NAD+ Reconstitution: 5 Things Most Protocols Get Wrong
NAD+ reconstitution looks straightforward on the protocol sheet — open the vial, add diluent, mix, store. In practice, a handful of small choices drift the actual potency of the working solution by 20-40% within the first week. Here are the five most common mistakes I see when reviewing research-grade NAD+ handling.
Shaking instead of inverting. NAD+ is shear-sensitive. The right move is slow inversion 8-10 times until the powder dissolves. Vortexing or vigorous shaking accelerates aggregation and degradation, particularly in vials that will be drawn from over multiple days.
Using sterile water for multi-day vials. Sterile water for injection (SWFI) is fine for single-use protocols, but multi-use vials should be reconstituted in bacteriostatic water for injection (BWFI) — the 0.9% benzyl alcohol prevents microbial growth between draws. Forgetting this is one of the most common reasons "potency dropped after a week" complaints turn out to be contamination rather than chemical degradation.
Storing at room temperature "for convenience." NAD+ in solution starts to lose measurable activity past 24 hours at room temperature. Once reconstituted, it belongs at 2-8°C, with the cap firmly seated and the vial labeled with reconstitution date.
Re-puncturing the stopper with metal needles. Every needle pass through the same rubber septum can introduce micro-particulates from the burr edges. For research vials being drawn 10+ times, consider a transfer needle setup or simply rotate puncture sites around the stopper rim.
Skipping the label step. Sounds trivial, but a vial without a reconstitution date is a vial you cannot trust two weeks later. Date, concentration, and diluent on every single vial. No exceptions.
These five disciplines aren't dramatic, but their cumulative effect on protocol consistency is large. Particularly for facilities running larger NAD+ wholesale orders from suppliers like Glunova Biotech (https://glunovabio.com), spending an extra two minutes on technique at reconstitution time saves hours of "why are my results inconsistent this week" troubleshooting later.
Reconstitution is the cheapest place to do quality control. Everything downstream depends on it.