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Analysis of hydroxyeicosatetraenoic acids (HETEs) in human whole blood by chiral ultrahigh performance liquid chromatgraphy (UHPLC)-electron capture atmospheric pressure chemical ionization/high resolution mass spectrometry (ECAPCI/HRMS).

Analysis of hydroxyeicosatetraenoic acids (HETEs) in human whole blood by chiral ultrahigh performance liquid chromatgraphy (UHPLC)-electron capture atmospheric pressure chemical ionization/high resolution mass spectrometry (ECAPCI/HRMS).
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Mazaleuskaya LL, Salamatipour A, Sarantopoulou D, Weng L, FitzGerald GA, Blair IA, Mesaros C,


Mazaleuskaya LL, Salamatipour A, Sarantopoulou D, Weng L, FitzGerald GA, Blair IA, Mesaros C, (click to view)

Mazaleuskaya LL, Salamatipour A, Sarantopoulou D, Weng L, FitzGerald GA, Blair IA, Mesaros C,

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Journal of lipid research 2018 01 04() pii jlr.D081414
Abstract

The biosynthesis of eicosanoids occurs enzymatically via lipoxygenases (LOXs), cyclooxygenases (COXs), and cytochrome P450, or through non-enzymatic free radical reactions. The enzymatic routes are highly enantiospecific. Chiral separation and high sensitivity detection methods are required to differentiate and quantify enantioselective HETEs (hydroxy-(5Z,8Z,11Z,13E)-eicosatetraenoic acids) in complex biological fluids. We report here a targeted chiral lipidomics analysis of human blood using ultrahigh performance liquid chromatgraphy (UHPLC)-electron capture atmospheric pressure chemical ionization/high resolution mass spectrometry (ECAPCI/HRMS). Monitoring the high-resolution (HR) ions formed by the fragmentation of pentafluorobenzyl derivatives of oxidized lipids during the dissociative electron capture, followed by in trap fragmentation, increased sensitivity by an order of magnitude when compared to the unit resolution mass spectrometry. 12(S)-HETE, 12(S)-HHT (12(S)-hydroxy-(5Z,8E,10E)-heptadecatrienoic acid) and 15(S)-HETE were the major hydroxylated, non-esterified chiral lipids in serum. Stimulation of whole blood with zymosan and LPS resulted in stimulus- and time-dependent effects. An acute exposure to zymosan induced ~80% of the chiral plasma lipids including 12(S)-HHT, 5(S)-HETE, 15(R)-HETE and 15(S)-HETE, while a maximum response to LPS was achieved after a long-term stimulation. The reported method allows for a rapid quantification with high sensitivity and specificity of enantiospecific responses to in vitro stimulation or coagulation of human blood.

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