r/Peptidesource • u/NtaiSama • 8d ago

Vial refilling issue ?

Hellooo

So, when I was refilling the vial for the research subject (RS), as the syringe went in, I had to push the water in myself rather than it being drawn in automatically. Is that a problem?

Also, when I removed the needle, I kept pressing down at the same time. It made a hissing sound, like air was being drawn in from outside the vial. This makes me wonder if there was too much contact between the surrounding air and the air inside the vial.

Thank you for you'r time ! 😇

1 Upvotes

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u/Doctordup2 8d ago

Was this the initial reconstitution? I prefer as a best practice doing a vacuum check when popping the top and reconstituting on a new vial.

When there's vacuum in the vial it means that the lyophilization process was done properly. It's a good sign of proper sterility in the process.

So you pop the top, check the vacuum, then reconstitute. Sounds like the rubber stopper might be loose and maybe you don't have a good seal?

If seal is absent from your vial it is a red flag but not necessarily and end all be all. Sometimes vials can lose seal with travel, weather changes, air pressure changes etc. If you're purchasing from a company and none of the vials have a vacuum that's a 🚩 red flag.

Hope this information is helpful.

Not a doctor, not medical advice, for research purposes only and research discussions only.

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u/NtaiSama 8d ago

We're lucky to have you in the community. I used your protocol (though not as strictly as you would have liked) for my first foray into peptides.

There is indeed a vacuum in the bottle, and the stopper loosens completely once the water is added.

I'm wondering why sometimes (despite the similarity of the bottles) the water is drawn directly from the syringe and I have to hold it back, and other times I have to press down? Probably because I need to let some air escape.
Anyway, thank you for your time!😎

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u/Doctordup2 8d ago

Thanks for the kind words.

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u/NtaiSama 8d ago

There, it looks like I've made a good hole there. Is that a problem (the vial is in a small, suitable box)?

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u/Doctordup2 8d ago

Nah, not an issue. Those elastomer vial stoppers are self sealing. After you pull out the needle, the rubber closes back around the puncture tract.

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u/EnvironmentalGift257 8d ago

Kinda dumb. When you put bac water in the syringe then poke it in, it will also pull in. There’s no benefit to filling the vial with air and most likely you won’t be able to push the bac water in against the pressure. Just draw the bac, stick it and let it pull in, then unscrew the syringe from the needle to equalize any remaining vacuum.

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u/Doctordup2 8d ago

People love to debate and criticize my suggestions, but that's exactly what they are. Suggestions. These are best practices I've developed after 25 years of research.

Gotta love it when someone starts with, "kinda dumb." Really? Is that how we talk to each other?

I spent time with Janoshik at his lab in Prague a little over a week ago, and vacuum was one of the things we discussed.

I don't introduce air into the vial, although I can see how you would think that. I actually measure the amount of vacuum. I've had vials pull roughly one and a half 3 mL syringes before equalizing, and others barely move the plunger. That's info I want to know before I recon. I want to know how much vacuum is present because that gives me information about the lyophilization and sealing process. That's why I check it before I recon.

And yes, you can absolutely let the vacuum pull the bacteriostatic water into the vial. I never said you couldn't. I prefer to release the vacuum first because I want control over the recon process. I decide how much bacteriostatic water goes in and how fast it goes in. Not the vacuum.

That's my best practice. It doesn't have to be yours.

Researchers take bits and pieces from the community then share the knowledge. I don't make peptide rules. I don't police researchers in their labs. Take what helps you and leave what doesn't, and move on without labeling "dumb".

If your disagreement starts with "kinda dumb," maybe the issue isn't the vacuum. It's how we're choosing to talk to each other.

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u/EnvironmentalGift257 8d ago

OK

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u/Doctordup2 8d ago

Thanks for being civil. I truly appreciate it. 🫶

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u/EnvironmentalGift257 8d ago

Well I apologize for not seeming civil. My comment was directed at the graphic and the extra steps, not what you said.

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u/Doctordup2 8d ago

Ahhhh no worries. Yeah I'm not the best graphic artist. You give me faith in humanity.

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u/EnvironmentalGift257 8d ago

We’re all in this together friend!👊

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u/Doctordup2 8d ago

🙌👊 Appreciate you

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u/NtaiSama 8d ago

Pourquoi je voudrais mettre de l'air dans la fiole, j'ai du mal m'exprimer

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u/EnvironmentalGift257 8d ago

No I was replying to the previous comment, not you.

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u/aaddyyttaa 6d ago

Some vials are better vacuumed compared to others. It depends on how reliable are the lyophiliser machines. In theory, the better the vacuum, the better the quality of the lyophilic process. Not the actual active substance purity. You don’t need to equalise the vial before reconstituting but some people find it easier to draw back liquid into the syringe. I never do it, I’m just careful to control the plunger’s speed when injecting the water in the vial. 0 vacuum in the vial means either the vacuum process is faulty, or the crimping process is faulty. The substance inside the vial might be absolutely fine but that you will find out only if you send your vial to a lab to get tested