Development of sensitive high-performance liquid chromatography with fluorescence detection using 4-(4,5-diphenyl-1H-imidazol-2-yl)-benzoyl chloride as a labeling reagent for determination of bisphenol A in plasma samples

A sensitive HPLC method for determination of bisphenol A (BPA) in plasma samples using 4-(4,5-diphenyl-1H-imidazol-2-yl)benzoyl chloride (DEB-Cl) as a fluorescence labeling reagent was developed. The fluorescence labeling reaction was completed within 10 min at room temperature. DEB-Cl reacts with the phenolic hydroxyl group of BPA in the presence of triethylamine (TEA). The DIB-Cl derivative of BPA (DIB-BPA) was separated within 30 min with an ODS column using acetonitrile-water (90:10, v/v) as the isocratic eluent, Calibration graphs were linear over the range of 1.0-100 ng/ml (r=0.999). The detection limit of DIB-BPA was 0.05 ng/ml (2.5 pg) at a signal-to-noise ratio of 3. The relative standard deviations (RSDs) of the method for between-run were 1.0-5.0%. The analytical recoveries of known amounts (1.0 and 100 ng/ml) of BPA-spiked rabbit plasma were around 95%. (C) 2001 Elsevier Science B.V. All rights reserved.