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Anabolic Steroids

Sterilizing Vials

master.on said:
bottomline:
don't bother sterilizing any vials

if anything, just sterilize receiver bottles in vacuum filtration units.
Click to expand...

You must have missed the first two sentences of my post. Did you just read the title, see your opportunity to sound experienced, andjump on it as fast as you could?

I haven't seen much, if anything, beneficial come from you. From the fucked up cycles you recommend, to the complete abortion of a PCT regiment you have set in stone, you aren't helpful.
You push a homebrew Bible, that I've read through, and did not pick up anything useful from. There's nothing new in it. Basic stuff.
Not to mention all the server space you occupy by posting your ass clown threads in the political discourse, which for whatever reason even exists on a steroid forum.
 
Leancuisine said:
If you leave the location of your AAS Stash to me, I'll bring a pan of brownies to your grave every week so your spirit can smell the aromas.
I think that's A thing, right?
I can pull up a chair and tell you all about my week while enjoying a grilled cheese sandwich with mozzarella sticks.
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I will come out of my grave to shit on your sandwich and sticks. Then ill take the brownies and eat them
 
Docd187123 said:
Lmao!!! Curse my name like I do yours :p

500deg bc dry heat oven tunnels used in pharmaceutical companies use clean and unidirectional air. Your oven has food residue and crap in it withou having constant fresh clean air being drawn over them. Also, a dry heat tunnel has multiple temp sensors to ensure its actually at required temps. Your ovens thermometer can be off an easy 10-15deg or more and there are hot and cold spots in the ovens that can alter actual temps considerably more. It's better safe than sorry IMO to use more heat and more time bc after a 3-log reduction in endotoxins, they don't die off linearly after that. It's a biphasic reduction so again, better safe than sorry.




It's preferred but that depends upon the application. You cannot use wet heat sterilization for oil based compounds, water sensitive materials, etc and it's generally accepted that dry heat or terminal sterilization is much better and should be used whenever possible over wet heat/autoclave.



I'm glad you posted that bc I think it's the same reference I will talk about here. FDA stops short of recommending time and temps for dry heat tunnels. What they do state is that a 3-log reduction in endotoxin quantity must be accomplished. In that study, posted here A Comparative Study of Different Methods for Endotoxin Destruction , autoclaves were unable to achieve a 3-log reduction in endotoxin quantity. Only dry heat was able to do that.

As for why I suggest high temps and long times, see my response to LeanCusiine.
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Thanks for sharing all this information! I'm going to reevaluate my approach and probably ditch the pressure cooker for glass now since I already do 500+ for 60 minutes or more.

When I was in school and during training for my job I remember seeing 500+ degrees for 3 or more hours being needed to guarantee destruction of all spores, yet I can't find anything about that now. Maybe it's because they recommend two 1 hour sessions now?

In our lab we use an autoclave for all glassware but we only care about living organisms that can contaminate cultures, so this is probably why we don't use the dry heat method.
 
Well for aqueous solutions, an autoclave is most definitely going to be the recommended method of sterilization, since you aren't dealing with anything that is typically water sensitive, as @Docd187123 already mentioned. We are dealing with oil based solutions here, so using a method that sterilizes using humidity, regardless of how much pressure is applied, is still going to leave your glassware prone to contaminating the gear with moisture.

Dry heat is definitely the way to go here, and I don't see why you wouldn't rather be safe than sorry, so two separate 1 hour periods at 500 degrees is more than sufficient, assuming the rest of your procedure maintains sterility. I personally wash with a labware detergent, then distilled water, 91% alcohol rinse, air dry, then two cycles of 1 hour each. Never had a problem in a decade or more. I also have a separate dry heat sterilization chamber specifically used for my glass, I do not use my oven. You can find them pretty cheap on the internet.

Also, I have no clue why master.on thinks it would be wise to neglect sterilizing vials. Just because your gear contains BA does not mean that sterilizes the vial. That's asking for trouble IMO
 
How I approached sterilization has evolved over time. I suspect it will get tighter as I continue to grow.

