Skip to main content

Clinical Assessment of the NADome as Biomarkers for Healthy Aging

  • Protocol
  • First Online:
Clinical and Preclinical Models for Maximizing Healthspan

Abstract

Nicotinamide adenine dinucleotide (NAD+) and its related metabolites (NADome) are important endogenous analytes that are thought to play important roles in cellular metabolism, inflammation, oxidative stress, cancer, neurodegeneration, and aging in mammals. However, these analytes are unstable during the collection of biological fluids, which is a major limiting factor for their quantitation. Herein, we describe a highly robust and quantitative method using liquid chromatography coupled to tandem mass spectrometry to quantify the NADome in whole blood, plasma, mononuclear cells, platelets, cerebrospinal fluid (CSF), and urine. This methodology represents a “gold standard” of measure for understanding biological pathways and developing targeted pharmacological interventions to modulate NAD+ biosynthesis and NAD-dependent mediators in health and disease.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Massudi H, Grant R, Guillemin GJ, Braidy N (2012) NAD+ metabolism and oxidative stress: the golden nucleotide on a crown of thorns. Redox Rep 17(1):28–46

    Article  CAS  Google Scholar 

  2. Braidy N, Guillemin GJ, Mansour H, Chan-Ling T, Poljak A, Grant R (2011) Age related changes in NAD+ metabolism oxidative stress and Sirt1 activity in wistar rats. PLoS One 6(4):e19194. https://doi.org/10.1371/journal.pone.0019194

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Braidy N, Poljak A, Grant R, Jayasena T, Mansour H, Chan-Ling T et al (2014) Mapping NAD(+) metabolism in the brain of ageing Wistar rats: potential targets for influencing brain senescence. Biogerontology 15(2):177–198

    Article  CAS  Google Scholar 

  4. Braidy N, Poljak A, Grant R, Jayasena T, Mansour H, Chan-Ling T et al (2015) Differential expression of sirtuins in the aging rat brain. Front Cell Neurosci 9:167. https://doi.org/10.3389/fncel.2015.00167

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Braidy N, Berg J, Clement J, Khorshidi F, Poljak A, Jayasena T et al (2018) Role of nicotinamide adenine dinucleotide and related precursors as therapeutic targets for age-related degenerative diseases: rationale, biochemistry, pharmacokinetics, and outcomes. Antioxid Redox Signal 30(2):251–294

    Article  Google Scholar 

  6. Trammell SA, Brenner C (2013) Targeted, LCMS-based metabolomics for quantitative measurement of NAD(+) metabolites. Comput Struct Biotechnol J 4:e201301012. https://doi.org/10.5936/csbj.201301012

    Article  PubMed  PubMed Central  Google Scholar 

  7. Clement J, Wong M, Poljak A, Sachdev P, Braidy N (2018) The plasma NAD(+) metabolome is dysregulated in “normal” aging. Rejuvenation Res 22(2):121–130

    Article  Google Scholar 

  8. Seyedsadjadi N, Berg J, Bilgin AA, Braidy N, Salonikas C, Grant R (2018) High protein intake is associated with low plasma NAD+ levels in a healthy human cohort. PLoS One 13(8):e0201968. https://doi.org/10.1371/journal.pone.0201968

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Long AN, Owens K, Schlappal AE, Kristian T, Fishman PS, Schuh RA (2015) Effect of nicotinamide mononucleotide on brain mitochondrial respiratory deficits in an Alzheimer’s disease-relevant murine model. BMC Neurol 15:19. https://doi.org/10.1186/s12883-015-0272-x

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Trammell SA, Schmidt MS, Weidemann BJ, Redpath P, Jaksch F, Dellinger RW et al (2016) Nicotinamide riboside is uniquely and orally bioavailable in mice and humans. Nat Commun 7:12948. https://doi.org/10.1038/ncomms12948

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Casabona G, Sturiale L, L’Episcopo MR, Raciti G, Fazzio A, Sarpietro MG et al (1995) HPLC analysis of cyclic adenosine diphosphate ribose and adenosine diphosphate ribose: determination of NAD+ metabolites in hippocampal membranes. Ital J Biochem 44(5):258–268

    CAS  PubMed  Google Scholar 

  12. Bernofsky C, Swan M (1973) An improved cycling assay for nicotinamide adenine dinucleotide. Anal Biochem 53(2):452–458

    Article  CAS  Google Scholar 

Download references

Acknowledgments

N.B. is the recipient of the Australian Research Council Discovery Early Career Research Award at the University of New South Wales, Sydney, Australia.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Nady Braidy .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2020 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Jayasena, T. et al. (2020). Clinical Assessment of the NADome as Biomarkers for Healthy Aging. In: Guest, P. (eds) Clinical and Preclinical Models for Maximizing Healthspan. Methods in Molecular Biology, vol 2138. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0471-7_13

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-0471-7_13

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0470-0

  • Online ISBN: 978-1-0716-0471-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics