An End-to-End Workflow Solution For Quick and Easy Quantitative Analysis of Multiclass Veterinary Drug Residues in Meat Using LC-MS/MS

Significance of the Topic

The monitoring of veterinary drug residues in meat products is critical to protect human health and comply with global regulatory limits. A single, streamlined workflow capable of detecting and quantifying hundreds of veterinary drugs across multiple chemical classes can reduce analysis time, labor and operational costs, while ensuring food safety and regulatory compliance.

Objectives and Study Overview

This study presents an end-to-end LC-MS/MS workflow for rapid, sensitive and reproducible screening and quantitation of over 200 multiclass veterinary drug residues in meat matrices. The method integrates sample preparation, targeted dynamic multiple reaction monitoring (dMRM) and optimized instrument parameters to achieve comprehensive coverage under a single analytical platform.

Methodology

The workflow begins with 2 g of homogenized meat (chicken, beef or pork) spiked or blank, followed by liquid extraction using EDTA and acidified acetonitrile. After centrifugation, lipid removal and cleanup are achieved with Agilent Captiva EMR-Lipid cartridges and PPM-48 plates. The purified extract is analyzed by LC-MS/MS using a Poroshell 120 EC-C18 column and a custom dMRM database for targeted acquisition. Calibration curves span 0.1 to 100 µg/kg, covering typical regulatory maximum residue limits (MRLs).

Instrumentation

• Agilent 6470 and 6495C Triple Quadrupole LC/MS systems
• Agilent InfinityLab Poroshell 120 EC-C18 column (p/n 695575-302)
• Captiva EMR-Lipid cartridge (p/n 5190-1003) and PPM-48 cleanup plate (p/n 5191-4101)
• Agilent MassHunter software with VetDrug dMRM database

Main Results and Discussion

• Limits of detection ranged from 0.1 to 10 µg/kg; calibration linearity (R²>0.99) was confirmed across 0.25–100 µg/kg.
• Accuracy of 70–120% and precision (%RSD) below 20% for response and 0.5% for retention time.
• Recovery in chicken muscle at 10 µg/kg exceeded 80% for over 98% of analytes.
• Intrabatch repeatability showed RSD <15% for most compounds; interbatch reproducibility remained below 32% RSD, with 194 targets under 15%.
• Method performance was consistent across chicken, beef and pork matrices.

Benefits and Practical Applications

The comprehensive dMRM workflow simplifies multiclass veterinary drug screening in routine food safety laboratories. It enables high-throughput analysis, reduces the need for multiple compound-class methods and supports compliance with AOAC, FDA and EU regulations. The integrated dMRM database allows rapid customization of sub-methods tailored to specific regulatory lists.

Future Trends and Applications

Further applications may extend this workflow to seafood, dairy and processed foods. Advances in automation of sample handling, expansion of compound libraries and integration with high-resolution MS platforms could enhance sensitivity, throughput and adaptability to emerging residues of concern. Continuous software updates will support evolving regulatory requirements and data analysis workflows.

Conclusion

This study validates a robust, end-to-end LC-MS/MS solution for rapid quantitation of over 200 veterinary drug residues in meat. The method combines efficient sample preparation, sensitive dMRM acquisition and reproducible performance, offering a practical tool for routine food safety monitoring and regulatory compliance.

References

1. AOAC Official Method Performance Requirements, Appendix F, 2016
2. Code of Federal Regulations Title 21, Part 556, US FDA, 2019
3. US FSIS Chemical Contaminants of Public Health Concern, 2017
4. Commission Regulation (EU) No 37/2010, Official Journal of the European Union
5. Agilent Application Note 5994-1932EN, 2020