Journal
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
Volume 50, Issue 23, Pages 6675-6682Publisher
AMER CHEMICAL SOC
DOI: 10.1021/jf025703+
Keywords
bromophos-ethyl; insecticide; immunoassay; ELISA
Funding
- NIEHS NIH HHS [P30ES05707, P42 ES04699] Funding Source: Medline
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A competitive enzyme-linked immunosorbent assay (ELISA) was developed for the quantitative detection of the organophosphorus insecticide bromophos-ethyl. Four bromophos-ethyl derivatives (haptens) were synthesized and were coupled to carrier proteins through the pesticide thiophosphate group to use as an immunogen or as a coating antigen. Rabbits were immunized with either one of two haptens coupled to bovine serum albumin for production of polyclonal antibodies, and the sera were screened against one of the haptens coupled to ovalbumin. Using the serum with highest titer, an antigen-coated ELISA was developed, which showed an IC50 of 3.9 ng/mL with a detection limit of 0.3 ng/mL (20% inhibition). An antibody-coated ELISA using an enzyme tracer was also developed, which showed an IC50 of 6.5 ng/mL with a detection limit of 1.0, ng/mL (20% inhibition). The antibodies, showed negligible cross-reactivity with other organophosphorus pesticides except with the insecticides bromophos-methyl and chlorpyrifos in the antibody-coated assay only. Recoveries of bromophosethyl from fortified crop and water samples ranged from 82 to 128% and from 95 to 17%; respectively.
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