Abstract
A competitive enzyme-linked immunosorbent assay (ELISA) was developed for the quantitative detection of the diethylstilbesterol (DES). Polyclonal rabbit antisera, raised against protein conjugate diethylstilbesterol-mono-caroxyl-propyl-ethyl-bovine-serum-albumin (DES-MCPE-BSA), were utilized in immobilized antibody-based and competitive immunoassays. Assay conditions, including concentrations of antisera and horseradish peroxidase, (HRP)-DES, were optimized. The effects of incubation time, surfactant concentration, ionic strength and pH of the medium were also investigated. The typical calibration curve gave an average IC(50) value of 2.4 ng/mL, calibration range from 0.2 to 30.5 ng/mL and a detection limit of 0.07 ng/mL. The specificity of the assay was tested against DES structurally related compounds, and the assay proved highly selective for DES. Assay performance was validated using spiked chicken meat and liver tissue samples. Moreover, it was compared with liquid chromatography-tandem mass spectrometry. The ion pair for quantification of DES was m/z 267.4/251.4, and the linear equation of DES was y = 0.1033x + 0.0126 (r = 0.9960). The two analytical methods can be applied to monitor DES and other steroid residues in foods.
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