NAD+ Dosing Protocol: Injection, Pen Vial, Nasal Spray & Reconstitution (2026)

NAD+ Quick Start

NAD+ (nicotinamide adenine dinucleotide) is a coenzyme present in every cell, central to energy metabolism, mitochondrial function, and DNA repair. It is a substrate for sirtuins and PARP enzymes, and cellular NAD+ declines with age. Note: NAD+ is a coenzyme, not a peptide. It is studied via injection, IV, and intranasal routes, alongside oral precursors (NMN/NR).

Quick reference Coenzyme · research-use

Class

Coenzyme (dinucleotide)

Formats

Vial · Pen vial · Nasal spray

Research focus

Cellular energy, longevity

Schedule shape

2–3× weekly

Vial size

1000 mg

This guide is an educational research reference. It does not diagnose, treat, or prescribe, and is not medical advice. Consult a licensed clinician before considering any compound.

NAD+ Dosing Protocols

The injectable (subcutaneous), pen-vial and nasal-spray formats are documented as separate protocols. NAD+ doses are in the milligram range and are commonly researched a few times weekly. Ranges are research-planning references, not personal dosing recommendations. NAD+ is notably prone to flushing and nausea when delivered too quickly, so slow administration is standard.

Injection — Subcutaneous

Reconstituted 1000 mg vial, U-100 insulin syringe; inject slowly.

Band	Per dose	Frequency
Low	50 mg	2–3× weekly
Standard	100 mg	2–3× weekly

1000 mg + 10 mL BAC → 100 mg/mL · 50 mg = 0.5 mL = 50 units

Nasal Spray (≈2× the injection mg)

Pre-mixed nasal spray, ready to use.

Band	Per dose	# of sprays	Frequency
Low	100 mg	5 sprays	2–3× weekly
Standard	200 mg	10 sprays	2–3× weekly

Each 0.1 mL spray delivers 20 mg

Pen Vial — Pre-Reconstituted 3 mL

Supplied already reconstituted to 3 mL — no mixing step. Doses are given in units, the scale a reusable pen dials in (300 units per vial). Pens step in whole or half units depending on the model — round any fractional figure to the nearest increment your device supports.

Band	Per dose (units)	Delivers	Frequency
Low	15 units	50 mg	2–3× weekly
Standard	30 units	100 mg	2–3× weekly

1000 mg in 3 mL → 3.33 mg per unit · 300 units per pen · ~20 doses at 50 mg

The pen is roughly 3× more concentrated than the 10 mL reconstitution described above (333 mg/mL vs 100 mg/mL), so draw volumes are proportionally smaller. The slow-administration convention still applies — concentration does not change the flushing and nausea profile.

Bioavailability adjustment: nasal NAD+ absorption is roughly 50–75% of the subcutaneous route, because NAD+ is a large, charged molecule that crosses the nasal mucosa inefficiently. To deliver a comparable systemic amount, the spray bands are set at about double the injection milligrams (100–200 mg nasal vs 50–100 mg injected). This is an approximation to offset lower absorption, not a precise equivalence — the two routes remain separate protocols.

NAD+ Reconstitution Guide

NAD+ ships as a lyophilized powder. The BAC water volume sets the concentration and draw volume for the injectable vial. Nasal sprays and pen vials both ship ready to use — the pen at a fixed 3 mL fill, so no water is added to it at all. NAD+ solutions are often slightly colored and very water-soluble.

Injection

BAC	Conc.	50 mg
10 mL	100 mg/mL	0.5 mL · 50 u
5 mL	200 mg/mL	0.25 mL · 25 u

The NAD+ vial holds up to ~10 mL; more dilution (10 mL) is gentler for slow SubQ. Units are U-100.

Pen Vial — 3 mL

Format	Conc.	50 mg
3 mL (as supplied)	333 mg/mL	0.15 mL · 15 u

Ships pre-reconstituted; no BAC water is added. 3.33 mg per unit on a U-100 scale.

