Development and validation of a multiclass method for the determination of antibiotic residues in honey using liquid chromatography-tandem mass spectrometry

• Published in: Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment Vol. 34; no. 4; pp. 582 - 597
• Main Authors: El Hawari, Khaled, Mokh, Samia, Doumyati, Samah, Al Iskandarani, Mohamad, Verdon, Eric
• Format: Journal Article
• Language: English
• Published: England Taylor & Francis 03-04-2017; Taylor & Francis Ltd
• Subjects: Acidification; Aminoglycosides; Aminoglycosides - analysis; Animals; Anti-Bacterial Agents - analysis; Antibiotics; Bees - physiology; Chromatography, Liquid - standards; Cleanup; Cleavage; Decision analysis; Dispersion; Drug and Narcotic Control - legislation & jurisprudence; Erythromycin; European Union; Extraction; Food Contamination - analysis; Honey; Honey - analysis; Humans; Hydrochloric acid; LC-MS/MS; Lincosamides; Lincosamides - analysis; Linkages; Liquid chromatography; Macrolides - analysis; Markers; Mass spectrometry; Mass spectroscopy; Methanol; Multi-class analysis; Neomycin; Reproducibility; Reproducibility of Results; Residues; Sample preparation; Separation; Shaking; Solid Phase Extraction - methods; Streptomycin; Sulfonamides; Sulfonamides - analysis; Tandem Mass Spectrometry - standards; Tetracyclines; Tetracyclines - analysis; Tylosin; Ultrasonic processing; validation; veterinary drugs; Veterinary Drugs - analysis; Water analysis; LC-MS/MS; Multi-class analysis; veterinary drugs; honey; validation
• ISSN: 1944-0049; 1944-0057
• DOI: 10.1080/19440049.2016.1232491

A new, simple and fast method was developed for the determination of multi-class antibiotic residues in honey (sulfonamides, tetracyclines, macrolides, lincosamides and aminoglycosides). Separation and determination were carried out by LC-MS/MS. During sample preparation, various parameters affecting extraction efficiency were examined, including the type of solvent, pH, efficiencies of cleavage of N-glycosidic linkages by hydrochloric acid, ultrasonic extraction and its duration compared with shaking, along with dispersive SPE clean-up. Experiments with fortified samples demonstrated that 10 min of ultrasonic treatment with acidified methanol (HCl 2 M) followed by dispersive SPE clean-up with 50 mg PSA gave an effective sample preparation method for several classes of antibiotics in honey. Anhydroerythromycin A, erythromycin A enol ether and desmycosin were used as markers for the presence of erythromycin A and tylosin A in honey samples. The method was validated according to European Commission Decision (EC) No. 2002/657. The recoveries of analytes ranged from 85% to 111%. Repeatability and intra-laboratory reproducibility were < 20.6% and 26.8%, respectively. Decision limit (CCα) and detection capability (CCβ) ranged from 6 to 9 µg kg
-
1
and from 7 to 13 µg kg
-
1
, respectively, except for streptomycin and neomycin, which showed slightly higher CCα at 25 µg kg
-
1
and CCβ at 34 µg kg
-
1
. Finally, the method was applied to the honey test material 02270 through a FAPAS proficiency test (PT) for the determination of tetracyclines. PT results were obtained within a z-score range of ±2, proving that the validated method is suitable for routine analysis to ensure the quality of honey.