Confirmatory analysis of β‐lactam antibiotics in kidney tissue by liquid chromatography/electrospray ionization selective reaction monitoring ion trap tandem mass spectrometry

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Abstract
Eleven β-lactam antibiotics were analyzed in fortified and incurred beef kidney tissue using high-performance liquid chromatography/electrospray ionization/selective reaction monitoring-ion trap tandem mass spectrometry (LC/ESI-SRM-MSn). The analytes included: deacetylcephapirin, amoxicillin, cephapirin, desfuroylceftiofur cysteine disulfide (DCCD, a biomarker of ceftiofur), ampicillin, cefazolin, Pen G, oxacillin, cloxacillin, naficillin and dicloxicillin. Analytes were extracted with acetonitrile and water. Clean-up was performed by solid-phase extraction. Limits of confirmation in fortified tissue are as follows (tolerances or target levels in parentheses): deacetylcephapirin: 10–50 ng/g (100 ng/g); amoxacillin: 50–100 ng/g (10 ng/g); cephapirin: 10 ng/g (100 ng/g); DCCD: 500 ng/g (8000 ng/g); ampicillin: 10 ng/g (10 ng/g); cefazolin: 10 ng/g (10–50 ng/g); Pen G: 10 ng/g (50 ng/g); oxacillin: 10 ng/g (10–50 ng/g); cloxacillin: 10 ng/g (10 ng/g); naficillin: 10 ng/g (10–50 ng/g); dicloxacillin: 100–500 ng/g (10–50 ng/g). The present method was also tested on incurred kidney tissue that had previously been analyzed using a microbial assay. Good correspondence was found between the results from this new method and the bioassay. However, the present method is much more specific and, in several cases, more sensitive than the bioassay. In addition, the time of analysis is significantly shorter than the bioassay. We also found that SRM MSn was superior in the analysis of unknown incurred tissue than full spectrum MSn. We also obtained an MS/MS spectrum of DCCD that is significantly at variance with previously published fragmentation spectra. Published in 2003 by John Wiley & Sons, Ltd.

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