Help for start conditions

[revere_ir]

I have a question:
I have to study a steroid (Acid cholic derivate and high insoluble in water) by HPLC-UV.
Can you tell me the best mobile phase and colum with which I can start?
I Have done some runs but I have had troubles related to insoluble in water

thanks for your answer

[danko]

Dear revere_ir,

First of all you should examine, or do some research on the solubility of your compound in water/ACN or MeOH mixtures and when you know it you will be far better off than you are now.

Also, you write:

I Have done some runs but I have had troubles related to insoluble in water

There is a lot of expertise to find on this site, but I don’t think this includes mind-reading.
Maybe you should tell us, what you have tried (stationary phase, mobile phase, flow rate, temperature etc.) and what were the results.

Best Regards

[Mark Tracy]

There is a ton of literature on the analysis of steroids, as a class or individually. A few minutes on Google and a trip the the library will save you an immense amount of time and frustration. For example see McWhinney, et. al. Clin. Chem. 1996 42:979-981 that many commercial labs use for hydrocortisone and related steroids.