Development Of A Biomimetic Enzyme-Linked Immunosorbent Assay For The Determination Of Tribenuron-Methyl

The wide use of pesticides is harmful to human health, and more strict demand of maximum residue limit of pesticide needs more sensitive determination methods. In this paper, we used molecular imprinting technique in immunoassay, and developed a biomimetic enzyme-linked immunosorbent assay for the detection of trace tribenuron-methyl (TBM) in environmental samples.With TBM as the template molecule, methacrylic acid as the functional monomer, ethylene glycol dimethacrylate as the cross-linker, a highly selective imprinted membrane was synthesized on the well surface of MaxiSorp polystyrene96-well plate by bulk polymerization method. And it was characterized by fourier transform infrared (FT-IR), static, and kinetic adsorption experiments after optimalized the synthesis conditions. The results showed that the molecularly imprinted membrane exhibit higher adsorption capacity and specificity. In the same conditions, the saturated adsorption capacity of the imprinted film was higher than that of the non-imprinted film (1.28times).64.8%of the saturated binding capacity was obtained within a short period of10min,96.2%of the saturated binding capacity was achieved within40min, and the adsorption equilibrium was reached within only60min.A direct competitive biomimetic enzyme-linked immunosorbent assay was developed using the molecularly imprinted film as artificial antibody. Under the optimum assay conditions, the sensitivity expressed by IC50and the limit of detection expressed by IC15was found to be19.73μg L-1and0.30μg L-1, respectively. The cross-reactivity (CR) with three sulfonylurea herbicides ranged from3.17to13.10%, which proved that the developed method had a good selectivity.The method was successfully applied to the determination of TBM in spiked environmental water and soil samples with recoveries from93.6to96.8%and87.5to97.5%, respectively. The accuracy of the results achieved by using ELISA method was validated by comparison of the results obtained by the method of traditional HPLC. Correlation coefficient (r2) obtained by ELISA and HPLC was0.9923for river water and0.9877for soil. The results indicated that the developed method was suitable for the fast detection of trace TBM in environmental samples.