Quantitation of spectinomycin residues in bovine tissues by ion-exchange high-performance liquid chromatography with post-column derivatization and confirmation by reversed-phase high-performance liquid chromatography–atmospheric pressure chemical ionization tandem mass spectrometry

• Published in: Journal of Chromatography A Vol. 812; no. 1; pp. 123 - 133
• Main Authors: Hornish, Rex E, Wiest, John R
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 03.07.1998
• Subjects: Aminocyclitols; Aminoglycosides; Animals; Antibiotics; Cattle; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Indicators and Reagents; Mass Spectrometry; Meat - analysis; Solutions; Spectinomycin; Spectinomycin - analysis; Spectinomycin; Aminoglycosides; Antibiotics; Aminocyclitols
• ISSN: 0021-9673
• DOI: 10.1016/S0021-9673(98)00368-9

Determinative and confirmatory methods of analysis for spectinomycin residue in bovine kidney, liver, muscle and fat have been developed. The determinative method is a single-column HPLC ion-exchange procedure that incorporates a two-step post-column oxidation of the secondary amines to primary amines followed by derivatization with o-phthalaldehyde. The method was validated in all tissues to a low-end concentration of 0.10 μg/g (limit of quantitation) and to a high-end of 10 μg/g for kidney, which is the rate-limiting tissue for residues of spectinomycin. The recovery of spectinomycin from all tissues was >80% and the variability (R.S.D.) was generally <10%. For liver, an alternative reversed-phase HPLC separation was required for incurred-residue samples. The confirmatory method employed an atmospheric pressure chemical ionization–MS–MS approach utilizing a rapid reversed-phase HPLC system with a mobile phase of methanol and 1% acetic acid. The protonated molecular ion for spectinomycin at m/ z 333 produced four diagnostic reaction–product ions at 98, 116, 158 and 189 for confirmation. The method was validated to a lower limit of confirmation of 0.10 μg/g.