半定量法检测结核DNA疫苗中SDS残留量的方法学验证

Validation of Semi-quantitative Method for Determination of SDS Residues in TUBERCULOSIS DNA Vaccine

目的建立半定量法检测十二烷基硫酸钠(sodium dodecyl sulfate,SDS)残留量并进行方法学验证,用于结核DNA疫苗的质量控制。方法样本中残留的SDS与吖啶橙形成复合物,用Spectra Max 340PC微孔板检测器测定该复合物的吸光度。通过定量法测定SDS的残留量时,样品的吸光度不在标准曲线线性范围内,采用对比供试品与对照溶液的吸光度的半定量法,判断供试品中SDS的残留量。结果该方法回收率范围为102%~105%,不同人员、不同时间测定结果一致,该法受辅料、测试温度、萃取时间的影响较小,测定的3批供试品SDS残留量均小于0.02%。结论建立的方法准确度、中间精密度、专属性和耐用性较好,可用于结核DNA疫苗中SDS残留量的快速检测。

OBJECTIVE To establish a semi-quantitative method for the determination of SDS residues and verify the method for the quality control of TUBERCULOSIS DNA vaccine.METHODS A Spectra Max 340PC microporous plate detector was used to determine the absorbance of the complex formed by residual SDS and acridine orange.The absorbance of the sample is not within the linear range of standard curve when the SDS residue is determined by quantitative method.The semi-quantitative method is used to judge the SDS residue in the tested sample by comparing the absorbance of the tested sample and the control solution.RESULTS The recovery range of the method was 102%~105%,and the results were consistent with those of different people and different time.The method was less affected by excipients,test temperature and extraction time,and the SDS residues of the three batches of tested products were all less than 0.02%.CONCLUSION The established method has good accuracy,intermediate precision,specificity and durability,and can be used for rapid determination of SDS residues in TUBERCULOSIS DNA vaccine.