Synthesis and NMR characterization of the methyl esters of eicosapentaenoic acid monoepoxides

The activities of cytochrome P450-derived epoxide metabolites of omega-6 polyunsaturated fatty acids (PUFAs) in cellular homeostasis have generated considerable topical interest, but there is less information on the effects of omega-3 PUFA epoxides. Mass spectroscopic data on the epoxides of the omega-3 PUFA eicosapentaenoic acid (EPA) have been reported but the absence of corresponding NMR data currently hinders their biological assessment. In the present study five monoepoxy derivatives of EPA methyl ester were synthesized by treating EPA methyl ester with m-chloroperbenzoic acid. The individual regioisomers were purified by normal-phase chromatography and characterized by LC-MS/MS and a combination of NMR approaches including H-1-, C-13-, H-1-H-1-COSY, H-1-C-13-HSQC, and H-1-C-13-HMBC. The chromatographic properties for these monoepoxides were studied in normal-phase and reverse phase-H PLC systems and the MS/MS fragmentation patterns using electrospray ionization were established. This paper also focuses on the NMR characterization of epoxide. olefinic and methylenic moieties and the complete assignments of the isomers. (C) 2009 Elsevier Ireland Ltd. All rights reserved.