Metabolomics & Nutrition NAD

NAD+ Index.

Comprehensive assessment of cellular NAD+ metabolism from a dried blood spot. Six analytes measured by LC-MS/MS, yielding a composite Niacin Number reflecting mitochondrial function and metabolic resilience.

Quick Reference
Method
LC-MS/MS
Sample Types
DBS
Analytes

6 analytes + composite index

Turnaround

3–5 working days

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What does this test assess?

The NAD+ Index is a comprehensive panel that quantifies the key molecules of NAD+ metabolism from a single dried blood spot. It measures both oxidised and reduced forms of the two principal nicotinamide coenzymes (NAD+ / NADH and NADP+ / NADPH), along with two vitamin B3 precursors (nicotinamide and nicotinic acid), producing a composite Niacin Number that reflects overall cellular energy status.

Clinical indications include:

  • Assessment of mitochondrial function and cellular energy production capacity
  • Evaluation of age-related NAD+ decline — tissue levels may halve between ages 20 and 60
  • Monitoring the efficacy of NAD+ precursor supplementation (NMN, NR, niacin)
  • Investigation of chronic fatigue, exercise intolerance, and metabolic resilience
  • Assessment of redox balance in chronic inflammation, where CD38 and PARP enzymes consume NAD+
  • Evaluation of DNA repair capacity and sirtuin-mediated longevity pathways
  • Screening for niacin (vitamin B3) insufficiency affecting NAD+ biosynthesis

Measured analytes

AnalyteRoleClinical Significance
NAD+ Nicotinamide adenine dinucleotide (oxidised) Central coenzyme in catabolic energy metabolism; electron carrier in mitochondrial respiration
NADH Reduced NAD+ Delivers electrons to the electron transport chain for ATP production; elevated levels may indicate metabolic bottleneck
NADP+ Nicotinamide adenine dinucleotide phosphate (oxidised) Coenzyme in anabolic biosynthetic pathways including fatty acid and cholesterol synthesis
NADPH Reduced NADP+ Essential for glutathione regeneration, antioxidant defence, and reductive biosynthesis
Nicotinamide (NAM) Vitamin B3 form; NAD+ precursor (salvage pathway) Elevated levels may indicate recent B3 supplementation; very high levels may inhibit sirtuin activity
Nicotinic Acid (NA) Niacin; NAD+ precursor (Preiss–Handler pathway) Found in meat, fish, and fortified foods; supports NAD+ synthesis via an alternative biosynthetic route

The Niacin Number (NAD/NADP Index) is a composite score calculated as (NAD+/NADH × NADP+/NADPH) × 100. This integrates the catabolic (energy production) and anabolic (biosynthetic) redox systems into a single metric reflecting overall NAD+ metabolic health. The optimal range is 90–350.

About NAD+ metabolism

Nicotinamide adenine dinucleotide (NAD+) is one of the most important molecules in human biology. Present in every living cell, it serves as a critical coenzyme in over 500 enzymatic reactions, functioning as an electron carrier in mitochondrial energy production and as a co-substrate for key regulatory enzymes including sirtuins, PARPs, and CD38.

500+ reactions

NAD+ participates in over 500 enzymatic reactions across all major metabolic pathways

Age-related decline

Tissue NAD+ levels decline significantly with age, contributing to metabolic dysfunction

DNA repair

PARP enzymes consume NAD+ during DNA repair; chronic damage accelerates depletion

In its role as an electron shuttle, NAD+ accepts electrons during glycolysis and the citric acid cycle (becoming NADH), which then delivers them to the mitochondrial electron transport chain to drive ATP synthesis. This redox cycling between NAD+ and NADH is fundamental to cellular energy production, and the NAD+/NADH ratio serves as a key indicator of catabolic metabolic health.

The phosphorylated counterpart, NADP+/NADPH, operates in parallel but supports anabolic processes — fatty acid synthesis, cholesterol biosynthesis, and critically, the regeneration of glutathione, the cell's primary antioxidant defence molecule. The balance between these two redox couples provides a comprehensive picture of cellular metabolic function.

