荧光定量免疫层析法测定鸡肉和牛奶中的氯霉素

Determination of chloramphenicol in chicken and milk by fluorescence quantitative immunochromatography

目的建立一种荧光定量免疫层析法测定动物源性食品鸡肉与牛奶中氯霉素残留的分析方法。方法将氯霉素鼠单克隆抗体与羧基化铕微球进行偶联标记。经过荧光微球活化、探针偶联等一系列条件优化确定最佳反应体系条件,根据纸条测试线(T线)信号值与质控线(C线)信号值的比值(T/C)和样本中氯霉素浓度建立定量标准曲线。针对鸡肉和牛奶样品,对荧光免疫层析检测方法的灵敏度、准确度和精密度等进行评价,并与液相色谱-串联质谱法(liquid chromatography-tandem mass spectrometry,LC-MS/MS)标准仪器分析方法进行比较。结果本研究建立的荧光免疫层析检测方法对鸡肉和牛奶中氯霉素残留的检出限分别为0.041μg/kg和0.040μg/kg,定量限分别为0.079和0.100μg/kg,添加回收率在76.26%~112.92%之间,变异系数均小于15%。结论本研究建立的荧光定量免疫层析方法灵敏度高、稳定性好,且与使用仪器方法得到的检测结果一致性较高,适用于鸡肉和牛奶中氯霉素残留的现场快速检测。

Objective To establish a method for the detection of chloramphenicol residues in chicken and milk by fluorescence quantitative immunochromatography.Methods Anti-chloramphenicol monoclonal antibody was conjugated to carboxylated europium microspheres.After a series of conditions of fluorescent microspheres activation and fluorescent probes coupling were optimized,the optimal reaction system conditions were determined.The quantitative standard curve was established according to the ratio(T/C)of the signal value of the test line(T line)to the signal value of the quality control line(C line)and the concentration of chloramphenicol in the sample.For chicken and milk samples,the sensitivity,accuracy and precision of the fluorescence immunochromatography method were evaluated,and compared with the liquid chromatography-tandem mass spectrometry(LC-MS/MS)standard method.Results The limits of detection of the fluorescence immunochromatography method for chloramphenicol residues in chicken and milk were 0.041μg/kg and 0.040μg/kg,respectively,and the limits of quantification were 0.079μg/kg and 0.100μg/kg,respectively.The recoveries were 76.26%‒112.92%,and the coefficients of variation were all less than 15%.Conclusion The fluorescence quantitative immunochromatography method established in this study has high sensitivity,good stability,and high consistency with detection results obtained by instrument method.It is suitable for on-site rapid detection of chloramphenicol residues in chicken and milk.