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J Lipid Res. 2017 Nov;58(11):2229-2237. doi: 10.1194/jlr.D077123. Epub 2017 Sep 5.

Comprehensive analyses of oxidized phospholipids using a measured MS/MS spectra library.

Aoyagi R1,2, Ikeda K1,2, Isobe Y1,2, Arita M3,2,4.

Author information

1
Laboratory for Metabolomics, RIKEN Center for Integrative Medical Sciences (IMS), Tsurumi, Yokohama, Kanagawa 230-0045, Japan.
2
Cellular and Molecular Epigenetics Laboratory, Graduate School of Medical Life Science, Yokohama City University, Tsurumi, Yokohama, Kanagawa 230-0045, Japan.
3
Laboratory for Metabolomics, RIKEN Center for Integrative Medical Sciences (IMS), Tsurumi, Yokohama, Kanagawa 230-0045, Japan makoto.arita@riken.jp.
4
Division of Physiological Chemistry and Metabolism, Graduate School of Pharmaceutical Sciences, Keio University, Minato-ku, Tokyo 105-0011, Japan.

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.

KEYWORDS:

broad-targeted lipidomics; lipidomics; lipoxygenase; mass spectrometry; oxidized lipids; tandem mass spectrometry; untargeted lipidomics

PMID:
28874441
PMCID:
PMC5665662
[Available on 2018-11-01]
DOI:
10.1194/jlr.D077123
[Indexed for MEDLINE]

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