Antibiotics | Application of a Multi-Antibiotic Residue Detection Method in Honey

This study developed and validated a multi-class antibiotic residue detection method based on UHPLC-TOF-MS, compliant with EU Regulation (EU) 2021/808, applicable to complex food matrices like honey. The method demonstrates high sensitivity and accuracy, with detection limits as low as 0.09 µg·kg⁻¹, providing a reliable tool for food safety monitoring.

 

Literature Overview
This paper, 'Multi-Analyte Method for Antibiotic Residue Determination in Honey Under EU Regulation 2021/808', published in the journal Antibiotics, reviews and summarizes a multi-antibiotic residue detection method compliant with EU regulations. By optimizing QuEChERS sample preparation combined with UHPLC-TOF-MS analysis, the study successfully detected 15 antibiotics, offering scientific support for food safety assessments of antibiotic residues in honey.

Background Knowledge
Antibiotics are extensively used in apiculture to prevent and treat bee infections. However, their residues may trigger antimicrobial resistance and health risks, making antibiotic monitoring in honey critical. Currently, no established maximum residue limits (MRLs) exist for antibiotics in bee products, posing regulatory and risk-assessment challenges. Honey, a complex matrix characterized by high viscosity and sugar content, complicates antibiotic extraction and detection, necessitating a universal, highly sensitive detection method. While multi-residue methods (e.g., UHPLC-MS/MS and HRMS) are widely applied in food analysis, technical challenges remain in antibiotic detection within honey. This study validates a multi-class antibiotic detection method for honey samples based on Commission Implementing Regulation (EU) 2021/808, aiming to provide standardized tools for food safety regulation.

 

 

Research Methods and Experiments
The study employed acetonitrile-water (80:20, v/v) containing 0.1% formic acid for honey sample extraction, followed by modified QuEChERS purification (adding 1g NaCl and 2g MgSO₄) for sample preparation. Subsequent analysis used UHPLC-TOF-MS, with compound identification based on accurate mass deviation (≤5 ppm) and relative retention time deviation (≤1.0%). Method validation included parameters such as linearity, recovery, repeatability, reproducibility, limit of detection (LOD), and limit of quantification (LOQ), with final application in screening 10 commercially available Portuguese honey samples.

Key Conclusions and Perspectives

• The method demonstrated excellent performance for identifying 15 antibiotics, achieving mass accuracy within 5 ppm and relative retention time deviation below 1.0%, meeting regulatory requirements.
• Recovery rates ranged from 80.1% to 117.6%, with repeatability of 0.5%–32.2%, reproducibility of 2.3%–31.6%, and R² values between 0.9429 and 0.9982, indicating strong linear relationships.
• LOD values were 0.09–6.19 µg·kg⁻¹, LOQ values 0.29–18.77 µg·kg⁻¹, and CCβ values 3–15 µg·kg⁻¹, confirming high sensitivity.
• No target antibiotic residues were detected in the 10 commercial Portuguese honey samples, suggesting minimal antibiotic contamination or absence of usage in current honey production.

Research Significance and Prospects
This study provides an efficient and reliable method for antibiotic residue detection in honey, supporting food safety regulation and risk assessment. Future work should expand sample sizes and incorporate geographical and seasonal factors to comprehensively evaluate global antibiotic contamination in honey. Additionally, the method can be extended to other bee products, such as propolis and royal jelly, for broader application.

 

 

Conclusion
This study successfully developed and validated a multi-class antibiotic residue detection method for honey samples in compliance with EU Regulation 2021/808. Through optimized extraction and purification protocols combined with high-resolution mass spectrometry, the method demonstrates high sensitivity, selectivity, and reproducibility, making it suitable for food safety testing. Results indicate no detectable antibiotic residues in Portuguese commercial honey, though large-scale investigations remain necessary to assess global antibiotic use in bee products. This method offers scientific support for future food regulation, antibiotic resistance studies, and apiculture management, particularly providing reliable reference for risk assessment in the absence of established MRL standards.

 

Helena Rodrigues, Marta Leite, Maria Beatriz P P Oliveira, and Andreia Freitas. Multi-Analyte Method for Antibiotic Residue Determination in Honey Under EU Regulation 2021/808. Antibiotics.