目的 以蛹虫草发酵液虫草素粗提物为原料，对高速逆流色谱（HSCCC）分离制备虫草素条件进行研究。方法 利用HPLC测定分配系数法结合分析型HSCCC对分离虫草素的溶剂体系进行筛选，确定虫草素分离的最佳溶剂体系为醋酸乙酯-正丁醇-0.5%氨水（2∶3∶5），并运用此溶剂体系，上相作为固定相，下相作为流动相，利用制备型HSCCC分离制备蛹虫草发酵液中虫草素。结果 400 mg虫草素粗产品通过制备型HSCCC一次分离制备获得质量分数为98.7%的虫草素产品43.8 mg。结论 该法效率高，操作简单，为虫草素的大量制备提供了重要参考。

Objective To develop a new method for the isolation and preparation of cordycepin by high speed counter-current chromatography (HSCCC) with Cordyceps militaris fermentation broth as raw material. Methods The solvent systems for cordycepin separation were assessed and selected by HPLC partition coefficient method and analytical HSCCC. A solvent system that consisted of ethyl acetate-n-butanol-0.5% ammonia water (2∶3∶5) was applied to the separation. The upper phase was used as the stationary phase, while the lower phase was used as the mobile phase for the cordycepin separation by HSCCC. Results A high efficiency of HSCCC separation was achieved. Finally, cordycepin (43.8 mg) was obtained from 400 mg crude extract of C. militaris fermentation broth in one-step separation with a purity of 98.7%. Conclusion HSCCC is an efficient and simple method for the large- scale preparation of cordycepin from C. militaris fermentation broth.

国家科技支撑计划（2012BAD33B11）；湖南省教育厅重点项目（11A048）；湖南省研究生科研创新项目（CX2011B305）