正相色谱 埃博霉素B 

分 类 号：O652.63[理学—分析化学] TQ460.1[理学—化学]