Liquid chromatographic determination of ethambutol in serum samples based on intramolecular excimer-forming fluorescence derivatization

Anal Sci. 2004 Mar;20(3):489-93. doi: 10.2116/analsci.20.489.

Abstract

A selective liquid chromatographic method has been developed for the assay of ethambutol in serum samples. The assay involves intramolecular excimer-forming derivatization with 4-(1-pyrene)butanoyl chloride (PBC) and isocratic reversed-phase chromatography with fluorescence detection. After acetonitrile-deproteinization of the serum sample, the derivatization reaction of ethambutol with PBC was completed within 30 min at 50 degrees C. N,N'-Diethylethylenediamine was used as an internal standard. The detection limit of ethambutol was 23 ng/ml serum, corresponding to 180 fmol on column at a signal-to-noise ratio of 3. The present method was selective enough to analyze ethambutol in rabbit serum without any tedious sample clean-up procedure because biogenic monoamines gave no peak in the chromatogram. The method was applicable to drug monitoring in rabbit serum.

MeSH terms

  • Animals
  • Antitubercular Agents / blood*
  • Antitubercular Agents / pharmacokinetics
  • Ethambutol / blood*
  • Ethambutol / pharmacokinetics
  • Ethylenediamines
  • Fluorescent Dyes
  • Indicators and Reagents
  • Rabbits
  • Reference Standards
  • Reproducibility of Results
  • Solutions
  • Spectrometry, Fluorescence

Substances

  • Antitubercular Agents
  • Ethylenediamines
  • Fluorescent Dyes
  • Indicators and Reagents
  • Solutions
  • N,N-diethylethylenediamine
  • Ethambutol