I used ethanol (50%) stock solution. It was dissolved properly but after further dilution it showed some haziness.
Dear, either increase the concentration of ethanol up to 100% or use DMSO. TQ
Dear Anil,
I think indomethacin is difficult to dissolve. Not in water, so in ethanol and possibly in methanol. You have to study the solubility of the product in some books like the Meck Index.
So good for dissolution of hydrophobic solutions are methanol , DMSO or ethanol ,(intermediate one) , either , for HPLC technic, acetonitril . And as said by Rahman, you can use first 100%,essentail is that after the dilution, the first high concentration is no more than 1% not to be toxic for cells.
But take care when you further dilute if it is directly in phosphate buffered saline solutions in example , because it precipitates (the solvants) with the phosphates of the solution.
So post dilution has to be made in mediums without PO4 ions; if it was in HBSS or RPMI, to verify also that there is no possibilities for precipitation.
And ia point has to be discussed: the final concentration you want to use: because often in research we begin at high concentrations to be sure to have an effect, but which are not compatible with blood concentrations you obtain ie when you treat a patient. You have to know these concentrations during treatments in human.
If not, you wouln't be able to apply your discoveries (so financial support by industrial before) if you have not these data (see bibliography).
I hope usefull.
Regards
Didier
Make a stock in DMSO and freeze, or try dissolving in 100mM sodium carbonate (Na2CO3) at 0.1mg/ml with warming. It should go in with a little shaking and warming, then you can dilute it into you final buffer.
Dear Didier sir,
Thank you for your valuable suggestion, as i told initially i disolved it in 100% of EtOH after that i made it 50% all was going well, it was properly dissolved a clear yellow color as described in sigma product specification. But the dilution to my desired concentration was problem as it was showing some milky precipitates. If i use 50% of EtOH again to dilute it will be surly toxic to cell viability (Hepatocytes in my case). Well thanks again for your valuable suggestions i'll try that.
Dear Neil Bradbury sir,
Thank you for your valuable suggestion. I'll try the way you suggested.
Can you give me some advice? I tried dissolving indomethacin in 96% EtOH overnight as it was still not properly dissolved after 2-3 hours.
Overnight it still hadn't dissolved. Does heating help dissolve the precipitates, and will I have further problems again when adding to my medium for cell culture after aliquoting?
Kind Regards,
Elaine
Dear Elaine,
As already said, to my mind, ethanol is not sufficient.
More usefull is DMSO or solutions used for lipid dissolution.
At a high concentration stock solution (with the maximal limit of solubility to be known), then dilution in culture media.
Regards
Didier
I agree with Didier, and as I mentioned before, go with DMSO. Make a stock in DMSO and freeze, or try dissolving in 100mM sodium carbonate (Na2CO3) at 0.1mg/ml with warming. It should go in with a little shaking and warming, then you can dilute it into you final buffer.
hello
i am trying to dissolve 0.357 gms of indomethacin in 0.1% of dmso . making final volume 100 ml. but it is not getting dissolved.
Following Hui Wang's instructions, 100% EtOH heated to 75°C dissolved indomethacin crystals instantly.
I would take care if you need to freeze aliquots however, for me they have reverted back slightly and have had to be heated again before use.
Hello,
If your protocol allows why do not you try to dissolve in ethanol, methanol, ether or acetone? As I know ethanol, methanol, ether and acetone are good choices for solvents since they have low toxicity over DMSO. Not sure, but some compounds better dissolve in vegetable oils such as sesame oil, castor oil. Again, if you are doing cell line work, oil might not be good selection. This is frustrating but we keep doing hit and trial :)
Note: I am concerned about indomethacin stability and effectiveness after heating at 75°C for 10min. The best way is to contact supplier/manufacturers who can help you out of this.
Good luck!!
Hello,
If your protocol allows why do not you try to dissolve in ethanol, methanol, ether or acetone? As I know ethanol, methanol, ether and acetone are good choices for solvents since they have low toxicity over DMSO. Not sure, but some compounds better dissolve in vegetable oils such as sesame oil, castor oil. Again, if you are doing cell line work, oil might not be good selection. This is frustrating but we keep doing hit and trial :)
Note: I am concerned about indomethacin stability and effectiveness after heating at 75°C for 10min. The best way is to contact supplier/manufacturers who can help you out of this.
Good luck!!
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