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Simultaneous Analysis of Multiple Lipid Oxidation Products In Vivo by Liquid Chromatographic-Mass Spectrometry (LC-MS)

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Free Radicals and Antioxidant Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 610))

Abstract

Free radical-induced oxidation of polyunsaturated fatty acid (PUFAs) has been linked to a number of human diseases including atherosclerosis and neurodegenerative disorders. Oxidation of PUFAs generates hydroperoxides and cyclic peroxides that are reduced to lipid alcohol, such as hydroxyeicosatetraenoic acid (HETEs), and isoprostanes (IsoPs) respectively. The IsoPs are isomers of prostaglandins that are generated from autoxidation of arachidonic acid (C20:4). Quantification of F2-IsoPs has been regarded as the “gold standard” to assess oxidative stress status in various human diseases. We herein report the protocol of analyzing HETEs and F2-IsoPs using a triple quadrupole mass spectrometer coupled to reverse phase liquid chromatography. The selected reaction monitoring (SRM) mode selects the parent ion of interest in the first Quad (m/z 319 for HETE and m/z 353 for F2-IsoPs) and fragments it in the second while an ion characteristic of the analyte of interest is monitored in the third Quad. This highly selective technique permits the simultaneous analysis of multiple oxidation products such as the HETEs and F2-IsoPs. This LC-MS technique can be applied to study the free radical oxidation mechanism in vitro and assess the oxidative stress status in biological tissues and fluids.

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Abbreviations

APCI:

atmospheric pressure chemical ionization

BHT:

butylated hydroxytoluene

CID:

collision induced dissociation

ESI-MS:

electrospray mass spectrometry

GC:

gas chromatography

HPLC:

high performance liquid chromatography

IsoP:

isoprostane

IPA:

isopropanol

LC:

liquid chromatography

MS:

mass spectrometry

PG:

prostaglandin

PGF2 α :

Prostaglandin F2 α

HpETE:

hydroperoxyeicosatetraenoate

SRM:

selective reaction monitoring.

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Acknowledgments

We thank Dr. David Hachey, Ms. Lisa Mannier, and Mrs. Betty Fox of the Mass Spectrometry Research Center of Vanderbilt University for assistance with the MS analysis. We acknowledge discussion with Dr. Jason Morrow in the Division of Clinical Pharmacology at Vanderbilt University. Financial support from NIEHS P01 ES013125, NIH grants DK 48831, RR00095, CA 77839, HL17921, GM15431, P30 ES00267 (HY, pilot grant), and NSF CHE 0412493 is gratefully acknowledged.

Author information

Authors and Affiliations

  1. Division of Clinical Pharmacology, Departments of Medicine and Chemistry Vanderbilt University, Nashville, TN, USA

    Huiyong Yin

  2. Department of Chemistry, Idaho State University, Pocatello, ID, 83209, USA

    Todd Davis

  3. Department of Chemistry, Center in Molecular Toxicology, and Vanderbilt Institute of Chemical Biology, Vanderbilt University, Nashville, TN, USA

    Ned A. Porter

Authors
  1. Huiyong Yin
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  2. Todd Davis
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  3. Ned A. Porter
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Editor information

Editors and Affiliations

  1. A& M College, Southern University &, James L. Hunt Street, Baton Rouge, 70813, U.S.A.

    Rao M. Uppu

  2. School of Medicine, Tulane University, Tulane Ave. 1430, New Orleans, 70112, U.S.A.

    Subramanyam N. Murthy

  3. Biodynamics Inst., Louisiana State University, Baton Rouge, 70808, U.S.A.

    William A. Pryor

  4. Dorothy M. Davis Heart &, Ohio State University, West 12th Ave. 473, Columbus, 43210, U.S.A.

    Narasimham L. Parinandi

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© 2010 Humana Press, a part of Springer Science+Business Media, LLC

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Yin, H., Davis, T., Porter, N.A. (2010). Simultaneous Analysis of Multiple Lipid Oxidation Products In Vivo by Liquid Chromatographic-Mass Spectrometry (LC-MS). In: Uppu, R., Murthy, S., Pryor, W., Parinandi, N. (eds) Free Radicals and Antioxidant Protocols. Methods in Molecular Biology, vol 610. Humana Press. https://doi.org/10.1007/978-1-60327-029-8_22

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  • DOI: https://doi.org/10.1007/978-1-60327-029-8_22

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-710-5

  • Online ISBN: 978-1-60327-029-8

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