Study of Oxidation of Ciprofloxacin and Pefloxacin by ACVA: Identification of Degradation Products by Mass Spectrometry and Bioautographic Evaluation of Antibacterial Activity

Abstract

The new RP-HPLC-DAD method for the determination of ciprofloxacin and pefloxacin, next to their degradation products after the oxidation reaction with 4,4′-azobis(4-cyanopentanoic acid) (ACVA) was developed. The method was validated according to the guidelines of the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH) and meets the acceptance criteria. The experimental data indicate that the course of the oxidation process depends on the type of fluoroquinolone (FQ), the incubation time and temperature. The performed kinetic evaluation allowed us to state that the oxidation of FQs proceeds according to the second-order kinetics. The degradation products of the FQs were identified using the UHPLC-MS/MS method and their structures were proposed. The results obtained by the TLC-direct bioautography technique allowed us to state that the main ciprofloxacin and pefloxacin oxidation products probably retained antibacterial activity against Escherichia coli.