Chromatography Forum

Hi
I noticed raising of pressure in the column during separation (usually 110 bar now around 155); except that I have peak splitting - all of my compound are flushed from the column at same time (around 3 minutes) and looks like "five peaks in the one". I changed frit on purge valve; try to clean the column with 100 ACN (my solvent was 80:20 ACN:water); I tried also equilibrate column but its still doesnt working. Can anyone give me some advice? I'm thinking about back flush of column. I'm working on Agilent system; binary pump

Clogged precolumn ? (if present).

To diagnose the problem remove the column and check pressure. You will find out where pressure builds (on the column or in the system before the column).

Replace Column with a new one. If the problem persist then try with a new column on another system.

If the problem still persists, then possibly you have made an error in your solvent preparation.

What column are you using and what are the dimensions?

It sounds like a bad contamination, but without knowing what the column material and size is and what you are injecting on it, it is difficult to trouble shoot the problem.