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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter / Tate, Jillian R.

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

Laura Guerrero1 / Pilar Martínez-Olondris2 / Montserrat Rigol3 / Mariano Esperatti2 / Cristina Esquinas2 / Néstor Luque2 / Raquel Piñer2 / Antoni Torres2 / Dolors Soy1

1Pharmacy Service, Hospital Clínic de Barcelona, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Universidad de Barcelona, Spain

2Pneumology Service, Institut Clínic del Tòrax, Hospital Clínic de Barcelona, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Universidad de Barcelona, Spain

3Cardiology Service, Institut Clínic del Tòrax, Hospital Clínic de Barcelona, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Universidad de Barcelona, Spain

Corresponding author: Dolors Soy, PharmD, PhD, Pharmacy Service, Hospital Clínic de Barcelona, C/Villarroel, 170 – 08036, Barcelona, Spain Phone: +34 93 2275479, Fax: +34 93 2275457,

Citation Information: Clinical Chemistry and Laboratory Medicine. Volume 48, Issue 3, Pages 391–398, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: 10.1515/CCLM.2010.078, February 2010

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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×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 HClO4 3% (1/1, v/v). The injection volume was 100 μL. Ofloxacin was used as an internal standard.

Results: The assay was linear over a linezolid concentration range: 1.6–100 μg/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 μg/mL) and quantitation (1.6 μg/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.

Clin Chem Lab Med 2010;48:391–8.

Keywords: analytics; chromatography; high pressure liquid; linezolid; tissue; validation

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