NAD+ levels decline with age through multiple mechanisms: increased consumption by CD38 (an enzyme upregulated by chronic inflammation), increased PARP activation due to accumulated DNA damage, and reduced biosynthetic capacity. This decline has been implicated in mitochondrial dysfunction, metabolic disease, neurodegeneration, and accelerated ageing. Restoring NAD+ levels through precursor supplementation (NMN, NR, niacin) has emerged as a major area of longevity and metabolic research.

Reference ranges

NAD+ levels decline with age, making age-stratified interpretation essential for meaningful results. Key points to understand about NAD+ reference ranges:

  • There are no universally standardised clinical reference ranges for NAD+ from dried blood spots yet, as this is an emerging biomarker
  • Masdiag provides population-based percentile ranges with each report
  • Higher NAD+ levels are consistently associated with better mitochondrial function, DNA repair capacity, and metabolic health
  • NAD+:NADH ratio is a key indicator of cellular energy metabolism efficiency
Interpreting your results

As NAD+ testing is a relatively new clinical biomarker, reference ranges are continually being refined. Your report includes percentile-based interpretation against our tested population.

Who should test their NAD+ levels?

NAD+ is one of the most important molecules in human biology — essential for energy production, DNA repair, and cellular signalling. Testing is particularly relevant for:

  • Adults over 40 — NAD+ levels can decline by up to 50% between ages 40 and 60
  • People interested in longevity and biological ageing — NAD+ is a core biomarker in ageing research
  • Anyone supplementing with NMN (nicotinamide mononucleotide), NR (nicotinamide riboside), or niacin who wants objective evidence that supplementation is raising NAD+ levels
  • Individuals experiencing chronic fatigue, brain fog, or reduced exercise capacity
  • People with metabolic conditions such as obesity, insulin resistance, or type 2 diabetes
  • Athletes and biohackers tracking mitochondrial performance and recovery
  • Patients undergoing or recovering from illness, surgery, or high physiological stress
  • Anyone using NAD+ IV therapy who wants to track sustained levels via a simple blood spot test

How to support healthy NAD+ levels

Declining NAD+ is one of the hallmarks of ageing, but research shows it can be meaningfully increased through targeted strategies:

Supplementation (most direct approach)

  • NMN (nicotinamide mononucleotide): a direct NAD+ precursor; typical doses 250–1000 mg/day
  • NR (nicotinamide riboside): another well-studied precursor; typical doses 300–600 mg/day
  • Niacin (vitamin B3): the original NAD+ precursor, though higher doses can cause flushing
  • Tryptophan: an amino acid that contributes to de novo NAD+ synthesis via the kynurenine pathway

Diet & Nutrition

  • Foods rich in niacin: chicken breast, tuna, turkey, mushrooms, green peas, and liver
  • Fermented foods may support NAD+ metabolism through gut microbiome modulation
  • Adequate protein intake ensures sufficient tryptophan for de novo NAD+ synthesis

Lifestyle

  • Exercise — both aerobic and resistance training stimulate NAMPT, the rate-limiting enzyme in NAD+ salvage pathway
  • Caloric restriction and intermittent fasting have been shown to upregulate NAD+ levels in animal studies
  • Heat stress (sauna) may activate sirtuins and support NAD+-dependent pathways
  • Prioritise quality sleep — circadian rhythm disruption impairs NAD+ metabolism
  • Limit alcohol — alcohol metabolism directly consumes NAD+
Track your progress

Re-test your NAD+ index after 8–12 weeks of intervention to objectively measure the impact of supplementation or lifestyle changes.

Analytical technique

All six analytes are measured simultaneously from a single DBS sample using liquid chromatography with tandem mass spectrometry (LC-MS/MS), the gold standard method for NAD+ biomarker measurement. Isotope-labelled internal standards are employed for each analyte to ensure accurate and reproducible quantification across the full dynamic range.

LC-MS/MS provides the sensitivity and specificity required to resolve structurally similar nicotinamide coenzymes (NAD+, NADH, NADP+, NADPH) within a single chromatographic run, while simultaneously quantifying the B3 precursors nicotinamide and nicotinic acid. The Niacin Number composite score is calculated from the measured ratios.

Sample information

The DBS format enables convenient home collection for longitudinal NAD+ monitoring, making it well suited for supplementation tracking and ageing research. Samples are stable at ambient temperature during postal transit.

