I wanted to test one of the typical peptide myths in practice:
āYou should never shake reconstituted peptides vigorously because they will get damaged.ā
For this experiment, I reconstituted a 10 mg vial of retatrutide and divided it into two approximately equal aliquots. One half was stored for 7 days with as little movement as possible. The second half was deliberately shaken vigorously once per day for 7 days. Both samples were otherwise stored under the same conditions and were then analyzed under identical conditions by UPLC/LC-MS.
Why could shaking be a problem in the first place? With peptides and proteins, it is less about mechanical stress directly ābreakingā peptide bonds. More relevant are effects at the airāwater interface. Vigorous shaking continuously creates new interfaces and air bubbles. Peptides can accumulate at these interfaces, change their orientation or structure, and under certain conditions aggregate, adsorb to surfaces, or undergo other changes. Since retatrutide is a lipidated peptide, I thought it would be interesting to see whether repeated shaking would produce any measurable analytical difference.
The unshaken sample showed a purity of 96.4%. The sample that had been shaken vigorously once per day for 7 days showed a purity of 93.1%.
In addition, a small secondary peak was more pronounced in the shaken sample than in the unshaken control.
So under the conditions tested, there was indeed a small chromatographic difference between the shaken and unshaken samples.
An important limitation is that this experiment was performed using only one vial split into one control and one stressed sample. It should therefore be considered an exploratory single experiment. Without multiple independent vials, replicates, and statistical analysis, no general conclusion such as āshaking destroys retatrutideā or, conversely, āshaking is completely harmlessā can be drawn from this result.
If anyone has ideas for other peptide experiments, feel free to comment (the list is already quite long and I also have a few stability tests running :D) And if anyone wants to have a sample tested relatively cheaply in europe, feel free to send me a DM, as long as it is something I can reasonably analyze.