Determination of ofloxacin in human aqueous humour by high-performance liquid chromatography with fluorescence detection

• Published in: Journal of pharmaceutical and biomedical analysis Vol. 15; no. 5; pp. 663 - 666
• Main Authors: Basci, N.E., Hanioglu-Kargi, S., Soysal, H., Bozkurt, A., Kayaalp, S.O.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01.02.1997; Elsevier Science
• Subjects: Analysis; Anti-Infective Agents - analysis; Aqueous Humor - chemistry; Aqueous humour; Biological and medical sciences; Chromatography, Liquid - methods; General pharmacology; High-performance liquid chromatography; Humans; Medical sciences; Ofloxacin; Ofloxacin - analysis; Pharmacology. Drug treatments; Sensitivity and Specificity; Spectrometry, Fluorescence - methods; High-performance liquid chromatography; Ofloxacin; Aqueous humour; Human; Fluorescence detector; Intravenous administration; Oral administration; HPLC chromatography; Aqueous humor; Biological material; Direct method; Eye; Local administration; Reversed phase chromatography; Fluoroquinolone derivatives; Antibacterial agent; Pharmacokinetics; Quinolone derivatives; Quantitative analysis
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/S0731-7085(96)01889-4

A reversed-phase high-performance liquid chromatographic method is described for the determination of ofloxacin in human aqueous humour; the method involves fluroescence detection (excitation at 290 nm; emission at 500 nm) after direct injection of samples. The method utilized a 100 mm × 8 mm i.d. cartridge column packed with 4 μm Novapak C 18 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 μg ml −1 ( n = 6). The within-day and day-to-day RSDs at 0.1 μg ml −1 and 1 μg 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.