Chromatography Forum

Hi! I'm looking for someone who has experiences in the separation of Sugars. I'm working with a LC-MS (UPLC) with the "SeQuant® ZIC®-HILIC 3.5µm,100Å 100 x 2.1 mm" column from Merck. I try to separate fructose, glucose, sorbose, psicose and sucrose. But unfortunately it's quite hard to get good results.

Anybody out there with practical knowledge?

Thanks!

Aylin

What appear to be the main problems with the analysis?

I've conducted analysis of sugars successfully in the past using NH2 based columns under reverse phase conditions with a MeCN:Water mobile phase (75:25) and refractive index detection. This was for the separation of sugars in foods and beverages.

First of all, any analysis that involves 75% ACN is using HILIC conditions, not reversed-phase. Second, different types of HILIC columns differ substantially in their selectivity. A "zwitterionic" coating will behave differently than will a neutral coating (diol; amide; etc.) or an amino (NH2) coating. I recommend you look up papers on the use of HILIC to separate the particular sugars you're interested in and then use the column that they used.

Does Merck-SeQuant claim that the column you have can separate the particular sugars you've listed?

First of all, any analysis that involves 75% ACN is using HILIC conditions, not reversed-phase

I stand corrected