High performance liquid chromatography of fatty acid metabolites - Improvement of sensitivity by radiometric, fluorimetric and mass spectrometric methods

To get abetter understanding and knowledge of microsomal cytochrome P450 involvement in the metabolism of fatty acids, e.g. lauric and oleic acids, high performance liquid chromatography (HPLC) was used to resolve the main metabolites. It was coupled with three detection techniques, i.e. radiometry, fluorimetry and mass spectrometry (MS), to compare their respective sensitivity. They all showed a high sensitivity in the investigations carried out; none of them destroy the substrate, which allowed recovery for further analysis by gas chromatography (GC)-MS for a complete identification. Their application to the study of fatty acid metabolism by liver microsomes from human and rats (control, ethanol- or clofibrate-treated) led to nearly alike results with significant correlation coefficients between the various methods in both human and rat liver microsomes. (C) 2002 Elsevier Science B.V. All rights reserved.