Determination of 68 Veterinary Drugs in Marine Products by Ultra High Performance Liquid Chromatography/Triple Quadrupole Mass Spectrometry

Significance of the Topic

Monitoring residues of veterinary drugs in seafood is critical due to potential human health risks and strict regulatory limits imposed worldwide. The development of rapid and sensitive analytical methods supports food safety, regulatory compliance, and public health protection.

Study Objectives and Overview

This work describes a method for simultaneous screening and quantification of 68 veterinary drugs across 12 chemical classes in marine products. The goal is to achieve high sensitivity, selectivity, and throughput using ultra high performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC-MS/MS).

Methodology and Instrumentation

Sample Preparation
Marine specimens were homogenized and extracted with acetonitrile. Following centrifugation, concentration, and filtration, extracts were directly injected into the UHPLC-MS/MS system for analysis.

Used Instrumentation

• Shimadzu Nexera UHPLC system with LC-30AD pumps, CTO-30A column oven, DGU-30A3 degasser, and SIL-30AC autosampler
• Shim-pack XR-ODSIII column (50 mm length × 2.0 mm i.d., 1.6 µm)
• Shimadzu LCMS-8040 triple quadrupole mass spectrometer with electrospray interface

Chromatographic Conditions
Mobile phase A consisted of 0.1% formic acid and mobile phase B was acetonitrile. A flow rate of 0.4 mL/min at 40 °C was used with a gradient elution program. Injection volume was 10 µL.

Mass Spectrometry Conditions
The mass spectrometer operated in both positive and negative ionization modes with fast polarity switching (15 ms) and a scan speed of 15,000 u/s. Nebulizing gas flow and drying gas pressure were set to 1.5 L/min and 10 L/min respectively. Interface temperatures were optimized at 250 °C (DL) and 400 °C (heater block).

Results and Discussion

Calibration curves for representative compounds exhibited excellent linearity (correlation coefficients > 0.998) over the range of 1–200 µg/L. Method repeatability assessed at 1–10 µg/L showed relative standard deviations below 5% for peak areas and below 0.9% for retention times. Recovery experiments in fish matrices spiked at 0.1–2 µg/kg yielded recoveries between 74% and 120% for 52 analytes, demonstrating accuracy and robustness of the method.

Benefits and Practical Applications

• The method enables simultaneous detection of a broad panel of veterinary drugs in seafood, reducing analysis time and costs.
• High sensitivity and precise quantification support compliance with regulatory requirements.
• The approach is suitable for routine monitoring in food safety laboratories and quality control departments.

Future Trends and Applications

• Integration of automated sample preparation to enhance throughput and reproducibility.
• Expansion of target analyte scope to include emerging contaminants and metabolites.
• Utilization of high resolution mass spectrometry and advanced data processing for improved confirmation and quantification.
• Development of multi-residue methods for other food matrices and environmental samples.

Conclusion

The described UHPLC-MS/MS method provides a rapid, cost-effective, and validated solution for the simultaneous analysis of 68 veterinary drug residues in marine products. Its high precision, accuracy, and sensitivity make it a valuable tool for food safety monitoring and regulatory compliance.