A validated UPLC-MS/MS method for the analysis of linezolid and a novel oxazolidinone derivative (PH027) in plasma and its application to tissue distribution study in rabbits

Mohsen A Hedaya; Vidhya Thomas; Mohamed E Abdel-Hamid; Elijah O Kehinde; Oludotun A Phillips

doi:10.1016/j.jchromb.2016.11.034

A validated UPLC-MS/MS method for the analysis of linezolid and a novel oxazolidinone derivative (PH027) in plasma and its application to tissue distribution study in rabbits

J Chromatogr B Analyt Technol Biomed Life Sci. 2017 Jan 1:1040:89-96. doi: 10.1016/j.jchromb.2016.11.034. Epub 2016 Nov 27.

Authors

Mohsen A Hedaya¹, Vidhya Thomas², Mohamed E Abdel-Hamid³, Elijah O Kehinde⁴, Oludotun A Phillips⁵

Affiliations

¹ Department of Pharmaceutics, Faculty of Pharmacy, Kuwait University, P.O. Box 24923, Safat 13110, Kuwait. Electronic address: mhedaya@hsc.edu.kw.
² Department of Pharmaceutics, Faculty of Pharmacy, Kuwait University, P.O. Box 24923, Safat 13110, Kuwait. Electronic address: vidhya@HSC.EDU.KW.
³ Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Kuwait University, P.O. Box 24923, Safat 13110, Kuwait. Electronic address: abdel-hamid@HSC.EDU.KW.
⁴ Department of Surgery, Faculty of medicine, Kuwait University, P.O. Box 24923, Safat 13110, Kuwait. Electronic address: ekehinde@HSC.EDU.KW.
⁵ Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Kuwait University, P.O. Box 24923, Safat 13110, Kuwait. Electronic address: dphillips@HSC.EDU.KW.

PMID: 27978473
DOI: 10.1016/j.jchromb.2016.11.034

Abstract

Objectives: Linezolid is the first approved oxazolidinone antibacterial agent, whereas PH027 is a novel compound of the same class that exhibits good in vitro antibacterial activity. The objective of this study was to develop an UPLC-MS/MS assay for the analysis of linezolid and PH027 in plasma and to apply the method for comparative pharmacokinetic and tissue distribution studies of both compounds.

Method: Plasma samples and calibrators were extracted with diethyl ether after addition of the internal standard solution. After evaporation of the ether layer, the residue was reconstituted in mobile phase and injected into UPLC-MS/MS. The mobile phase consisted of 2mM ammonium acetate buffer solution and acetonitrile (70:30) at a flow rate of 0.2ml/min. Separation was achieved using UPLC BEH C₁₈ column, and quantitative determination of the analytes was performed using multiple-reaction monitoring (MRM) scanning mode. The method was validated by analyzing quality control tissue homogenate samples, and was applied to analyze tissue homogenate samples obtained following IV injections of linezolid and PH027 in rabbits.

Results: The developed UPLC-MS/MS method was linear in the concentration range of 50-5000ng/ml. Validation of the method proved that the method's precision, selectivity and stability were all within the acceptable limits. Linezolid and PH027 concentrations were accurately determined in the quality control tissue homogenate samples, and analysis of samples obtained following IV administration of the two compounds showed that the tissue to plasma concentration ratio of PH027 was higher than that of linezolid probably due to its higher lipophilicity.

Conclusions: The developed UPLC-MS/MS method for the analysis of linezolid and PH027 in rabbit's plasma can accurately determine the concentrations of these compounds in different tissues.

Keywords: Linezolid; Oxazolidinone antibacterial agents; Tissue analysis; Tissue distribution; UPLC–MS/MS method validation.

Publication types

Validation Study

MeSH terms

Animals
Anti-Bacterial Agents / blood
Anti-Bacterial Agents / pharmacokinetics*
Chromatography, High Pressure Liquid / methods*
Drug Monitoring / methods
Limit of Detection
Linezolid / blood
Linezolid / pharmacokinetics*
Oxazolidinones / blood
Oxazolidinones / pharmacokinetics*
Rabbits
Tandem Mass Spectrometry / methods*
Tissue Distribution
Triazoles / blood
Triazoles / pharmacokinetics*

Substances

Anti-Bacterial Agents
Oxazolidinones
PH 027
Triazoles
Linezolid