NAD+ Reconstitution: 250mg + 2.5mL Chart

How to Reconstitute NAD+

NAD+ for subcutaneous injection comes as a lyophilized (freeze-dried) white powder in a sealed vial. It is reconstituted with bacteriostatic water before injection. The process is identical to other peptides with one key difference: NAD+ is dosed in milligrams (not micrograms), so the math works a little differently.

This guide covers mixing, measuring doses, and storing NAD+ so nothing goes to waste.

Research-context information only. NAD+ is a research peptide. Protocols, doses, and reactions reported below come from published research and self-reported community sources. This article reports what has been documented, not what should be done. Consult a licensed physician for personal medical decisions.

New to bacteriostatic water? Start with what it is, how much to use per vial, and how long it lasts — or see the full Bacteriostatic Water Guide.

What You Need

Documented protocols call for the following:

• NAD+ lyophilized vial (typically 100mg, 250mg, or 500mg)
• Bacteriostatic water (BAC water) — not sterile water, not saline
• Insulin syringes — 1mL (100 unit), 29-31 gauge
• Alcohol swabs — for cleaning vial tops
• A clean, flat workspace

Why bacteriostatic water? It contains 0.9% benzyl alcohol that prevents bacteria from growing in the solution. Regular sterile water has no preservative, so the vial must be used within 24 hours. BAC water allows up to 28 days refrigerated.

Note on NAD+ stability: NAD+ is less stable in solution than many peptides. Prompt refrigeration after reconstitution is especially important. Some users reconstitute smaller vials (100-250mg) to use the solution within 2-3 weeks rather than pushing the full 28-day window.

Step-by-Step Reconstitution

Step 1: Clean Everything

Protocols describe wiping the tops of both vials (NAD+ and BAC water) with alcohol swabs, then letting them air dry for 10 seconds. This prevents contamination — a step community resources flag as essential.

Step 2: Draw Your Bacteriostatic Water

Protocols describe using a fresh insulin syringe to draw the desired amount of BAC water. The amount added determines the concentration (see the dilution charts below).

Community resources commonly document 2.5mL into a 250mg vial as the sweet spot. It gives 100mg/mL — clean, round numbers for standard doses.

Step 3: Add Water to the NAD+ Vial

Protocols describe inserting the needle into the NAD+ vial at an angle, aiming at the glass wall — not directly at the powder — and letting the water trickle down the side of the vial gently.

Water is not squirted directly onto the powder. While NAD+ is more robust than many peptides, gentle technique preserves molecular integrity.

Step 4: Let It Dissolve

Protocols describe swirling the vial gently with a slow rotating motion rather than shaking it. NAD+ typically dissolves quickly — within 1-2 minutes into a perfectly clear, colorless solution.

If particles remain after 5 minutes of gentle swirling, the NAD+ may be degraded. A properly manufactured NAD+ dissolves easily.

Step 5: Store Correctly

Reconstituted vials are refrigerated immediately at 36-46°F (2-8°C). The reconstituted solution is stable for up to 28 days with bacteriostatic water, though using it within 21 days is ideal given NAD+'s relative instability in solution.

Dilution Charts

Why these water amounts? They all produce a 100mg/mL concentration, making dose calculation simple: 1 unit on the syringe = 1mg of NAD+. A 50mg dose = 50 units. A 100mg dose = 100 units (full 1mL syringe).

The Math (So You Can Do It Yourself)

Here's the formula for any vial size and any amount of water:

Concentration = Vial Size (mg) ÷ Water Added (mL)

Then to find the injection volume:

Units to inject = Desired Dose (mg) ÷ Concentration (mg/mL) × 100

Example: 250mg vial + 2.5mL water = 100mg/mL. For a 50mg dose: 50 ÷ 100 × 100 = 50 units.

For variable vial sizes or doses, the Reconstitution Calculator applies this formula automatically. Vendors selling NAD+ as a research compound are compared on the best NAD+ sources page, including current coupon codes.

