Site-heterologous haptens and competitive monoclonal antibody-based immunoassays for pyrimethanil residue analysis in foodstuffs

Pyrimethanil is a widely used post-harvest fungicide whose residues are commonly found in fruits and vegetables. We herein report the synthesis of two new functionalized derivatives of pyrimethanil carrying an equivalent spacer arm at different positions of the molecule. The influence of the linker tethering site on the affinity of monoclonal antibodies was shown. Moreover, the development of competitive immunoassays in different formats and with high sensitivity to the target analyte - IC50 values below 0.3μg/L - is described. Optimized assays were characterized by the determination of the limit of quantification, trueness, and precision using water-diluted QuEChERS extracts of fruits and vegetables fortified with pyrimethanil. Finally, in-field treated and blind spiked strawberry, tomato, and cucumber samples were analyzed by the developed immunoassays and a reference chromatographic method. Using Deming regression analysis, we show that results from both methodologies were statistically comparable.This work was supported by the Spanish Ministerio de Ciencia e Innovación (AGL2009-12940-C02-01-02/ALI) and cofinanced by FEDER funds. J.V.M. and F.A.E.-T. were hired by the Consejo Superior de Investigaciones Científicas (CSIC), the former under a Ramón y Cajal contract and the latter under a JAE-doc contract, both of them financed by Ministerio de Ciencia e Innovación and the European Social Fund.Peer Reviewe