Sanitizing Yeast Storage Bottles

[InspectorJon]

My son came home with these little bottles of Immunity Defense shots. It's mostly fruit juice and root extracts. It has bacillus something or other in it, some kind of probiotic bacteria. I don't know if it would cause souring or something like that but I don't want to try. The bottles look perfect for storing yeast samples, 60 ml size. Is a soak in Starsan adequate to sanitize them after a good wash? What recommendations are there for sanitizing plastic for yeast storage containers? I don't think they would survive my pressure cooker.

 

[brewkinger]

They appear to be plastic and they actually look A LOT like the vials that White Labs yeast used to come in.
I prefer glass mason jars for all my yeast harvesting, so I know that my process works and YMMV.
I suppose you could just sanitize them in boiling water and then cap them after they cool (keeping sanitized water inside of them).

 

[dwhite60]

I like mason jars too.

I fill them about halfway with water then bring to a rapid boil in the microwave. Once there I put the lid on tight. I will hold it with a thick towel and shake it to get the inside of the lid too. I don't open it until it's time to use it.

All the Best,
D. White

 

[RPh_Guy]

Is a soak in Starsan adequate to sanitize them after a good wash? What recommendations are there for sanitizing plastic for yeast storage containers? I don't think they would survive my pressure cooker.

I suppose you could just sanitize them in boiling water and then cap them after they cool (keeping sanitized water inside of them).

Assuming they're PET, you cannot sanitize them with heat.

They need to first be properly cleaned with a PBW/Oxiclean soak, rinsed, and then sanitized before use.
This process is no different than what you should be doing for all your cold side equipment.

 

[InspectorJon]

These are plastic bottles. I don't think boiling would work. I use mason jars for storing in the refrigerator up to a few generations. I put them and the lids in my Instant Pot for 10 minutes. That should be pretty close to sterile. That won't work with plastic I don't think.

I want to freeze a few samples of some yeast I harvested and grew up from bottles that is not available for purchase. These seemed like the right size to freeze. Seeing as they will be used to preserve early generation samples for future propagation I want to be sure they are as sterile as possible. So, is Starsan enough after a good cleaning or should I use something else? Chlorine solution and then rinse with boiled then cooled water maybe?

 

[RPh_Guy]

Star San is good.

 

[Deadalus]

Iodophor. I use that on glass then Star San when storing yeast. I save 2-3 bottles with about 200 ml of starter when I buy a new liquid yeast this way.

I like the Instapot idea too I may try that.

 

[RPh_Guy]

Iodophor.

Any no-rinse sanitizer is fine.

 

[Deadalus]

Any no-rinse sanitizer is fine.

Probably, but for long term storage I like to hit the storage vessel twice. They are sanitizers not sterilizers right? I used to make slants a long time ago and tried my best to keep things as "sterile" as possible (in my home kitchen of course) but once in a while I would have an obviously infected slant. I suspect it may have been the caps as I pressure cooked the vials. Either I couldn't afford star san back then or it wasn't readily available (20+ years ago). I stopped making them and just decided it was easier to harvest some yeast from starts and buy a new pack of yeast every 1-2 years. What I don't want to happen is to blow a whole day all grain brewing over such as easy step. Besides, I still a have a big bottle of Iodophor hanging around which I only use occasionally to switch up the sanitizer type.

 

[InspectorJon]

Thanks for the replies. The yeast I am trying to preserve is the house yeast at Lone Pint brewery in Magnolia, TX. I like it and I can't buy their beer here in California so I am going to a little more effort than I would otherwise to preserve what I have. I got it from a pils/mosaic SMASH they call Yellow Rose. My daughter and SIL brought me some when they visited last fall and I brought some home in February. I have used it in pale ale, IPA and a big stout. It worked great for all these. Ferments fast without giant krausen, around 80% attenuation and has a mild ester character.

I have been overbuilding starters but would like to preserve some before I get too many generations into it.

 

[RPh_Guy]

They are sanitizers not sterilizers right?

This is a reason to use glass that can be heat sanitized, not a reason to use two sanitizers. If there's a biofilm that wasn't properly cleaned, you could throw a dozen sanitizers at it and it wouldn't help.

