HPLC · CoA
How Do You Reconstitute AOD-9604?
Guide · 8 min read · Updated 30 September 2026
AOD-9604 is reconstituted by adding a measured volume of bacteriostatic water down the inside wall of the vial and swirling gently until the solution is clear and colourless. Adding 1 mL to a 2mg vial yields 2 mg per mL, and 2.5 mL in a 5mg vial gives the same 2 mg per mL, so one set of calculations covers both sizes. Dilute acetic acid is reserved for lots that resist dissolving, and the finished solution is held at 2 to 8 °C.
What Is AOD-9604 and Why Does Its Chemistry Matter for Reconstitution?
AOD-9604 is a synthetic 16 amino acid peptide, and its charge and disulfide ring decide how it behaves once diluent reaches the powder. The sequence is Tyr-Leu-Arg-Ile-Val-Gln-Cys-Arg-Ser-Val-Glu-Gly-Ser-Cys-Gly-Phe. It mirrors the 177 to 191 stretch of the human growth hormone chain, with one tyrosine added at the amino end, giving a molecular weight near 1815 g/mol. A disulfide bond between the two cysteines closes part of the chain into a small ring.
Two arginines give the chain a net positive charge at neutral pH, offset by a single glutamic acid, so most lots dissolve directly in bacteriostatic water. A mildly acidic diluent adds further positive charge and keeps individual molecules apart, which is why some protocols specify acetic acid. The disulfide ring is the vulnerable feature: heat, vigorous agitation and repeated freeze and thaw cycles all put it at risk.
What Equipment Is Needed to Reconstitute AOD-9604?
A standard reconstitution needs the peptide vial, a preserved diluent, measuring syringes, swabs and a record sheet, all set out on a clean bench before any stopper is pierced.
- A sealed vial of AOD-9604 research peptide in 2mg or 5mg fills, labelled with lot number and storage temperature.
- Bacteriostatic water in multi-draw vials, the default diluent for most peptide work.
- Optional: dilute acetic acid for acid-soluble peptides, held in reserve in case the cake turns cloudy.
- U-100 insulin syringes for measuring diluent, plus alcohol swabs for the stoppers.
- A laboratory notebook to record lot number, diluent, volume and date.
The AOD-9604 starter kit supplies the vial together with diluent and consumables in a single order.
What Are the Steps to Reconstitute AOD-9604?
The procedure takes about five minutes and follows the same seven steps for the 2mg and the 5mg vial:
- Take the vial from frozen storage and allow 10 to 15 minutes on the bench so no condensation forms inside.
- Flip off the plastic cap and wipe the rubber stopper with a fresh alcohol swab. Swab the diluent vial in the same way.
- Draw the planned volume of bacteriostatic water. A full 1 mL fill reads as 100 units on the U-100 scale.
- Pierce the stopper at an angle and direct the stream against the glass wall, not straight onto the powder.
- Swirl or roll the vial slowly. Never shake it. Most lyophilised cakes clear within one or two minutes.
- Inspect the solution against a light. It should be clear and colourless. If haze or fine particles remain after five minutes of gentle swirling, move to the acetic acid method.
- Label the vial with concentration and date, then return it to the refrigerator without delay.
AOD-9604 Reconstitution Chart for 2mg and 5mg Vials
Concentration equals vial milligrams divided by diluent millilitres. On a U-100 syringe one unit is 0.01 mL, so ten units hold one tenth of the concentration figure in milligrams.
| Vial | Diluent added | Concentration | Peptide in 10 units (0.1 mL) | Peptide per unit |
|---|---|---|---|---|
| 2mg | 1 mL | 2 mg per mL | 200 mcg | 20 mcg |
| 2mg | 2 mL | 1 mg per mL | 100 mcg | 10 mcg |
| 5mg | 1 mL | 5 mg per mL | 500 mcg | 50 mcg |
| 5mg | 2 mL | 2.5 mg per mL | 250 mcg | 25 mcg |
| 5mg | 2.5 mL | 2 mg per mL | 200 mcg | 20 mcg |
| 5mg | 5 mL | 1 mg per mL | 100 mcg | 10 mcg |
Round figures make later arithmetic safer, which is why 2.5 mL in the larger vial is so widely used: it matches the 2 mg per mL of the small vial with 1 mL. For any other combination, the peptide reconstitution calculator performs the division and converts the result to syringe units.
