We performed mtt test for both two compounds soluable in medium and DMSO for cell culture applications. When we repeated mtt 2 weeks later, we did not see any effect compared to the control group, even though we used the same stock solution. Is it necessary to prepare the substances that we want to see the effect of fresh in solution, that is, to prepare them on the day of the application? The substances that we could not see the effect of were prepared 2 weeks ago, but unfortunately we could not see it.
Without knowing the identity of your compounds no one can give you a definitive answer. Have you looked up the solubility and stability characteristics in a CRC Handbook or the Beilstein database? It is not uncommon for poorly soluble compounds to precipitate out in cold storage on DMSO. They may degrade if you warm them to redissolve the precipitate.
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