The products I suggested are percarbonate-based cleansers, which release hydrogen peroxide. Therefore they not only remove organic matter, but they are also powerful antimicrobials. The sanitizing step is just to kill any strays that may have landed on the surface during storage, a job easily accomplished by any of the no-rinse sanitizers available.

If you really feel like you need to step up your anti-microbial game, you should use PBW for cleansing followed by an acid rinse to remove inorganic residue. For sanitizing make a vinegar + bleach no-rinse sanitizer because it actually does sterilize (a clean surface). Or use glass and get an autoclave.

There's no reason to switch to sanitizers because microbes don't become resistant to any of what we use.

Hope this makes sense.

 

[Deadalus]

This is a reason to use glass that can be heat sanitized, not a reason to use two sanitizers. If there's a biofilm that wasn't properly cleaned, you could throw a dozen sanitizers at it and it wouldn't help.

The products I suggested are percarbonate-based cleansers, which release hydrogen peroxide. Therefore they not only remove organic matter, but they are also powerful antimicrobials. The sanitizing step is just to kill any strays that may have landed on the surface during storage, a job easily accomplished by any of the no-rinse sanitizers available.

If you really feel like you need to step up your anti-microbial game, you should use PBW for cleansing followed by an acid rinse to remove inorganic residue. For sanitizing make a vinegar + bleach no-rinse sanitizer because it actually does sterilize (a clean surface). Or use glass and get an autoclave.

There's no reason to switch to sanitizers because microbes don't become resistant to any of what we use.

Hope this makes sense.

First off, please don't selectively quote me. I say that because what doesn't make sense is why you quoted me in the first place since Iodophor is a no rinse sanitizer. If you think any no rinse sanitizer was good then you could have just said that first correct? Maybe you thought because I posted after you I was addressing you? Common forum problem I wasn't.

Thank you for the suggestions about upping my game, although I wasn't asking. While informative, your solution doesn't cover all the bases. My statement is true, there is a difference between sanitizers and sterilizers, in fact a numerical difference by definition. So while very small, a sanitizer is not 100% effective. By definition it may miss something. Will it? Probably not, which is what I said. However, human beings make mistakes. Clearly homebrewers make mistakes. Sooner or later we get an infected batch. We don't always know why but somewhere along the way there was a sanitation snafu. A lot of times it's operator error. Maybe you don't get the lip of the storage vessel sanitized or the very top. Maybe you mix the sanitizing solution up wrong. Maybe your wife walks in and washes her hands over your starter wort cooling in the sink (ok different problem). How many times do you think the average homebrewer can sanitize something before screwing up? Perhaps what I'm doing is redundant but it's insurance for 2 or 3 batches of beer plus it helps to remove a potential possibility if troubleshooting a sanitation issue. I don't suggest it for everybody but the OP was looking for a specific solution beyond Star San.

Hope that makes sense for you too.

 

[RPh_Guy]

The OP asked about Star San specifically, asking if it is enough, and it is (when combined with proper cleaning).

Iodophor is not superior to Star San in this context.

Your preference to use multiple sanitizers is not based on science.

 

[Deadalus]

The OP asked about Star San specifically, asking if it is enough, and it is (when combined with proper cleaning).

Iodophor is not superior to Star San in this context.

Your preference to use multiple sanitizers is not based on science.

I never said either was more effective, nor am I arguing one over the other. You seem to still have an issue that my post mentioned Iodophor right after you.

Also, you are ignoring the point that Star San, as scientifically determined, is only a sanitizer. That means it is only effective at a specific rate. The rate is not 100%. That means there's a chance that it won't work. It's small (1 in 100,000). (see https://www.sterilex.com/interpreting-an-epa-label-sanitizer-vs-disinfectant/) However it exists. Since both Star San and Iodophor are sanitizers, by using both I reduce the risk to 1 in 10,000,000,000, as we need both to fail so we multiply the probabilities (math, an integral part of science). I have to say, that is super-effective. Better even than the numbers for a disinfectant.

Predominately though, as I said, even with "proper cleaning" and proper scientific approaches, humans make mistakes. You can't take the human out of the equation. Scientists build redundancies into systems all the time for the event of failure. Suppose you have a 1 in 1000 chance of improperly applying the sanitizer. (You mess up one bottle in a thousand.) That's less than the effectiveness of the sanitizer itself. Apply it twice, that error rate becomes 1 in a 1,000,000.

