NAD+ (500 mg vial)

Protocol Overview

Concise summary of the regimen.

Dosing & Reconstitution

Frequency: Inject once daily subcutaneously. This gradual titration begins at 50 mg daily to assess tolerance, as starting too high can produce adverse reactions such as insomnia, anxiety, or fatigue. Most individuals find doses in the 50 to 100 mg range well-tolerated after ramp-up.

Reconstitution Steps

1. Allow the lyophilized vial to reach room temperature before opening to minimize moisture condensation.
2. Draw 3.0 mL bacteriostatic water (0.9% benzyl alcohol) with a sterile syringe.
3. Inject slowly down the vial wall to avoid foaming; do not aim directly at the powder.
4. Gently swirl until the powder fully dissolves (do not shake vigorously). Solution should be clear and colorless.
5. Label with the reconstitution date and refrigerate at 2 to 8 °C, protected from light.
6. Use within 14 days; inspect before each use for clarity. Discard if discoloration or precipitate appears. At 166.7 mg/mL, 1 unit = 0.01 mL = 1.67 mg on a U-100 insulin syringe. 50 mg = 30 units; 75 mg = 45 units; 100 mg = 60 units.

Storage Instructions

Proper storage preserves peptide quality.

• Lyophilized (unopened): store at -20 °C or below (ideally -80 °C for multi-year storage) in a dry, dark

environment. NAD+ powder is hygroscopic.

• Reconstituted: refrigerate at 2 to 8 °C and use within approximately 14 days. Protect from light (UV

exposure degrades NAD+).

• Avoid freeze-thaw cycles: do not repeatedly freeze and thaw reconstituted NAD+ solution.
• Inspect before each use: fresh NAD+ solution should be clear and colorless.

Supplies Needed

Plan based on an 8 to 16 week daily protocol with gradual titration.

Peptide Vials (NAD+, 500 mg each):

• 8 weeks: ~11 vials (total 5,075 mg used).
• 12 weeks: ~16 vials (total 7,875 mg used).
• 16 weeks: ~22 vials (total 10,675 mg used).

Insulin Syringes (U-100, 1 mL):

• Per week: 7 syringes (1/day).
• 8 weeks: 56. 12 weeks: 84. 16 weeks: 112.

Bacteriostatic Water (10 mL bottles):

• 8 weeks (11 vials): 4 bottles. 12 weeks (16 vials): 5 bottles. 16 weeks (22 vials): 7 bottles.

Alcohol Swabs:

• Per week: 14 swabs.
• 8 weeks: 112 (2 x 100-count). 16 weeks: 224 (3 x 100-count).

Important Notes

Practical considerations for consistency and safety.

• Use new sterile insulin syringes for each injection; dispose in a sharps container.
• Rotate injection sites systematically (abdomen at least 2 inches from navel, outer thighs, back of upper

arms) to reduce soreness.

• Clean both the vial stopper and injection site with alcohol swabs; allow skin to air-dry before injecting.
• Inject slowly over 5 to 10 seconds to minimize tissue irritation; wait a few seconds before withdrawing the

needle.

• Document daily dose, injection site, and any observed effects to maintain consistency and track

tolerance.

• If persistent redness or a lump develops at injection sites, apply over-the-counter hydrocortisone cream

and rotate to a different location.

• Starting at the low end (50 mg) is recommended to assess individual tolerance before escalating dose.

How This Works

NAD+ is a coenzyme central to redox reactions, energy metabolism (glycolysis, TCA cycle, oxidative phosphorylation), and cellular maintenance pathways including DNA repair and mitochondrial biogenesis. Cellular NAD+ levels decline with age and metabolic stress, which may contribute to reduced mitochondrial function and impaired cellular resilience. Clinical research on NAD+ therapy has primarily used intravenous infusions at high doses (500 to 1,000 mg) for applications such as addiction treatment and acute metabolic support. A pilot metabolic study demonstrated that a 750 mg NAD+ IV infusion over 6 hours was well-tolerated in humans, with rapid metabolic clearance and no acute toxicity. Subcutaneous (SC) or intramuscular (IM) injections at lower doses (tens to low hundreds of milligrams) are emerging as practical alternatives for maintenance therapy. Conservative protocols start around 50 to 100 mg per injection a few times per week; the present protocol uses daily SC administration with gradual titration to optimize individual tolerance.

Benefits & Side Effects

Observations from preclinical models and clinical case reports.

• May support cellular energy production and mitochondrial function through replenishment of NAD+ pools.
• Human case reports and pilot studies suggest benefits at 100 to 300 mg/day ranges for cognitive support

and metabolic health.

• High-dose IV protocols (500 to 1,500 mg daily for ~10 days) have been used in open-label studies for

substance use disorder, with reports of reduced cravings and improved mood in follow-ups.

• No severe adverse events have been reported in published NAD+/NADH trials; side effects are generally

mild.

• Potential side effects: high doses can produce adverse reactions such as insomnia, anxiety, or fatigue

if escalated too quickly; gradual titration is advised.

• Mild injection-site reactions (redness, itching, soreness) may occur with subcutaneous administration;

typically resolve with site rotation.

• Some individuals report transient headache or flushing; these effects are generally dose-dependent.
• Doses exceeding ~200 to 300 mg/day should generally be reserved for supervised therapeutic use.

Lifestyle Factors

Complementary strategies for best outcomes.

• Nutrition: consume a balanced diet rich in NAD+ precursors (niacin/vitamin B3, tryptophan) and support

nutrients (B-vitamins, magnesium).

• Exercise: regular physical activity (resistance and aerobic) naturally upregulates cellular NAD+

biosynthesis.

• Sleep: prioritize 7 to 9 hours of quality sleep to support cellular repair and optimize circadian NAD+

metabolism.

• Stress management: chronic stress depletes NAD+ pools through activation of poly(ADP-ribose)

polymerases (PARPs).

• Avoid excessive alcohol: alcohol metabolism consumes NAD+; moderate or eliminate alcohol intake

during NAD+ protocols.

Injection Technique

General subcutaneous guidance from clinical best-practice resources.

1. Wash hands thoroughly with soap and water.
2. Clean the vial stopper with an alcohol swab and allow to air-dry.
3. Select injection site (abdomen at least 2 inches from navel, outer thigh, or back of upper arm); clean and allow to dry.
4. Use a fine, short needle: 28 to 31 gauge insulin syringe; 5/16 to 1/2 inch (8 to 12 mm) length.
5. Pinch a fold of skin; insert the needle at approximately 45 degrees into the pinched subcutaneous layer.
6. Do not aspirate for subcutaneous injections. Depress the plunger slowly over 5 to 10 seconds.
7. Wait a few seconds before withdrawing the needle at the same angle.
8. Apply gentle pressure with a clean swab; do not rub vigorously.
9. Dispose of the used syringe immediately in a sharps container. 10 For doses above 150 mg, consider splitting into two injections at different sites. .

References

Source citations for further reading.

1. Clinical evidence for targeting NAD+ therapeutically: metabolic pathways and therapeutic potential (Pharmaceuticals, Basel).
2. NAD+ and enkephalinase infusions in substance use disorder: pilot of 50 cases (Current Psychiatry Research and Reviews).
3. Pilot study: plasma and urine NAD+ metabolome during 6-hour IV infusion (750 mg) (Frontiers in Aging Neuroscience).
4. Comprehensive review of NAD+ metabolism, aging, and therapeutic applications (PMC).