高速逆流色谱分离制备蛹虫草中虫草素和N^6-(2-羟乙基)-腺苷

Isolation and purification of cordycepin and N^6-(2-hydroxyethyl)-adenosine from Cordyceps militaris fruit bodies by high speed counter-current chromatography

采用高速逆流色谱(HSCCC)技术从蛹虫草子实体粗提物中分离制备高纯度虫草素和N6-(2-羟乙基)-腺苷。利用高效液相色谱(HPLC)测定目标产物在溶剂体系中的分配系数,优化HSCCC分离虫草素和N6-(2-羟乙基)-腺苷的溶剂体系,确定了以乙酸乙酯-正丁醇-1.5%氨水(1:4:5,V/V/V)为HSCCC的两相溶剂体系,并运用此溶剂体系,上相为固定相,下相为流动相,主机转速850r/min,流动相流速为1.5m L/min,检测波长为254nm条件下进行分离制备,在250min内从200mg蛹虫草子实体粗提物中一步分离得到10.8mg纯度99%的虫草素和6.1mg纯度98%的N6-(2-羟乙基)-腺苷。该方法简便、快速,为虫草素和N6-(2-羟乙基)-腺苷的大量制备建立了基础。

Cordycepin and N6-（2-hydroxyethyl）-adenosine were separated and purified from Cordyceps militaris fruit bodies by high speed couter-current chromatogrphy（HSCCC）. The partition coefficient was assessed by HPLC method to select the suitable solvent systems. A solvent system that consisted of ethyl acetate, n-butanol, 1.5% ammonia in water（1：4：5, V/V/V） was applied to the separation system. The upper phase was used as the stationary phase, while the lower phase was the mobile phase. The apparatus was rotated at 850r/min and the flow rate was operated at 1.5m L/min. The detection wave lengh was set at 254 nm. A high efficiency of separation and purification by HSCCC was achieved. The target compounds, cordycepin（10.8mg） with a purityof 99% and N6-（2-hydroxyethyl）-adenosine（6.1mg） with a purity of 98%, were obtained from 200 mg crude sample of Cordyceps militaris fruit bodies in one-step separation. The established method is simple and suitable for large-scale separation of cordycepin and N6-（2-hydroxyethyl）-adenosine from C. militaris fruit bodies, which could provide the material basis for the pharmacological research of cordycepin and N6-（2-hydroxyethyl）-adenosine.