Sterilising vials the pedantic way

[Drugsarefun]

Ok so as a carry over from a previous thread I thought I'd share my technique for sterilising vials.

I see alot of practices that claim sterile but are far from sterile, if you want actual sterile vials this is one way to achieve that, although I have yet to swab my vials and put it on agar but I trust the process.

I pulled alot of this from growing mushrooms, adapted and used equipment I had on hand, it's much the same process but you have to be alot more sterile with mushroom growing than you do bottling gear.

Clean your vials as per lab glass cleaning instructions which you can find online:

Wash and rinse both vials and rubber stoppers in tap water and dish soap then rinse 3 - 4 times in distilled water, place in the oven to completely dry, usually 20 mins or so, wait for them to cool to the touch.

Place them into autoclave bags with the rubber stopper just above the vial, do not seal the vials, it helps if you have self sealing bags but the non sealing ones are cheaper and painters tape is fine to seal them with.

Place the packets into a PP5 bucket, this is important, it must be PP5, it'll have a recyclable symbol with a 5 in the centre, these can with stand the heat of a pressure cooker, everything else will melt, the one I have is 5 litres, cut a hole in the lid of the bucket and stuff with poly fill, again must be poly fill as that handles the heat, the hole stuffed with polyfill allows pressure relief while reducing the moisture inside the bucket, your bucket will most likely implode without it.

Cover the top with tinfoil to protect any water drops dripping onto the polyfill.

Place into the pressure cooker with 2.5 quarts of water at the bottom.
Turn on high and allow steam to escape for 10mins, place the weight on and bring the pressure up to a minimum 15psi, turn it down and hold it steady, I run mine at 18psi.
20mins is probably fine at 15psi but I like to run it for 45 just to be sure.

When it's cooled down you can remove the bucket, it's contents will be sterile untill you open it, so could sit on a shelf for months and be completely sterile inside.

Once everything has cooled down you can manouver the rubber stoppers onto the vials through the plastic without opening them, they should stay sterile in the packet for upto a year.

You can also fill them while still in the packet if using a syringe filter, just peirce right through the plastic and rubber stopper and fill them up, once full remove them from the packet and cap them.

If using a bottle top filter you'd need a flow hood or at a minimum a still air box to remain sterile while filling.

 

[testocu]

Dude thank you for your sharing, I sterilized my vials in a similar way to you. First, I washed the vials and stoppers with distilled water, then I baked them at 140 degrees and dried them, then I took the stoppers out of the oven and cooked the vials at high temperature for half an hour. Finally, I sterilized it in an instant pot with sterilization pouch and autoclave bags.
I mixed mct oil and raw material for 15 minutes at 130 degrees, then I added benzyl benzoate and cooked for another 15 minutes at 130 degrees. After cooling to 40 degrees, I filtered it with 0.22 µm. I didn't use benzyl alcohol,
Result : zero pip, smooth testosterone prop

 

[OffendHand]

I bought a stove top autoclave a long time ago for... Reasons. I would recommend using an actual autoclave over a pressure cooker; the metal-on-metal seal is going to last longer (lubricate it with a little mineral oil), and you get a much more certain pressure. The weights you see on most stove top canners and pressure cookers aren't necessarily as reliable as you might want for something that you're going to be injecting. If you fuck up canning something, well, you'll probably figure it out once the lid starts bulging. If you fuck up sterilizing your vials and stoppers (and any glassware that's coming in contact with your oil after it's been filtered), you likely won't be able to tell until you end up with a nasty infection.

You MUST get a full head of steam before you allow the autoclave/pressure canner to start building pressure; the steam is what sterilizes, and unless it's penetrated every nook and cranny, the contents won't be sterile.

I would also suggest not using your autoclave for food.
If you rinse with distilled water, you can go directly into the bags from the rinse, which minimizes the odds that you can get some contamination during drying. The pouches are going to be saturated with steam anyways, and you're going to have to let them dry after they come out, so it's not a big deal.

Depending on what bags you get, they should have some kind of indicator that will change color in the presence of steam or sufficient heat; those are not a reliable way to know if the contents are sterile, it's just a quick way to know if you got up to temperature and pressure or not. You can also get indicator tape.

You should consider spore testing your autoclave.

Sterilized items should be used within 4 months, or re-sterilized. Autoclave bags and pouches are not air impermeable. Ideally, you'd sterilize just what you need prior to brewing.

If you want to get really spendy, look for a vacuum (Statim) autoclave on eBay. They have very fast cycle times; they're often used in tattoo studios as on-demand sterilizers.

 

[klokke23]

In your opinion; clean with alcohol, rinse in distilled water, tinfoil as cover + heat in oven for an hour doesn't sterilize?

 

[OffendHand]

It's not an opinion, but fact.

Dry heat can sterilize, but it takes a longer time overall, and is at a higher temperature than steam sterilization. Static heat takes longer than forced air (i.e., a convection oven will keep things at a sterilizing temperature more effectively than non-circulating air). Things like rubber stoppers can start breaking down at the temperatures required for sterilization (source: Dry Heat Sterilization - Principle and Uses - Laboratoryinfo.com). There are guidelines for how long things need to be held at specific temperatures in order to sterilize; you need to get your items to be sterilized up to 170C/340F, and then keep it there for an hour. Home ovens tend to have inaccurate thermometers, so you may want to hold it at temperature for as long as two hours, in case it's running cold. As long as you follow those guidelines, everything in the oven should be sterilized.

Even if you have sufficient heat and time for dry heat sterilizing, you should be bagging everything, because as soon as it's out of the oven and in contact with air, you have the potential for contamination. Plus, most home ovens are gross, and you don't want aerosolized grease and carbon particles ending up in your vials somehow.

