I tred to dissolve vinclozolin for cell culture use. I dissolved it in DMSO. There was no problem in this step. For further dilution, I was used full medium (DMEMF12+FBS), and water, seperatly. Howewer, it precipitated. So, I can not use it. Dose anyone have suggestion for further dilution step?Thanks
What final concentration do you require? What dilution factor are you using from your stock to your final?
Water solubility (media should be quite similar) is 2.6 mg/L @ 20 degC (Tomlin, C.D.S. (ed.). The Pesticide Manual - World Compendium, 11 th ed., British Crop Protection Council, Surrey, England 1997, p. 1267)
I'm not sure if there's a standard formulation for TC use in the literature for vinclozolin, if so you should obviously go with that. Remember to add the stock dropwise into a stirring or vortexing vial of warmed media to ensure maximum solubility.
In general I've used the following aqueous-miscible organic solvents either alone or as a 50:50 mixture with aqueous solution as the initial solvent to enable dilution into culture medium:
- DMSO (Dimethylsulfoxide)
- DMF (Dimethyl formamide)
- THF (Tetrahydrofuran)
- NMP (N-Methyl-2-pyyrolidone)
- DMA (Dimethylacetimide)
- Acetonitrile
- Ethanol
- Methanol
My advice is to prepare a small amount of your drug in each of these solvents (or whichever of them you have easily available to start) perhaps in a 96 well plate, and then try diluting each into media at the desired final concentration as described above. If none work, you could expand your selection of solvents and also try a 50:50 mix of each with media or PBS. I've found rare compounds for which a stock prepared in either DMSO or PBS couldn't be dissolved into media but a stock from a mixture of the two worked fine.
Once you figure out what solvent you want to use, make sure to determine the tolerance of your cells/assay for that solvent. For example, perhaps your cells are fine with 1% DMSO but 2% begins to alter growth or differentiation or signaling in your hands. If so you need to limit the final solvent concentration to 1%. Usually, the maximum tolerable solvent in the final solution gives most robust and reproducible solubility. This is achieved by having a low dilution factor of your stock into the final solution (within the limits allowed by your system).
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