DEVELOPMENT AND VALIDATION OF A LIQUID CHROMATOGRAPHY TANDEM MASS SPECTROMETRY METHOD FOR THE ANALYSIS OF CHLORAMPHENICOL IN HONEY

Chloramphenicol (CAP) is a broad spectrum antibiotic with excellent antibacterial and pharmacokinetic properties. Because of its side effects (e.g. aplastic anemia and hypersensitivity) the European union banned its use in food producing animals. The aim of this study was to develop and validate method for qualification and quantification of CAP in honey based on liquid chromatography – tandem mass spectrometry with negative electrospray ionisation. The target antibiotic was separated using reversed-phase liquid chromatography on chromatographic column Zorbax SB C18 (150 mm × 2.1 mm, 3.5 μm) with a gradient elution using acetonitrile – 0.1% formic acid mobile phase at a flow rate of 0.3 mL min-1, with column temperature 35 oC for CAP and 5D-CAP as internal standard. Homogenised honey samples were diluted with acetate buffer solution at pH 6 and extracted on Oasis HLB sorbens. Recoveries for real (acacia, chestnut, linden and flower) honey samples (n = 36) were 102% with RSD 8.4%. Method quantification limit for CAP was 0.3 µg kg-1. For detection and quantitative determination selected reaction monitoring was used. The method was applied for analyses of honey samples collected from whole Croatian area.