I am observing a high UV260 absorbance plateau during the cleaning in place of my HIC column using 1 M NaOH. This is giving me confounding results in regards to mass balance. The HIC column uses a butyl ligand. Could this suggest that the integrity of the ligand is poor or could it be leaching out because of the high concentration of NaOH? The UV260 peak appears with or without loading sample.
Dear Jomar Difuntorum sorry to see that your interesting technical question has not yet received any expert answers. Unfortunately I'm not an expert either because we work in synthetic inorganic chemistry. However, I just found out that a closely related technical question has been asked several years ago on RG. Perhaps it might be helpful reading the answers given to that question:
Does anyone know why my 1N NaOH solutions have a high absorbance at 280nm during chromatography runs?
https://www.researchgate.net/post/Does_anyone_know_why_my_1N_NaOH_solutions_have_a_high_absorbance_at_280nm_during_chromatography_runs
(7 answers)
Also please check the following ppotentially useful link:
Sodium hydroxide sanitization of AxiChrom™ columns packed with chromatography resin
https://www.cytivalifesciences.com/en/de/solutions/bioprocessing/knowledge-center/Cleaning-sanitization-of-mixed-mode-resin-in-AxiChrom-columns
I hope this helps. Good luck with your work and best wishes!
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