Oil will be at high concentrations of 10000ppm. The aim is to enhance the bioavailability of microorganisms. I had thought dissolve in hexane or add Tween 100x to the medium. Thank You.
Hi Juan Carlos,
The oil is petroleum (density 0.93 g/cm3). If It is in small amounts, that can be homogenized with the agitator, but at higher concentrations, It forms large micelles so bioavility and real concentration decreases.
At first, my plan was to use a surfactant for it, but I read in the literature that these compounds can dissolve in nonpolar substance and It could be another option, hence my doubt if anyone has used quimical solvents. Also If bioabilidad or cytotoxicity increases?
Thank you very much!
Hi Borja, I agree with Juan Carlos in that the most promising way is to sonicate the oil with detergents to form small micelles that are stable in aqueous solutions .
Tween-20 (or 80), SDS or cholates can be worth trying.
Yes Steingrimur, It seems the most promising. Is a good idea to sonicate the oil with surfactant.
Thank you too
Depending on your oil, you can try a micelle formation protocol. I've used that with some luck for making cholesterol compounds available to microorganism. They were all over it in that form. Try:
- Mix oil, and tween20 in varying ratios 0.1x - 10x in methylene chloride
- Evaporate to dryness
- Dissolve the resulting paste in culture medium
I would try using using living organisms strategy: proteins. You can test solving your oil in medium containing 1 to 10 % BSA. For this simply put the BSA on the medium and wait for 15 min without shaking. It will solve. It may not work but I would give it a try.
There are a number of different strategies for presenting small amounts of insoluble materials to cells in culture. Crude oil is such a heterogeneous mixture that it is unlikely that any solvent would completely dissolve all of its components. Those that come closest (hexane, for instance) are themselves water insoluble and just present another oil-water interface in the culture. Detergents can disperse the components but the oil may be present in several different physical states (micellar, but more likely multilamellar). In any case, your bacteria are going to see an oil-water interface. Perhaps you should define the state of the system that you want to work with and go from there. What kind of interface do you wish to present to your bacteria?
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