Drug recrystallize when i added to DMEM - (Aug/24/2012 )

Dear All,

I just received a compound. In its brochure, it is mentioned that it is soluble in Chloroform. I made 1mM solution in Chloroform. If I add this to water/Media, chloroform won't dissolve (Stupid me!)

The trick that I did was to dissolve it in DMSO (100uM solution) and use that as stock solution and make 100nM working solution in cell culture media. I can't see anything precipitates and it seems to be quite stable.

I am wondering if my way of doing this will be OK. I thank you if you could comment on this.

Thanks,

I used to work with PAHs (phenanthrene, pyrene,...) and we disolved the compound in acetone. However if you added to a medium without a surfactant (I used Tween80), it appeared as cloudy due to thin precipitation but concentration was 100 uM in the media, 1000 times higher than yours. It also depends on your compound, medium,...

If you give details of compound, culture media,... would be easier to give a proper response

hello every one,

i am working on organic drug which could be dissolve in DMSO. After i added the drug which is dissolved in DMSO stock, to the cells in DMEM for MTT assay, i have observed that the drug was recrystallized.

i have tried strong vortexing at 37 degree C, but it didnt dissolved completely.

I request you to please help me to figured it out..

Looking for your kind reply.

Thank you,

micronagu.

Guys, I merged your two questions because they are very similar (and came one after the other, please check the forum before posting).

Andreea

for cell culture assays, the DMSO concentration should be same or below 0.01 %, hence i dissolve the drug in DMSO in higher molar concentration like 40 mM, and added to the DMEM medium that DMSO concentration should be prescribed. but even though the drug concentration was very low, it recrystallize, and i can see the crystals under microscope, when i was performing MTT assay.

I wish you to help in this issue and kindly help me suggest me how to dissolve the drug completely in the DMEM.

my drug was Piperine and i used this in microbiological and cell culture assays..

thank you,

micronagu.

sometimes disolving in FBS or BSA will help

DMSO is ok to make stocks but if you have to add it to a aqueous medium it will happen the same as with the PAHs. To keep hydrophobic compounds/compounds with very low water solubility away to precipitate you will need something else: a surfactant, or any other substance that can increase the solubility.

A cyclodextrin may be one suitable surfactant for this, specially the cell cultures, as some are already in use in drug delivery. As DRT said, some sera can also help to keep it soluble.

thank you Xabi, and DRT.

so, i would like to dissolve with surfactant such tween 20... so please tell me the concentration of DMSO and TWEEN ... it would be very useful to me..

looking for your kind reply,

thank you,

micronagu

If you use Tween20, you don't need to add DMSO by itself, it's added because you dissolve your compound on it. But you may want to add some extra DMSO to compensate the controls, so the DMSO in all cultures is exactly the same.

I used Tween80 at 1% but my compounds final concentration were 100uM and Tween20 is different. You may need test it and remember to add the same concentration to all media to control the possible toxic effects by the surfactant

Thank you dear Xabi..

so, as you suggested me that, I will going to use 1 % Tween 80..

From stock I will dilute my drug to 1 % tween 80 for preparing working stock.. from that I will add the drug dissolved in Tween to the cells. for control, same concentration of DMSO and Tween should be added. can you please tell the maximum percentage of Tween 80 could be not toxic to the cells.. ?

thank you,

micronagu