Multi-residue confirmation of aminoglycoside antibiotics and bovine kidney by ion spray high-performance liquid chromatography/tandem mass spectrometry

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Abstract
An ion spray high-performance liquid chromatographic/tandem mass spectrometric (HPLC/MS/MS) method capable of determining the following six aminoglycosides in bovine kidney is presented: spectinomycin, hygromycin B, streptomycin, dihydrostreptomycin, gentamicin C complex and neomycin B. Tobramycin was used as an internal standard. This method uses an improved matrix solid-phase dispersion (MSPD) method for tissue extraction. A gradient HPLC separation was developed with mobile phases consisting of aqueous 20 mM pentafluoropropionic acid and acetonitrile. Protonated molecules served as precursor ions for collision-induced dissociation (CID) and three product ions were chosen for each analyte for selected reaction monitoring (SRM) where possible. A validation study was conducted for the confirmation of dihydrostreptomycin, neomycin B and four major components of the gentamicin C complex throug SRM HPLC/MS/MS analysis of negative control, fortified and incurred bovine kidney samples. All of the samples analyzed could be confirmed with ion ratios within 15% of the daily mean of fortified standards and 90% of the samples had ion ratios within 10%. All compounds except spectinomycin could be detected (while monitoring three ions by SRM) in bovine kidney tissue at or below the regulatory level of concern. MSPD recoveries were aceptable with the exception of the 27% value observed for spectinomycin.