Fer Maude, Goulitquer Sophie, Dréano Yvonne, Berthou François, Corcos Laurent, Amet Yolande
Laboratoire de Biochimie EA948, Faculté de Médecine, CS 93837, 29238-Brest-Cedex 3, France.
J Chromatogr A. 2006 May 19;1115(1-2):1-7. doi: 10.1016/j.chroma.2006.02.060. Epub 2006 Mar 20.
Despite the implication of polyunsaturated fatty acid monoepoxides in a large panel of biological effects, few methods allowing their separation in a single run are available. We describe here a simple method based on reversed-phase ion-pair high-performance liquid chromatography (RP-HPLC) and developed to successfully separate the various monoepoxides of eicosatrienoic, arachidonic, eicosapentaenoic and docosahexaenoic acids. These compounds were easily identified by liquid chromatography-mass spectrometry (LC-MS) with atmospheric pressure chemical ionisation owing to the volatility of counter-ion species. Compared to established methods, this new protocol proved its ability to totally resolve, in a single run, all of the different regioisomeric epoxides. In the long run, this method will demonstrate its efficacy to give insights into the cytochrome P450-dependent metabolism of polyunsaturated fatty acids (PUFAs) and the generation of physiologically active epoxy-derivatives.
