Determination of polyunsaturated fatty acid monoepoxides by high performance liquid chromatography-mass spectrometry.
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.
Fer
Maude
M
Laboratoire de Biochimie EA948, Faculté de Médecine, CS 93837, 29238-Brest-Cedex 3, France.
Goulitquer
Sophie
S
Dréano
Yvonne
Y
Berthou
François
F
Corcos
Laurent
L
Amet
Yolande
Y
D001189:Aryl Hydrocarbon Hydroxylases / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D065731:Cytochrome P-450 CYP2C19; D004281:Docosahexaenoic Acids / Q000378:metabolism; D004852:Epoxy Compounds / Q000032:analysis* / Q000302:isolation & purification; D005231:Fatty Acids, Unsaturated / Q000032:analysis*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D006899:Mixed Function Oxygenases / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism
D004852:Epoxy Compounds; D005231:Fatty Acids, Unsaturated; D011994:Recombinant Proteins; D004281:Docosahexaenoic Acids; D006899:Mixed Function Oxygenases; D001189:Aryl Hydrocarbon Hydroxylases; C045793:CYP2C19 protein, human; D065731:Cytochrome P-450 CYP2C19