Solid-phase fluoroimmunoassay for the determination of mesotrione —a novel triketone herbicide— in water with direct measurement of the fluorescence onto the solid support

Abstract

A straightforward, low-cost fluoroimmunoassay (FIA) for the determination of the new triketone herbicide mesotrione has been developed and optimized. The protein–mesotrione conjugate, immobilized on white opaque microtitration wells competes with the mesotrione in the sample or standard for the limited binding sites of a liquid phase anti-mesotrione antibody. The assay is based on the measurement of fluorescence intensity directly onto the solid support, using a fluorescein labeled second antibody and a fluorescence plate reader. To stabilize and enhance the fluorescence signal a glycerine-based treatment of the microtitration wells was included in the protocol. The detection limit of the assay is 40 ng l⁻¹ (4 pg per well), the working range extends up to 9 µg l⁻¹, whereas the within and between run CVs are 0.7–4.2% and 2.1–5.5%, respectively. To evaluate the assay specificity, the cross-reactivities of two mesotrione metabolites: 4-methylsulfonyl-2-nitrobenzoic acid and 2-amino-4-methylsulfonyl-benzoic acid and several other compounds similar in structure to mesotrione such as: fomesafen, prosulfocarb, fluazinam, sulcotrione, 1,2-cyclohexanedione, 1,3-cyclohexanedione, 2-acetyl-1,3-cyclohexanedione were assessed. Most of the substances tested presented very low (<0.05%) cross-reactivity values with the exception of sulcotrione that cross-reacted 23% in the mesotrione assay. The assay was used for the determination of mesotrione in bottled natural waters fortified with the analyte and in a commercial herbicide formulation, namely CALLISTO™.