Optimization and validation of an enzyme immunoassay for the insect growth regulator fenoxycarb |
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Authors: | András Székács Hong TM Le Bruce D Hammock |
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Institution: | a Plant Protection Institute, Hungarian Academy of Sciences, Herman Otto ut 15, P.O. Box 102, H-1525 Budapest, Hungary b Department of Entomology and Cancer Research Center, University of California, 1 Shields Avenue, Davis, CA 95616, USA |
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Abstract: | A competitive enzyme-linked immunosorbent assay (ELISA) was developed for the quantitative detection of the insect growth regulator fenoxycarb. Polyclonal rabbit antisera, raised against protein conjugates of four haptenic derivatives of fenoxycarb, were utilized in immobilized antigen-based, competitive immunoassays. With ELISA systems that were both hapten- and carrier-heterologous, most antiserum titers fell in the range of 1:1000-1:30,000. Assay conditions, including concentrations of antisera and coating antigens, were optimized. The effect of pH, organic solvents, and various blocking agents was also investigated. A hapten-homologous and two hapten-heterologous indirect ELISAs allowed fenoxycarb determination in the range of 0.1-85 ng ml−1 with apparent IC50 values of 1.2-2.8 ng ml−1. Cross-reactivities with a number of compounds (e.g. pesticides of related structure, hapten synthesis intermediates, fenoxycarb metabolite, photodegradation products) were determined, and the assay proved highly selective for fenoxycarb. In particular, no significant interference was found with selected pyrethroid and juvenile hormone analog insecticides, phenoxyacetic acid herbicides, and photodegradation products of fenoxycarb. Using spiked water samples, assay performance was validated by SPME/GC-MS. |
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Keywords: | Fenoxycarb ELISA Immunoassay optimization Characterization and validation GC-MS SPE SPME |
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