I start with "clean vials", not sterilized. They are supposed to be dust free and clean and are sealed from outside air pretty well. First thing I do take those vials, maybe 20/25 at a time, and soak/wash them in a labware detergent bath. They soak for a few minutes.

Directly next to the detergent bath is a distilled water rinse bath.

After the distilled water bath is a second distilled water bath.

They are placed upside down on a clean and sanitized disposable baking pan to drain for a short time.

After this short time they are capped with a small tin foil square and a small hole is poked in the top of the tin foil square. The tin foil is dust free and disinfected prior to use.

At this point I have whatever quantity of vial I washed, for conversation sake let's say 100 vials. I take them all and put them in a washed and disinfected disposable baking pan and cover it loosely with cleaned and disinfected tin foil. I put the oven on 450 degrees and start the time for 90 minute's when it reaches temp.

To recap:

1. I start with clean and dust free vials.
2. I wash each vial individually with labware detergent.
3. I rinse with distilled water.
4. I rinse with distilled water, again.
5. I cap vials with a piece of tin foil and make it 'breathable'
6. I bake at 450 for 90 minutes.



Some things to note...

When I write disinfectant I am specifically referring to cavicide.

I stand each vial up in the detergent and distilled water baths so they are completely exposed at all points to the cleaner and water baths.

The vials are agitated in each bath.

I shake them as they empty. Not sure if this helps?

I do not care how the vials are arranged in the baking pan. They get tossed in one after the next.

I cap each vial individually to help maintain sterility post dry heat. The pin hole is to release the remaining moisture in each vial. I've found that 450 for 90 minutes will guarantee all moisture is gone.

I do cover my baking pan full of vials loosely with tin foil. This tin foil is clean and disinfected.

When baking media bottles I find that standing them up yields better results with my method; laying them down retains moisture. I think this is due to size maybe?

For stoppers is I simply let them soak in 70% rubbing alcohol for a small amount of time, maybe 10 minutes? I take them out of the rubbing alcohol l and place them on cleaned and disinfected tin foil, upside down. I use them as I clean them.
I have yet to have the stoppers come out of my autoclave or oven looking even remotely close to usable. I have never had an issue soaking/washing with 70% ISO.

When filling vials I use gloves, a body suit and a respirator. It can get a little warm so be prepared to have the air conditioning ripping and tearing! I do not have any fans or other form of air movement while doing this.

I do use an autoclave for some basic tools and lab glassware, stiring rods etc. It's something I have access to and for some stuff it does the job just fine. For larger media bottles and larger quantities of vials it sucks balls. Rather use dry heat.

Well guys, that's how I've been doing it. Take it fwiw, if I can improve I'm all ears. I'll field any questions you guys have.
 
Homebrew Homeboy said:
Well for aqueous solutions, an autoclave is most definitely going to be the recommended method of sterilization, since you aren't dealing with anything that is typically water sensitive, as @Docd187123 already mentioned. We are dealing with oil based solutions here, so using a method that sterilizes using humidity, regardless of how much pressure is applied, is still going to leave your glassware prone to contaminating the gear with moisture.

Dry heat is definitely the way to go here, and I don't see why you wouldn't rather be safe than sorry, so two separate 1 hour periods at 500 degrees is more than sufficient, assuming the rest of your procedure maintains sterility. I personally wash with a labware detergent, then distilled water, 91% alcohol rinse, air dry, then two cycles of 1 hour each. Never had a problem in a decade or more. I also have a separate dry heat sterilization chamber specifically used for my glass, I do not use my oven. You can find them pretty cheap on the internet.

Also, I have no clue why master.on thinks it would be wise to neglect sterilizing vials. Just because your gear contains BA does not mean that sterilizes the vial. That's asking for trouble IMO
Click to expand...

Very well said señor
 
Ickyrica said:
I cap each vial individually to help maintain sterility post dry heat. The pin hole is to release the remaining moisture in each vial. I've found that 450 for 90 minutes will guarantee all moisture is gone.
Click to expand...

Ickyrica said:
When baking media bottles I find that standing them up yields better results with my method; laying them down retains moisture. I think this is due to size maybe?
Click to expand...

