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.
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