A simple, rapid and high-throughput fluorescence polarization immunoassay for simultaneous detection of organophosphorus pesticides in vegetable and environmental water samples

[Display omitted] ► A broad-specificity FPIA for organophosphorus pesticides (OPs) was developed. ► The effect of tracer structure and several physicochemical parameters were studied. ► The FPIA can detect 5 OPs simultaneously with a limit of detection below 10 ng mL −1. ► Recovery tests from vegetable and environmental water samples were studied. ► Good accuracy and reproducibility were obtained. A simple, rapid and high-throughput fluorescent polarization immunoassay (FPIA) for simultaneous determination of organophosphorus pesticides (OPs) using a broad-specificity monoclonal antibody was developed. The effects of tracer structure, tracer concentration, antibody dilution, methanol content and matrix effect on FPIA performance were studied. The FPIA can detect 5 OPs simultaneously with a limit of detection below 10 ng mL −1. The time required for the equilibrium of antibody–antigen interaction was less than 10 min. The recovery from spiked vegetable and environmental samples ranged from 71.3% to 126.8%, with the coefficient of variations ranging from 3.5% to 14.5%. The developed FPIA was applied to samples, followed by confirmation with high-performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS) analysis. The developed FPIA demonstrated good accuracy and reproducibility, and is suitable for rapid and high-throughput screening for OP contamination with high-efficiency and low cost.