Development of an Accelerated Solvent Extraction Approach for Quantitative Analysis of Chloramphenicol, Thiamphenicol, Florfenicol, and Florfenicol Amine in Poultry Eggs

Abstract

This manuscript describes a simple, reliable, and sensitive approach that utilizes high-performance liquid chromatography-electrospray ionization tandem triple quadrupole mass spectrometry (HPLC-ESI/MS/MS) for the selective determination of chloramphenicol, thiamphenicol, florfenicol, and florfenicol amine in poultry eggs (hen eggs, duck eggs, and goose eggs). Samples were extracted with methanol-ammonium hydroxide-ultrapure water (97:2:1, v/v) via accelerated solvent extraction (ASE) at 80 °C and 1500 psi and defatted with hexane saturated with acetonitrile. The experimental method uses ESI (+) and ESI (−) combination mode, full-scan qualitative analysis, and multiple reaction monitoring (MRM) combined with an internal standard for quantitative analysis. This optimized method was validated according to the requirements defined by the European Union and the Food and Drug Administration. The poultry eggs were spiked with the target compounds at four levels, and the average recoveries were all higher than 88.3%, and the relative standard deviations did not exceed 3.9%. The limits of detection and limits of quantification were 0.04–0.5 μg/kg and 0.1–1.5 μg/kg, respectively. The decision limits were 0.37–102 μg/kg, and the detection capabilities were 0.44–103 μg/kg. Finally, the new approach was successfully applied to the quantitative determination of these analytes in 120 commercial poultry eggs from local supermarkets.