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PMID: 21317107 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

LC/MS/MS method for analysis of E₂ series prostaglandins and isoprostanes.

Journal of lipid research ·Vol. 52 ·No. 4 ·2011-04-00 ·Pages 850-9

Brose SA, Thuen BT, Golovko MY

Abstract

15-series prostaglandins (PGE₂s) and isoprostanes (isoPGE₂s) are robust biomarkers of oxidative stress, possess potent biological activity, and may be derived through cyclooxygenase or free radical pathways. Thus, their quantification is critical in understanding many biological processes where PG, isoPG, or oxidative stress are involved. LC/MS/MS methods allow a highly selective, sensitive, simultaneous analysis for prostanoids without derivatization. However, the LC/MS/MS methods currently used do not allow for simultaneous separation of the major brain PGE₂/D₂) and isoPGE₂ without derivatization and multiple HPLC separations. The developed LC/MS/MS method allows for the major brain PGE₂/PGD₂/isoPGE₂ such as PGE₂, entPGE₂, 8-isoPGE₂, 11β-PGE₂, PGD₂, and 15(R)-PGD₂ to be separated and quantified without derivatization. The method was validated by analyzing free and esterified isoPGE₂ in mouse brains fixed with head-focused microwave irradiation before or after global ischemia. Using the developed method, we report for the first time the esterified isoPGE₂ levels in brain tissue under basal conditions and upon global ischemia and demonstrate a nonreleasable pool of esterified isoPG upon ischemia. In addition, we demonstrated that PGE₂s found esterified in the sn-2 position in phospholipids are derived from a free radical nonenzymatic pathway under basal conditions. Our method for brain PG analysis provides a high level of selectivity to detect changes in brain PG and isoPG mass under both basal and pathological conditions.

MeSH Terms
Animals Chromatography, High Pressure Liquid/methods Chromatography, Liquid Dinoprostone/analysis In Vitro Techniques Isoprostanes/analysis Male Mice Molecular Structure Tandem Mass Spectrometry/methods
Chemicals
Isoprostanes Dinoprostone
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Brose Stephen A
Department of Pharmacology, Physiology, and Therapeutics, University of North Dakota, Grand Forks, ND 58202-9037, USA.
Thuen Brock T
Golovko Mikhail Y
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Article Info
Journal
Journal of lipid research
Abbr.
J Lipid Res
ISSN
1539-7262
DOI
10.1194/jlr.D013441
Published
2011-04-00
Epub
2011-00-10
Pages
850-9
Language
English
Region
United States
NLM ID
0376606
PMCID
PMC3284174
Subset
IM
Grants
NCRR NIH HHS · P20 RR017699 · United States
NINDS NIH HHS · R21 NS064480 · United States
NINDS NIH HHS · 5R21NS064480-02 · United States
NCRR NIH HHS · 5P20RR017699 · United States
External Links
PubMed source DOI: 10.1194/jlr.D013441 PMC Full Text View journal details
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