title: Determination of ofloxacin in human aqueous humour by high-performance liquid chromatography with fluorescence detection.
abstract: A reversed-phase high-performance liquid chromatographic method is described for the determination of ofloxacin in human aqueous humour; the method involves fluorescence detection (excitation at 290 nm; emission at 500 nm) after direct injection of samples. The method utilized a 100 mm x 8 mm i.d. cartridge column packed with 4 microns Novapak C18 with a mobile phase methanol-acetonitrile-0.4 M citric acid (3:1:10, v/v/v) and a flow rate of 1 ml min-1 at ambient temperature. The retention times for the internal standard pipemidic acid and for ofloxacin were 4.82 and 7.32 min respectively. The mean recovery (+/- ISD) from human aqueous humour was 103.24 +/- 4.45% for ofloxacin at 1 microgram ml-1 (n = 6). The within-day and day-to-day RSDs at 0.1 microgram ml-1 and 1 microgram ml-1 were less than 6.71% (n = 6) and the lower limit of reliable determination corresponding to a signal-to-noise ratio of 2.5:1 was 20 ng ml-1. The assay was shown to be suitable for measuring ofloxacin levels in human aqueous humour samples after topical, oral and intravenous administration.
authors.affiliation: Department of Pharmacology, Faculty of Medicine, Hacettepe University, Ankara, Turkey.
authors.forename: N E
authors.initials: NE
authors.lastname: Basci
authors.forename: S
authors.initials: S
authors.lastname: Hanioglu-Kargi
authors.forename: H
authors.initials: H
authors.lastname: Soysal
authors.forename: A
authors.initials: A
authors.lastname: Bozkurt
authors.forename: S O
authors.initials: SO
authors.lastname: Kayaalp
mesh_terms: D000890:Anti-Infective Agents / Q000032:analysis*; D001082:Aqueous Humor / Q000737:chemistry*; D002853:Chromatography, Liquid / Q000379:methods; D006801:Humans; D015242:Ofloxacin / Q000032:analysis*; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods
chemical_list: D000890:Anti-Infective Agents; D015242:Ofloxacin