赖脯胰岛素制剂中四环素残留检测研究

Detection of tetracycline residue in insulin preparation of lispro

目的:将酶联免疫法应用于重组赖脯胰岛素制剂中检测四环素残留。方法:本研究采用竞争性酶联免疫ELISA方法测定四环素含量,以赖脯胰岛素不同剂型(R型、25R型、50R型)为研究对象,建立了四环素残留检测方法,进行了准确度、精密度等各项参数的方法学验证,并对赖脯胰岛素制剂中的四环素残留进行了检测。结果:质控品回收率均在80%~120%范围内,且相对标准偏差(RSD)<9%;精蛋白赖脯胰岛素混合注射液50R、精蛋白赖脯胰岛素混合注射液25R、赖脯胰岛素注射液四环素残留量分别为0.37,0.24,0.22 ppm;精蛋白锌重组赖脯胰岛素混合注射液50R、精蛋白锌重组赖脯胰岛素混合注射液25R、赖脯胰岛素注射液加标回收率分别为95%,83%,113%,均在80%~120%范围内;使用四环素国家标准品进行准确度分析,回收率均在80%~120%范围内,该方法准确度较好;对赖脯胰岛素注射液进行高、中、低加标浓度回收率测定,回收率均在80%~120%范围内,RSD<15%;使用四环素国家标准品进行重复性验证,RSD≤9.5%。结论:该方法操作简便、准确度好、重复性好、灵敏度高,可以满足四环素残留检测需求,可用于系列赖脯胰岛素及其他重组人胰岛素类似物制剂中的四环素工艺残留检测。

Objective:An enzyme-linked immunosorbent assay was applied to the detection of tetracycline residues in recombinant insulin lispro preparations.Methods:A competitive enzyme-linked immunosorbent assay(ELISA)was used to determine the content of tetracycline in different dosage forms of insulin lispro(R,25 R,50 R),by which a method of detection of tetracycline residues was established.Key parameters of methodology such as accuracy and precision were validated and tetracycline residues in insulin lispro formulations were detected.Results:The recovery rate of the quality control products was in the range of 80%to 120%,and the relative standard deviation(RSD)was less than 9%.For protamine lispro insulin mixture injection 50 R,protamine lispro insulin mixture injection 25 R,and insulin lispro injection,the detection results of the residual tetracycline amount were 0.37,0.24,and 0.22 ppm,respectively.The detection results of the spiked recovery rates of protamine zinc recombinant insulin lispro mixed injection 50 R,protamine zinc recombinant insulin lispro mixed injection 25 R,and insulin lispro injection were 95%,83%,and 113%,respectively,within the range of 80%to 120%.Using the national standard of tetracycline to analyze the accuracy,the recovery rate was in the range of 80%to 120%,and the accuracy of this method was good.The recovery rates of high,medium and low standard additions were determined for insulin lispro injection,and the recoveries were all within the range of 80%to 120%,and the RSD was less than 15%.Using tetracycline national standard for repeatability verification,RSD≤9.5%was obtained.It shows that the method has good repeatability and high precision.Conclusion:The method has the advantages of simple operation,good accuracy,good repeatability and high sensitivity,meeting the detection requirements of tetracycline residues.It can be used for the detection of tetracycline residues in a series of insulin lispro and other recombinant human insulin analog preparations.