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高速逆流色谱法分离纯化内吗啡肽-1

许旭旭1,毛 政2,吴 斌1,孙洪林1,何冰芳1   

  1. 1南京工业大学生物与制药工程学院,江苏 南京 210009;2浙江省食品药物监督管理局培训中心,浙江 杭州 310012
  • 出版日期:2011-09-05 发布日期:2011-09-05

Preparative isolation and purification of endormophin-1 byhigh-speed counter-current chromatography

XU Xuxu1,MAO Zheng2,WU Bin1,SUN Honglin1,HE Bingfang1   

  1. 1College of Biotechnology and Pharmaceutical Engineering,Nanjing University of Technology,Nanjing 210009,Jiangsu,China;2Zhejiang Food and Drug Administration Training Center,Hangzhou 310012,Zhejiang,China
  • Online:2011-09-05 Published:2011-09-05

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(31.53,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。

Abstract:

Endormophin-1 synthesized by chem-biological method was purified by high-speed counter-current chromatograph technologyHSCCC. The optimum separation conditions were as followsA two-phase solvent system was ethyl acetate-methanol-water31.53v/v/v. The lower phase as the mobile phase was operated at a flow rate of 1.0 mL /minwhile the apparatus was rotated at 900 r/minand detection were length was at 280 nm. Each time 120 mg of the crude sample was loaded under these conditionsand 71 mg of endorphin-1 with the purity of 99.1% was obtained. The recovery was 91.5%. The method was relatively simplefastand applicable to the large-scale isolation and separation of endormophin-1.

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