Identification of metabolites of ampicillin using liquid chromatography/thermospray mass spectrometry and fast atom bombardment tandem mass spectrometry

Abstract

Mass spectrometry is combined with liquid chromatography (LC/MS) and mass spectrometry (MS/MS) to identify ampicillin and two known metabolites--ampicillin penicilloic acid and ampicillin piperazine-2,5-dione--in human urine samples. Identifications were based on the fact that the metabolites or degradation products contain a substructure of ampicillin. In addition, two previously unidentified components in human urine samples were detected, corresponding to newly discovered metabolites or degradation products of ampicillin. Different chromatographic retention times in the LC mass spectra indicated two different compounds. However, the tandem mass spectra of these two components were similar, suggesting that they are stereoisomers. The use of LC/MS and MS/MS confirmed that the unknown components are the (3S,5R) and (3S,5S) epimers of ampicillin penilloic acid. Further study showed that only one of the components was eliminated from the body; the other arises from interconversion to the epimer in a urine sample. It is speculated that the component produced in vivo is the (3S,5R) epimer of ampicillin penilloic acid.