NAD+ and Senolytics: Laboratory Assessment of Anti-Aging Interventions

NAD+ metabolism and senolytics — laboratory assessment

NAD+ and Senolytics: Laboratory Assessment of Therapeutic Interventions

Restoring NAD+ (nicotinamide adenine dinucleotide) levels and eliminating senescent cells with senolytics are two of the most promising directions in anti-aging therapy. Clinical trials of NMN, NR (nicotinamide riboside), dasatinib + quercetin and fisetin require objective laboratory monitoring.

For an anti-aging laboratory, this means mastering specific methods for measuring NAD+ and senescence markers — with equipment that partly overlaps with the LC-MS/MS oxidative stress panel.

Methods for Measuring NAD+ and Its Metabolites

Comparison of NAD+/NADH measurement methods:

Method	Sensitivity	Advantages	Limitations
LC-MS/MS	pmol	Maximum accuracy, multiplexing	Equipment cost
Enzymatic assay	nmol	Simplicity, accessibility	Interference
Fluorimetry	nmol	High throughput	Requires calibration

Markers of Senolytic Therapy Efficacy

Evaluating the efficacy of senolytics requires monitoring SASP factors and markers of cellular senescence:

SASP factors — GDF15, IL-6, MCP-1, PAI-1 — a decrease after a course of senolytics indicates elimination of senescent cells.
SA-β-galactosidase — cytochemical and fluorimetric determination in skin cells (biopsy) or blood.
LC3-II/p62 — markers of autophagy and mitophagy. Western blot or flow cytometry.
[Epigenetic clocks](/blog/epigenetic-clocks-biological-age) — DunedinPACE and GrimAge as an integrated measure of efficacy.
[Telomeres](/blog/telomeres-cellular-aging-research-equipment) — telomere length as an additional marker of cellular senescence.

Laboratory Equipment

Equipment set for monitoring NAD+ and senolytic therapy:

LC-MS/MS — the gold standard for NAD+, NADH, NMN and NR. Triple quadrupole with a HILIC column.
Fluorimeter — Tecan Infinite, BMG CLARIOstar — for enzymatic NAD+ and SA-β-gal assays.
Flow cytometer — for LC3-II, autophagy and SA-β-gal in cells.
Western blot station — for LC3-II and p62/SQSTM1. Automated (ProteinSimple Jess/Wes) or conventional.
Multiplex platform — MSD V-PLEX, Luminex — for simultaneous measurement of a SASP panel.

KombiMED: Equipment for NAD+ and Senolytic Monitoring

KombiMED GmbH supplies equipment for laboratories that monitor NAD+ therapy and senolytic interventions. We will select a configuration that integrates with your existing equipment. Contact KombiMED for a consultation.