Multi-class determination of antimicrobials in meat by pressurized liquid extraction and liquid chromatography–tandem mass spectrometry

A multi-residue method using pressurized liquid extraction (PLE) and liquid chromatography–tandem mass spectrometry (LC–MS/MS) has been developed for determining trace levels of 31 antimicrobials, including β-lactams, lincosamides, macrolides, quinolones, sulfonamides, tetracyclines, nitroimidazoles and trimethoprim. The extraction method required pre-homogeneization of the meat with EDTA-washed sand and subsequent one-static-cycle extraction for 10 min with 40 ml of water at 1500 psi and 70 °C. The effect of operation temperature, pressure, flush volume, and static cycles on PLE performance was studied. Average recoveries ranged from 75 to 99% with relative standard deviations <18%. The method was validated according to the European Union requirements (2002/657/EC). In addition to the quality parameters included in that decision, the limits of detection (LODs) and quantification (LOQs) were determined. The use of LC–MS/MS provided LODs (between 3 and 15 μg kg−1) and LOQs (between 10 and 50 μg kg−1), by far lower than half of their maximum residue limits (MRLs) (between 50 and 1200 μg kg−1). Confirmation of the presence of any of the studied compounds was accomplished in 1 h after sample receipt. This methodology has been successfully applied to the analysis of cattle and pig tissue samples from local markets and slaughterhouses of the Valencian Community (Spain). The results showed the presence of some antimicrobials at different concentrations. Quinolones and tetracyclines were the antimicrobials most detected in cattle and pig samples, respectively. Sulfonamides were also frequently detected in both types of samples.