Abstract:

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.

摘要：

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。