I am working on Tanshinone I, a Chinese herbal drug known for its anti-cancer properties. As this drug is highly hydrophobic in nature I can dissolve this drug only in the DMSO. This drug gets easily dissolved in dmso, but when I tried to make serial dilution from the stock solution(let’s say 10mg/200ul of DMSO) in the RPMI media or PBS, the homogenous mixture is not being formed. I want 0.2%v/v final dmso concentration to rule out its toxicity and to perform my MTT assay. Is there anyone who can help me out of this mess……. Plz do reply to this post
Over what concentration would you like to test? Seems to me that you are trying to dissolve at 181mM concentration. Is nt it too high? Is that your working solution in rpmi?
For in assay concentration of 100microM, pls follow this.
1) Prepare 100mM soln in DMSO (I hope its soluble at that levels).
2) Pipette 5microL of the above soln into 4995microL of RPMI. This will give you a concentration of 100microM in RPMI. Add 200microL of this (if your cell incubation volume is 200microL/well). This will give you a DMSO concentration of 0.1%. Run a DMSO blank and DMSO control at 0.1%. For lower concentrations, use this as working solution.
Hope this helps.
Dear sudeep indeed it is sigma product but i m not able to dissolve this product in the methanol even at 1mg/ml level.
One way may be dissolving the drug in PEG solution in water/buffer.
The other possibility is to dossolve it in human serum albumin solution - then you simulate blood as the hydrophobic drugs commonly stick to albumin.
I use to dissove hydrophobic drugs on Kolliphor EL (formerly known as Cremophor EL)
thanks to all for their kind suggestions....... :) i thing DMSO will be the best option for me as the drug is readily soluble in dmso. dilution will be in complete RPMI which contains protein like albumin.
Apurva sir, actually i want 1mg/ml of working soln. with 0.1% dmso v/v. this is possible only if i prepare approx. 200mM of conc. with dmso. even i can strt my MTT gradient with 0.5mg/ml. the drug actually function at 50ug/ml...
Apurva sir, actually i want 1mg/ml of working soln. with 0.1% dmso v/v. this is possible only if i prepare approx. 200mM of conc. with dmso. even i can strt my MTT gradient with 0.5mg/ml. the drug actually function at 50ug/ml...
Since in vivo your compound would bind to some carrier protein, e.g., serum albumin, I would suggest you dissolve your drug in solution of human serum albumin. It worked in my practice for dissolution studies...
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