Abstract: A method was developed for simultaneous qualitative and quantitative analysis of five metabolites of nitrofuran antibiotics, including 3-amino-2-oxazolidinone (AOZ), 5-morpholino-methyl-3-amino-2-oxazolidinone (AMOZ), semicarbazide (SEM), 1-aminohydantoin (AHD) and 3,5-dinitrosalicylic acid hydrazide (DNSH) in aquatic products by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The samples were hydrolyzed with HCl, and derivatized with 2-nitrobenzaldehyde at 37 ℃ for 16 hours. The derivative solutions were adjusted to pH 7.0-7.5, and the analytes were extracted by ethyl acetate. The separation was based on Thermo Aquasil C18 column (150 mm×4.6 mm, 3.0 μm). The analytes were detected by tandem mass spectrometry with electrospray ionization source with multiple reaction monitoring (MRM) mode. The developed method showed good linear correlation between the peak area ratios of the analyte and the internal standard and the concentration of the analyte with the correlation coefficients all above 0.99 over the dynamic range of 0.5-10 μg/kg. The limits of quantitation (LOQs) of AOZ, AMOZ, SEM, AHD and DNSH were 0.5 μg/kg. The average recoveries of all the compounds at four spiked levels of 0.5, 1.0, 2.0 and 4.0 μg/kg ranged from 81.3% to 100.5% with the RSDs between 3.4% and 10.0% (n=6). The method is proved to be fast and effective for simultaneous qualitative and quantitative analysis of the metabolites of the nitrofuran antibiotics in aquatic products.

Key words: aquatic products, metabolites, nitrofuran, liquid chromatography-tandem mass spectrometry (LC-MS/MS)