A Rapid UPLC-MS/MS Assay for Eicosanoids in Human Plasma: Application to Evaluate Niacin Responsivity

Abstract A rapid and sensitive method using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed to simultaneously quantify hydroxyeicosatetraenoic (HETE), dihydroxyeicosatrienoic (DiHETrE), epoxyeicosatrienoic acid (EET), and prostaglandin metabolites of arach...

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Bibliographic Details
Published in:Prostaglandins, leukotrienes and essential fatty acids leukotrienes and essential fatty acids, 2018-09, Vol.136, p.153-159
Main Authors: Miller, Tricia M, Poloyac, Samuel M, Anderson, Kacey B, Waddell, Brooke L, Messamore, Erik, Yao, Jeffrey K
Format: Article
Language:English
Subjects:
Advanced Basic Science
Arachidonic acid
Eicosanoids
Endocrinology & Metabolism
Mass spectrometry
Niacin
Plasma, Schizophrenia
Prostanoids
Ultra performance liquid chromatography
UPLC-MS/MS
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Summary:Abstract A rapid and sensitive method using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed to simultaneously quantify hydroxyeicosatetraenoic (HETE), dihydroxyeicosatrienoic (DiHETrE), epoxyeicosatrienoic acid (EET), and prostaglandin metabolites of arachidonic acid in human plasma. Sample preparation consisted of solid phase extraction with Oasis HLB (30 mg) cartridges for all metabolites. Separation of HETEs, EETs, and DiHETrEs was achieved on an Acquity UPLC BEH C18, 1.7 µm (100 × 2.1 mm) reversed-phase column (Waters Corp, Millford, MA) with negative electrospray ionization mass spectrometric detection. A second injection of the same extracted sample allowed for separation and assessment of prostaglandin metabolites under optimized UPLC-MS/MS conditions. Additionally, the endogenous levels of these metabolites in five different matrices were determined in order to select the optimal matrix for assay development. Human serum albumin was shown to have the least amount of endogenous metabolites, a recovery efficiency of 79 − 100% and a matrix effect of 71 – 100%. Linear calibration curves ranging from 0.416–66.67 ng/mL were validated. Inter-assay and intra-assay variance was less than 15% at most concentrations. This method was successfully applied to quantify metabolite levels in plasma samples of healthy control subjects receiving niacin administration to evaluate the association between niacin administration and eicosanoid plasma level response.
ISSN:0952-3278
1532-2823
DOI:10.1016/j.plefa.2017.01.003