Carpenters have a much easier way of saying it, "Measure twice cut once."

 

[RPh_Guy]

by using both I reduce the risk to 1 in 10,000,000,000

You're making the assumption that the 1 in 100,000 microbes that slips by Star San is killed by Iodophor and the 1 in 100,000 that slips by Iodophor is killed by Star San.
Are you basing this on some kind of data?

 

[Deadalus]

You're making the assumption that the 1 in 100,000 microbes that slips by Star San is killed by Iodophor and the 1 in 100,000 that slips by Iodophor is killed by Star San.
Are you basing this on some kind of data?

Sanitizer effectiveness is measured by load reduction. The microbial load reduction for the first one will be log 5 and the second will also be log 5 for a total reduction of log 10. That's all the data you have on whatever might be residing on your storage vessel. It's relative to what you are starting out with and my eyesight isn't that good to see to or count it. The claim is made by the producers that the microbial load will be reduced by that amount. It's a fair assumption to make. If there are superbugs resistant to both then I guess we are screwed anyway. Technically it could just be one single organism that might infect it. On the flip side, you are assuming it kills nothing else. I seriously doubt that would be the case, where's your data for that? That's not even the driving part of my argument though.

How often do you mistakenly measure things in general? Misplace a decimal? Divide wrong? Because you have to measure the sanitizer to use it effectively. What if you accidentally use the wrong amount? What if you don't apply it correctly? And what do a lot of people do when making beer? They drink it. How often do you think you might mess up the sanitation of a yeast storage vessel. It's not washing bottles repeatedly either. This usually will have a few days in between, a cold start so to speak. I am not of the opinion that a human could do it correctly 1000 times in a row. Perfectly. Every time.

Again though, you are just being selective in what you are choosing to discuss. Measure twice cut once.

 

[RPh_Guy]

We know that both Star San and Iodophor exceed their labeling with regard to efficacy.
Your mathematical argument for separate log 5 reductions is nonsense because you're assuming that the log reduction is the same even after application of a different sanitizer, which is pretty unlikely. You're also forgetting that the EPA label requirements with regard to log reduction are from testing against specific microbes, and not the microbes we actually care about or even a random assortment.

I perform QA checks with my single-sanitizer process by incubating wort for weeks in Star San sanitized vessels, and there's no microbial growth. These sanitizers are quite effective.

I can't really argue against building in extra redundancies if you want, but with proper cleaning and the intrinsic anti-microbial properties of beer, it's certainly not needed.

Cheers

 

[Deadalus]

I have no problem conceding that a second sanitizer may not achieve the same log reductions. But nonsense means no validity at all. They are not the same chemicals. To say that there would be no further effect that would be nonsense. Log 3, log 2, C'mon man. Zip, zero, nada? Not helpful in the least. Do you own Star San stock?

Your QA approach sounds scientifically developed but I suspect your methods aren't typical of your average homebrewer, could be professional even. The majority of homebrewers are hobbyists. Not to say at all that they can't get really good at this. I think you should mix it up a little bit. A couple of runs where you drink 2, 4, or 6 beers, a couple of experiments with kids running through the kitchen, some where you work on the car all day maybe not fix it and sanitize a few bottles, leave the news on the TV in the other room, have somebody vacuum the room overhead or play lousy music in the room next door. That sort of thing. Everybody is freakin' home too right now in the house. Nobody leaves.

Cheers.

 

[TennBrewer]

I thought I would just toss in my 2 cents here as I have dealt with this issue before. I make starters from liquid yeast and pour off 500ml to start the next batch. I usually washed the containers with PBW, rinsed and used StarSan to sanitize before storing off some of the starter. Every once in awhile, I would get a bad yeast contamination that showed itself in the primary fermenter, had to dump it. Lately, I have been sterlizing the jars in the oven, covering the opening with aluminum foil and heating at 350 degrees for 1 hour. When I need a jar, I peel off the foil and pour it in. I use StarSan on the lids, tops, and stoppers. Haven't had a bad batch yet.

