An adaptable HPLC method for the analysis of frequently used antibiotics in ocular samples

Lindsay T. Davis, Neeru Kumar, Lisa M. Nijm, Lawrence J. Ulanski, Elmer Y. Tu, Richard G. Fiscella, Randal J. Peterson, Randolph D. Glickman

Research output: Contribution to journalArticlepeer-review

20 Scopus citations


Four different antibiotics, delivered individually to rabbit eyes via hydrophilic intraocular lenses soaked in the drug solution prior to implantation, were measured in aqueous and vitreous humor samples from the eyes. To meet this analytical need, we developed a sensitive, high performance liquid chromatographic (HPLC) method for measuring the concentrations of moxifloxacin, gatifloxacin, linezolid, and cefuroxime in the ocular tissue. Separations were carried out on a LichroSpher RP-18 column, maintained at room temperature. The fluoroquinolones were eluted with a mobile phase consisting of 20% acetonitrile, in 0.1% trifluoroacetic acid (pH 3.0) with 30. mM tetrabutylammonium chloride. Linezolid and cefuroxime were eluted with 25% acetonitrile in 25. mM Na acetate buffer, pH 5.0. All elutions were isocratic. With ultraviolet detection, the lower limit of quantitation (LLOQ) for these compounds approached 1. ng (on-column injection). By using fluorescence detection, the LLOQ for the fluoroquinolones improved to 200. pg. The overall accuracy of the method was ≥90%. With minor modifications, the method was optimized for each of the agents, and the resulting analytical sensitivity made the method suitable for clinical investigations of the ocular penetration of these drugs.

Original languageEnglish (US)
Pages (from-to)2421-2426
Number of pages6
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Issue number26
StatePublished - Sep 2010


  • Antibiotics
  • Fluoroquinolones
  • HPLC
  • Ocular tissue
  • Oxazolidinones
  • β-Lactams

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Cell Biology


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