Protocol Online logo
Top : New Forum Archives (2009-): : General Lab Techniques

HPLC column cleaning - (Jan/30/2011 )

Hi everyone,

I am about to start a new HPLC assay using a different mobile phase from what I have used before. The new mobile phase will be
acetonitrile:water, 50:50 v/v, with 25mM citric acid and 10mM sodium dodecyl sulfate added. I have never added citric acid or SDS to my mobile phase, so I am not sure if there are any precautions I need to take in order to protect my column. The pH of the mobile phase is expected to be approximately 2.0.

The column I will be using is an Apollo C18, 5 um (250mm x 4.6 ID), with an optiguard C18 pre-column.

I am not sure if I need to clean/rinse my column with anything after using it.

Any thoughts would be appreciated.



adding SDS to a RP column can be tricky, it can end up spoiling the column so be cautious.



Thanks for your response. When you say SDS can spoil the column, does that mean that I may not be able to use that column for other assays, but it will still be ok for the assay using SDS? I have read some information that alludes to this, but nothing concrete.

Thanks again.