Assessment of NAD+ Metabolism in Human Cell Cultures, Erythrocytes, Cerebrospinal Fluid and Primate Skeletal Muscle

Demarest, T.G.; Truong, G.T.D.; Lovett, J.; et al.

Given the importance of the NAD⁺ metabolome in aging and cellular health, Demarest et al. developed a novel isotope dilution mass spectrometry method (utilizing HILIC-MRM/MS) for their quantification in a range of biosamples (ranging from cultured cells to clinical tissues). The optimized method is both rapid (14 min runs) and sensitive (e.g., NAD⁺ and NMN quantified for first time in CSF), while utilizing a U-¹³C yeast extract (ISO1) for absolute quantification. A breadth of cofactors (e.g., NAD⁺, NADH, NMR) and AcCoA are successfully quantified via standard curves. This method should prove useful in better understanding the metabolic alterations arising from normal processes and disorders.