硅胶柱层析法分离普那霉素  

Separation of pristinamycin by silica gel column chromatography

作　　者：贾波[1] 金志华[1] 张达[2] 梅乐和[1] 

机构地区：[1]浙江大学化学工程与生物工程学系,浙江杭州310027 [2]浙江大学宁波理工学院生物与化学工程分院,浙江宁波315100

出　　处：《浙江大学学报（工学版）》2008年第5期895-899,共5页Journal of Zhejiang University：Engineering Science

基　　金：浙江省重大科技攻关资助项目(2004C13007)

摘　　要：通过硅胶柱层析法在等度洗脱方式下对始旋链霉菌发酵产生的普那霉素进行分离纯化．结果表明，以体积比为90：10的二氯甲烷和甲醇或者体积比为60：35：5的二氯甲烷、乙酸乙酯和乙醇作为展开剂，采用硅胶GF254单向一次薄层层析（展开距离为5．O～6．0cm，展开时间为6～7min）即可对普那霉素作初步定性分析．柱层析操作的适宜条件为：25℃，以100-200目的硅胶为固定相，洗脱剂为体积比为80：35：5的二氯甲烷、乙酸乙酯和乙醇，流速为2．0mL／min，普那霉素的进样质量浓度为2．26g／L，硅胶的总负载量约为0．81mg／g．在此条件下能够分离得到纯度在95％以上的普那霉素，产品回收率保持在95％以上．The separation and purification of pristinamycin from the fermentation broth of Streptomyces pristinaespiralis was investigated by using silica gel column chromatography under isocratic elution. The results showed that qualitative analysis could be elementarily made by the method of thin layer chromatog- raphy （TLC） with silica GF254 （developing distance of 5.0-6.0 cm and developing time of 6-7 min）, under the developing agent composed either of methylene chloride and methanol with their volume proportions of 90 and 10, or of methylene chloride, ethyl acetate and ethanol with their volume proportions of 60, 35 and 5. The column chromatography was preferably conducted under such conditions： 25 ℃, 100-200 mesh silica gel as the stationary phase, solvent system of methylene chloride, ethyl acetate and ethanol with their volume proportions of 80, 35 and 5 as the eluent, the eluent flow rate of 2.0 mL/min, pristina- mycin feeding concentration of 2.26 g/L, and the total loading amount of silica gel of about O. 81 mg/g. Under these operational conditions, pristinamycin with purity of more than 95% was recovered, and the product recovery was over 95 % simultaneously.

关 键 词：普那霉素 薄层层析 硅胶 柱层析 分离 

分 类 号：TQ465.9[化学工程—制药化工]