I have a problem with solvent for extract for MTT assay. This extract made from n-hexane solvent. I have used DMSO for solvent before it mixed with culture medium, but the extract didnt dissolve. I have tried to use Tween 80 but it's too toxic for the cell line (HepG2). Are there suggestion for me what is the best solvent for my extract? thanks
Hello, Mia!
I use isopropanol containing 0.04 M HCl to dissolve in the MTT test on cell cultures
Try to use ultrasonic bath for extraction process ( sonication technique ) which makes the material dissolve in the solvent ( n- hexane or isopropanol).
Try find out the nature of your extract (lipophilic, Hydrophilic, ...). Then, you will be able to find the appropriate solvent, accordingly. As you may know, n-hexane is also toxic for the cells. As Faiq H S Hussain said, ultrasonication is working in many cases, but you must be alert whether your extract is sensitive to the heat produced by the sonication or not.
Hello Mia
As Arman mentioned n-hexane is also toxic to the cells. Is it a herbal extract ? If yes, then try making your extract in aqueous alcohol (70% or 50% ethanol). It is less toxic to the cells. You can also use acetone in varying percentages. It is a polar solvent and easily miscible in water and other organic substances. It does not affect the viability of the cells.
Hi Mia,
I had the same problem with some natural compounds such as monoterpenes. Employing ethanol instead of DMSO worked better in some cases, particularly for high concentrations of the compounds. In HepG2 cells, concentrations up to 0.5% v/v ethanol work well (no cytotoxic). Good luck!
Thank you for all. I just try to dissolve my extract with ethanol 0,5% and it doesnt have a cytotoxic effect. but ethanol has volatile effect, is it will influence the concentration of the extract on culture medium? Boris Rodenak Kladniew
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