Development of a sensitive and rapid fluorescence polarization immunoassay for high throughput screening eight glucocorticoids in beef

Abstract

In this study, a sensitive, rapid, homogeneous, and high-throughput fluorescence polarization immunoassay (FPIA) for the rapid screening of eight glucocorticoids (GCs) in beef samples was successfully established. Two tracers including 5-aminofluorescein-labeled dexamethasone (5-AF-DMS) and fluorescein isothiocyanate-labeled dexamethasone (FITC-DMS) were studied to select appropriate antibody-tracer pairs using four previously produced broad-specific monoclonal antibodies. An optimal combination of the antibody 12D9 and the tracer FITC-DMS was selected. Under optimal detection conditions, the half inhibitory concentrations of dexamethasone (DMS), betamethasone (BMS), prednisolone (PNS), hydrocortisone (HCS), beclomethasone (BCMS), cortisone (CS), 6-α-methylprednisone (6-α-MPNS), fludrocortisone acetate (HFCS) were 1.00, 2.17, 3.49, 12.45, 1.20, 5.66, 6.85 and 3.45 ng/mL, respectively. The average recoveries of the proposed method in beef samples ranged from 77.3-91.7% with the coefficient of variation less than 12%. The developed FPIA was time-saving that could be completed within 10 min. The FPIA was applied to beef samples and showed a good correlation with liquid chromatography-tandem mass spectrometry (LC-MS/MS) (R² = 0.9894). Thus, the proposed method provides a rapid, reliable, sensitive, and high-throughput screening tool for the simultaneous screening of eight GCs in beef, which shows great potential in the food safety analysis.

Keywords: Beef; Fluorescence polarization immunoassay; Glucocorticoids; High-throughput.