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Assessment of NAD+metabolism in human cell cultures, erythrocytes, cerebrospinal fluid and primate skeletal muscle
- Demarest, Tyler G., Truong, Gia Thinh D., Lovett, Jacqueline, Mohanty, Joy G., Mattison, Julie A., Mattson, Mark P., Ferrucci, Luigi, Bohr, Vilhelm A., Moaddel, Ruin
- Analytical biochemistry 2019 v.572 pp. 1-8
- NAD (coenzyme), NADP (coenzyme), autoxidation, cell culture, cerebrospinal fluid, erythrocytes, humans, liquid chromatography, mass spectrometry, metabolites, metabolome, skeletal muscle
- The reduction-oxidation state of NAD+/NADH is critical for cellular health with NAD+ and its metabolites playing critical roles in aging and pathologies. Given the inherent autooxidation of reduced dinucleotides (i.e. NADH/NADPH), and the well-established differential stability, the accurate measurement of NAD+ and its metabolites is technically challenging. Moreover, sample processing, normalization and measurement strategies can profoundly alter results. Here we developed a rapid and sensitive liquid chromatography mass spectrometry-based method to quantify the NAD+ metabolome with careful consideration of these intrinsic chemical instabilities. Utilizing this method we assess NAD+ metabolite stabilities and determine the presence and concentrations of NAD+ metabolites in clinically relevant human samples including cerebrospinal fluid, erythrocytes, and primate skeletal muscle.