Glucose and Fructose separation and detection

[patomb]

Hi folks!

I've to develop a protocol to separate sugars by LC and detected by MS.
To this, I'm using an Agilent LC 1200 coupled to a Q-TOF 6520 mass spectrometer.
For separation the column used was a Zorbax HILIC Plus (2.1x100mm, 3.5 um) for small polar compounds
and LC conditions were:

Solvent A: water + 0.1% Formic acid.
Solvent B: Acetronitrile + 0.1% Formic acid.
Flux: 0.6 ml/min
Injection: 5ul (50ppm each sugar)
Column temp: 40`C

Proportions tested:
80,82,85,87,90,92,95 and 98% of B separated.

The best condition seems to be 87%B but glucose and fructose peaks were overlapped in all test.

I changed the formic acid porcentage to 10mM and made a test without it, but the results still been similar.

Can you recommend me some alternatives?

I'm thinking to change buffer, amonium formate or amonium acetate.
I will be very grateful with your help.

Regards!

[yangz00g]

HILIC is not a good choice if you want base-line separation of sugars.

You may want to try amine or other sugar-specified columns. There are lot of them out there.

[patomb]

HILIC is not a good choice if you want base-line separation of sugars.

You may want to try amine or other sugar-specified columns. There are lot of them out there.

Thanks for replay.

You right, I´m purchasing a Zorbax CH column to improve this, but my purpose to use HILIC are related with my samples, I´m trying to detect several small polar compounds (inner cell metabolites), so I like to have few protocols as possible, but I´m totally sure that in some cases I´ll need different ones to achieve my objectives.
Some people suggest HILIC for these approaches (metabolomics).

Thanks!