重组人胰岛素类似物的HPLC分析

The Analysis of Insulin Lispro by HPLC

目的 :探讨HPLC分析在重组人胰岛素类似物鉴别和杂质检测中的适用性。方法 :采用反相色谱柱(C18) ,以硫酸钠 -磷酸缓冲液 (0 2mol·L-1,pH 2 3)和乙腈为流动相 ,流速 0 8或 1 0mL·min-1,柱温40℃ ,检测波长 2 14nm ;用分子筛柱 (ZorbaxGF2 5 0 )以磷酸铵缓冲液 (0 1mol·L-1,pH 7 5 )和乙腈为流动相 ,流速 0 5mL·min-1,检测波长 2 14nm ;检测重组人胰岛素类似物及其酶解片段。结果 :获得胰岛素及其类似物的RP -HPLC图谱 ,肽图谱和高效体积排阻色谱 (HPSEC)图。结论 :本文方法能够有效地区别和检测胰岛素类似物及有关物质 ,为该类药物的质控提供了科学依据。

Objective:To study the suitability of HPLC method for determination of impurities Insulin lispro as well as identity of Insulin and Insulin analogs.Methods:Chromatographic conditions included Vydac C 18 column and the mobile phase consisting of a mixture of acetonitrile-sulfate solution(0 02 mol·L -1 ,pH 2 3).Gradient program elute isocratically or elute linearly.Detection wavelength was at 214 nm.The flow rate was 0 8 or 1 mL·min -1 .The temperature of column was 40 ℃.The Zorbax GF250 column was used,mobile phase:Ammonium phosphate buffer-acetonitrile(65∶35),flow rate: 0 5 mL·min -1 .Results:The peptide mapping,RP-HPLC and high performance size exclusive chromatography(HPSEC) chromatograms of Insulin and Insulin analogs were obtained.Conclusion:These methods are found to be effective for identity of Insulin and Insulin analogs,suitable for determination of impurities Insulin lispro.This paper provides the scientific basis for the quality control of Insulin analogs products.