ELISA法与LC-MS/MS法测定动物组织中克仑特罗残留

目的:比较酶联免疫法(ELISA)和液相色谱-串联质谱法(LC-MS/MS)测定动物组织中克仑特罗残留量的准确度、精密度及检测限。方法:样品经过处理后,分别采用ELISA法和LC-MS/MS法进行测定;用ELISA法对组织样品进行初筛,测出阳性样品利用LC-MS/MS法进行确证。结果:应用ELISA法测定样品猪肉样品中克仑特罗的回收率为67.0%～99.6%,批内变异系数3.4%～8.7%,批间变异系数6.1%～9.6%,灵敏度为0.025μg/L,最低检测限0.025μg/kg;LC-MS/MS检测方法回收率88.62%～111.43%,批内变异系数4.4%～7.4%,最低检测限0.5μg/kg;用ELISA法对50份猪肉和50份猪肝样品进行检测,筛选出3个阳性样品,经LC-MS/MS确证为阳性,两种方法检测结果一致。结论:ELISA法灵敏度和准确度较高,样品处理方法简单,成本低,适合组织中克仑特罗残留大规模筛查;LC-MS/MS准确度高,适合于阳性样品精确定量。

Enzyme-linked Immunosorbent Assay (ELISA) and LC-MS/MS for Determination of Clenbuterol Residue in Animal Tissue

Objectives： To compare the precision, accuracy and limit of detection of enzyme-linked immunosorbent assay （ELISA）and liquid chromatography tandem mass spectrometry （LC-MS/MS） in determining clenbuterol residue in animal tissue. Methods： Samples were prepared and then determined by ELISA and LC-MS/MS, respectively. LC-MS/MS was used to confirm positive samples from ELISA analysis. Results： The ELISA recoveries across three spike levels, intra-batch and inter-batch coefficients of variation, sensitivity and limit of detection were 67.0% -- 99.6%, 3.4% -- 8.7%, 6.1% -- 9.6%, 0.025μg/L and 0.025 μg/kg, respectively. For LC-MS/MS, the recoveries across three spike levels were 88.62% -- 111.43%, the intra-batch coefficients of variation 4.4% -- 7.4%, and the limit of detection 0.5 μg/kg. Three positive samples were screened out of 50 pork samples and 50 pig＇s liver samples using ELISA, which was consistent with the results from LC-MS/MS analysis. Conclusion： ELISA is sensitive and accuracy and needs easy sample preparation and low cost, thus being suitable for large-scale screening of clenbuterol residue in animal tissue. In contrast, LC-MS/MS has higher accuracy and can therefore be used to quantify positive samples.