When Should Acetic Acid Be Used to Reconstitute AOD-9604?
Acetic acid is used only when an AOD-9604 solution stays hazy in bacteriostatic water, usually at high concentrations such as 5 mg per mL, or when a protocol specifies an acidic stock. A weak acid lowers pH gently, adds positive charge to the arginine side chains and holds the molecules apart so they remain in solution. Most lots never need it.
Laboratories use one of two approaches:
- Acid only: the full volume is dilute acetic acid instead of water. The method is simple, but the solution contains no preservative, so it suits single-session work or aliquots that are frozen promptly.
- Acid first, water second: a small portion of acid, for example 0.2 mL, wets the cake and is swirled until clear, then bacteriostatic water brings it to the final volume. Concentration is based on the total volume, so 0.2 mL of acid plus 1.8 mL of water in a 5mg vial still gives 2.5 mg per mL.
Both volumes belong in the notebook so the stock can be reproduced exactly.
How Should AOD-9604 Be Stored Before and After Mixing?
Sealed AOD-9604 powder is stored at -20 °C away from light, while the reconstituted solution is refrigerated between 2 and 8 °C, where laboratories commonly work within a 28-day window. Published stability data for this fragment in solution are limited, so these conditions follow general peptide practice and the in-use conventions for preserved multi-draw vials.
| Stage | Temperature | Typical laboratory window | Notes |
|---|---|---|---|
| Sealed powder, long term | -20 °C | Up to the date on the label | Keep in the box, away from light |
| Sealed powder, short term | 2 to 8 °C | A few weeks | Acceptable while a study is being set up |
| Mixed with a preserved diluent | 2 to 8 °C | Commonly up to 28 days | Mirrors the in-use convention for preserved multi-draw vials |
| Solution in acetic acid only | 2 to 8 °C | Days, or freeze aliquots | No preservative in the diluent |
| Frozen aliquots | -20 °C | Several weeks | Thaw once and do not refreeze |
The 28-day figure is a handling convention rather than a stability claim for this peptide, so any study that runs longer should confirm integrity by HPLC or open fresh vials.
Common Reconstitution Errors
- Shaking the vial to hurry the powder, which can fold or aggregate the chain.
- Spraying diluent directly onto the cake instead of down the wall.
- Omitting the diluent volume from the record, which turns every later calculation into an estimate.
- Leaving the mixed vial at room temperature on the bench between draws.
- Freezing and thawing the same vial more than once.
Which AOD-9604 Vial Size Suits a Research Study?
We recommend the 5mg vial reconstituted with 2.5 mL of bacteriostatic water for most studies, because it gives an even 2 mg per mL, leaves headspace for clean draws and provides more material from one lot for repeat experiments. The 2mg fill is the better fit for a short pilot. Every batch we supply reaches 98% or higher purity on HPLC, mass spectrometry confirms its identity, and the lot number on the label links to its certificate of analysis. The studies behind the fragment are collected in our AOD-9604 research library, and a design comparison with two related metabolic research peptides is in AOD-9604 vs tesamorelin vs MOTS-c. To order vials, diluent and syringes together, browse every research peptide we stock.
Questions about how do you reconstitute AOD-9604
Sources
- Metabolic studies of a synthetic lipolytic domain (AOD9604) of human growth hormone. Hormone Research. 2000.
- Safety and tolerability of the hexadecapeptide AOD9604 in humans. Journal of Endocrinology and Metabolism. 2013.
Written by the Macropus Peptides technical team for in-vitro laboratory research reference. Not medical advice; products are not for human or veterinary use and are sold to buyers aged 18 and over.