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Why back pressure increases 100%MeOH mobile phase
Posted: Mon Jan 21, 2013 8:01 pm
by montana
I have experienced that when I run on HPLC for several days that the back-pressure increases (4300 PSI) with 100% methanol as mobile phase and flow 1ml/min . I analyze serum samples ( Deproteinization with methanol)
What is the reason for that? and what is the solution
Thank you
Re: Why back pressure increases 100%MeOH mobile phase
Posted: Mon Jan 21, 2013 8:35 pm
by Vlad Orlovsky
if you are using silica column, there is a chance that you solubilize silica or "rip off" tiny particles of silica, thye travel to filter in the column and plug it.
Re: Why back pressure increases 100%MeOH mobile phase
Posted: Mon Jan 21, 2013 8:47 pm
by montana
if you are using silica column, there is a chance that you solubilize silica or "rip off" tiny particles of silica, thye travel to filter in the column and plug it.
I am using C18 column,so what is the solution that you suggest.
Re: Why back pressure increases 100%MeOH mobile phase
Posted: Mon Jan 21, 2013 9:00 pm
by Vlad Orlovsky
look at other brands, don't leave column in MeOH overnight, get away from MeOH (by using EtOH, for example), flip column from time to time, replace filter on the column (not a trivial task for many columns), redevelop method with ACN.
Re: Why back pressure increases 100%MeOH mobile phase
Posted: Mon Jan 21, 2013 9:23 pm
by montana
Thank you very much Vlad Orlovsky.
Re: Why back pressure increases 100%MeOH mobile phase
Posted: Tue Jan 22, 2013 2:10 am
by tom jupille
And you may still have residual protein in there (MeOH does not remove 100%).