Unauthorised antibiotic treatments in beekeeping: Development and validation of a method to quantify and confirm tylosin residues in honey using liquid chromatography–tandem mass spectrometric detection

• Published in: Analytica chimica acta Vol. 520; no. 1; pp. 87 - 92
• Main Authors: Benetti, C., Dainese, N., Biancotto, G., Piro, R., Mutinelli, F.
• Format: Journal Article Conference Proceeding
• Language: English
• Published: Amsterdam Elsevier B.V 23.08.2004; Elsevier
• Subjects: Analytical chemistry; Chemistry; Chromatographic methods and physical methods associated with chromatography; Exact sciences and technology; Honey; Mass spectrometry; Other chromatographic methods; Spectrometric and optical methods; Tylosin; Validation; Validation; Honey; Tylosin; Mass spectrometry; Ammonium; Internal standard; HPLC chromatography; Acetate; Antibiotic; Sample preparation; Residue; Mass spectrometry MS/MS; Acetonitrile; Solid phase; Quantitative analysis
• ISSN: 0003-2670; 1873-4324
• DOI: 10.1016/j.aca.2004.04.070

A liquid chromatographic–tandem mass spectrometric (HPLC-MS/MS) method is proposed for the identification and quantification of tylosin in honey. Sample treatment involves an extraction in a Tris buffer at pH 10.5, followed by a solid-phase clean up step on an Oasis HLB column. Roxithromycin was used as the internal standard. Chromatographic separation of tylosin and roxithromycin was performed on an XTerra MS C 18 column (100 mm × 2.1 mm i.d., 5 μm) using a gradient of aqueous 0.01 M ammonium acetate pH 3.5 and acetonitrile as the mobile phase, at a flow rate of 0.25 ml min −1. The method was validated according to the guidelines laid down by the Commission Decision 2002/657/EC. Tylosin residues were confirmed by MS/MS experiments considering the appropriate identification points. All validation parameters such as Ccα (lower than 3 ng g −1), Ccβ (lower than 5 ng g −1), recovery and precision were assessed on the basis of the “critical ion” (less intense ion permitting unambiguous identification of the analyte).