Targeted, LCMS-based metabolomics for quantitative measurement of NAD+ metabolites

TY - GEN

T1 - Targeted, LCMS-based metabolomics for quantitative measurement of NAD+ metabolites

AU - Trammell, Samuel A.J.

AU - Brenner, Charles

PY - 2013/1/1

Y1 - 2013/1/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84888626285&partnerID=8YFLogxK

U2 - 10.5936/csbj.201301012

DO - 10.5936/csbj.201301012

M3 - Short survey

AN - SCOPUS:84888626285

SN - 2001-0370

VL - 4

SP - e201301012

JO - Computational and Structural Biotechnology Journal

JF - Computational and Structural Biotechnology Journal

IS - 5

M1 - e201301012

ER -