Development and validation of a high performance liquid chromatography method to determine linezolid concentrations in pig pulmonary tissue

Clin Chem Lab Med. 2010 Mar;48(3):391-8. doi: 10.1515/CCLM.2010.078.

Abstract

Background: Linezolid is the first synthetic compound of a new group of antimicrobials, the oxazolidinones, which inhibit protein synthesis. It shows a broad spectrum of activity against Gram positive organisms. With respect to its pharmacokinetics, linezolid shows a relatively high volume of distribution and good penetration into inflammatory fluids, bone, fat and muscle.

Methods: A reversed-phase isocratic high-performance liquid chromatographic method for linezolid analysis in piglet pulmonary tissue is described. Tissue samples and controls were prepared in 1 x TBE (1 M Tris, 0.9 M boric acid, 0.01 M EDTA). The mobile phase consisted of 20% ultrafiltered water and 80% of (A) 15 mM potassium monohydrogen phosphate buffer (pH = 5) with (B) acetonitrile (80%/20%; v/v). Samples were homogenized and precipitated with HClO(4) 3% (1/1, v/v). The injection volume was 100 microL. Ofloxacin was used as an internal standard.

Results: The assay was linear over a linezolid concentration range: 1.6-100 microg/mL. The method provided good validation data (n = 15): inaccuracy (3.6%), intra and inter-day variability (4.2% and 5.2%, respectively), recovery (91.8%), limit of detection (0.8 microg/mL) and quantitation (1.6 microg/mL) and acceptable stability within 24 h in the auto-sampler.

Conclusions: The method offers a fast and simple approach to determine linezolid in pulmonary tissue which could be of use in pharmacokinetic studies.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Acetamides / analysis*
  • Acetamides / pharmacokinetics
  • Animals
  • Anti-Infective Agents / analysis*
  • Anti-Infective Agents / pharmacokinetics
  • Calibration
  • Chromatography, High Pressure Liquid / methods*
  • Chromatography, High Pressure Liquid / standards
  • Drug Stability
  • Limit of Detection
  • Linezolid
  • Lung / chemistry*
  • Oxazolidinones / analysis*
  • Oxazolidinones / pharmacokinetics
  • Reproducibility of Results
  • Swine

Substances

  • Acetamides
  • Anti-Infective Agents
  • Oxazolidinones
  • Linezolid