Reconstitution steps

Inspect the vial. Confirm label and intact powder.
Wipe the stoppers. Alcohol swab on both vials.
Draw BAC water. 10 mL into the vial for injection (the NAD+ vial holds up to ~10 mL).
Inject down the wall. Release water slowly down the inside wall, not onto the powder.
Swirl, do not shake. Roll gently until fully dissolved.
Refrigerate. Store at 2–8 °C; do not freeze.

How to use the pen vial

Inspect the vial. Confirm the label and strength. The solution should be clear and colorless.
Never add water. A pen vial ships at its final 3 mL volume. Adding bacteriostatic water would dilute it and invalidate every unit figure on this page.
Load the vial into the pen. Unscrew the pen's cartridge holder, seat the vial, and screw the holder back on until firm.
Attach a fresh needle tip. Screw a new disposable pen needle onto the holder, then pull off the outer and inner caps. Use a new tip for every dose.
Prime the pen. Dial 2 units, hold the pen needle-up, tap the holder so any air rises to the top, and press the button until a drop appears at the tip. Repeat until it does.
Spin to the dose. Turn the dial to the unit figure from the table above. If you overshoot, spin back — never press the button to correct an overshoot.
Press slowly. NAD+ is notably prone to flushing and nausea when delivered quickly. Push the button down at a deliberate pace rather than in one motion.
Inject and hold. Press the button all the way down, then keep the needle in place for about ten seconds before withdrawing so the full dose is delivered.
Remove and dispose of the needle. Unscrew the tip into a sharps container. Never store the pen with a needle attached — it lets solution leak out and air in.
Cap and refrigerate. Store the pen with the vial still seated at 2–8 °C. Do not freeze.

How to use the nasal spray

Prime first use. Pump 2–3 sprays into a tissue until a fine, even mist appears.
Position. Tilt the head slightly forward; insert the tip just inside one nostril, aimed slightly outward toward the ear — not at the septum.
Spray and breathe. Press once while breathing in gently; do not sniff hard, which sends the solution down the throat instead of onto the mucosa.
Alternate nostrils. For multi-spray doses, switch nostrils each spray to spread absorption and limit irritation.
Count per the protocol. Use the sprays-per-dose shown above; if a dose isn't a whole number, round up.
Between uses. Wipe the tip, recap, and refrigerate.

How NAD+ Works

NAD+ is a coenzyme that shuttles electrons in the redox reactions of cellular energy production (glycolysis, the TCA cycle, and oxidative phosphorylation). Beyond metabolism, it is the required substrate for sirtuins (linked to stress resistance and longevity signaling), PARP enzymes (DNA repair), and CD38. Because tissue NAD+ declines with age and metabolic stress, raising NAD+ availability is a central theme in longevity research.

Energy metabolism

Essential electron carrier for ATP production in mitochondria.

Sirtuins

Required substrate for sirtuin enzymes implicated in stress resistance and aging.

DNA repair

Substrate for PARP enzymes involved in repairing DNA damage.

Open question

Whether raising NAD+ delivers durable clinical benefits in humans is unsettled.

Who Should Avoid NAD+

Pregnancy & lactation

No human safety data in this context.

Active cancer

NAD+ metabolism intersects with cell proliferation; discuss with a clinician.

Cardiovascular sensitivity

Rapid delivery can cause flushing and chest tightness; caution with cardiac conditions.

Anyone seeking a treatment

NAD+ is a research compound, not a medical treatment.

NAD+ Side Effects & Safety

Rate-dependent reactions

Flushing, nausea, cramping, and chest tightness are classic with fast IV/injection; slowing delivery reduces them.

Injection-site stinging

SubQ NAD+ can sting; more dilution and slow injection help.

Nasal irritation

The concentrated spray can irritate nasal mucosa; alternate nostrils.

Quality-control risk

Verify identity and purity against a Certificate of Analysis.

Timeline & What to Monitor

Timeframe	Commonly tracked	Notes
During dosing	Flushing/nausea, injection-site response	Slow the delivery rate if reactions occur.
Week 1–4	Energy/wellbeing self-report	Subjective; no validated NAD+-specific marker.
Ongoing	General metabolic context	Interpret with a clinician; routine labs aren't NAD+-specific.

Research Evidence Context

Precursor trials

Most human data involves oral precursors (NR/NMN) raising blood NAD+ markers; clinical-endpoint data is mixed.

