Development of a UPLC–MS/MS method for the determination of lomefloxacin in rabbit aqueous humor and its application to a pharmacokinetic study

Abstract

Lomefloxacin is a kind of synthetic fluoroquinolone antibiotic, which is used for the treatment of infectious diseases. In this study, a rapid and efficient liquid chromatography-tandem mass spectrometric assay was developed to determine the concentration of lomefloxacin in rabbit aqueous humor quantitatively. Aqueous humor samples were extracted by protein precipitation. Ofloxacin was chosen as internal standard. The chromatographic separation was achieved on a Kinetex C18 (50mm×2.10mm, 2.6μm, Phenomenex Corp, USA) column, with a gradient of methanol (0.1% formic acid) and water (0.1% formic acid). Multiple reaction monitoring (MRM) with positive ionization mode was used for the mass analysis. The validation of this method was based on the European Medicines Agency (2011) [1] and US FDA Guidelines (2001) [2]. The calibration range of aqueous humor samples was 5-1200ng/mL with r=0.9990 (n=6). For all QC samples, Inter-and intra-run precisions were less than 15% and accuracies were between 80%-120%. In conclusion, the assay was rapid, sensitive and able to determinate the lomefloxacin in rabbit aqueous humor accurately. At the same time, this method was successfully applied to study the pharmacokinetics of lomefloxacin hydrochloride eye drops and lomefloxacin hydrochloride ophthalmic gel in rabbit aqueous humor.