Characterization of a new norfloxacin metabolite monitored during a bioequivalence study by means of mass spectrometry and quantum computation

A bioequivalence study of two formulations containing norfloxacin was used for identification and assay of the metabolite of norfloxacin in human serum samples. The bioequivalence study was based on an analytical method using liquid chromatography with fluorescence detection. The plasmatic profile of metabolite was similar to norfloxacin for both formulations. Three plasma fractions of norfloxacin metabolite from a volunteer were isolated by liquid chromatography and investigated by atmospheric-pressure chemical ionization mass-spectrometry. The structure of norfloxacin metabolite (7-aminoethylenamino-6-fluoro-4-hydroxy quinoline-3-carboxylic acid) was identified taking into account also the mass spectrometry investigations achieved for norfloxacin and ciprofloxacin (used as internal standard for the analytical method). A theoretical procedure based on quantum chemical calculations has been also used to explain the mass fragmentation in molecules of norfloxacin metabolite that differ from the molecule of norfloxacin.