高速逆流色谱分离制备蛹虫草中虫草素和N6-(2-羟乙基)-腺苷

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1.5mL/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。

Isolation and purification of cordycepin and N6-(2-hydroxyethyl)-adenosine from Cordyceps militaris fruit bodies by high speed counter-current chromatography

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 1.5mL/min. The detection wave lengh was set at 254nm. A high efficiency of separation and purification by HSCCC was achieved. The target compounds, cordycepin (10.8mg) with a purity of 99% and N⁶-(2-hydroxyethyl)-adenosine (6.1mg) with a purity of 98%, were obtained from 200mg 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.