Development and Validation of a Method for the Determination of Aminoglycosides in Foods using LC-MS/MS with a Zwitterionic HILIC Stationary Phase

Significance of the Topic

Aminoglycosides are critical veterinary antibiotics widely used as therapeutics and growth promoters in food animals. Monitoring their residues ensures food safety, prevents antimicrobial resistance and enforces regulatory standards for consumer protection.

Objectives and Study Overview

This work aimed to develop and validate a sensitive and robust LCMSMS method employing a zwitterionic HILIC stationary phase to determine eight aminoglycosides in milk, eggs and honey. The study evaluated analytical performance against EU 2021/808 criteria.

Methodology and Used Instrumentation

Samples were extracted with ammonium acetate, EDTA, NaCl and TCA, then cleaned by SPE on Oasis HLB with tobramycin as internal standard. Chromatography used an Atlantis Premier BEH Z-HILIC column on ACQUITY I-Class with BSM and FTN sample manager. Detection was by Xevo TQ-XS with UniSpray ion source and MS friendly mobile phase.

• HPLC column Atlantis Premier BEH Z-HILIC
• UHPLC system ACQUITY I-Class with BSM and FTN
• Mass spectrometer Xevo TQ-XS with UniSpray
• Solid phase extraction Oasis HLB

Results and Discussion

Validation covered identification, trueness, repeatability (RSDr), reproducibility (RSDRL) and decision limit (CCa). Recovery ranged 80-120 % at 0.5x to 2x MRL levels, with precision below 22 %. Decision limits met EU thresholds for milk, eggs and the MMPR for honey.

Benefits and Practical Applications

The method delivers reliable quantitative analysis for compliance testing of regulatory residue limits in diverse food matrices and supports detection of unauthorised aminoglycoside use.

Future Trends and Opportunities

Potential developments include extending the approach to other polar antibiotic classes, integrating high resolution MS for non-target screening and advancing automation for high throughput analysis.

Conclusion

A robust, sensitive and regulatory compliant LCMSMS method using a zwitterionic HILIC column was established for routine monitoring of eight aminoglycosides in milk, eggs and honey.

References

• Analysis of Aminoglycosides in Foods Using a Zwitterionic Stationary Phase and Liquid Chromatography Tandem Mass Spectrometry Waters App Note 720007442