I am trying to observe antimicrobial activity of Tween 80 on certain clinical isolates of E. coli and P. aeruginosa that I have collected.
I need to sterilize Tween for this. However, right now my lab is not providing me a sterilizing filter. So I want to autoclave Tween 80.
My current plan is to prepare various conc, of Tween 80 solutions (I am doing broth macrodilution) and then autoclave the solutions.
Is that scientifically sound?
Autoclaving of solutions is generally not advised. Sterile filtration is more easily
done if the liquid is warmed to about 40 °C and alternate portions of hot distilled water and TWEEN 80 are poured through the 0.22 µm filter. The TWEEN 80
will blend and remain in solution.
Membrane filters are used for sterilization of biological solutions which can be degraded by heating. Tween is not reported to be degraded by heating and so it can and alwaays sterilized by autoclaving.
Its not favorable to sterilize it in autoclave, however you can sterilized in autoclave at 105 C for 5 min.
According to the Sigma website you should not autoclave it but rather filter sterilise it (easier said than done though). https://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Product_Information_Sheet/1/p6224pis.pdf
Tween 80 is very viscous and thus will be difficult to filter sterilise especially with a small pore size filters.
I mix it first before filter sterilizing to reduce viscosity. I am skeptical of the concentration of most things that are autoclaved, because water is lost. I like the suggestion (above) that you heat it up first and filter it warm by alternating water and tween.
If you autoclave tween it will degrade.
I would filter. If you filter tweens, it can adsorb to filters. And of course, filtration will be difficult due to viscosity.
You may add the tween 80 with sterilized tips...it won't bring much contamination, don't worry
What is the evidence that Tween 80 will degrade if autoclaved? I have used it a lot in culture media and I am pretty sure it was added before autoclaving.
Autoclaving of solutions is generally not advised, so sterile filtration is more easily done if the liquid is warmed to sbout 40 c and alyernate its portions. Tween 80 have to sterlize by filtration through millipore filter.
Hans-Christian Mahler: The paper that you showed the link for supports the view that Tween 80 can be autoclaved: from the Figures in the paper, nothing much happens until temperatures of approx 150-200deg C are reached.
Bahgat Gerges: but Tween 08 is growth enhancer for many bacteria, especially many lipolytic ones
I agree with Dr. Roger about the using of tween 80 at 0.1
% in bacterial media for increasing the growth.
As far as I know, tween 80 (0.01%) is also used to make polysorbate solutions that is used in extracting microbial cells from materials when doing bioburden counting. It helps in separating cells from each other and from surfaces.
It will degrade. The only question is if you see it depending on if you measure it.
Hanns-Christian Mahler
Can you share data showing important degradation? I too read the paper cited and did not find helpful. The metrics were peripheral to the question. Do you have analytical data to share?
In addition to OP's inquiry, context is the common and compendial use of Tweens/polysorbates (esp. tween 80) @ 1-5% in autoclave sterilized quality control media used for QC testing of productrs from drugs to liquid laundry detergents. Largely academic as your answer will not change the practice.
Phil, thanks for asking. I understand the frustration with somewhat limited information in the paper, and unfortunately I cannot share further data given data confidentiality. my suggestion for you would be to test control vs autoclaved using polysorbate content/degradation methods incl ELSD, CAD and FL methods plus specific fatty acid/FAE methods, complemented by assessing parameters like particulates (SbVP and VP). You will find it.
You will likely see a how panel of components incl fatty acids (who have lower solubility and hence occur as particles) but also aldehydes, acids, ketons, alkans and FA esters possibly too
2. Can Tween-80 be autoclaved?
It is perfectly fine. It can improve the liquid culture single colony. However, after sterilization, when the temperature is high, Tween-80 is turbid, but the liquid is clarified when it cools down.
https://www.irochemical.com/product-detail/tween-80/
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