摘要
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%。
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%.
出处
《生物加工过程》
CAS
CSCD
2010年第3期6-12,共7页
Chinese Journal of Bioprocess Engineering
关键词
环孢菌素A
大孔吸附树脂
硅胶
柱层析
纯化
cyclosporin A
macroporous resin
silica gel
column chromatography
purification