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dc.contributor.authorRøst, Lisa Marie
dc.contributor.authorShafaei, Armaghan
dc.contributor.authorFuchino, Katsuya
dc.contributor.authorBruheim, Per
dc.identifier.citationJournal of chromatography. B. 2020, 1144 1-12.en_US
dc.description.abstractThe pyridine nucleotides nicotineamide adenine dinucleotide (NAD) and nicotineamide adenine dinucleotide phosphate (NADP) are conserved coenzymes across all domains of life, and are involved in more than 200 different hydride transfer reactions supporting essential catabolic and anabolic functions. The intracellular levels of these metabolites, and the ratio of their oxidized to reduced forms regulate an extensive network of reactions ranging beyond metabolism. Hence, monitoring their intracellular levels provides information about, but not limited to, the metabolic state of a cell or tissue. Interconversion between oxidized and reduced forms, varying pH liability and varying intracellular concentrations of the different species leaves absolute quantification of the pyridine nucleotides analytically challenging. These polar metabolites are poorly retained on conventional reverseed-phase stationary phases without ion-pair reagents that contaminates the LC-system. Herein we demonstrate that zwitterionic HILIC-tandem mass spectroemtry can be applied to successfully resolve the pyridine nucleotides in biological extracts in a fast, robust and highly sensitive way. The presented method applies isotope dilution to compensate potential loss of these labile metabolites and is validated for low, medium and high biomass samples of two popular biological model systems; Escherichia coli and the human cell line JJN-3. High stability and rapid sample preparation without solvent removal allows for long sequence runs, making this method ideal for high-throughput analysis of biological extracts.en_US
dc.rightsNavngivelse 4.0 Internasjonal*
dc.titleZwitterionic HILIC tandem mass spectrometry with isotope dilution for rapid, sensitive and robust quantification of pyridine nucleotides in biological extractsen_US
dc.typePeer revieweden_US
dc.typeJournal articleen_US
dc.source.journalJournal of chromatography. Ben_US
dc.relation.projectNorges forskningsråd: 258657en_US
dc.relation.projectNorges forskningsråd: 269432en_US
dc.relation.projectNorges forskningsråd: 248885en_US
dc.description.localcodeThis is an open access article distributed under the terms of the Creative Commons CC-BY license, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.en_US

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