Rapid, Simple, and Effective Cleanup of Seafood Extracts Prior to UPLC-MS/MS Multiresidue Veterinary Drugs Analysis

Importance of the Topic

Reliable monitoring of veterinary drug residues in seafood is essential to protect public health and meet regulatory standards. Seafood tissues present analytical challenges due to high fat and phospholipid content, which can interfere with UPLC-MS/MS performance and instrument stability.

Objectives and Study Overview

This study aimed to design a rapid, unified sample preparation and cleanup workflow to quantify a broad spectrum of veterinary drugs in shrimp and salmon using a single UPLC-MS/MS method, maximizing throughput and minimizing costs.

Methodology and Instrumentation

• Sample Extraction: Homogenized tissue (2.5 g) was spiked, then extracted with 0.2 % formic acid in 80:20 acetonitrile/water, precipitating proteins and co-extracting lipids.
• Cleanup: A pass-through SPE cleanup with Oasis PRiME HLB cartridges (3 cc, 60 mg) removed over 90 % of fats and ≥ 95 % of phospholipids without conditioning, pressure set at 1–2 psi.
• UPLC-MS/MS Analysis: Waters ACQUITY UPLC I-Class with CSH C18 column (1.7 µm, 100 × 2.1 mm) and Xevo TQ-S mass spectrometer. Mobile phases were 0.1 % formic acid in water (A) and acetonitrile (B), with ESI positive mode (negative for chloramphenicol).

Key Results and Discussion

The workflow yielded overall recoveries above 70 % for most analytes across tetracyclines, fluoroquinolones, sulfonamides, macrolides, beta-lactams, steroids, NSAIDs, and beta-agonists. SPE cleanup recoveries exceeded 80 % in shrimp and 90 % in salmon, except for phenylbutazone (≈55–60 %). Matrix effects averaged 40 % but remained consistent across matrices. Chromatograms confirmed effective removal of lipids, preserving chromatographic integrity and reducing maintenance.

Benefits and Practical Applications

• Streamlined, single-step lipid and phospholipid removal without hexane or lengthy defatting procedures.
• Particle-free extracts eliminate additional filtration, speeding analysis.
• High-throughput multiclass screening suitable for regulatory testing, QA/QC, and routine monitoring of seafood safety.

Future Trends and Opportunities

Advances may include automated SPE platforms for higher throughput, mixed-mode sorbents for broader analyte coverage, and miniaturized UPLC-MS/MS systems to reduce solvent consumption and expand applicability to diverse food matrices.

Conclusion

The proposed protocol provides a rapid, robust, and sensitive solution for multiresidue veterinary drug analysis in seafood, balancing efficient cleanup with reliable UPLC-MS/MS performance.

Reference

• Young MS, Tran KV. Oasis PRiME HLB Cartridge for Effective Cleanup of Meat Extracts Prior to Multi-Residue Veterinary Drug UPLC-MS Analysis. Waters Application Brief, 2015.
• Lehotay SJ. High-Throughput Screening Analysis by UHPLC-MS/MS of >60 Veterinary Drugs in Animal Tissues. 125th AOAC Annual Meeting, 2011.
• Young MS, Tran KV. Optimized Extraction and Cleanup Protocols for LC-MS/MS Multiresidue Determination of Veterinary Drugs in Edible Muscle Tissues. Waters Application Note, 2011.