A rapid UPLC-MS/MS assay for eicosanoids in human plasma: Application to evaluate niacin responsivity. (September 2018)

• Record Type: Journal Article
• Authors: Miller, Tricia M.; Poloyac, Samuel M.; Anderson, Kacey B.; Waddell, Brooke L.; Messamore, Erik; Yao, Jeffrey K.
• Is Part Of: Prostaglandins, leukotrienes, and essential fatty acids. Volume 136(2018)
• Page Start: 153
• Page End: 159
• Publication Date: 2018-09
• Subjects: AA arachidonic acid -- BHT butylated hydroxytoluene -- BSA bovine serum albumin -- CBF cerebral blood flow -- COX cyclooxygenase -- CSF cerebrospinal fluid -- CYP cytochrome P-450 -- DiHETrE dihydroxyeicosatrienoic acid -- DiHOME dihydroxyoctadecenoic acid -- EDTA Ethylenediaminetetraacetic acid -- EET epoxyeicosatrienoic acid -- Epome epoxyoctadecanoic acid -- ESI electrospray ionization -- HETE hydroxyeicosatetraenoic acid -- HSA human serum albumin -- LOX lipoxygenase -- ME matrix effect -- MS Mass spectrometry -- RE recovery efficiency -- SAH subarachnoid hemorrhage -- SPE solid phase extraction -- SRM selected reaction monitoring -- PG prostaglandins -- QC quality control -- UPLC Ultra performance liquid chromatography; Arachidonic acid -- Eicosanoids -- Prostanoids -- Mass spectrometry -- Ultra performance liquid chromatography -- UPLC-MS/MS -- Plasma, Schizophrenia -- Niacin; Lipids -- Periodicals; Unsaturated fatty acids -- Periodicals; Prostaglandins -- Periodicals; Leukotrienes -- Periodicals; Fatty Acids, Unsaturated -- Periodicals; Acides gras insaturés -- Périodiques; Prostaglandines -- Périodiques; Leucotriènes -- Périodiques; Lipides -- Périodiques; 612.01577
• DOI: 10.1016/j.plefa.2017.01.003
• Languages: English
• ISSNs: 0952-3278

Abstract

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 to 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.