Compound-specific isotope analysis reveals no retroconversion of DHA to EPA but substantial conversion of EPA to DHA following supplementation: a randomized control trial

Am J Clin Nutr. 2019 Oct 1;110(4):823-831. doi: 10.1093/ajcn/nqz097.


Background: It has long been believed that DHA supplementation increases plasma EPA via the retroconversion pathway in mammals. However, in rodents this increase in EPA is likely due to a slower metabolism of EPA, but this has never been tested directly in humans.

Objective: The aim of this study was to use the natural variations in 13C:12C ratio (carbon-13 isotopic abundance [δ13C]) of n-3 PUFA supplements to assess n-3 PUFA metabolism following DHA or EPA supplementation in humans.

Methods: Participants (aged 21.6 ± 2.2 y) were randomly assigned into 1 of 3 supplement groups for 12 wk: 1) olive oil control, 2) ∼3 g/d DHA, or 3) ∼3 g/d EPA. Blood was collected before and after the supplementation period, and concentrations and δ13C of plasma n-3 PUFA were determined.

Results: DHA supplementation increased (P < 0.05) plasma EPA concentrations by 130% but did not affect plasma δ13C-EPA (-31.0 ± 0.30 to -30.8 ± 0.19, milliUrey ± SEM, P > 0.05). In addition, EPA supplementation did not change plasma DHA concentrations (P > 0.05) but did increase plasma δ13C-DHA (-27.9 ± 0.2 to -25.6 ± 0.1, P < 0.05) toward δ13C-EPA of the supplement (-23.5 ± 0.22). EPA supplementation increased plasma concentrations of EPA and docosapentaenoic acid (DPAn-3) by 880% and 200%, respectively, and increased plasma δ13C-EPA (-31.5 ± 0.2 to -25.7 ± 0.2) and δ13C-DPAn-3 (-28.9 ± 0.3 to -25.0 ± 0.1) toward δ13C-EPA of the supplement.

Conclusions: In this study, we show that the increase in plasma EPA following DHA supplementation in humans does not occur via retroconversion, but instead from a slowed metabolism and/or accumulation of plasma EPA. Furthermore, substantial amounts of supplemental EPA can be converted into DHA. δ13C of n-3 PUFA in humans is a powerful and underutilized tool that can track dietary n-3 PUFA and elucidate complex metabolic questions. This trial was registered at as NCT03378232.

Keywords: carbon-13 isotopic abundance; docosahexaenoic acid; eicosapentaenoic acid; humans; isotope ratio mass spectrometry; metabolism; polyunsaturated fatty acid.

Publication types

  • Randomized Controlled Trial
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbon Isotopes
  • Dietary Supplements
  • Docosahexaenoic Acids / administration & dosage*
  • Docosahexaenoic Acids / chemistry
  • Docosahexaenoic Acids / metabolism*
  • Double-Blind Method
  • Eicosapentaenoic Acid / administration & dosage*
  • Eicosapentaenoic Acid / chemistry
  • Eicosapentaenoic Acid / metabolism*
  • Female
  • Humans
  • Male
  • Young Adult


  • Carbon Isotopes
  • Docosahexaenoic Acids
  • Eicosapentaenoic Acid

Associated data