大孔吸附树脂结合硅胶柱层析纯化环孢菌素A

采用大孔吸附树脂层析结合硅胶柱层析,对环孢菌素A的分离纯化进行研究,确定了最佳层析条件,建立了工业化制备环孢菌素A的工艺。大孔吸附树脂层析选用D101树脂作为吸附介质,提取液丙酮含量控制在50%,最大吸附量为35 mg/g湿树脂,洗脱剂选用丙酮;硅胶柱层析选用42~64μm硅胶作为层析介质,最优层析条件为柱床高径比10∶1,流动相配比V(石油醚)∶V(丙酮)=70∶30,流速80 mL/m in,环孢菌素A上样质量浓度100 g/L,硅胶层析平均收率为84.2%,环孢菌素A纯度可达到97%以上,整个工艺总收率为65%~70%。

Purification of cyclosporin A by using macroporous resin and silica gel column chromatography

The purification process of cyclosporin A by using macroporous resin and silica gel column chromatography was studied.In the first step,D101 resin was used to separate CysA.The optimal conditions were as follows： the concentration of acetone in the mobile phase was 50%.The D101 resin showed better property with the absorption capacity of 35 mg/g resin.Acetone was selected as the eluant.In the next step of silica gel column chromatography,42～64 μm mesh silica gel was used as the stationary phase.The mobile phase was comprised of petroleum ether and acetone（70∶30）.The flow rate was 80 mL/min.The loading concentration of cyclosporin A on silica gel was 100 g/L.The average recovery of cyclosporin A with purity of more than 97% and the average yield of silica gel column chromatography was over 84.2%.The total recovery of the process ranged from 65% to 70%.