Direct NAD+ routes

IV/SubQ NAD+ is widely used in wellness settings but has limited controlled outcome data.

Aging biology

Strong preclinical rationale via sirtuin/PARP biology and age-related NAD+ decline.

Open question

Optimal route, dose, and durable benefit in humans remain unresolved.

Storage & Handling

State	Storage	Notes
Lyophilized (powder)	−20 °C long-term; fridge short-term	More stable than reconstituted solution.
Reconstituted (liquid)	2–8 °C	Use within ~3–4 weeks; do not freeze. NAD+ degrades faster in solution than many peptides.
Appearance	Clear, may be faintly colored	Discard if cloudy or heavily discolored.

Mistakes & Troubleshooting

Flushing / nausea. Almost always rate-related — slow the injection or reduce per-dose amount.
Injection-site stinging. Use the 10 mL (100 mg/mL) reconstitution and inject slowly; the spray is a gentler-feeling alternative for some researchers.
Spray vs injection dose. The spray runs at ~2× the injection mg to offset lower nasal absorption (≈50–75%); still treat the two routes as separate protocols.
Left out overnight. Treat reconstituted solution as compromised and discard.

NAD+ vs Precursors

Feature	NAD+ (direct)	NMN / NR (precursors)
Form	The coenzyme itself	Building blocks converted to NAD+
Route	IV / SubQ / intranasal	Usually oral
Evidence	Limited controlled outcome data	More human data on raising NAD+ markers

Frequently Asked Questions

What is NAD+?

A coenzyme central to energy metabolism, DNA repair, and sirtuin activity. It is not a peptide.

How is the injection dosed vs the nasal spray?

Separate protocols. Injection commonly references 50–100 mg SubQ, 2–3× weekly. Because nasal absorption is roughly 50–75% of injection, the spray runs at about double — 100–200 mg per dose. At 20 mg per spray (5 mL fill) that is 5–10 sprays, 2–3× weekly, to deliver a comparable systemic amount.

Why does NAD+ cause flushing?

Flushing, nausea, and chest tightness are typically rate-related; delivering NAD+ slowly markedly reduces them.

How is NAD+ reconstituted?

For injection, 1000 mg in 10 mL BAC water (the NAD+ vial holds up to ~10 mL) gives 100 mg/mL, so 50 mg = 0.5 mL = 50 units; the extra dilution is gentler for slow SubQ. The spray ships pre-mixed and delivers 20 mg per spray.

What is the pen-vial format?

A 1000 mg pen vial supplied already reconstituted to 3 mL — 300 units at 3.33 mg per unit. A 50 mg reference dose is 15 units. Note this is a far more concentrated solution than the 10 mL reconstitution shown above, so the same milligram dose is a much smaller volume.

Is this page medical advice?

No. It is an educational research reference and does not diagnose, treat, or prescribe. Consult a licensed clinician before considering any compound.

NAD+ Formats & Sourcing

The protocols above are written against the formats NAD+ is actually supplied in. Batch-specific Certificates of Analysis are issued per lot; always verify the strength on the label against the figures used here before calculating anything.

Format	Supplied as
Vial	1000 mg lyophilized powder
Pen vial	Pre-reconstituted, 3 mL
Nasal spray	Pre-mixed, 20 mg per spray
Nasal spray — 5-pack	Bulk: five nasal spray units
Vial — 5-pack	Bulk: five vial units

5-packs are available for the nasal spray and vial formats. Strengths and formats are listed as sold; confirm against the product page and the batch COA.

Reference material for this protocol, with batch COA:

View NAD+ on Spyro Peptides →

Purity 99+% HPLC Batch COA available Research use only

References

Rajman L, Chwalek K, Sinclair DA. Therapeutic potential of NAD-boosting molecules. Cell Metab (2018).
Covarrubias AJ, et al. NAD+ metabolism and its roles in cellular processes during ageing. Nat Rev Mol Cell Biol (2021).
Martens CR, et al. NR supplementation and NAD+ metabolome. Nat Commun (2018).
Conlon N, et al. Intravenous NAD+ in humans: pharmacokinetics. Front / J (review).