Affiliate disclosure: vendor links in this article are affiliate links — The Peptide Catalog may earn a commission if you buy through them, at no additional cost to you.

Common Mistakes

Shaking the Vial

While NAD+ is more structurally robust than large peptide chains, shaking creates foam that makes accurate volume measurement difficult and can introduce air bubbles that interfere with injection. Protocols describe swirling gently rather than shaking.

Using Too Little Water

Adding 0.5mL to a 250mg vial gives 500mg/mL. That means a 50mg dose is only 10 units — harder to measure accurately on an insulin syringe. Protocols describe using enough water to keep doses above 25 units for accurate measurement.

Leaving It at Room Temperature

NAD+ degrades faster in solution than most peptides. Every hour at room temperature reduces potency. Protocols describe pulling the vial out, drawing the dose, and returning it to the refrigerator rather than leaving it on the counter.

Reusing Needles

Each puncture through the rubber stopper dulls the needle and increases contamination risk. Protocols describe a fresh syringe every time — they cost pennies.

Freezing Reconstituted NAD+

Freezing a liquid solution creates ice crystals that can damage the molecular structure. Protocols describe freezing only lyophilized (powder) NAD+. Once reconstituted, the solution is refrigerated — never frozen.

Not Accounting for Larger Volumes

NAD+ doses (50-200mg) require larger injection volumes than typical peptides. A 200mg dose at 100mg/mL = 2mL, which exceeds a standard 1mL insulin syringe. Protocols describe either using a 3mL syringe with a fine gauge needle or splitting into two 1mL injections at different sites.

How Many Doses Per Vial?

This depends on the dose and vial size:

100mg vial at 50mg/dose = 2 doses 100mg vial at 100mg/dose = 1 dose 250mg vial at 50mg/dose = 5 doses 250mg vial at 100mg/dose = 2.5 doses 500mg vial at 50mg/dose = 10 doses 500mg vial at 100mg/dose = 5 doses 500mg vial at 200mg/dose = 2.5 doses

Community protocols commonly documenting 100mg three times weekly would exhaust a 250mg vial in under a week. A 500mg vial lasts about 1.5 weeks. NAD+ protocols consume vials faster than most peptide protocols.

Storage Quick Reference

Pro tip: With multiple vials, unopened ones are kept frozen and only what will be used within 2-3 weeks is reconstituted. NAD+ lyophilized powder is much more stable than the reconstituted solution.

Related Reading

• NAD+ Side Effects: Flushing, Nausea, Infusion — adverse event profile from clinical trials and community reports
• NAD+ Dosing Guide — protocols, injection sites, and cycling
• NAD+ Bloodwork Guide — what labs to track and optimal ranges
• NAD+ Benefits — what the research shows
• NAD+ Results Timeline — week-by-week expectations
• SS-31 Dosing Guide — mitochondrial peptide often stacked with NAD+
• MOTS-c Reconstitution Guide — another longevity peptide with mg-level dosing
• Calculate Your NAD+ Cycle Cost — estimate vials needed and total cost for your protocol

References

1. Martens, C.R., et al. (2018). Chronic nicotinamide riboside supplementation is well-tolerated and elevates NAD+ in healthy middle-aged and older adults. Nature Communications, 9(1), 1286. PubMed
2. Grant, R., et al. (2019). A pilot study investigating changes in the human plasma and urine NAD+ metabolome during a 6 hour intravenous infusion of NAD+. Front Aging Neurosci, 11, 257. PubMed
3. USP General Chapter 797: Pharmaceutical Compounding — Sterile Preparations. Storage and beyond-use dating guidelines for reconstituted compounds.

This guide is for educational and informational purposes only. It is not medical advice. NAD+ is sold as a research compound and is not FDA-approved for human use as an anti-aging therapy. Reconstituting and self-administering injectable compounds carries inherent risks including infection, contamination, and dosing errors. Always use proper sterile technique. Consult a qualified healthcare provider before starting any NAD+ protocol. The Peptide Catalog is not responsible for any adverse effects resulting from the use or misuse of information presented here.