Chromatography Forum

Hi

i have been doing certain comparison studies…recently I analyzed a protein in porous (300Å) column and a non-porous column….
In non porous column under same gradient % per minute (around 1% ) the protein gave very wide peak..under lower % gradient the peak width went on increasing

Non porous column peak width 1.503 min (1% gradient ), porous column :- 0.647 min (1% gradient )..to achieve peak width of porous column I have to run under 3% gradient

Why is there such a drastic difference in behavior of non-porous and porous columns???

Thanks

The most likely cause of your observation is the fact that you overloaded the non-porous column more than the porous column. A non-porous column has a very small surface area compare to a 300 A column.