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Anal Biochem. 2019 Feb 27;572:1-8. doi: 10.1016/j.ab.2019.02.019. [Epub ahead of print]

Assessment of NAD+metabolism in human cell cultures, erythrocytes, cerebrospinal fluid and primate skeletal muscle.

Author information

1
Biomedical Research Center, National Institute on Aging, National Institutes of Health, Baltimore, MD, USA.
2
Biomedical Research Center, National Institute on Aging, National Institutes of Health, Baltimore, MD, USA. Electronic address: moaddelru@mail.nih.gov.

Abstract

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.

KEYWORDS:

LC-MS/MS; Metabolomics; NAD+ metabolome; NAD+/NADH metabolism

PMID:
30822397
DOI:
10.1016/j.ab.2019.02.019

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