An Immunoassay Based on the Antibody Specific for Herbcide Metolachlor
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Abstract:
An antibody-based ic-ELISA was established to detect the pesticide residue of metolachlor in food and reduce its potential harm to human. Metolachlor hapten was synthesized from herbicide metolachlor and 3-mercaptopropionic acid, and identified by 1H-NMR and mass spectrometry. The artificial antigen of metolachlor was prepared by coupling the hapten with carrier protein using EDC method. Antibody against metolachlor was raised by rabbit. The optimal dilution times of antigen and antibody were determined by chessboard titration. The antibody dilution solution for ELISA reaction was PBS (1% PEG6000), and the drug dilution solution was PBS (10% methanol). On this basis, the established ic-ELISA calibration curve for metolachlor was conducted by antibody, which exhibited IC50 value of 14.64ng/mL, detection limit (LOD) of 0.8 ng/mL and linear detection range (IC20-IC80) of 1.51~141.92 ng/mL. The cross-reactivity with 14 structural analogues was less than 3%. The recoveries of spiked samples were in the range of 86.60%~102.16% with RSD ranging from 7.05%~13.30%. This antibody with high sensitivity and specificity can be used to detect metolachlor in the food or environment.
