Hi Marina,

First, chloroform and water do not mix together, so if you disolve your drug in chloroform, it will remain in choloroform, and won't be solubilized in water.

Second, chloroform has a volumic mass superior to water, so you disolved drug will sink into your culture medium.

Third chloroform is cytotoxic. It can dissolve lipids such as membranes and induces cell death.

If your drug has a carboxyllic or hydroxyle fucntion, maybe you can produce a salt of it by disolving it in NaOH or KOH. It is commonly used to solubilize highly hydrophobic compounds in plant cell biology.

Hi Marina,

First, chloroform and water do not mix together, so if you disolve your drug in chloroform, it will remain in choloroform, and won't be solubilized in water.

Second, chloroform has a volumic mass superior to water, so you disolved drug will sink into your culture medium.

Third chloroform is cytotoxic. It can dissolve lipids such as membranes and induces cell death.

If your drug has a carboxyllic or hydroxyle fucntion, maybe you can produce a salt of it by disolving it in NaOH or KOH. It is commonly used to solubilize highly hydrophobic compounds in plant cell biology.

Its not soluble in methanol, ethanol, isopropanol, PEG, or glycerol either? Formalin even? You'need to find something miscible with water. Otherwise I agree with Olivier, try producing a salt of it.

Dear Marina, maybe this answer is already extemporaneous.

Chloroform has a solubility of 0.8 g per 100 mL of water (at 20 °C), or approximately 0.5% v / v. In culture medium this solubility could be somewhat lower. So in a MTT cytotoxicity assay in 96-well microplates, if the final mixing volume is 250 uL, I would recommend 249 uL of cells and 1 uL of chloroform (control) or 1 uL of drug in chloroform (250X solution). In theory, this would avoid the problems of immiscibility.

Naturally, this proposal is feasible if drug solutions can be prepared at 250X concentration in chloroform. In addition, It is important to keep in mind that, before doing the above, it would be necessary to verify that chloroform does not significantly affect cell viability in the absence of the drug.