Magnetic beads-based immunoassay as a sensitive alternative for atrazine analysis |
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Authors: | Madalina Tudorache Anca Tencaliec Camelia Bala |
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Affiliation: | Laboratory of Quality Control and Process Monitoring, Department of Analytical Chemistry, University of Bucharest, 4-12 Regina Elisabeta, 030018 Bucharest, Romania |
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Abstract: | A new immunoassay strategy for sensitive atrazine determination based on magnetic beads is reported. The immuno-method is a competitive solid-phase immunoassay where the anti-atrazine antibody is immobilized on the magnetic beads surface and fixed at the reaction cell bottom using a simple magnet, which generates a magnetic field. Analyte and HRP (horseradish peroxidase) tracer compete for active sites of antibody. After the immunointeractions antibody-analyte and antibody-tracer, atrazine quantification from the sample is performed by injection of the chemiluminescence substrate (luminol, hydrogen peroxide and p-iodophenol). Different antibodies (polyIgG anti-atrazine Ab I and affinity purified polyIgG anti-atrazine AbI) were tested in this configuration. Also, optimum concentration of antibody-covered magnetic beads was set up (8 mg/l Ab II). Finally, the performance of magnetic beads-based immunoassay for atrazine determination was evaluated demonstrating that the magnetic beads-based immunoassay is one of the most sensitive method for atrazine determination (LoD = 3 pg/l, IC50 = 37 pg/l, DR = 10-1000 pg/l). |
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Keywords: | Immunoassay Magnetic beads Atrazine Pesticides Sensitivity Hook effect |
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