Oxidized fatty acid analysis by charge-switch derivatization, selected reaction monitoring, and accurate mass quantitation

Xinping Liu, Sung Ho Moon, David J. Mancuso, Christopher M. Jenkins, Shaoping Guan, Harold F. Sims, Richard W. Gross

Research output: Contribution to journalArticle

31 Scopus citations

Abstract

A highly sensitive, specific, and robust method for the analysis of oxidized metabolites of linoleic acid (LA), arachidonic acid (AA), and docosahexaenoic acid (DHA) was developed using charge-switch derivatization, liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI MS/MS) with selected reaction monitoring (SRM) and quantitation by high mass accuracy analysis of product ions, thereby minimizing interferences from contaminating ions. Charge-switch derivatization of LA, AA, and DHA metabolites with N-(4-aminomethylphenyl)-pyridinium resulted in a 10- to 30-fold increase in ionization efficiency. Improved quantitation was accompanied by decreased false positive interferences through accurate mass measurements of diagnostic product ions during SRM transitions by ratiometric comparisons with stable isotope internal standards. The limits of quantitation were between 0.05 and 6.0 pg, with a dynamic range of 3 to 4 orders of magnitude (correlation coefficient r2 > 0.99). This approach was used to quantitate the levels of representative fatty acid metabolites from wild-type (WT) and iPLA 2γ-/- mouse liver identifying the role of iPLA 2γ in hepatic lipid second messenger production. Collectively, these results demonstrate the utility of high mass accuracy product ion analysis in conjunction with charge-switch derivatization for the highly specific quantitation of diminutive amounts of LA, AA, and DHA metabolites in biologic systems.

Original languageEnglish
Pages (from-to)40-50
Number of pages11
JournalAnalytical Biochemistry
Volume442
Issue number1
DOIs
StatePublished - Jan 1 2013

Keywords

  • Docosanoids
  • Eicosanoids
  • LC-MS/MS
  • Metabolite determination
  • Phospholipase

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