EuroResidue: DEVELOPMENT OF A METHOD FOR THE DETERMINATION OF AMINOGLYCOSIDES IN FOODS USING LC-MS/MS WITH A NOVEL ZWITTERIONIC HILIC STATIONARY PHASE

Importance of the Topic

Aminoglycosides are widely used veterinary antibiotics that may leave residues in food products, posing health risks and regulatory concerns. Sensitive and selective analytical methods are essential to ensure compliance with maximum residue limits in milk, meat, liver, and honey.

Objectives and Study Overview

The study aimed to develop and validate a robust LC-MS/MS method for simultaneous determination of seventeen aminoglycoside antibiotics in various food matrices. A zwitterionic HILIC stationary phase was evaluated to overcome limitations of reversed-phase and conventional HILIC separations.

Methodology and Instrumentation

Sample preparation involved extraction with an aqueous solution containing trichloroacetic acid, ammonium acetate, EDTA, and NaCl, followed by solid-phase extraction on Oasis HLB cartridges. Chromatographic separation was performed on an Atlantis Premier BEH Z-HILIC column using a gradient of 20 mM ammonium formate (pH 3.0) and acetonitrile with 0.1 % formic acid. Detection was carried out by electrospray ionization triple quadrupole mass spectrometry in positive mode, employing matrix-matched calibration to correct for ion suppression or enhancement.

Used Instrumentation

• LC System: Waters Arc Premier with MaxPeak High Performance Surfaces technology
• Column: Atlantis Premier BEH Z-HILIC, 2.1×150 mm, 2.5 µm
• MS System: Xevo TQ-S micro triple quadrupole
• Ionization: Electrospray, positive mode
• Key parameters: desolvation temperature 600 °C, capillary voltage 1.5 kV, desolvation gas flow 1000 L/h

Main Results and Discussion

Under optimized conditions, all seventeen aminoglycosides were well separated with retention time RSDs below 1 %. Sensitivity met or exceeded required limits for regulatory monitoring. Matrix effects ranged from suppression to enhancement across milk, muscle, liver, and honey, necessitating the use of matrix-matched calibration. Initial recovery studies at 200–250 µg/kg showed recoveries from 101 % to 134 % and repeatability below 10 % RSD for most analytes.

Benefits and Practical Applications

The method eliminates the need for ion-pair reagents and high buffer concentrations, reducing system contamination and ion suppression. It delivers reliable quantification of aminoglycoside residues at or below established maximum residue limits, supporting routine compliance testing in food safety laboratories.

Future Trends and Opportunities

Further validation should explore additional isotopically labeled internal standards to improve accuracy for compounds with high recovery. The zwitterionic HILIC approach can be extended to other polar veterinary drugs. Advances in high-performance surface technologies and faster gradients may enhance throughput in regulatory and quality control settings.

Conclusion

A novel zwitterionic HILIC-based LC-MS/MS method was developed for sensitive and robust determination of seventeen aminoglycosides in diverse food matrices. Its performance supports regulatory monitoring of antibiotic residues with high specificity and reproducibility.

Reference

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