Hi,

Can anyone tell me how you address the problem of peak splitting due to alpha and beta forms of the saccharide when you seperate sugars using Reversed phase?

And how will the operating temp of the HPLC run affect the seperation and what is the basis of this?

Thank You

Increase the temperature. Speeds equilibration between alpha and beta. On Ca-form cation exchange columns, the standard temp is 65 degrees for just that reason.

Increasing the temperature generally decreases retention time someshat. Selectivity may also change (can either improve or deteriorate), depending on excactly what you're separating and how you're separating it.

Another solution to this is to use an alkaline pH. This speeds up the interconversion between the anomers. This is the reason that you get a single peak with an amino column. Of course, you need to use a column that is compatible with alkaline pH, if you want to do this.