Chromatography Forum

Dear all,
I am a newer in HPLC and unluckily, I made big mistake today. I am very worried at the moment. By mistake, I put wrong channel for my pump, the solvent in the bottle run out and I pump bubbles to my column for more than 1 hour! I am really shameful for my stupidity. Could you please help me the way to remove the bubbles form the column? I took the column out and use the syringe to remove the bubbles from the lines but when I put the column back, the pressure jump up and down. I am too worried because it is the new column, I think I will losse the job if it is damaged! Please, please help me! Thanks in advance!

I seriously doubt that you actually forced air into the column. Typically, a pump that runs dry will just cavitate the heads and introduce a little bit of air ito the lines just outside the pump heads (the compressibility of air and its propensity for leaking back into the heads serves the chromatographer well in these situations). Pull the column, purge the pump with some mobile phase and then put the column back. The pressure should settle down after a little while.

Depending on the system you have, you can actually damage newer membrane/vacuum degassers by trying to draw mobile phase through them with a syringe. It's better to use a dry prime routine (Waters systems) or push mobile phase through to prime the lines.

See the discussion "RP-HPLC Baseline drift", and do what wayne did: Run MeOH or ACN through the column (both are not necessary) until the spikes and or baseline drift, etc., stops.
(I havn´t worked with conventional piston pumps for a long time, but the old ones didn´t pump air, maybe some bubbles as you say, by "sucking" some gas together with the mobile phase. Can´t imagine that one can ruin the column that way, maybe the pump?)

VIT,

I have never seen a column destroyed due to air bubbles. The up and down in your pressure is because you still have some remaining bubbles in your LC system.

As HWM mentioned just flash the system with an organic solvent (if the LC system is completly dry I prefer to use ACN due to it's low viscosity, followed by isopropanol).

Dear all,
Thank you very much for all replies. I followed your suggetions, run with MeOH and with ACN, the pressure is stable now and I tried to inject my sample, everything is OK, my column is working well. It is a good experience for me, you can't imagine how worried I was! Thanks again!