Targeted, LCMS-based metabolomics for quantitative measurement of NAD+ metabolites
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Nicotinamide adenine dinucleotide (NAD+) is a coenzyme for hydride transfer reactions and a substrate for sirtuins and other NAD+-consuming enzymes. The abundance of NAD +, NAD+ biosynthetic intermediates, and related nucleotides reflects the metabolic state of cells and tissues. High performance liquid chromatography (HPLC) followed by ultraviolet-visible (UV-Vis) spectroscopic analysis of NAD+ metabolites does not offer the specificity and sensitivity necessary for robust quantification of complex samples. Thus, we developed a targeted, quantitative assay of the NAD+ metabolome with the use of HPLC coupled to mass spectrometry. Here we discuss NAD+ metabolism as well as the technical challenges required for reliable quantification of the NAD+ metabolites. The new method incorporates new separations and improves upon a previously published method that suffered from the problem of ionization suppression for particular compounds.
Originalsprog | Engelsk |
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Artikelnummer | e201301012 |
Tidsskrift | Computational and Structural Biotechnology Journal |
Vol/bind | 4 |
Udgave nummer | 5 |
ISSN | 2001-0370 |
DOI | |
Status | Udgivet - 2013 |
Eksternt udgivet | Ja |
ID: 220855687