大孔树脂富集发酵液中9α-羟基双氢青蒿素的工艺研究

Enrichment of 9α-Hydroxyl-Dihydroartemisinin from Fermentation Broth with Macroporous Resin

9α-羟基双氢青蒿素由双氢青蒿素生物转化发酵而得,该研究考察了大孔吸附树脂法富集发酵液中9α-羟基双氢青蒿素的工艺,为9α-羟基双氢青蒿素的转化制备提供参考。静态吸附法考察4种型号大孔吸附树脂对目标产物的吸附及解吸性能,优选出AB-8树脂作为9α-羟基双氢青蒿素吸附树脂;动态吸附法进一步考察目标产物最佳分离条件,HPLC-ELSD法检测9α-羟基双氢青蒿素的含量。结果表明:当发酵液上样体积流量为25 BV/h时,最大上样量为5 BV,以8 BV水洗除去水溶性杂质,再以4 BV 95%乙醇洗脱,9α-羟基双氢青蒿素回收率可达87.46%,按照最佳工艺重复操作3次,9α-羟基双氢青蒿素回收率稳定在87%左右。该工艺操作简单,稳定性好,可应用于9α-羟基双氢青蒿素大规模生产制备。

9α-hydroxyl-dihydroartemisinin was obtained by biotransformation fermentation of dihydroartemisinin,and the enrichment of 9α-hydroxyl-dihydroartemisinin from fermentation broth with different macroporous adsorption resins were studied,which could provide a reference for its biotransformation preparation. The adsorption properties and desorption properties of four types of macroporous adsorption resins on target product were investigated by static adsorotion method,and AB-8 macroporous adsorption resin was selected as the optimal adsorption resin. Dynamic adsorption method was furthur studied to optimize the 9α-hydroxyldihydroartemisinin separation conditions,and the content of 9α-hydroxyl-dihydroartemisinin was detected by HPLC-ELSD. The results showed that the optimal sample flow rate of fermentation broth was 25 BV/h,and the maximum loading amount of AB-8 macroporous adsorption resin was 5 BV,95% ethanol eluted 4 BV after water was eluted 8 BV to remove the aqueous impurity,the recovery rate of 9α-hydroxyl-dihydroartemisinin reached 87. 46%. Repeated experiments were carried out three times based on the optimum process condition,the recovery rate of 9α-hydroxyl-dihydroartemisinin was around 87%. This process is easy to operate and has good stability,so it can be applied for large-scale production of 9α-hydroxyl-dihydroartemisinin.