酶联免疫分析方法测定动物性食品中西马特罗残留

Determination of cimaterol residue in animal-orgin foods by enzyme-linked immunosorbent assay

中文关键词:  动物性食品  酶联免疫分析方法  西马特罗

英文关键词:animal-orgin foods  enzyme-linked immunosorbent assay  cimaterol

中文摘要:

      目的 建立检测西马特罗药物残留的酶联免疫分析方法(enzyme-linked immunosorbent assay, ELISA)。方法 对主要影响因素, 包括包被原稀释倍数、抗体稀释倍数、竞争时间、标准品稀释液pH等参数进行优化, 对样品进行前处理, 经提取净化后进行检测, 并对ELISA检测结果与仪器方法进行对比。结果 西马特罗ELISA方法的最佳反应条件为: 抗原稀释倍数为100000, 抗体稀释倍数为150000, 标品稀释液pH为7.5, 竞争反应时间为20 min。在最佳反应条件下, 绘制标准曲线, 西马特罗的IC50为1.32 μg/L, 线性范围为0.16~6.95 μg/L, 检出限为0.09 μg/L。西马特罗在0.16、0.32、1.6 μg/kg添加水平的回收率分别为80.31%~113.81%, 变异系数低于10%, 且实际样品的测定结果与超高效液相串联质谱法的相关性良好(r2=0.9962)。结论 本研究建立的ELISA方法具有较高的灵敏性, 适合西马特罗残留检测。

英文摘要:

      Objective To establish a method for the detection of cimaterol drug residues by enzyme-linked immunosorbent assay (ELISA). Methods The main influencing factors, including coating original dilution multiple, antibody dilution multiple, competition time, pH of standard diluent and other parameters were optimized. The samples were pretreated, extracted and purified, and detected. The ELISA test results were compared with the instrument method. Results The optimal reaction conditions for the detection of cimaterol by ELISA method were as follows: dilution ratio of antigen was 100000, dilution ratio of antibody was 150000, pH of standard diluent was 7.5, and competitive reaction time was 20 min. Under the optimal reaction conditions, the standard curve was drawn. The IC50 of cimaterol was 1.32 μg/L, the linear range was 0.16?6.95 μg/L, and the limit of detection was 0.09 μg/L. The recoveries of cimaterol at the addition levels of 0.16, 0.32, and 1.6 μg/kg were 80.31%?113.81%, respectively, and the coefficient of variation was less than 10%. The correlation between the results of the actual samples and UPLC-MS/MS was good (r2=0.9962). Conclusion The ELISA method established in this study has high sensitivity and is suitable for the detection of cimaterol residues.