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

J Pharm Biomed Anal. 1997 Feb;15(5):663-6. 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 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Infective Agents / analysis*
  • Aqueous Humor / chemistry*
  • Chromatography, Liquid / methods
  • Humans
  • Ofloxacin / analysis*
  • Sensitivity and Specificity
  • Spectrometry, Fluorescence / methods


  • Anti-Infective Agents
  • Ofloxacin