FK-520分离纯化工艺的研究

Separation and purification of the FK-520 from fermented broth

目的开发出一种FK-520分离纯化方法。方法本研究主要采用薄层层析-硅胶柱层析法考察了不同展开剂的层析效果,以洗脱液的平均含量和收率作为评价指标,考察了流速和进样量对层析分离的影响。结果实验结果表明,硅胶柱层析方法,结合以萃取和结晶工艺,可以从FK-520发酵液中分离纯化出高纯度的FK-520。硅胶柱层析分离FK-520的最佳操作条件为:洗脱剂乙酸乙酯-石油醚(80:20),洗脱剂流速300m L/h,上样量10mg/m L柱体积。洗脱液脱色结晶后,FK-520纯度达到95%以上,收率超过80%。结论建立了一种新的FK-520分离纯化方法,该方法简单、易行,为该品种工业化生产提供一定的技术参考。

Objective A method for separation and purification of FK-520 was developed. Methods In this study, thin-layer chromatography and silica gel column chromatography were used to investigate the chromatographic effects of different eluents. In order to investigate the effects of different flow rates and loading capacities for chromatographic separation, the average content of eluent and yield were utilized as the evaluation index. Results Experimental results showed that the high purity FK-520 could be separated from fermented broth through method of silica gel column chromatography combined with extraction and crystallization processes. The optimal operating conditions for silica gel column chromatography separating FK-520 were flow rate 300mL/h, loading capacity 10mg/mL column volume. After decolored by activated carbon the decolorizing solution was crystallized by cyclohexane. The purity of FK-520 was more than 95%, and the yield was over 80%. Conclusion The method for separation and purification of FK-520 was established. It was simple and easy, and provided a certain technical reference for FK-520 industrial production.