大孔吸附树脂AB-8和十八烷基硅胶色谱分离纯化蛹虫草发酵液中虫草素工艺研究  

Purification of cordycepin from fermentation broth of Cordyceps militaris by use of macroporous resin AB-8 and octadecyl bonded silica chromatography

作　　者：张虎成[1] 范海涛[1] 王晓杰[1] 张征田[2] 杨军[1] 杨国伟[1] 王维彬[1] 李佳[1] 丁乾[1] 李华鑫[1] 王策[1] 

机构地区：[1]北京电子科技职业学院生物工程学院,北京100090 [2]南阳师范学院生命科学与技术学院,河南南阳473000

出　　处：《菌物学报》2015年第3期490-498,共9页Mycosystema

基　　金：北京市教委面上项目(SQKM201210858002)

摘　　要：主要研究了从蛹虫草发酵液中提取虫草素的工艺,确定了大孔吸附树脂AB-8及十八烷基键合硅胶反相层析柱对蛹虫草发酵液中虫草素的分离条件;吸附最佳工艺条件为上样液p H6,上样浓度0.4mg/m L,上样量5BV,吸附流速1.0BV/h;解吸最佳工艺条件包括解吸液20%乙醇,解吸体积15BV,解吸流速4BV/h。得到的解吸液进一步以十八烷基键合硅胶分离,条件为上样液p H6,梯度洗脱,流动相为p H6的水和p H6的乙醇。洗脱液通过结晶和重结晶得到精制虫草素。三步得到的虫草素纯度分别达到40%、90%和99%以上。The purification technique of cordycepin from the fermentation broth of Cordyceps militoris was investigated Cordycepin was adsorbed and desorbed by using macroporous resin AB-8 column chromatography and purification of cordycepin was carried out by reversed-phase chromatography. When pH value was 6.0 and concentration of cordycepin was 0.4mg/mL, the AB-8 column had a good adsorption capacity for sample loading amount of 5BV at 1.0BV/h loading flow rate Using 20% ethanol as desorption solution, elution flow rate of 4BV/h and elution amount of 15BV were optimal for AB-8columndesorption. The elution sample obtained was then loaded onto reversed-phase chromatography with octadecyl bonded （ODS） as stationary phase and ethanol （95%, pH6.0）-acetic acid solution （pH6.0） as mobile phase. Gradient elution with 95% ethanol was carried out in 40min at crystallized and recrystallized. Under these a flow rate of three processes, 1.0mL/min. The elution solution containing cordycepin was the purity of cordycepin obtained could reach 40%, 90% and silica 5% to then 99%,respectively.

关 键 词：虫草素 大孔吸附 十八烷基键合硅胶 纯化 蛹虫草发酵液 

分 类 号：TQ461[化学工程—制药化工]