title: A UPLC-MS Method for the Determination of Ofloxacin Concentrations in Aqueous Humor.
abstract: A rapid, simple, and specific method based on ultra performance liquid chromatography (UPLC) with mass spectrometry detection has been developed for quantitative analysis of ofloxacin in human aqueous humor using tobramycin as internal standard (IS). Chromatographic separation was achieved on a Waters Acquity UPLC BEH C18 Shield column (150 × 2.1 mm, 1.7 μm) eluted with 95:5 water: acetonitrile (v/v) containing 0.1% formic acid and a flow rate of 0.3 mL/minute. The total analysis time was three minutes with ofloxacin eluting at 1.67 ± 0.03 minutes. The linearity of the method ranged from 0.1 to 8 μg/mL with r2 = 0.998. The method was validated according to FDA guidelines with respect to linearity, accuracy, precision, specificity, and stability. The limits of detection and quantification were 0.03 and 0.10 μg/mL, respectively. The developed method was successfully applied to the analysis of samples that have been obtained from patients.
authors.affiliation: Department of Ophthalmology, Medical School, University of Patras, Patras, Greece. ; Laboratory of Public Health, School of Medicine, University of Patras, Patras, Greece.
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Plotas
authors.affiliation: Department of Chemistry, University of Patras, Patras, Greece.
authors.forename: Charalampos
authors.initials: C
authors.lastname: Anastasopoulos
authors.affiliation: Department of Ophthalmology, Medical School, University of Patras, Patras, Greece.
authors.forename: Olga E
authors.initials: OE
authors.lastname: Makri
authors.affiliation: Laboratory of Public Health, School of Medicine, University of Patras, Patras, Greece.
authors.forename: Michalis
authors.initials: M
authors.lastname: Leotsinidis
authors.affiliation: Department of Ophthalmology, Medical School, University of Patras, Patras, Greece.
authors.forename: Constantine D
authors.initials: CD
authors.lastname: Georgakopoulos
keywords: UPLC; aqueous humor; mass spectrometry; ofloxacin