A Chemiluminescent Magnetic Enzyme Immunoassay Method for 2 Triazole Pesticide Detection in Wheat

We developed an alkaline phosphatase (AP) chemiluminescence immunoassay method by combining the superparamagnetic magnetic beads and the biotin–streptavidin signal amplification system to detect the triazolone and tebuconazole in wheat. Through optimization of the extraction solution and extraction time, acetonitrile–PBS was selected as the extraction solution with an extraction time of 5 min as the optimal pretreatment condition. Optimizing the dilution ratio of antigen antibodies, the optimal detection conditions were selected as the dilution ratios of 1:8000 and 1:20,000 for the triazolone monoclonal antibody solution and biotinylated triazolone solution, and 1:4000 and 1:20,000 for the tebuconazole monoclonal antibody solution and biotinylated tebuconazole solution, respectively. Under the optimal conditions, the method demonstrated that the limits of detection (LOD) of triazolone and tebuconazole were 0.002835 μg·mL−1 and 0.00064 μg·mL−1, respectively. The recovery rate was between 90.1% and 103.6%, and the relative standard deviation (RSD) was lower than 10%. The cross-reaction rates for structural analogs were all less than 0.1%, showing good specificity. In actual sample detection, this method did not detect triazolone and tebuconazole, and the results were consistent with UHPLC-MS/MS.