 

[RPh_Guy]

To say that there would be no further effect that would be nonsense. Log 3, log 2, C'mon man. Zip, zero, nada? Not helpful in the least. Do you own Star San stock?

Do we know that Iodophor/Star San/etc actually leave viable Saccharomyces cells (for example) on a clean non-porous surface? Nope, they very well may be "disinfectants" with regard to this particular genus and the small microbial load present after a good cleaning. Without those data, arguments about how much microbial reduction a second sanitizer might contribute are just hypothetical, predicated on vague and untested assumptions. Empirical results are the only resource we have for determining the best use of these products in our brewing context.

I favor Star San for no other reason than because it's shelf stable, unlike everything else.

Your QA approach sounds scientifically developed but I suspect your methods aren't typical of your average homebrewer, could be professional even.

My cleaning process is more advanced than most because my risk of contamination is about as high as it can get. I use one set of equipment for both clean (Sacc-only) beers and also things with all manner of wild microbes (Brett, Pediococcus, etc), and everything is bottled.
Sanitation is the easy part -- a spray bottle with Star San.

QA testing is simple and it can be a useful tool. See how long you can keep unhopped wort in a vessel without it growing something (i.e. becoming turbid).

 

[Deadalus]

Do we know that Iodophor/Star San/etc actually leave viable Saccharomyces cells (for example) on a clean non-porous surface? Nope, they very well may be "disinfectants" with regard to this particular genus and the small microbial load present after a good cleaning. Without those data, arguments about how much microbial reduction a second sanitizer might contribute are just hypothetical, predicated on vague and untested assumptions. Empirical results are the only resource we have for determining the best use of these products in our brewing context.

I favor Star San for no other reason than because it's shelf stable, unlike everything else.


My cleaning process is more advanced than most because my risk of contamination is about as high as it can get. I use one set of equipment for both clean (Sacc-only) beers and also things with all manner of wild microbes (Brett, Pediococcus, etc), and everything is bottled.
Sanitation is the easy part -- a spray bottle with Star San.

QA testing is simple and it can be a useful tool. See how long you can keep unhopped wort in a vessel without it growing something (i.e. becoming turbid).

So let me get this straight. You don't really know for certain that any microbial organisms may be left after using Star San one way or the other. Like from a real controlled experiment? You were pretty adamant about telling me otherwise, that your way is sufficient for you. Your "empirical results" are what are called anecdotal evidence absent a well designed experiment. The level of certainty you have been projecting is unsupported, it's anecdotal. Personally I find my method sufficient for me. I presented it to the OP as there was some concern evident with the use of PET. But I know there's enough uncertainty that I wouldn't claim it completely necessary to follow for everybody, as you shouldn't either for your methods since we also talked about the human factor for which your only response was perfectly done proper cleaning every time.

Also, if I were to run your QA procedure on my methods you should expect that I would get the same results. I will properly clean everything and perform to perfection. Your QA is not sufficient to give you the level of confidence you are projecting. It has no control, one treatment, and no randomization.

 

[day_trippr]

Academic exercises aside, one could whack said vials with a sequence of detergent, acid, and whatever sanitizer - or sanitizers - one has on hand, and be done with it.

There's no harm in overkill...

Cheers! (...unless nukes are involved )

 

[RPh_Guy]

You don't really know for certain that any microbial organisms may be left after using Star San one way or the other. Like from a real controlled experiment?

Right, no such data exist.

You were pretty adamant about telling me otherwise, that your way is sufficient for you. Your "empirical results" are what are called anecdotal evidence absent a well designed experiment.

Exactly. But I'm not just looking at my results, but the entire world of brewing. Pretty much no one uses or recommends using multiple sanitizers. Whether you consider that a strong argument or not is just opinion. It is what it is.

The level of certainty you have been projecting is unsupported, it's anecdotal. Personally I find my method sufficient for me.

That's fine. This whole time I've only been pointing out that there's no evidence to support the use of multiple sanitizers.

Your QA is not sufficient to give you the level of confidence you are projecting. It has no control, one treatment, and no randomization.

I'm not projecting any confidence because I'm not trying to prove anything. I'm just making observations about the lack of any data supporting your claims that using multiple sanitizers is somehow beneficial.