Evaluation of rapid and standard tandem mass spectrometric methods to analyse veterinary drugs and their metabolites in antemortem bodily fluids from food animals

ABSTRACT

Antemortem bodily fluids can serve as an indicator of veterinary medicine exposure prior to food animal slaughter. A multi-residue, rapid screen electrospray ionisation mass spectrometric (RS-ESI-MS) method was developed to analyse 10 veterinary drugs or metabolites (clenbuterol, erythromycin, flunixin, 5-hydroxyflunixin, meloxicam, ractopamine, ractopamine-glucuronide, salbutamol, tylosin, and zilpaterol) in hog oral fluid and bovine urine. Simple acetonitrile extraction with salting-out was employed to remove the analytes from matrices in less than 30 minutes. Instrumental analysis time was < 1 min/injection. Regression coefficients of matrix-matched calibration curves ranged 0.9743–0.9999 across all compounds with limits of detection ranging from 0.46–108 ng mL⁻¹ for cattle urine and 0.19–64.4 ng mL⁻¹ for hog oral fluid across all analytes. Except for ractopamine-glucuronide, analyte recoveries ranged from 92.7–106% for oral fluid and urine fortified at 30, 100, and 300 ng mL⁻¹, with inter-day variations of < 25%. Ractopamine-glucuronide recovery was 93.3% for oral fluid fortified at 300 ng mL⁻¹. The RS-ESI-MS method accurately identified ractopamine and/or ractopamine-glucuronide in incurred cattle urine with results correlating well with traditional LC-MS/MS and HPLC fluorescence methods. As far as we are aware, this is the first report of the direct quantification of ractopamine-glucuronide from biological matrices without lengthy hydrolysis and cleanup steps.

KEYWORDS:

• ESI-MS
• LC-MS/MS
• cattle urine
• drug residue
• hog oral fluid
• multi-residue method
• rapid analysis
• veterinary drugs

Acknowledgments

We are thankful to Isaac Courneya, and Amy McGarvey for their help with sample preparation and analysis. The authors wish to acknowledge Mr. Robert Elder at Seaboard Foods for providing the previously screened hog oral fluid containing incurred residues. Graphical abstract was made using smart servier medical art templates licensed under a creative common attribution 3.0 unported license.

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