mobile phase, HPLC
Posted: Fri Feb 14, 2014 5:58 pm
by sophieUMB
May I use more aqueous phase than organic phase for reverse phase HPLC column, such as C18 column?
like 60% aqueous phase, 40% organic phase?
I am an newer for HPLC. Hope it is not silly question.
Thanks,
Sophie
Re: mobile phase, HPLC
Posted: Fri Feb 14, 2014 7:26 pm
by Consumer Products Guy
May I use more aqueous phase than organic phase for reverse phase HPLC column, such as C18 column?
like 60% aqueous phase, 40% organic phase? I am an newer for HPLC. Hope it is not silly question.
Thanks, Sophie
Yes. We go low organic-high aqueous frequently as starting point for gradient elutions for polar species.
There are some C18 columns that cannot well tolerate less than 5% organic, so to go that low, check with supplier.
Re: mobile phase, HPLC
Posted: Mon Feb 17, 2014 11:36 am
by Gerhard Kratz
For a standard RP column I would recommend to follow the 5% rule. Keep always minimum 5% aqueous phase or on the other side 5% organic phase.