Chromatography Forum

Could you advise on best method to condition a column.
I am about to start using the new Amino Apex column, to detect tocopherols (vitamin E) in food oils.
Mobile phase is hexane: ethyl acetate 80:20, flow rate 1ml/min, please advice how I should begin as the specification sheet states that the shipping solvent is IPA?
What steps should I take before using the mobile phase? One method that I have been given uses methanol, followed by ethyl acetate then the mobile phase.
Is this the best way to condition the column??

is your HPLC conditionned to work under normal phase conditions?

It's really dosen't matter, just looking for the best way to condition it.

I was to keep the flow rate the same as the run, so not to build up to much pressure on it.

is your HPLC conditioned to work under normal phase conditions?
If you've been running reverse phase conditions then it really will matter!
You will need to check the mobile phase to be used is miscible with IPA.
You will need to take out reverse phase column and flush the system with IPA for few hours at 1ml/min.
Then connect column and flush with IPA (and therefore through detector).
Then purge pump to waste with mobile phase to be used.
Then pump mobile phase to column.
WK

to rinse with isopropanol at 0.5-0.7ml/min (isopropanol is miscible with both hexane and acetonitrile).
Due to its viscous nature the backpressure when pumping isopropanol will still be quite high (hence the lower flowrate of 0.5-0.7ml/min).
Afterwards, you can use the mobile phase hexane/ethyl acetate again.
As a storage solvent, I would recommend hexane.