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Lipidomic profiling of targeted oxylipins with ultra-performance liquid chromatography-tandem mass spectrometry

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Abstract

Oxylipins are bioactive mediators that play diverse roles in (patho)physiology. We developed a sensitive and selective ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous profiling of 57 targeted oxylipins derived from five major n-6 and n-3 polyunsaturated fatty acids (PUFAs) that serve as oxylipin precursors, including linoleic (LA), arachidonic (AA), alpha-linolenic (ALA), eicosapentaenoic (EPA), and docosahexaenoic (DHA) acids. The targeted oxylipin panel provides broad coverage of lipid mediators and pathway markers generated from cyclooxygenases, lipoxygenases, cytochrome P450 epoxygenases/hydroxylases, and non-enzymatic oxidation pathways. The method is based on combination of protein precipitation and solid-phase extraction (SPE) for sample preparation, followed by UPLC-MS/MS. This is the first methodology to incorporate four hydroxy-epoxy-octadecenoic acids and four keto-epoxy-octadecenoic acids into an oxylipin profiling network. The novel method achieves excellent resolution and allows in-depth analysis of isomeric and isobaric species of oxylipin extracts in biological samples. The method was quantitatively characterized in human plasma with good linearity (R = 0.990–0.999), acceptable reproducibility (relative standard deviation (RSD) < 20% for the majority of analytes), accuracy (67.8 to 129.3%) for all analytes, and recovery (66.8–121.2%) for all analytes except 5,6-EET. Ion enhancement effects for 28% of the analytes in tested concentrations were observed in plasma, but were reproducible with RSD < 17.2%. Basal levels of targeted oxylipins determined in plasma and serum are in agreement with those previously reported in literature. The method has been successfully applied in clinical and preclinical studies.

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Acknowledgments

This work was supported by the Intramural Programs of the National Institute on Aging (NIA), National Institute on Alcohol Abuse and Alcoholism (NIAAA), NIH Clinical Center Department of Perioperative Medicine, and the Mayday Fund. The authors would like to thank Dr. Charles N. Serhan for sharing methods and providing resolvin and protectin standards, the research participants who provided plasma samples, the many NIH co-investigators and staff who contributed to collection and processing of these plasma samples.

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Authors and Affiliations

  1. Lipid Mediators, Inflammation, and Pain Unit, Laboratory of Clinical Investigation, National Institute on Aging/NIH, Baltimore, MD, USA

    Zhi-Xin Yuan, Gregory S. Keyes & Christopher E. Ramsden

  2. Section of Nutritional Neuroscience, Laboratory of Membrane Biochemistry and Biophysics, National Institute on Alcohol Abuse and Alcoholism/NIH, Bethesda, MD, USA

    Sharon Majchrzak-Hong & Christopher E. Ramsden

  3. Department of Perioperative Medicine, Clinical Center, NIH, Bethesda, MD, USA

    Michael J. Iadarola & Andrew J. Mannes

  4. Department of Physical Medicine and Rehabilitation, School of Medicine, Chapel Hill, NC, USA

    Christopher E. Ramsden

  5. School of Agriculture, Food and Wine, University of Adelaide, Adelaide, Australia

    Christopher E. Ramsden

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  1. Zhi-Xin Yuan
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  2. Sharon Majchrzak-Hong
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Correspondence to Zhi-Xin Yuan.

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Protocol 11-AA-0028 was reviewed and approved by an NIH Institutional Review Board (FWA# 00005897), and informed consent was obtained from all study participants.

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Yuan, ZX., Majchrzak-Hong, S., Keyes, G.S. et al. Lipidomic profiling of targeted oxylipins with ultra-performance liquid chromatography-tandem mass spectrometry. Anal Bioanal Chem 410, 6009–6029 (2018). https://doi.org/10.1007/s00216-018-1222-4

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