I have isolated some compounds from a medicinal plant. The compounds are very soluble in DCM but not soluble in DMSO. I want to carry out MTT assay using these compounds against selected cancers. What alternative solvent can I use other than DMSO?
The solvent/vehicle should be chosen to optimize the solubility of the test agent without adversely impacting the assay conduct, i.e., cell growth, integrity of the test material, reaction with culture vessels, metabolic activation system, etc. (Ref. to add). It is recommended that, wherever possible, the use of an aqueous solvent should be considered first. Well established solvents/vehicles are for example water, cell culture medium, dimethyl sulfoxide). Generally organic solvents should not exceed 1% (v/v) and aqueous solvents should not exceed 10% (v/v) in the final treatment medium.
If other than well established solvents are used, their use should be supported by data indicating their compatibility with the test substance and their lack of genetic toxicity. In the absence of that supporting data, it is important to include untreated controls (see Glossary) to demonstrate that no deleterious or mutagenic effects are induced by the chosen solvent. "-- So, DMSO still may be the stock solvent of choice, despite its immunomodulatory effects (it quenches free radicals, hence is anti-inflammatory).
Before selecting an alternative solvent, you should test by using low percentages if these solvents may impact your MTT assays.
You might be able to get it to dissolve a little in ethanol or N-methylpyrrolidone, both of which are miscible with water, but it is likely that the substance will precipitate once the solvent is diluted. The aqueous solubility will be very limited, but may be sufficient to test in culture if it is very potent. You could consider using a carrier, such as a cyclodextrin or a Pluronic detergent, or liposomes.
These 2 papers might help
http://pubs.rsc.org/en/content/articlehtml/2016/ob/c6ob01850e
http://s3.amazonaws.com/academia.edu.documents/46257857/Ebada_S.S._Edrada_R.A._Lin_W.__Proksch_P20160605-24497-21w791.pdf?AWSAccessKeyId=AKIAIWOWYYGZ2Y53UL3A&Expires=1489457378&Signature=4ddtxYIb8xCLgJiJ2AqayWMlVUM%3D&response-content-disposition=inline%3B%20filename%3DMethods_for_isolation_purification_and_s.pdf
Dear Moses
Can you make it a salt form? It will be best technique if you have sufficient amount in hand.
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