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

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An Erratum to this article was published on 24 February 2015

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 PGF2α 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.

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Abbreviations

AA:

Arachidonic acid

COX:

Cyclooxygenase

DHA:

Docosahexaenoic acid

EPA:

Eicosapentaenoic acid

HETE:

Hydroxyeicosatetraenoic acid

IS:

Internal standard

IsoPs:

Isoprostanes

LLOQ:

Lower limit of quantification

LOX:

Lipoxygenase

LT:

Leukotriene

LX:

Lipoxin

MaR:

Maresin

PD1:

Protectin D1

PG:

Prostaglandin

PUFA:

Polyunsaturated fatty acid

SPE:

Solid-phase extraction

SPM:

Specialized pro-resolving mediator

SRM:

Selected reaction monitoring

Rv:

Resolvin

TX:

Thromboxane

UPLC/MS/MS:

Ultra-performance liquid chromatography/tandem mass spectrometry

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Acknowledgments

We thank volunteers for donating urine samples. This work was supported in part by the Japan Society for the Promotion of Science (JSPS) KAKENHI grant number 25460184 (HY), and by the Towa Foundation for Food Research (HY).

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Authors and Affiliations

  1. Laboratory of Clinical Pharmaceutics and Therapeutics, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Hokkaido, 060-0812, Japan

    Ai Sasaki, Narumi Shiida, Ayako Furugen & Jiro Ogura

  2. Laboratory of Organic Chemistry for Drug Development, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Hokkaido, 060-0812, Japan

    Hayato Fukuda & Satoshi Shuto

  3. Laboratory of Clinical Pharmacy, Graduate School of Pharmaceutical Sciences, Tohoku University, Aoba-ku, Sendai, 980-8578, Japan

    Nobuaki Tanaka, Nariyasu Mano & Hiroaki Yamaguchi

  4. Department of Pharmaceutical Sciences, Tohoku University Hospital, 1-1 Seiryo-machi, Aoba-ku, Sendai, 980-8574, Japan

    Nariyasu Mano & Hiroaki Yamaguchi

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  1. Ai Sasaki
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  2. Hayato Fukuda
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Correspondence to Hiroaki Yamaguchi.

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Sasaki, A., Fukuda, H., Shiida, N. et al. Determination of ω-6 and ω-3 PUFA metabolites in human urine samples using UPLC/MS/MS. Anal Bioanal Chem 407, 1625–1639 (2015). https://doi.org/10.1007/s00216-014-8412-5

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