# Basic Filtration Items



## powders101 (May 22, 2012)

Posted by: Dat

*WHEATEN* or *PYREX* Glass Media Bottles

Sterilize them with dry heat in the oven using established guidelines w/ tinfoil covering the opening. (NOTE: See next post for Dry Heat Sterilization of glassware)

NOTE: The media bottles Wheaton versus Pyrex will vary by neck size (33mm or 45mm). The neck size needs to be matched up with the proper size caps (either 33mm or a reusable combo of 45mm and disposable septa insert). These two items need to be matched up w/ a sterile filter which will fit the neck size. So glass media bottle, cap and filter need to be matched by the neck size.


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## powders101 (May 22, 2012)

*Dry Heat Sterlization*

Steam sterilization uses pressurized steam at 121-132° C (250-270° F) for 30 or 40 minutes. This type of heat kills all microbial cells including spores, which are normally heat resistant. In order to accomplish the same effect with dry heat in an oven, the temperature needs to be increased to 160-170° C (320-338° F) for periods of 2 to 4 hours. 5

A table of most studies conducted up to 1961 indicates that they are in enough of an agreement to support the above statements from the Berkley Lab Manual indicating that 170° C (338° F) for at least 2 hours is sufficient to sterilize objects.6 (See table below)

Using more current references such as the Dental Technician Manual, Volume 1 form the U.S. Navy we find that for sterilization of metal instruments "a typical dry heat cycle is 90 minutes at 320-345°F, plus the time required to preheat the chamber." A current microbiology text book states that "the standard settings for a hot air oven sterilization are the preheat time plus 1.5 to 2 hours at 160 °C (320 °F)."7

Based on the conformity in scientific research and medical statements we should feel comfortable relying on the following information from EngenderHealth (a leading international health organization working in third world countries).
* Sterilizing by dry heat (i.e. oven). From: EngenderHealth

The temps and times to sterilize:

Place instruments and other items in the oven, and heat to the designated temperature. The oven must have a thermometer or temperature gauge to make sure the designated temperature is reached.

Use the list here to determine the appropriate amount of time to sterilize instruments and other items for different temperatures. (do not begin timing until the oven reaches the desired temperature, and do not open the oven door or add or remove any items). The times shown here represent the amount of time that items must be kept at the desired temperature to ensure that sterilization is achieved. Keep in mind that the total cycle time--including heating the oven to the correct temperature, sterilization, and cooling--is usually twice as long as the time noted here.

Temperature
170 degrees C (340 degrees F) - 1 hour
160 degrees C (320 degrees F) - 2 hours
150 degrees C (300 degrees F) - 2.5 hours
140 degrees C (285 degrees F) - 3 hour

I suggest that as long as your glassware is capable, that the highest temperature (170 degrees C (340 degrees F) ) be used for twice the stated time... making the safe time 2 hours.
REFERENCES

1 - Development of a Multidose Formulation for a Humanized Monoclonal Antibody Using Experimental Design Techniques, Supriya Gupta, AAPS PharmSci 2003; 5 (2) Article 8
2 - A Contribution to our Knowledge of Disinfectant Action. III. Unsaturated Compounds as Germicides, H. D. Cheeseworth, E. A. Cooper, J. Phys. Chem., 1929, 33 (5), pp 720–728
3 - The veterinary formulary By Yolande M. Bishop, British Veterinary Association Pharmaceutical Press; 6 edition (October 30, 2004) page 359
4 - Disinfection, sterilization, and preservation By Seymour Stanton Block, Lippincott Williams & Wilkins; Fifth Edition edition (December 15, 2000)
5 - Biosafety Manual, Berkley National Laboratory http://www.lbl.gov/ehs/biosafety/Bio...lization.shtml
6 - Sterilization by dry heat, E. M. Darmady, K. E. A. Hughes, J. D. Jones, D. Prince, and Winifred Tuke, J Clin Pathol 1961; 14: 38-44
7 - Textbook of Microbiology by Ananthanarayan and Panikar, Orient Longman; 7Rev Ed edition (December 6, 2006)


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## powders101 (May 22, 2012)

Continued:


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## powders101 (May 22, 2012)

*Benzyl alcohol (BA) as a disinfectant*

Implements of sanitation for home brewing injectables

    Benzyl alcohol (BA)

    Benzyl alcohol (BA) is used as a bacteriostatic preservative at low concentration in intravenous medications. It exhibits low toxicity in low concentrations. The FDA has approved a medication with 5% Benzyl alcohol so a 5% concentration is our standard for the term "low concentration".

    As a bacteriostatic preservative BA prevents bacteria from multiplying. However its preservative activity is pH specific and is only effective in solutions with a pH range of 4 - 7. 1

    In addition, due to its polarity BA is an adequate solvent (capable of dissolving another solid).

