Chromatography Forum

Hi
I am pretty new to HILIC techniques and I was trying to figure out if I can device some useful method for my compounds (anionic sugars and glycoconjugates mostly).

From googling around I have found that amide and silica columns could be suitable for my purposes. I was wandering if there is any difference between HILIC silica columns and regular bare silica columns in terms of packing material. I have found in my lab a brand new Alltech absorbosphere XL Silica, and before buying any new column I was wandering if I can just convert this in my HILIC column or it will bleed to death in my mobile phase...

Any help will be appreciated...
Thanks
Serena

You can use is as initial screening tool. Run a wide gradient first using ACN/water/Ammonium acetate pH 4.5-5 for example. keep buffer constant and go from 855 ACN to 405 ACN in 10 minutes with 5 minutes hold. If retention is too long or too short change initial and end point of gradient.

Just make sure to use a suitable transition solvent first, as your "normal-phase silica" will be shipped in hexane, heptane or some other water-insoluble solvent, whereas a "HILIC-silica" is already "water-compatible".

Thank you. I was thinking of just flushing extensively with Acetone (it's in Hexane/Ethanol 95/5), then AcCN and then introduce water or buffer gradually. Do you think it's OK?

we usually use IPA to go from NP to HILIC.

Ethanol, acetone, and 2-propanol are all good solvents, in my experience, to switch between non-polar and HILIC normal phase modes.