Comprehensive analyses of oxidized phospholipids using a measured MS/MS spectra library[S]
- Laboratory for Metabolomics,* RIKEN Center for Integrative Medical Sciences (IMS), Tsurumi, Yokohama, Kanagawa 230-0045, Japan
- Cellular and Molecular Epigenetics Laboratory,† Graduate School of Medical Life Science, Yokohama City University, Tsurumi, Yokohama, Kanagawa 230-0045, Japan
- Division of Physiological Chemistry and Metabolism,§ Graduate School of Pharmaceutical Sciences, Keio University, Minato-ku, Tokyo 105-0011, Japan
- ↵1To whom correspondence should be addressed. e-mail: makoto.arita{at}riken.jp
Abstract
Oxidized phospholipids (OxPLs) are widely held to be associated with various diseases, such as arteriosclerosis, diabetes, and cancer. To characterize the structure-specific behavior of OxPLs and their physiological relevance, we developed a comprehensive analytical method by establishing a measured MS/MS spectra library of OxPLs. Biogenic OxPLs were prepared by the addition of specific oxidized fatty acids to cultured cells, where they were incorporated into cellular phospholipids, and untargeted lipidomics by LC-quadrupole/TOF-MS was applied to collect MS/MS spectra for the OxPLs. Based on the measured MS/MS spectra for about 400 molecular species of the biogenic OxPLs, we developed a broad-targeted lipidomics system using triple quadrupole MS. Separation precision of structural isomers was optimized by multiple reaction monitoring analysis and this system enabled us to detect OxPLs at levels as low as 10 fmol. When applied to biological samples, i.e., mouse peritoneal macrophages, this system enabled us to monitor a series of OxPLs endogenously produced in a 12/15-lipoxygenase-dependent manner. This advanced analytical method will be useful to elucidate the structure-specific behavior of OxPLs and their physiological relevance in vivo.
- lipidomics
- lipoxygenase
- mass spectrometry
- oxidized lipids
- untargeted lipidomics
- broad-targeted lipidomics
- tandem mass spectrometry
Footnotes
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- Abbreviations:
- AA
- arachidonic acid
- AA+O
- mono-oxidized arachidonic acid
- CE
- collision energy
- DC
- dendritic cell
- EET
- epoxyeicosatrienoic acid
- EpDPE
- epoxydocosapentaenoic acid
- EpETE
- epoxyeicosatetraenoic acid
- HDoHE
- hydroxydocosahexaenoic acid
- HEPE
- hydroxyeicosapentaenoic acid
- IDA
- information-dependent acquisition
- LC-QTOF-MS
- LC-quadrupole/TOF-MS
- LOX
- lipoxygenase
- MRM
- multiple reaction monitoring
- OxPL
- oxidized phospholipid
- PC
- phosphatidylcholine
- PE
- phosphatidylethanolamine
- PG
- phosphatidylglycerol
- PI
- phosphatidylinositol
- PL
- phospholipid
- PS
- phosphatidylserine
- tripleQ
- triple quadrupole
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This work was supported by Japan Society for the Promotion of Science Grants 15H05898, 15H05897, and 15H04648; the Science and Technology Research Promotion Program for Agriculture, Forestry, Fisheries, and Food Industry; the Japan Agency for Medical Research and Development, and the Japan Science and Technology Agency Core Research for Evolutional Science and Technology. Additional support was provided by the RIKEN Junior Research Associate Program (to R.A.).
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↵[S] The online version of this article (available at http://www.jlr.org) contains a supplement.
- Received April 26, 2017.
- Revision received August 26, 2017.
- Copyright © 2017 by the American Society for Biochemistry and Molecular Biology, Inc.
