Determination of ω-6 and ω-3 PUFA metabolites in human urine samples using UPLC/MS/MS

Ai Sasaki, Hayato Fukuda, Narumi Shiida, Nobuaki Tanaka, Ayako Furugen, Jiro Ogura, Satoshi Shuto, Nariyasu Mano, Hiroaki Yamaguchi

Research output: Contribution to journalArticlepeer-review

41 Citations (Scopus)

Abstract

The ω-6 and ω-3 polyunsaturated fatty acids (PUFAs) such as arachidonic acid (AA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) are the precursors of various bioactive lipid mediators including prostaglandins, thromboxanes, leukotrienes, hydroxyeicosatetraenoic acid, isoprostanes, lipoxins, and resolvins (Rvs). These lipid mediators play important roles in various physiological and pathological processes. The quantitative determination of PUFA metabolites seems necessary for disease research and for developing biomarkers. However, there is a paucity of analytical methods for the quantification of ω-6 and ω-3 PUFA metabolites - the specialized pro-resolving mediators (SPMs) present in the human urine. We developed a method for the quantification of ω-6 and ω-3 PUFA metabolites present in human urine using ultra-performance liquid chromatography/tandem mass spectrometry (UPLC/MS/MS). The developed method shows good linearity, with a correlation coefficient >0.99 for all of the analytes. The validation results indicate that our method is adequately reliable, accurate, and precise. The method was successfully used to examine urine samples obtained from 43 healthy volunteers. We could identify 20 PUFA metabolites, and this is the first report of the quantitative determination of RvD1, 17(R)-RvD1, 11-dehydro thromboxane B3, RvE2, and 5(S)-HETE in human urine. The urinary 8-iso PGF and PGE2 levels were significantly higher in the men smokers than in the men nonsmokers (p<0.05). In this study, we developed an accurate, precise, and novel analytical method for estimating the ω-6 and ω-3 PUFA metabolites, and this is the first report that the SPMs derived from EPA and DHA are present in human urine.

Original languageEnglish
Pages (from-to)1625-1639
Number of pages15
JournalAnalytical and Bioanalytical Chemistry
Volume407
Issue number6
DOIs
Publication statusPublished - 2015 Feb 17

Keywords

  • Eicosanoids
  • Polyunsaturated fatty acids
  • Specialized pro-resolving mediators
  • UPLC/MS/MS
  • Urine

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

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