Abstract
An ultrasensitive indirect competitive enzyme-linked immunosorbent assay (ic ELISA) using monoclonal antibodies (mAbs) was developed for the specific detection of diethylstilbestrol (DES) residues. To establish an ELISA based on mAbs, hapten diethylstilbestrol mono-carboxypropyl-ether (DES-MCPE) was chemically synthetized and then conjugated to bovine serum albumin (BSA) for immunization in mice. This ic ELISA was further optimized for DES determination. The sensitivity of the ic ELISA was found to be 0.49 μg/kg and the limit of detection was 0.075 μg/kg. DES residues in salmon meat and pork were tested with the recovery range from 74.0 to 85.2% and the coefficient of variation (CV) was less than 10%. Parallel analysis of DES samples from salmon meat showed comparable results from the ic ELISA with high-performance liquid chromatography. The ic ELISA provides a useful screening method for the quantitative detection of DES residues in animal-derived food.
Highlights
Specific and sensitive methods for detecting DES have been developed
We described the development of an ic enzyme-linked immunosorbent assay (ELISA) for the rapid detection of DES residues in animal muscle tissues using a monoclonal antibodies (mAbs) 4C7, specific for DES
Fifty-four milligrams of DES was dissolved in 2 mL of anhydrous dimethyl sulfoxide (DMSO), 27.5 mg of potassium carbonate was added, and the mixture was stirred at room temperature for 1.5 h in the dark
Summary
Classical methods for the determination of DES include high-performance liquid chromatography (HPLC)[10], ultra-HPLC tandem mass spectrometry (UHPLC- MS/MS)[11,12], and gas chromatography tandem mass spectrometry (GC-MS/MS)[13,14]. These methods are highly accurate, they do require extensive sample preparation, expensive instruments, and professionals to operate. The immunoassay is suitable for the screening of large numbers of animal-derived food samples
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