利用高速逆流色谱技术对化学酶法合成的内吗啡肽-1粗样进行分离。分析了内吗啡肽-1粗样杂质的分配系数,选择乙酸乙酯-甲醇-水(3∶1.5∶3,体积比)作为两相溶剂系统,上相为固定相,下相为流动相,在主机转速为900 r/min,流速为1.0 mL/min,检测波长为280 nm条件下分离制备。从120 mg粗样中分离得到71 mg样品,内吗啡肽-1的纯度达99.1%,样品回收率为91.5%。该方法简便、快速,为高纯度内吗啡肽-1少量制法提供了技术,并可为高速逆流分离小肽合成产物提供指导。
Endormophin-1 synthesized by chem-biological method was purified by high-speed counter-current chromatograph technology(HSCCC).The optimum separation conditions were as follows:A two-phase solvent system was ethyl acetate-methanol-water(3∶1.5∶3,v/v/v).The lower phase as the mobile phase was operated at a flow rate of 1.0 mL /min,while the apparatus was rotated at 900 r/min,and detection were length was at 280 nm.Each time 120 mg of the crude sample was loaded under these conditions,and 71 mg of endorphin-1 with the purity of 99.1% was obtained.The recovery was 91.5%.The method was relatively simple,fast,and applicable to the large-scale isolation and separation of endormophin-1.