A simplified procedure for semi-targeted lipidomic analysis of oxidized phosphatidylcholines induced by UVA irradiation
- Florian Gruber*,
- Wolfgang Bicker†,
- Olga V. Oskolkova§,
- Erwin Tschachler*,** and
- Valery N. Bochkov1,§
- *Department of Dermatology, Medical University of Vienna, Vienna, Austria
- †FTC-Forensic-Toxicological Laboratory Ltd., Vienna, Austria
- §Department of Vascular Biology and Thrombosis Research, Center for Physiology and Pharmacology, Medical University of Vienna, Vienna, Austria; and
- **CE.R.I.E.S, Neuilly sur Seine, France
Abstract
Oxidized phospholipids (OxPLs) are increasingly recognized as signaling mediators that are not only markers of oxidative stress but are also “makers” of pathology relevant to disease pathogenesis. Understanding the biological role of individual molecular species of OxPLs requires the knowledge of their concentration kinetics in cells and tissues. In this work, we describe a straightforward “fingerprinting” procedure for analysis of a broad spectrum of molecular species generated by oxidation of the four most abundant species of polyunsaturated phosphatidylcholines (OxPCs). The approach is based on liquid-liquid extraction followed by reversed-phase HPLC coupled to electrospray ionization MS/MS. More than 500 peaks corresponding in retention properties to polar and oxidized PCs were detected within 8 min at 99 m/z precursor values using a single diagnostic product ion in extracts from human dermal fibroblasts. Two hundred seventeen of these peaks were fluence-dependently and statistically significantly increased upon exposure of cells to UVA irradiation, suggesting that these are genuine oxidized or oxidatively fragmented species. This method of semitargeted lipidomic analysis may serve as a simple first step for characterization of specific “signatures” of OxPCs produced by different types of oxidative stress in order to select the most informative peaks for identification of their molecular structure and biological role.
Footnotes
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↵1 To whom correspondence should be addressed. e-mail: valery.bochkov{at}meduniwien.ac.at
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- Abbreviations:
- BHT
- butylated hydroxytoluene
- CV
- coefficient of variation
- DNPC
- 1,2-dinonanoyl-sn-glycero-3-phosphocholine
- ESI
- electrospray ionization
- LLE
- liquid-liquid extraction
- OxPC
- oxidized phosphatidylcholine
- OxPL
- oxidized phospholipid
- PAzPC
- 1-palmitoyl-2-azelaoyl-sn-glycero-3-phosphocholine
- PAPC
- 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine
- PARP
- poly ADP-ribose polymerase
- PGPC
- 1-palmitoyl-2-glutaroyl-sn-glycero-3-phosphocholine
- PLPC
- 1-palmitoyl-2-linoleoyl-sn-glycero-3-phosphocholine
- PONPC
- 1-palmitoyl-2-(9-oxo)nonanoyl-sn-glycero-3-phosphocholine
- POVPC
- 1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphocholine
- SAPC
- 1-stearoyl-2-arachidonoyl-sn-glycero-3-phosphocholine
- SLPC
- 1-stearoyl-2-linoleoyl-sn-glycero-3-phosphocholine
- SRM
- selected reaction monitoring
- UVA
- ultraviolet A
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The work was supported by grants from Österreichische Forschungsförderungsgesellschaft (project 815445 to V.N.B.) and Fonds zur Förderung wissenschaftlicher Forschung (P20801-B11 to V.N.B. and P22267-B11 to O.V.O.).
- Received February 9, 2012.
- Revision received March 1, 2012.
- Copyright © 2012 by the American Society for Biochemistry and Molecular Biology, Inc.
