Development and validation of a novel UPLC-MS/MS method for quantification of delafloxacin in plasma and aqueous humour for pharmacokinetic analyses

Abstract

Acute bacterial skin and skin structure infections are one of the most frequent infectious disease requiring hospitalization for treatment. Delafloxacin is a clinically approved fluoroquinolone antibiotic for the treatment of ABSSSIs. In spite of being marketed since 2017, there is no published analytical method for quantification of delafloxacin in biological samples. Herein, a selective and sensitive UPLC-MS/MS method was developed and validated for quantitative analysis of delafloxacin in rat plasma and rabbit aqueous humour samples. The liquid liquid extraction (using ethyl acetate) was used for analyte extraction form rat plasma, whereas protein precipitation (acetonitrile) was used for aqueous humour samples preparations. An Acquity UPLC BEH C₁₈ column was used for chromatographic separation of delafloxacin and internal standard (rivaroxaban). The mobile phase composition of acetonitrile (containing 0.1% formic acid) and 10 mM ammonium acetate in ratio of 60:40 were used for sample elution at 300 µL/min flow rate. The electrospray ionization operated in positive mode was used for sample ionization and detection of analyte and internal standard were performed by multiple reaction monitoring (MRM) mode. The MRM transitions were set to 441.14 > 379.09 and 436.89 > 144.87 for delafloxacin and internal standard, respectively. The method was validated as per USFDA guideline for bioanalytical method and all the evaluated parameters were within the acceptable ranges. The developed method in plasma was successfully used to analyze samples in pharmacokinetic study of newly developed stearic acid-chitosan solid lipid nanoparticles formulation of delafloxacin in rat.

Keywords: Aqueous humour; Delafloxacin; Nanoparticles; UPLC-MS/MS.