Questions about sterility

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  1. #1
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    Questions about sterility

    Hey everyone, I wanted to start a conversation about sterility of home-brewed vials. I'm sure it's been discussed, but as a Registered Nurse, I don't think we can discuss it enough due to its importance. Here's what I've gathered so far in my research:

    1) Measure your powder, BA, BB, and oil per one of the hundreds of recipes available.

    2) Mix in beaker over low heat with glass mixing rod.

    3) Draw up solution in syringe.

    4) Inject through filter into sterile vial with a second 25g hypo stuck in the rubber stopper to ventilate.

    There are more details than this, obviously, and this also isn't the ONLY way to brew your own gear, but it seems to be the simplest and most common for those only brewing small batches for themselves.

    Here come the questions...... First of all, none of these items are sterile. I'm a work in surgery, btw, and I know a thing or two about sterile processes and aseptic technique (however, the processes of steam sterilization for surgical instruments is vastly different than on injectable medications). I'm assuming it would be wise to clean your beaker and glass stirring stick with alcohol prior to mixing? Autoclaving it seems like it would work better, but I don't know a lot of people with an autoclave hanging around....

    Am I to assume that the BA that I mixed in with the solution is enough to kill any microorganisms that may have come intact with it? The raw powder isn't sterile, nor is the oil or BA/BB mixed in, so contamination with some kind of organism is probable.

    Does the filter help with sterilization too, or is that just to remove dust/debris?

    Lastly, working in healthcare in the United States, we're taught that any medication in a multidose vial (like the 10ml vials we most commonly use) is only to be considered "sterile" for 28 days after its first puncture unless the manufacturer states otherwise. The question is, by including BA in the mixture, how long can we somewhat "guarantee" sterility? That 28 day timeframe would begin as soon as the vial is first filled. Maybe the BA can help with sterility beyond that point, but I'd like to see the scientific research that proves it before mixing my own. The other alternative then, would be filling vials without tops, then capping them yourself and sterilizing the solution.... now we need an autoclave again..... or does that destroy the compound we just mixed?

    What are everyone's thoughts on this? As stated above, I'm sure there has already been a discussion about this that I couldn't find, so I'm sorry if I'm rehashing a previously finalized discussion, but I'm sure there are others who have the same questions and are searching for answers.

    Thank you for your thoughts and input.


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  2. #2
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    Follow-on question: using the syringe filters, how many mL can we put through them before we need to grab a new one?


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  3. #3
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    The sterilization process occurs during the filtering, both media and bacterial/bio (.22 mic sterile whatman or inline). You need to use the correct material as well or you'll degrade the filter. I haven't used whatman filters for a long time, but I can easily pass 100ml through an inline and peristaltic pump. I push into sterile 100ml vials and transfer to sterile 10's as needed. The only time I could see an autoclave being needed is if your capping/crimping your own vials. The bb increases solubility and ba is a preservative (antiseptic to inhibit bacterial growth, not to actually sterilize)
    Last edited by thewaterboy; 12-24-2016 at 10:46 AM.

  4. #4
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    Quote Originally Posted by thewaterboy View Post
    The sterilization process occurs during the filtering, both media and bacterial/bio (.22 mic sterile whatman or inline). You need to use the correct material as well or you'll degrade the filter. I haven't used whatman filters for a long time, but I can easily pass 100ml through an inline and peristaltic pump. I push into sterile 100ml vials and transfer to sterile 10's as needed. The only time I could see an autoclave being needed is if your capping/crimping your own vials. The bb increases solubility and ba is a preservative (antiseptic to inhibit bacterial growth, not to actually sterilize)
    Thanks for your input! All this stuff seems so foreign until you really learn it. As of now, I've decided to go with a 500ml .22 "complete filtration system" with a handheld vacuum pump. I'm going to aseptically draw it from the filter after the entire batch has ran through it, and inject into sterile vials. After doing a lot more research as well, I've decided to purchase pre-sterilized vials instead of doing it on my own. They cost a little more, but I'm only brewing enough for myself, so it's not THAT much more $$$.... instead of injecting the filtered gear through the rubber stopper of the sterile vials, in going to use a pair of decrimping pliers to remove the top. I'll probably remove the tops of 30 vials and put them in a rack so they remain upright (I don't want them falling over and getting contaminated). Once I fill the syringes, I'm going to place sterile rubber stoppers and flip top caps on them (I'll be wearing sterile gloves while I do this.... I'm going to put the rubber stoppers and flip top caps into a bowl of 70% alcohol and let them soak for a day, then remove them from the alcohol and, using sterile gloves, place them on a sterile drape to dry... perks of being an Operating Room Nurse), then I'll use a pair of crimpers to crimp the flip top vials on. This way, all the gear has been filtered, everything has been sterilized, and the integrity of the rubber stopper has not been compromised (by puncturing it with a needle or two to fill it). I would feel confident that my gear would remain sterile for a long, long time. Anyone have any thoughts on this technique?


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