As far as how you sterilize, it depends on what level of risk you find acceptable, and how much money you have to throw at the problem. Unless you are working inside of a laminar flow hood with HEPA air filters when you are filling vials, you're going to have some kind of non-sterile exposure, and the potential for some kind of contamination. Even if you have exposure to non-sterile air between when you sterilize your vials, and when you put the cap on, the benzyl alcohol in your mixture should prevent bacterial or fungal growth, so that you aren't going to give yourself a nasty infection.

Because I already had an autoclave from a previous life, I'm more comfortable using that than I would be using an oven.

 

[tuffie]

//off-topic
This reminds me of that fly episode from the great protagonist walter white, best episode imo.

 

[Drugsarefun]

//off-topic
This reminds me of that fly episode from the great protagonist walter white, best episode imo.

And here I was thinking that episode was universally hated

 

[Cdtoth666]

Why is anyone doing this? There is absolutely no need to autoclave your vials lol 90% of the sources don’t even do it. I am familiar with growing mushrooms and the same sterilization practices absolutely do not apply.

 

[clearheaded]

yes you need to in order to have 100% and sterile viles that are dry... yes in mushroom growing you autoclave/PC.. you have no idea what your talking about.

 

[Cdtoth666]

Why? Where did you read this?

 

[klokke23]

I also think it's overkill. I'm doing just fine with dry heat in oven.

 

[clearheaded]

sterile is not overkill..people are "fine' reusing syringes too.. doesnt mean its smart. why even bother bake in ur oven? just fills back up with bacteria.

 

[Cdtoth666]

I also think it's overkill. I'm doing just fine with dry heat in oven.

sterile is not overkill..people are "fine' reusing syringes too.. doesnt mean its smart. why even bother bake in ur oven? just fills back up with bacteria.

Linking back to microbiology, i’m not here to issue be a contrarian. Are you using an FDA certified laminar flow hood for all procedures? I m going to say no. The process of filling vials is not “sterile” you break the chain of sterility once and it’s no longer a sterile procedure. Autoclaving or dry heat or whatever is completely unnecessary. Are you using a bacteriostatic agent in your formula and are you filtering it? Literally all that matters.

There is this whole thing about some compounding pharmacy in MA where a bunch of vials were contaminated with some fungus and lawsuits etc. you can read all the discovery related to the case and guess what? If they had just properly filtered everything the whole debacle would have been avoided. Like to make a point, you could lick all of your glassware and spit into your carrier oil but as along as you filter to .13 microns, you will technically have “sterile” gear and it will be safe to inject.

 

[pinkie]

hey man awesome setup, do you find the vials carry alot of condensated water after the autoclave?

 

[clearheaded]

lol filtering is different than having sterile glassware..

also clearly have 0 idea what your talking about as said dont need PC or autoclave for mushrooms bahahaha..

 

[Cdtoth666]

I mean I’m confused I’m trying to explain to you that what you are doing is not necessarily whatsoever to save you the hassle or you can just continue to waste your time doing something that absolutely does nothing and makes no difference if the only goal is to make a product that is safe to inject. It’s absolutely and completely a waste time and resources and energy.

 

[tuffie]

And here I was thinking that episode was universally hated

We are the 1% who appreciate

inb4 mushrooms in my diet

 

[Cdtoth666]

I bought a stove top autoclave a long time ago for... Reasons. I would recommend using an actual autoclave over a pressure cooker; the metal-on-metal seal is going to last longer (lubricate it with a little mineral oil), and you get a much more certain pressure. The weights you see on most stove top canners and pressure cookers aren't necessarily as reliable as you might want for something that you're going to be injecting. If you fuck up canning something, well, you'll probably figure it out once the lid starts bulging. If you fuck up sterilizing your vials and stoppers (and any glassware that's coming in contact with your oil after it's been filtered), you likely won't be able to tell until you end up with a nasty infection.

You MUST get a full head of steam before you allow the autoclave/pressure canner to start building pressure; the steam is what sterilizes, and unless it's penetrated every nook and cranny, the contents won't be sterile.

I would also suggest not using your autoclave for food.
If you rinse with distilled water, you can go directly into the bags from the rinse, which minimizes the odds that you can get some contamination during drying. The pouches are going to be saturated with steam anyways, and you're going to have to let them dry after they come out, so it's not a big deal.

Depending on what bags you get, they should have some kind of indicator that will change color in the presence of steam or sufficient heat; those are not a reliable way to know if the contents are sterile, it's just a quick way to know if you got up to temperature and pressure or not. You can also get indicator tape.

You should consider spore testing your autoclave.

Sterilized items should be used within 4 months, or re-sterilized. Autoclave bags and pouches are not air impermeable. Ideally, you'd sterilize just what you need prior to brewing.

If you want to get really spendy, look for a vacuum (Statim) autoclave on eBay. They have very fast cycle times; they're often used in tattoo studios as on-demand sterilizers.

what on earth are you doing. you do not need to sterilize ANYTHING in order to prepare a safe injectable product. as in nothing you do has to be sterile from start to finish. you can either research this for yourself and get third party accounts or even better just do it for yourself and it will be self evident. first hand knowledge is the best knowledge. actually its the only knowledge.

I am curious if you can explain to mean why you think you need to autoclave anything when preparing injectable steroids.

 

[klokke23]

sterile is not overkill..people are "fine' reusing syringes too.. doesnt mean its smart. why even bother bake in ur oven? just fills back up with bacteria.

It's a common method that is good enough. And I don't feel like discussing that with you. It's already been talked about a million times here.

 

[TestTrenPrimo]

I feel like this has devolved into almost a bad joke.. when preparing injectables why wouldn’t you want to be as sanitary as possible, extra work or not?