吡柔比星的分离与纯化

Separation and purification of pirarubicin

目的确定用硅胶柱层析纯化吡柔比星粗品的工艺条件。方法采用薄层层析-硅胶柱层析法考察不同展开剂的层析效果,最优洗脱剂为二氯甲烷-甲醇;以吡柔比星的回收率和平均含量为评价指标,考察了流速、进样量与层析柱再生条件对层析粗分和精分的影响。结果硅胶柱层析粗分吡柔比星的最佳工艺条件:以梯度洗脱方式洗脱,洗脱剂流速5.0mL.min-1,上样量16.6mg.mL-1柱体积,0.5g.L-1氢氧化钠溶液离线再生硅胶。吡柔比星质量分数经粗分后由原来的35.7%提高到90%左右,回收率为96.2%。硅胶柱层析精分吡柔比星的最佳工艺条件为洗脱剂二氯甲烷-甲醇比例为90∶10,流速3.5mL.min-1,上样量16.6mg.mL-1柱体积,甲醇再生3倍柱体积。吡柔比星质量分数经精分后由粗分的90%提高到99.6%左右,回收率为97.5%。结论硅胶柱层析纯化吡柔比星所得产品符合质量要求。

Objective To study the separation and purification of pirarubicin by silica gel column chromatography.Methods Thin-layer chromatography-silica gel column chromatography was used to analyze the effect of different mobile phases.The effects of flow rate,sample load,and the regenerated conditions were further researched on the purity and the recovery of pirarubicin in the first and second separation respectively.Results The optimal conditions for the first separation were gradient elution,flow rate 5.0 mL·min-1,sample load 16.6 mg·mL-1·CV-1.The silica gel might be regenerated by 0.5 g·L-1 sodium hydroxide solution.The purity of pirarubicin could be improved from 35.7% to about 90% after the first separation,and the recovery was 96.2%.And the optimal conditions for the second separation were mobile phase(dichloromethane-methanol=90∶10),flow rate 3.5 mL·min-1,sample load 16.6 mg·mL-1·CV-1.The silica gel column might be regenerated by 3 bed volumes of methanol.The purity of pirarubicin could be improved from 90% to 99.6% after the second separation,and the recovery was 97.5%.Conclusion After the two processes of silica gel column chromatography,the pirarubicin can meet the demand of the market.