Multiclass Residue Analysis of Veterinary Drugs in Pork and Hen Eggs Using an Agilent Ultivo Triple Quadrupole LC/MS System

Importance of the Topic

Veterinary drugs such as antimicrobials, hormones, and parasiticides are routinely used in livestock to prevent disease and improve growth. However, residues of these compounds in meat and eggs can lead to allergic reactions and contribute to antibiotic resistance. Regulatory agencies worldwide have established maximum residue limits to protect consumer health, creating a demand for robust analytical methods capable of detecting multiple drug classes at trace levels in complex food matrices.

Objectives and Study Overview

This study aimed to develop and validate a single-run, multiclass screening method for 151 veterinary drug residues spanning 27 chemical classes in pork and hen eggs. The method integrates Agilent’s EMR–Lipid sample preparation kit with an Ultivo triple quadrupole LC/MS system coupled to a 1290 Infinity II UHPLC, enabling high sensitivity and throughput in routine food safety testing.

Methodology and Applied Instrumentation

Sample preparation employed EMR–Lipid dispersive SPE to remove high lipid content, followed by cleanup with a polish pouch. Extracts were concentrated, reconstituted, and injected onto a ZORBAX Eclipse Plus C18 column. Chromatographic separation used a 26-minute gradient (0.2% formic acid in water/acetonitrile) at 0.5 mL/min and 40 °C. Detection was performed in dynamic MRM mode with fast polarity switching.

• UHPLC: Agilent 1290 Infinity II
• Column: ZORBAX Eclipse Plus C18, 3.0 × 150 mm, 1.8 µm
• Mass spectrometer: Agilent Ultivo triple quadrupole with dual hexapole and VacShield
• Ionization: Electrospray, positive/negative mode
• Sample kit: EMR–Lipid dSPE (p/n 5982-1010) and polish pouch (p/n 5982-0102)

Main Results and Discussion

Calibration curves (nine levels, 0.1–40 ng/g) achieved R2 ≥ 0.990 for over 87% of analytes in pork and 89% in eggs. At a 2 ng/g spiking level, 93% of compounds in eggs exhibited signal-to-noise ratios above 10 for both qualifier and quantifier transitions. Repeatability at 5 ng/g (n = 7) yielded %RSD ≤ 20% for over 92% of targets, demonstrating high precision.

Benefits and Practical Applications

This method allows comprehensive, sensitive, and reproducible monitoring of a wide range of veterinary drugs in lipid-rich food matrices within a single analytical run. Its streamlined workflow and minimal maintenance requirements suit high-throughput laboratories tasked with regulatory compliance and quality control.

Future Trends and Potential Applications

Further development may extend this platform to additional animal-derived foods such as milk and seafood. Integration with automated data processing and machine learning algorithms could improve identification confidence and reduce evaluation time. Adoption of microfluidic interfaces and higher-resolution MS technologies may further lower detection limits and expand analyte scope.

Conclusion

The proposed EMR–Lipid sample preparation combined with the Agilent Ultivo triple quadrupole LC/MS system provides a reliable, efficient solution for multiclass veterinary drug residue analysis in pork and eggs. It meets stringent regulatory requirements for sensitivity, accuracy, and precision, supporting food safety and public health initiatives.

References

1. Department of Health and Human Services, FDA, 21 CFR Parts 514 and 558, FDA-2010-N-0155.
2. Proposal for a Regulation of the European Parliament and of the Council on Veterinary Medicinal Products, COM(2014) 0257 (COD).
3. Announcement No. 235, Ministry of Agriculture of China, Maximum Residue Limits in Animal-Derived Foods, 2002.