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- Posts: 199
- Joined: Tue Feb 08, 2005 5:27 pm
I have a problem and I would like to know if some one can help me since I do not have to much experience with this kind of separation
I am trying to do an analysis of Galactose in urine samples using an Alliance 2795 with IRD using a Carobopac PA1.
My movil Phase is NaOH 20mM.
At the beginning the Rt of galactose is around 6 minutes but as the time pass the retention start to goes shorter and the peak overlaps with glucose. I read and I found this is caused because of CO2.
I regenerated the column passing 60 ml of HCl 1N follow by NaOH 200mM and after that RT got fine again, but the peak got broader, and after 1 hr Galactose started to elute earlier in the chromatogram.
I have two main question:
1. how can I remove CO2 from my MP? I was thinking to try to attach some kind of NaOH pellets filter on my bottle but if some one have any advice it would be welcome
2. How can I regenerate my column in order to get nice peaks again?
can this column only work with an IC provided with CO2 removal device or could work with a normal HPLC?
Thanks in advance for your help.
Oscar