Aslamo Alikom/Greetings everyone,
I've been working on MCF7 cancer cell line, and I would like to study their response to cyclophosphamide in vitro.
I was wondering what's the best solvent to use to solubilize it. Concentration to be prepared is 30 mg/mL.
As per the manufacturing company, water is used to solubilize it. Yet, upon googling the solvent, I found that DMSO is considered to be the best solvent.
I would highly appreciate your help as per your experience in helping me choose the best solvent. Also any tips for stability and storage will be highly appreciated.
Thank you in advance.
Hello Mariam Ahmed Galal,
The best solvent for cyclophosphamide monohydrate in cell culture would be DMSO. You may prepare a stock solution of cyclophosphamide in DMSO and store the stock at -20 degree C for many months. Further dilutions to be used in cell culture could be made in aqueous buffer in such a way that the final concentration of DMSO in culture is less than 0.1% so as to avoid toxic effects of DMSO on the cells.
On the other hand, cyclophosphamide monohydrate is more stable in water than in any other solvent. However, cyclophosphamide in water starts to degrade on storage either in the refrigerator at 4 degree C or when frozen at -20 degree C.
So, if you have to store the stock solution of cyclophosphamide monohydrate for future use, it would be better to use DMSO.
But if you use water to solubilize cyclophosphamide, then you will have to make it fresh each time you use in culture.
You may want to refer to the paper attached below for more information.
https://www.orientjchem.org/vol30no4/structural-and-stability-investigation-of-the-anticancer-drug-cyclophosphamide-via-quantum-chemical-calculations-a-nanotube-drug-delivery/
Please note: Since you need a concentration of 30mg/ml, you need to check the solubility of cyclophosphamide monohydrate in DMSO and water. For instance, cyclophosphamide monohydrate is soluble in water at 40mg/ml and with heat it is soluble at 100mg/ml in water, while the solubility of cyclophosphamide monohydrate in DMSO is approximately 5mg/ml.
Best.
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