Testing process

From enquiry to results in a few simple steps — no clinic visit required.

1
Get in touch
Contact us to discuss your testing requirements
2
Collect your sample
Simple finger-prick onto a dried blood spot card — at home or in clinic
3
Post to our lab
DBS cards are stable at room temperature — ship by regular post worldwide
4
Receive results
Results delivered within 3–5 working days of sample receipt

Where this test is available

This test is available to healthcare professionals, wellness brands, clinics, and research institutions worldwide. We currently serve partners in:

  • Europe (EU & non-EU)
  • United Kingdom
  • Asia & Southeast Asia
  • Australia & New Zealand
  • United States

Whether you need testing services for your patients, white-label kits for your brand, or method transfer to your own laboratory — get in touch to discuss how we can work together.

Literature

  1. Yoshino J, Baur JA, Imai S. “NAD+ intermediates: the biology and therapeutic potential of NMN and NR.” Cell Metabolism, 2018, 27(3):513–528. 10.1016/j.cmet.2017.11.002
  2. Covarrubias AJ, Perrone R, Grozio A, Verdin E. “NAD+ metabolism and its roles in cellular processes during ageing.” Nature Reviews Molecular Cell Biology, 2021, 22(2):119–141. 10.1038/s41580-020-00313-x
  3. Verdin E. “NAD+ in aging, metabolism, and neurodegeneration.” Science, 2015, 350(6265):1208–1213. 10.1126/science.aac4854
  4. Rajman L, Chwalek K, Sinclair DA. “Therapeutic potential of NAD-boosting molecules: the in vivo evidence.” Cell Metabolism, 2018, 27(3):529–547. 10.1016/j.cmet.2018.02.011
  5. Cantó C, Menzies KJ, Auwerx J. “NAD+ metabolism and the control of energy homeostasis: a balancing act between mitochondria and the nucleus.” Cell Metabolism, 2015, 22(1):31–53. 10.1016/j.cmet.2015.05.023

Frequently Asked Questions

What is the NAD+ index?

The NAD+ index is a comprehensive panel that measures NAD+, NADH, NADP+, NADPH, and the vitamin B3 precursors nicotinamide and nicotinic acid from a single dried blood spot. These six analytes reflect cellular energy production and mitochondrial function. NAD+ is essential for over 500 enzymatic reactions in the body, and the composite Niacin Number calculated from the results provides a single metric of overall NAD+ metabolic health.

Why do NAD+ levels matter for ageing?

NAD+ levels decline significantly with age, driven by increased consumption from CD38 and PARP enzymes, and reduced biosynthetic capacity. This decline is linked to mitochondrial dysfunction, impaired DNA repair, and cellular senescence. Testing reveals your baseline NAD+ status before starting supplementation, allowing you to measure whether interventions like NMN or NR are actually restoring your levels.

How is the sample collected?

The test uses a simple finger-prick to collect a few drops of blood onto a dried blood spot (DBS) card. You can do this at home or in a clinic — no venous blood draw is needed. The DBS card is stable at room temperature and can be posted to our laboratory by regular mail from anywhere in the world.

How long does it take to get results?

Results are typically delivered within 3 to 5 working days from the time your sample arrives at our laboratory. The analysis is performed by LC-MS/MS (liquid chromatography-tandem mass spectrometry), which provides highly accurate quantification of all six NAD+ metabolites simultaneously.

Can NAD+ levels be improved through supplementation?

NMN (nicotinamide mononucleotide) and NR (nicotinamide riboside) are the most common NAD+ precursors used in supplementation. Niacin and nicotinamide are also effective but may have different side effect profiles. Testing before and after a supplementation period allows you to track your response objectively rather than relying on subjective improvements alone.

Which countries is this test available in?

Masdiag's NAD+ index test is available worldwide through our partner network. We currently serve healthcare professionals, wellness brands, and clinics in Europe, the United Kingdom, Asia, Australia, New Zealand, and the United States. Contact us to discuss testing services, white-label kits, or method transfer to your laboratory.

Interested in this method?

Whether you need testing services, method transfer, or white-label kit development — we'd love to hear from you.