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

Abstract

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 and from 7 to 13 µg kg, respectively, except for streptomycin and neomycin, which showed slightly higher CCα at 25 µg kg and CCβ at 34 µg kg. 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.

Design for the transfer of a validated liquid chromatography/tandem mass spectrometry analytical method for the determination of antimicrobial residues in honey from low‐resolution to high‐resolution mass spectrometry

“…The extraction and clean‐up of antimicrobial residues in honey have been described in our previous report . In brief, 2 ± 0.05 g of honey were weighed into a 50‐mL disposable centrifuge tube and spiked with 100 μL of IS working solution and kept overnight in the dark.…”

“…To the best of our knowledge, there has been no published comparison of the analytical performance provided by an LC/MS/MS method and a LC/Orbitrap method for the determination of 23 antimicrobial residues in honey. The main objective of this paper is to transfer the analytical method previously developed and validated in honey by using LC/MS/MS technology to a LC/Orbitrap system in full scan mode. Our aim is to demonstrate that the LC/Orbitrap analytical technique provides as accurate results for quantification of antimicrobial residues as an LC/MS/MS system and also improves the degree of identification of the target compounds along with their metabolites in honey samples.…”

Trends in sensitive detection and rapid removal of sulfonamides: A review

“…As a confirmatory detector, MS can provide information about the molecular structures and quantitative data on the analytes with high sensitivity and selectivity. Furthermore, for the purpose of enhancing sensitivity and accuracy, MS/MS [50,51,53,55] has been utilized for the analyses of sulfonamides. With the advantages and disadvantages mentioned above, MS/MS is outstanding compared with the other detectors.…”

Determination of sulfonamides in milk by ID-LC-MS/MS

“…Matrix-matched calibration standards were prepared by spiking on blank extracts (10,15,20,30,40, 50 ng/g, each contained 20 ng/g of ISTD) to generate six-point calibration curve, which was plotted by peak area ratio (analyte/ISTD) versus concentration ratio (analyte/ISTD). The LOD and LOQ values of the individual analytes were estimated from the standard deviation of 10 samples prepared by spiking blank samples with sulfonamides (10 ng/g of analyte and 20 ng/g of ISTD).…”

“…The available methods provide only 1-3 isotopically labelled sulfonamides to be used as internal standards [25][26][27][28][29][30][31][32]. This study describes the development and validation of a full IDMS technique for all 14 sulfonamides in milk.…”