Chromatography Forum

I hv some unknown water soluble compound need to be analysed..but i hv no idea what shld i start with?can i use hplc and reverse phase C18 to separate them? what solvent system need to be used? thanks..

If they are unknows, what's the reason for you to seprarte them? For fraction and identification? preparative LC?

because they hv some antifungal activity and i wanna identify what is the compound present..

If you have MW more than 1000 Da and difference in MW is 500 Da you can separate them also with SEC on a silica based SEC column.

If they are unknown and you want to know what they are you probably need some mass spectrometry.

Alternatively, you might try RP HPLC. Run a wide slow gradient with low UV monitoring and see if you get some peaks. Collect each of the peaks you see and test each one for activity. If you see activity with one of the peaks you could look at ways of identifying or send out to a lab specializing in this.

Hi
I would start with reverse phase HPLC. You can either hire a contract research company (like mine! ) to do the preliminary method development for you.

Or, an affordable option would be to contact a column manufacturer, provide information about your compound, and use their suggestions. Column makers have very good knowledge about their product and can steer you in a helpful direction.