Analysis of Veterinary Drugs in Meat with the Agilent 6495 Triple Quadrupole LC/MS

Importance of the topic

The widespread use of veterinary drugs in food-producing animals requires rigorous monitoring to protect public health and prevent antimicrobial resistance. Rapid, sensitive methods capable of detecting multiple classes of residues at trace levels are vital for regulatory compliance and ensuring the safety of meat products.

Objectives and study overview

This study aimed to develop and validate a 12-minute UHPLC-MS/MS method for the simultaneous analysis of 120 veterinary drugs in bovine meat (kidney and liver). Key goals included achieving detection limits well below regulatory tolerances, using minimal internal standards, and demonstrating robust quantitative performance across a multiclass panel.

Methodology and sample preparation

Homogenized tissue samples (2 g) were extracted with acidified acetonitrile and cleaned using Agilent’s Enhanced Matrix Removal–Lipid (EMR–L) method to reduce lipid interferences. After centrifugation and lipid removal, extracts were diluted and analyzed by UHPLC. The method employed dynamic multiple reaction monitoring (dMRM) with three transitions per analyte to ensure selectivity and compliance with US and EU identification criteria.

Used instrumentation

• Agilent 1290 Infinity II UHPLC with a ZORBAX C-18 column (2.1×150 mm, 1.8 µm)
• Agilent 6495 Triple Quadrupole LC/MS with iFunnel and Jet Stream technology
• Simultaneous positive/negative electrospray ionization
• Agilent MassHunter software (Version 07.00)

Main results and discussion

Limits of detection ranged from 0.1 to 2 ng/mL and quantification limits from 0.1 to 5 ng/mL, all below established tolerance levels. Over 89% of compounds achieved calibration curve R² values above 0.99, both at tolerance-level calibrations and in the low-end 1–100 ng/g range. Intra- and interday precision tests showed relative standard deviations below 15% for more than 90% of analytes, confirming method repeatability and reproducibility. Matrix-interference checks identified a few transitions requiring alternate ions, demonstrating the value of multiple MRM transitions.

Benefits and practical applications

• High throughput: 12-minute runtime for 120 analytes
• Excellent sensitivity and selectivity with minimal internal standards
• Robust quantitation using matrix-matched calibration
• Cost-effective workflow with simplified sample cleanup

Future trends and opportunities for use

Advances may include expanding the analyte panel, integrating high-resolution mass spectrometry for non-target screening, automating sample preparation, and applying the approach to other food matrices. Enhanced data analysis tools and real-time monitoring may further streamline residue surveillance.

Conclusion

The developed UHPLC-MS/MS method delivers rapid, sensitive, and reliable analysis of multiclass veterinary drug residues in bovine meat, meeting regulatory requirements and supporting effective food safety monitoring.

Reference

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