If you'll allow the vials and media bottles to air dry, upside down before you bake them, the moisture will be less of a problem. This way, you won't have to poke holes in the tin foil. You want them as close to sealed off from outside air as possible, so poking holes in what would be acting as the barrier is counterproductive. Instead, try letting them air dry first, then cap with foil. Then, after two dry heat sessions of 60 minutes each, do not remove them from the heat source until the entire unit is cool.

If you take them out as soon as the timer goes off, and set them on a surface to cool in room temperature, they will cool too fast and you'll run into condensation issues inside, especially with larger stuff like media bottles. Then you'll have to start all over and you'll want to kill everyone on your block, so let the glassware cool before removing it.

Ickyrica said:
It can get a little warm so be prepared to have the air conditioning ripping and tearing!
Click to expand...

I wouldn't advise this, the amount of dust particles and various undesirables floating around and being blown through your A/C system is staggering. It might get warm, but any time you have an open vial or media bottle that is post sterilization and filtration, meaning it's exposed to the air between the heat/filter and your ass cheek, you want as close to zero air movement as possible. Turning the A/C and fans off at least 20 minutes prior to filling/capping gives the particles sufficient time to settle and greatly reduces the risk. It's not perfect, but without a laminar flow hood, it's the best chance you've got.
 
Homebrew Homeboy said:
If you'll allow the vials and media bottles to air dry, upside down before you bake them, the moisture will be less of a problem. This way, you won't have to poke holes in the tin foil. You want them as close to sealed off from outside air as possible, so poking holes in what would be acting as the barrier is counterproductive. Instead, try letting them air dry first, then cap with foil. Then, after two dry heat sessions of 60 minutes each, do not remove them from the heat source until the entire unit is cool.

If you take them out as soon as the timer goes off, and set them on a surface to cool in room temperature, they will cool too fast and you'll run into condensation issues inside, especially with larger stuff like media bottles. Then you'll have to start all over and you'll want to kill everyone on your block, so let the glassware cool before removing it.



I wouldn't advise this, the amount of dust particles and various undesirables floating around and being blown through your A/C system is staggering. It might get warm, but any time you have an open vial or media bottle that is post sterilization and filtration, meaning it's exposed to the air between the heat/filter and your ass cheek, you want as close to zero air movement as possible. Turning the A/C and fans off at least 20 minutes prior to filling/capping gives the particles sufficient time to settle and greatly reduces the risk. It's not perfect, but without a laminar flow hood, it's the best chance you've got.
Click to expand...

^^^^ this. Never ever have an AC or fan on and keep windows closed.
 
XmadXscientist said:
Lol this x 100. Your full of good advice brother!
Click to expand...
lol thanks bud, unfortunately I'm speaking from experience. I was so fuckin mad! You never see anyone post that advice, so I didn't know any better back then, and boy was I an angry motherfucker when I walked past 20 minutes into them cooling and saw all that condensation in the top lol

All that research and preparation and here we go back from square one thanks to some damn water drops haha at least I know to try and spread the word now, hopefully it will save someone's neighborhood.

Also, I gotta note here, knowing what I know about humidity, that this is likely something that occurs in similar atmospheric conditions to mine. I don't know this for a fact, so better safe than sorry, but I doubt someone in a very dry area would have this same issue, or at least not quite to the extent I see it here. I'm on the gulf coast in a relatively high humidity environment year round, for what it's worth.
 
Homebrew Homeboy said:
lol thanks bud, unfortunately I'm speaking from experience. I was so fuckin mad! You never see anyone post that advice, so I didn't know any better back then, and boy was I an angry motherfucker when I walked past 20 minutes into them cooling and saw all that condensation in the top lol

All that research and preparation and here we go back from square one thanks to some damn water drops haha at least I know to try and spread the word now, hopefully it will save someone's neighborhood.

Also, I gotta note here, knowing what I know about humidity, that this is likely something that occurs in similar atmospheric conditions to mine. I don't know this for a fact, so better safe than sorry, but I doubt someone in a very dry area would have this same issue, or at least not quite to the extent I see it here. I'm on the gulf coast in a relatively high humidity environment year round, for what it's worth.
Click to expand...

