A simple method for the determination of fluoroquinolone residues in tilapia (Oreochromis niloticus) and pacu (Piaractus mesopotamicus) employing LC-MS/MS QToF

Abstract

The use of antimicrobials in livestock production is a powerful resource applied throughout the world to guarantee high yield and control bacterial diseases in aquaculture. However, residues of these substances in animal products represent a potential risk to consumer health when residue levels are above the established maximum residue limits (MRLs). Fluoroquinolones (FQs) are antimicrobials commonly used worldwide in aquaculture. The aim of this work was to develop and validate a simple analytical method for the simultaneous determination of norfloxacin, danofloxacin, enrofloxacin and ciprofloxacin levels in tilapia (Oreochromis niloticus) and pacu (Piaractus mesopotamicus) fillets using liquid chromatography–tandem mass spectrometry (LC-MS/MS) quadrupole time of flight (QToF). The FQs were extracted from the fillets with 1% acetic acid–methanol and 1% acetic acid–acetonitrile solutions using ultrasonic assistance. The clean-up was performed with hexane. Chromatographic separation was conducted in an XTerra RP18 column (2.1 × 150 mm, 5 µm) at 25 °C with a flow of 0.2 mL min⁻¹. The mobile phase consisted of 0.1% aqueous formic acid and acetonitrile, with gradient elution. The validation parameters for all FQs were linearity (>0.99), intra-day precision (CV of 1%–9%), inter-day precision (CV of 3%–17%), decision limit (63–126 ng g⁻¹), detection capability (76 –152 ng g⁻¹) and accuracy (90%–111%). The limit of quantification was lower than the MRL for each FQ, indicating that the method is suitable for the determination of the FQ levels in the fish fillets. The mass analyser of the QToF type was able to confirm the identities of the FQs with an error of the accuracy of the mass (reasons m/z) of less than 10 ppm.

Keywords:

• veterinary drug residues
• fluoroquinolones
• antimicrobials
• food analysis
• tilapia
• pacu
• fish
• LC-MS/MS

Acknowledgements

The authors gratefully acknowledge the financial support received from São Paulo Research Foundation (FAPESP), the Coordination for the Improvement of Higher Level Education Personnel (CAPES), and the National Council for Scientific and Technological Development (CNPq), Brazil. The authors also thank Elsevier Language Editing Services for language assistance.