Targeted, LCMS-based metabolomics for quantitative measurement of NAD+ metabolites
Research output: Contribution to journal › Journal article › Research › peer-review
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.
Original language | English |
---|---|
Article number | e201301012 |
Journal | Computational and Structural Biotechnology Journal |
Volume | 4 |
Issue number | 5 |
ISSN | 2001-0370 |
DOIs | |
Publication status | Published - 2013 |
Externally published | Yes |
ID: 220855687