I did the same thing with media bottles and a bunch of vials. Brewed up two batches and got everything ready to filter and bottle, then went upstairs to grab all the glass and motherfucking fuck shit balls dammit Fuck! It was 1am so yeah finished that one the next day. I'm in the middle of the country in a spot that was never meant to be inhabited by humans due to extreme seasonal changes so it can probably happen anywhere.
 
Homebrew Homeboy said:
If you'll allow the vials and media bottles to air dry, upside down before you bake them, the moisture will be less of a problem. This way, you won't have to poke holes in the tin foil. You want them as close to sealed off from outside air as possible, so poking holes in what would be acting as the barrier is counterproductive. Instead, try letting them air dry first, then cap with foil. Then, after two dry heat sessions of 60 minutes each, do not remove them from the heat source until the entire unit is cool.

If you take them out as soon as the timer goes off, and set them on a surface to cool in room temperature, they will cool too fast and you'll run into condensation issues inside, especially with larger stuff like media bottles. Then you'll have to start all over and you'll want to kill everyone on your block, so let the glassware cool before removing it.



I wouldn't advise this, the amount of dust particles and various undesirables floating around and being blown through your A/C system is staggering. It might get warm, but any time you have an open vial or media bottle that is post sterilization and filtration, meaning it's exposed to the air between the heat/filter and your ass cheek, you want as close to zero air movement as possible. Turning the A/C and fans off at least 20 minutes prior to filling/capping gives the particles sufficient time to settle and greatly reduces the risk. It's not perfect, but without a laminar flow hood, it's the best chance you've got.
Click to expand...
I've played with baking times to get rid of the moisture and I think I've found a good heat range and time with 450 @ 90 minutes, but I understand the pun hole issue. I'll give your two sessions at 500 a shot. And yeah, lol, I've wanted to kill when they don't come out dry. It took me two times in the oven before they came out dry but my temp and time wasn't high enough or long enough I think. Nothing sucks worse than cleaning and rinsing and all that just to have to do it over. Worst feeling ever is when you anticipate filtering but end up cleaning and baking, again.

When you let the vials air dry I assume they are upside down? Prior to tin foil...

I use air conditioning, yes. I have a two room setup with double doors at the threshold of the rooms. I work in the second room with the air conditioning unit in the first. The second room is shrouded behind a sheet that I hung at the double doors. I did think about restricting as much of the particulate as I could. I get so hot with a suit and respirator on. HEPA filter the incoming air maybe and regulate air flow?

I looked up that flow hood, seeing a price tag on them that looks kind of high. Think that is something that can be homemade successfully? It seems like a self contained HEPA setup, correct? It filters air creating a positive pressure to keep contaminated air from entering the chamber?
 
Last edited:
http://m.ebay.com/itm/322200072433?_trkparms=aid=222007&algo=SIM.MBE&ao=1&asc=20161006002618&meid=37ece21f73794db89d438fcbfabb7bd4&pid=100694&rk=2&rkt=29&sd=321982077953&_trksid=p2385738.c100694.m4598&_mwBanner=1

Something like this seems reasonable for a small to medium size operation maybe. Between this unit (or similar) gloves, body suit, respirator and proper disinfecting I feel like the likely hood of contamination would be very low, provided your vials and oil are also sterilized/filtered. The unit is around a grand so it's not short money but it's not earth shattering cash either.

Thoughts?
 
Ickyrica said:
http://m.ebay.com/itm/322200072433?_trkparms=aid=222007&algo=SIM.MBE&ao=1&asc=20161006002618&meid=37ece21f73794db89d438fcbfabb7bd4&pid=100694&rk=2&rkt=29&sd=321982077953&_trksid=p2385738.c100694.m4598&_mwBanner=1

Something like this seems reasonable for a small to medium size operation maybe. Between this unit (or similar) gloves, body suit, respirator and proper disinfecting I feel like the likely hood of contamination would be very low, provided your vials and oil are also sterilized/filtered. The unit is around a grand so it's not short money but it's not earth shattering cash either.

