Rapid Detection of 7 Illegal Veterinary Additives in Animal Feed Using Oasis PRiME HLB Clean-up and UPLC-MS/MS

Importance of the Topic

The presence of illegal veterinary additives in animal feed represents a significant risk to food safety and public health. Rapid, reliable detection methods are essential to enforce regulations, prevent drug accumulation in edible tissues, and protect consumers from potential toxic and mutagenic effects.

Objectives and Overview of the Study

• Develop a streamlined multi-residue analysis for seven illicit veterinary drugs in animal feed.
• Implement an efficient cleanup protocol using Oasis PRiME HLB pass-through SPE.
• Employ UPLC-MS/MS for sensitive, accurate quantification that complies with regulatory criteria.

Methodology and Instrumentation

Sample preparation involves weighing 1 g of feed, extracting with 80% acetonitrile/20% water, and centrifugation. A two-step pass-through cleanup on an Oasis PRiME HLB cartridge removes matrix interferences without conditioning. The cleaned extract is diluted 1:3 with water and injected into the UPLC-MS/MS system.

• Chromatography: ACQUITY UPLC I-Class System, HSS T3 column (2.1 × 100 mm, 1.8 μm), gradient of 0.1% formic acid in water and methanol, 0.4 mL/min, 45 °C, 9 min runtime.
• Mass Spectrometry: Xevo TQ-S micro, ESI-positive mode, capillary voltage 3.0 kV, desolvation temperature 550 °C, optimized MRM transitions for each analyte.

Main Results and Discussion

The method achieved recoveries from 70.6% to 112% across low, medium, and high spike levels. Precision (RSD) ranged from 0.9% to 4.2% for medium/high and 4.3% to 8.8% for low spikes. Matrix effects remained within ±16%, and matrix-matched calibration curves showed correlation coefficients above 0.9991. In real samples, olaquindox was detected at concentrations between 1.9 and 18 mg/kg, demonstrating robust applicability to feed matrices.

Benefits and Practical Applications

• Fast and straightforward sample preparation with no cartridge conditioning.
• High sensitivity and reproducibility, meeting stringent regulatory limits.
• Effective matrix cleanup reduces analytical interferences.
• Suitable for routine quality control in feed safety laboratories.

Future Trends and Potential Applications

• Expansion to broader panels of veterinary drugs and growth promoters.
• Integration with high-resolution mass spectrometry for comprehensive non-targeted screening.
• Automation of sample preparation workflows to increase throughput.
• Development of novel sorbent materials for enhanced selectivity and cleaner extracts.

Conclusion

The combination of Oasis PRiME HLB cleanup and UPLC-MS/MS provides a validated, efficient workflow for the rapid detection of illegal veterinary additives in animal feed, delivering simplicity, speed, and reliable performance to support feed safety monitoring.

Instrumentation Used

• Oasis PRiME HLB Cartridge (6 cc, 200 mg)
• ACQUITY UPLC I-Class System with ACQUITY UPLC HSS T3 column
• Xevo TQ-S micro Mass Spectrometer

References

1. Announcement No. 2086-5-2014 of the Ministry of Agriculture of the People’s Republic of China. Determination of carbadox, mequindox, quinocetone and olaquindox in feeds by liquid chromatography–tandem mass spectrometry.