    Although it is not widely known BA is a disinfectant. 2 This means it is capable of destroy microorganisms. It is not a perfect disinfectant. However it is listed in the British Veterinary Association's veterinary formulary as being a capable disinfectant, with an indicated useage "Cleaning and disinfection of teats" with the further usage defined as "Post milking teat dip 4% Benzyl Alcohol for cattle, use undiluted." 3

    Because of BA's antibacterial and weak local anesthetic properties the British Pharmacopoeia recommends that a 1% concentration be used in injectable medicines.4

    At higher concentrations BA may contribute to the degradation of proteins in the form of aggregation (increasing the clinging of the protein chain to surfaces making them susceptible to breakage or change in 3-dimensional structure).1 This happens frequently in the reconstitution of IGF-1 LR3 and in the Supriya Gupta study occurred at concentrations greater than 1%.

    In the same Supriya Gupta study BA was tested against other preservatives for effectiveness at killing mircrobes.
    Preservative Screening Test (Bactericidal/Fungicidal Activity) 1

    The efficacy of the preservative against various microorganisms was measured using a modified USP/EP PET (referred to as preservative screening test in this document)... In the procedure, formulations were tested against the following microorganisms: Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, and Aspergillus niger. The 3 bacterial strains were inoculated together at a total concentration of ~105 cfu/mL, as were the 2 fungi. Samples were incubated for 7 days at room temperature (25°C), and the total bacterial and fungal counts were measured using a colony counter. The log reduction (LR) values for the bacterial and fungal counts were calculated as log (initial count/final count).
    ...

    Results of the preservative screening tests showed that the formulations containing 0.75% and 0.5% benzyl alcohol are potential candidates to meet the USP/EP criteria (Table 4). Both formulations demonstrated a complete kill of the tested bacterial and fungal species after 7 days.

      ...
    Benzyl alcohol caused significant aggregation at high concentrations (>=1.0%); however, it was the most effective preservative in maintaining antimicrobial efficacy against bacteria and fungi.


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## powders101 (May 22, 2012)

Continued:


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## cobraforce (Dec 5, 2013)




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## BuryMeBig (Oct 22, 2014)

Whats up broskies! Hopefully this may bring some life back to this thread asking this. So my autoclave has a liquids cycle which I know all about as far as what liquids are harmless or dangerous to autoclave, and which are okay. But what I dont know is whether the autoclave is bad for the liquid! My oils..... 

I filled up a 250ml media storage bottle about half way full tad less, with olive oil as a tester.... Put the cap on, tighten it, then unloosen it by half of a turn. Thats just perfect so bottles dont blow. Ran it through a liquids cycle which doesnt have a dry cycle that proceeds its obviously... But check this out, oil came out looking sooooo nice and clear and since it was hot as all hell god it was so nicely thinned out just looked beautiful and pure  hour later!!! olive oil in bottle became completly turbid.... it was the oddest thing ever and I really have no clue to why.... now get this, a few days later, I come to find the oil once again crystal clear! Still the color of olive oil of course but when compared to some that I did not autoclave, you could tell which had been sterilized and which didnt just by its shade of yellow and level of how crystal clear it looked... tasted completly normal and all..... I have photo's if anyone would like to see...   Anyhow whats your guys's take on that? It safe to autoclave your carrier and if so, is there any reason to why we shouldnt run it through a cycle?


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## Ironbuilt (Oct 23, 2014)

Why autoclave a carrier when you have to heat it to melt a compound in the first place?  You arent sterilizing any carrier  untill you add ba, bb or whatever chem and filtering it so its pointless..


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## BuryMeBig (Oct 24, 2014)

Ironbuilt said:


> Why autoclave a carrier when you have to heat it to melt a compound in the first place?  You arent sterilizing any carrier  untill you add ba, bb or whatever chem and filtering it so its pointless..



well lets think about that for a second bro... seriously.... why autoclave it, when im going to heat it to melt the compound in the first place? alright well first off my oils are not USP or Reagent grade, therefor I in reality, dont know what was used to filter, if it was even filtered, or what the oil was even stored in, tranfered to multiple times until reaching me, and god knows how sanitary the entire process was... keep in mind bro, this oil isnt being packaged NOR handled for injection... now when we heat the compound when mixing it, are you suggesting that that level of heat is sufficient enough to call the oil clean and gtg? I understand that in the end yea I filter my shit, but lol bro, how much do you know about brewing brother? 

" You arent sterilizing any carrier  untill you add ba, bb or whatever chem" 

okay well adding BB isnt for sanitizing anything, and neither is BA. Adding BA to a ALREADY sterile solution will help keep it sterile, it doesn't sterilize anything bro....   and BB lol, we use BB if we have to to keep the hormones suspended keeping them in solution until, and in the injection site. 

