Monoclonal antibodies (mAb) against the main photo-addition products of chlorothalonil with ole®nic compounds of plant cuticles were produced. An indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed for the detection of free and bound chlorothalonil and its derivatives. For the characterization of the binding properties of the mAb, derivatives of chlorothalonil (simulating structures of cuticle bound residues) were synthesized. The cross-reactivities of these products were determined by ELISA. The test system was employed to detect bound residues of chlorothalonil in enzymatically isolated tomato cuticles, which had been spiked with methanolic solutions of the compound, irradiated by simulated sunlight and extracted. The use of isolated cuticles allows work to be carried out with authentic material without disturbance by metabolic processes.
An indirect competitive enzyme-linked immunosorbent assay (ELISA) was used to determine photochemically cutin-bound residues of chlorothalonil in enzymatically isolated tomato and apple fruit cuticles. The samples were spiked, irradiated, exhaustively extracted, and depolymerized with boron trifluoride complex resulting in a soluble depolymerisate. With this procedure, the ELISA could be calibrated with free target molecules for the quantification of originally bound chlorothalonil residues. In fruit cuticles that were irradiated for 8 h by simulated sunlight, 0.030 and 0.068 mg/g photoinduced cutin-bound residues of wax-free cuticles (calculated as chlorothalonil) were determined for tomatoes and apples, respectively. For the used antibody mAb chl. 4/11, cross-reactivities with derivatives of chlorothalonil simulating different types of cuticle-bound residues are given and discussed.
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