Thoughts?
Click to expand...

Honestly, the medical/sterile response would obviously be yes, that would technically be the best case scenario for keeping contaminants out, but the real world response is that it may be a little excessive in terms of a homebrew op. A small UGL may benefit from it, but we all know those fuckers aren't dropping a grand on a LFH or substantially more than that on a class II biology safety cabinet.

If you're hell bent on using one, it's actually very easy to build one out of pressboard, plexiglass, a fan, and a serviceable 12"x12" HEPA filter for less than $200. If done correctly, you can achieve extremely sterile airflow and seriously reduce the chance of contamination. It may not be lab grade, but I'm willing to bet good money that it's more than what 99% of UGLs use, much less the typical hobbyist/homebrewer. You'd shit yourself if you saw some of the garage workbench setups I have seen. Fuckers brewing and filtering gear in their garage next to their filthy ass tools and shit on a plywood shelf with the garage door open, drinking beer and stopping to pump the handheld vacuum pump a few times in between sets on their Bowflex a few feet away.


For what you're trying to accomplish, if the airflow is really your concern and you want to be able to use your A/C in the process, I don't see much of an issue with just using a glove box. You can sterilize the inside of it, and don't uncap the foil until everything is sealed inside. There won't be any airflow in the box, so no chance of outside room air being a problem, since the doors are sealed with rubber gaskets.

I actually used a converted sandblasting glove box for a long time and had great success with it. It was an extra my brother had never used and he gave it to me, but to purchase one is super cheap, though you will likely have to change the gloves out to something a little thinner, as the ones that come with most sandblast boxes are very thick and cumbersome. Not exactly ideal for capping vials or pinching foil caps off lol.
 
1 buy brand-name sterile vials
i.e without too much looking in Ebay/amazon
Hospira vials are available
Products + Services
Hospira - Wikipedia
True big pharma, so why look further?
At about $2 per vial in Ebay/Amazon, Why even bother sterilizing your vials and risk your health? let's say you use an average of 3 vials per week = $6 per week, about $24 a month. Is money that tight? Is your health worth so little?



2 As for media bottles

2.1 see if a lab can sterilize them in an autoclave for a few bucks, if asked say you're growing yeast for your custom-brewed beer, or growing orchid seeds (they need sterility)

2.2 use chemicals proven for sterilization
Sterilization (microbiology) - Wikipedia

2.2.1 ethylene oxide is mostly gaseous, thus clumbersome and rarely available in Ebay/amazon, although some dentist and medical shops may sterilize bottles for you

2.2.2 Nitrogen dioxide also a gas

2.2.3 Ozone, a gas as well

2.2.4 Gultaraldehyde/Formaldehyde
Glutaraldehyde and formaldehyde solutions (also used as fixatives) are accepted liquid sterilizing agents, provided that the immersion time is sufficiently long. To kill all spores in a clear liquid can take up to 22 hours with glutaraldehyde and even longer with formaldehyde. The presence of solid particles may lengthen the required period or render the treatment ineffective. Sterilization of blocks of tissue can take much longer, due to the time required for the fixative to penetrate. Glutaraldehyde and formaldehyde are volatile, and toxic by both skin contact and inhalation. Glutaraldehyde has a short shelf life (<2 weeks), and is expensive. Formaldehyde is less expensive and has a much longer shelf life if some methanol is added to inhibit polymerization to paraformaldehyde, but is much more volatile. Formaldehyde is also used as a gaseous sterilizing agent; in this case, it is prepared on-site by depolymerization of solid paraformaldehyde. Many vaccines, such as the original Salk polio vaccine, are sterilized with formaldehyde.
Sterilization (microbiology) - Wikipedia

Premixed solutions are readily available in Ebay/amazon as 'MetriCide' 'Cavacide' about $20 to $25 a gallon
https://www.metrex.com/products/high-level-disinfectants/metricide
https://www.metrex.com/products/surface-disinfectants/cavicide
they even come with test strips to make sure they remain potent after opening.
Why even look further?


So Hospira vials + Metrex sterilizing solutions is the way to go.
 
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