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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 investigate the applicability of HPLC analysis in the identification of recombinant human insulin analogs and detection of impurities. METHODS: A reversed phase column (C18) was used with a mobile phase of 0 2 mol·L-1, pH 2 3 and acetonitrile at a flow rate of 0 8 or 10 mL · min-1 with a column temperature of 40 ℃, the detection wavelength was 2 14nm; the mobile phase was 50mL · min-1 with a molecular sieve column (Zorbax GF250) with ammonium phosphate buffer (0 1mol·L-1, pH7 5) Detection of recombinant human insulin analogs and their enzymatic fragments. Results: RP-HPLC, insulin and their analogues were obtained. Peptide mapping and high-performance size exclusion chromatography (HPSEC) images were obtained. Conclusion: This method can effectively distinguish and detect insulin analogs and related substances, and provide a scientific basis for the quality control of these drugs.