目的: 使用超高效液相色谱-四极杆飞行时间串联质谱法（UPLC-Q TOF MS/MS）对速效胰岛素制剂谷赖胰岛素注射液一级结构进行测定。方法: 使用二硫苏糖醇（DTT）和碘乙酰胺（IAM）对谷赖胰岛素注射液进行还原烷基化前处理,通过UPLC-Q TOF MS/MS分析其一级结构。氨基酸序列测定采用Waters ACQUITY UPLC CSH C18色谱柱（3 mm×150 mm,1.7 μm）,流动相为0.1%甲酸水溶液,流动相B为0.1%甲酸乙腈溶液,梯度洗脱,流速0.3 mL · min-1;采用电喷雾离子源正离子模式和MSE采集模式进行数据采集,通过优化质谱参数,对谷赖胰岛素注射液进行一级结构测定,同时考察该方法对精蛋白重组人胰岛素注射液的适用性。结果: 谷赖胰岛素A链与B链一级质量数准确,氨基酸序列覆盖率良好,通过b/y离子分析,A链和B链前5个强度高的b/y离子的离子强度RSD均在10.0%以内,专属性强,重复性好,对精蛋白重组人胰岛素注射液也具有良好的适用性。结论: 该方法操作简便快速、灵敏、准确,可用于谷赖胰岛素注射液及其他胰岛素类似物制剂的一级结构测定。
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