So my question was, is taking your carrier oil, and placing it in a media bottle, and running it through a liquids cycle in a autoclave, which its sole purpose being to sanitize a given liquid, at 121*C and at 18lbs of pressure, could that in fact have any negative effect on the carrier oil? im not asking whether its necessary or not, that changes from ones standards to another... some people also filter there gear 2 to 3 times, great bro, if it makes you happy, filter it 10 times, AS LONG AS THERE ISNT A NEGATIVE IMPACT once all said and done. and btw, yea I filter all my oils prior to adding it to my powders and solvents, then filtering them all together... try it, and see how much easier the final filtration is, and compare filters... then youll see why I pre filter my oils like many do.. thanks though bro.


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## BuryMeBig (Oct 24, 2014)

dont take that to offense either bro, but you honestly, although I appreciate you taking the time to doing so,  just didnt one bit address my question whatsoever...thats all...


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## Ironbuilt (Oct 24, 2014)

BuryMeBig said:


> dont take that to offense either bro, but you honestly, although I appreciate you taking the time to doing so,  just didnt one bit address my question whatsoever...thats all...



Ok , so no Is the answer.  My question Is why are you autoclaving an oil in a bottle? . Will it sterilize it? No it wont,  oils need media filtration, usually 22um ,not heat to sterilize .  If you want to warm it use a hotplate.  Thks


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## BuryMeBig (Oct 24, 2014)

Ironbuilt said:


> Ok , so no Is the answer.  My question Is why are you autoclaving an oil in a bottle? . Will it sterilize it? No it wont,  oils need media filtration, usually 22um ,not heat to sterilize .  If you want to warm it use a hotplate.  Thks



Okay got ya, I keep my oil in media bottles brotha thats why I would autoclave the oil in them for storage.. silicone septum tops for drawing out the oil, basic shit lol, but I was aiming at killing what ever microorganisms and there spores that may be in the oil ahead of time, it was more along the lines of, IF I could kill the shit ahead of time, why the hell not you feel me? But regardless yea I run the finished compounds through a .2um obviously, but I think your missing my point or what my goal was.... If I could have a sterile oil to mix into the hormone by simply autoclaving it, I would do it rather then pre-filtering my plain oils... (just oil, nothing else). Then let me say it again though, once all is brewed, I run it all through a .2 BT filter anyhow... Yea I use a hot plate though for sure..  

but hey I did find some interesting shit which did answer why its a waste of time pretty much, kinda a crap shoot because the guarantee that all microbial shit has been killed is pretty much impossible to predict, heres some facts that some may find useful: 

Steam Sterilization Disadvantages
"Unsuitable method for sterilization of anhydrous oils, greases and powders. 

The World of Autoclaves article gives a partial explanation why:
The time required to kill a known population of microorganisms in a specific suspension at a particular temperature is referred to as thermal death time (TDT). However, fats and oils slow heat penetration and increase TDT.

Fats and oils have a great protective effect on microorganisms and their spores by interfering with the penetration of wet heat. As has been noted, wet heat at a given temperature is more lethal than dry heat, because moisture is an effective conductor of heat and penetrates into microbial cells and spores. If microorganisms are trapped within fat globules, then moisture can less readily penetrate into the cells and heating becomes more like dry heat."

vhall.nl/International/C...reservation.pdf

Full article:
esf.uvm.edu/uvmsafety/labsaf...autoclaves.html

Hope that helps anyone else with the same question, thanks bro.


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## Ironbuilt (Oct 24, 2014)

Good find.  But actually the worse contaminents happen from human dander ( hair, daniels cheeto hands, etc if no vent hood is used ) after filtration during filling stage..thks.


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## Faily1976 (Jan 24, 2015)

These may be worst for human


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## Diesel250 (Jan 11, 2016)

powders101 said:


> Posted by: Dat
> 
> *WHEATEN* or *PYREX* Glass Media Bottles
> 
> ...



These media bottles are only rated for 220 degrees. ...how long should they be put in the oven to be considered sterile?


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## TeK5 (Jan 12, 2016)

Bump this ^^
Would like to know.  Thanks to anyone in advance.


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## Diesel250 (Jan 12, 2016)

I went ahead and put them for 285 for 4 hrs. ...they came out fine


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## Sdsnacks (Jan 13, 2016)

I notice that you post a PVDF and PES filter membrane for bottle top filtration. I just tried my first homebrew and everywhere I've read said that Nylon was the way to go. But I haven't had much luck filtering through a Nylon bottle top. Actually, it just sits there. When I look up the filter stability, PVDF seems good, but PES is sketchy with limited resistance at even 3% BA. Granted, I'm only using 2%, but why pin a polymer that may have dissolved with my gear? Any advice?


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