Detection of [U‐13C]eicosapentaenoic acid in rat liver lipids using13C nuclear magnetic resonance spectroscopy

Abstract

Fatty acid carbons are well-resolved in 13C nuclear magnetic resonance (NMR) spectra of lipid extracts, but application of this methodology to the metabolism of 13C-labelled fatty acids has not yet been reported. In the present study, 13C NMR was used to monitor the presence of 98% [U-13C]eicosapentaenoic acid (EPA) in liver and carcass lipids 24 h after it had been injected into the stomach of a rat. Natural abundance 13C NMR spectra of liver total fatty acid extracts were obtained from four control rats for comparison. At 24 h post-injection, quantitative high resolution 13C NMR showed 13C enrichment in liver fatty acid extracts was present mainly at olefinic and at the n-1 to n-4 carbons, but 13C signal intensities for C-1 to C-4 of [U-13C]EPA were markedly reduced or absent. Small 13C resonances, possibly indicative of some 13C incorporation into docosahexaenoic acid and saturated or monounsaturated fatty acids, were present in spectra of liver fatty acids. Liver and carcass fatty acid composition was similar in both the controls and the EPA-injected rat, suggesting little accumulation of the injected [U-13C]EPA after 24 h. We conclude that the carbon-specific data provided by 13C NMR of lipid extracts may be useful in monitoring the fate of individual carbons during tracer studies using 13C-labelled fatty acids.