Rapid quantitation of NAD⁺/NADH and NADPH/NADP⁺ with mass spectrometry by using calibration constants.

ABSTRACT: NAD⁺/NADH and NADPH/NADP⁺ are important indicators of cellular redox status, but they cannot be reliably calculated from the relative intensities of mass spectrometry signals alone. Establishing accurate redox ratios by mass spectrometry has historically required converting relative signal intensities into absolute concentrations, which is a time-consuming process that limits rapid analysis. Here, we describe a simpler strategy to determine NAD⁺/NADH and NADPH/NADP⁺ by using mass spectrometry. While ionization is strongly influenced by factors such as instrument drift and sample type, we discovered that the relative signal intensities of oxidized and reduced cofactors change at the same rate across experiments performed on the same mass spectrome

SUBMITTER: Guo Q 

PROVIDER: S-EPMC12765170 | biostudies-literature | 2025 Nov

REPOSITORIES: biostudies-literature