Development of an indirect competitive enzyme-linked immunosorbent assay for propiconazole based on monoclonal antibody

Propiconazole is a widely used fungicide in agricultural production, but it poses risks to human health. Here, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on a monoclonal antibody (mAb) was established to detect propiconazole residues. Three haptens of propiconazole were designed and synthesized to prepare antigens. Ten mAbs were obtained using hapten B (5-(4-(2-((1H-1,2,4-triazol-1-yl)methyl)-4-propyl-1,3-dioxolan-2-yl)-3-chlorophenyl) pentanoic acid). The ic-ELISA based on the combination of mAb 2G2E12 and hapten B-ovalbumin (coating antigen) showed the highest sensitivity. After optimization, the 50% inhibition concentration, limit of detection and linear range of the ic-ELISA were 2.33 μg/L, 0.26 μg/L and 0.26–21.28 μg/L, respectively. The cross-reactivities for the analogs of propiconazole were ≤1.9%. The average recoveries of the spiked samples were 75.7%–117.0% with relative standard deviations of ≤9.4%. The results of the ic-ELISA were consistent with those of gas chromatography-mass spectrometry in the analysis of blind samples. Therefore, we provide an alternative tool for the accurate and reliable determination of propiconazole. •Three haptens of propiconazole was designed and synthesized successfully.•Ten mAbs were prepared and the optimal combination of mAb and antigen was obtained.•An ic-ELISA for propiconazole with high sensitivity and specificity was developed.•The ic-ELISA was successfully used to detect propiconazole in agro-products.•The ic-ELISA had good correlation with GC-MS in the detection of propiconazole.