高效液相色谱法测定混合重组人胰岛素注射液中异天冬氨酸的含量  

Quantification of Isoaspartic Acid in the Mixture of Recombinant Human Insulin Injection by HPLC

作　　者：晏菊姣 王婧 张一唱 喻悦 周朝晖 李恒 YAN Jujiao;WANG Jing;ZHANG Yichang;YU Yue;ZHOU Chaohui;LI Heng(NMPA Key Laboratory for Quality Research and Control of Drug Products,Wuhan Institute for Drug and Medical Device Control,Wuhan 430075,China;Department of Chemistry and Chemical Biology,Barnett Institute of Chemical and Biological Analysis,Northeastern University,Boston,Massachusetts 02115,USA)

机构地区：[1]武汉药品医疗器械检验所,国家药品监督管理局药物制剂质量研究与控制重点实验室,武汉430075 [2]美国东北大学化学与生物化学系,巴奈特化学与生物分析研究所,波士顿02115

出　　处：《医药导报》2022年第2期231-234,共4页Herald of Medicine

基　　金：武汉市第五批“黄鹤英才计划”(2017);湖北省食品药品监督管理局科研项目(201801013)。

摘　　要：目的建立测定混合重组人胰岛素注射液中异天冬氨酸(isoAsp)含量的方法。方法采用ISOQUANT^(?)异天冬氨酸检测试剂盒结合高效液相色谱法(HPLC)测定,并对该方法进行加标回收率实验,对不同生产企业样品的isoAsp含量进行检测。结果S-腺苷高半胱氨酸(SAH,酶促反应产物)对照品进样量在5~75 pmol范围内与其峰面积积分值呈良好的线性关系(r=0.9993),3种浓度SAH的加标回收率在96%~116%;不同厂家不同批号样品中isoAsp含量为0.07%~0.31%。结论该文所建立的方法可作为此类胰岛素产品的isoAsp含量的常规检测方法,也为生产流通领域、新药研发领域生物技术药中isoAsp的含量检测提供研究基础。Objective To establish a method to determine the content of isoaspartic acid(isoAsp) in the mixture of recombinant human insulin injection.Methods The isoAsp content in the mixture of recombinant human insulin injection was determined by ISOQUANT^(?)(Isoaspartic Acid or isoAsp) Detection kit coupled with high performance liquid chromatography(HPLC).The method was verified by a standard recovery rate test,and used to quantify isoAsp in samples from different manufacturers.Results There was a good linear relationship between the injection amount of S-adenosyl-L-homocysteine(SAH,the enzymatic product from the IsoQuant assay) and its peak area integral in the range from 5 to 75 pmol(r=0.999 3).The recovery yields for the reference substances in three concentrations were all in the range from 96% to 116%.The percentages of isoAsp in samples from different manufacturers and batches ranged from 0.07% to 0.31%.Conclusion The quantification method for isoAsp is robust for the mixture of recombinant human insulin injection,and will also be useful for other peptide-and protein-pharmaceuticals in both quality control and development.

关 键 词：混合重组人胰岛素注射液 异天冬氨酸 色谱法 高效液相 

分 类 号：R977.6[医药卫生—药品] R927.2[医药卫生—药学]