Hello everyone,
I am wondering if anyone had previous succesful experiences with specific LC columns and chromatographic methods to separate sugars and if possible, other polar metabolites too. We are specially interested in Glucolisis intermediates.
I tried HILIC in the past but with not much success. Too much chromatographic background. Those columns are painful... ;)
Cheers!!
Albert.
You may try carbohydrate analysis columns, but their resolution and robustness may not be as good as what you get from your typical RP columns.
Check with these guys. You may find something useful.
https://www.shimadzu.com/an/hplc/support/lib/lctalk/49/49intro.html
Hi Albert,
you can use Phenyl isocyanate (PIC) for derivatization of carbohydrates. A very sensitive method is desribed here:
Article Determination of sorbitol and galactitol at the nanogram lev...
Regards
Markus
There are many 'carbohydrate analysis columns'. You can check the Restek, Phenomenex, Agilent, and Waters websites. However, you can use Biorad's Aminex columns which use both and organic and water mobile phase.
Dear Albert Rivas-Ubach
I agree, HILIC sometimes can be a pain in the ... neck.
In my team we had best experiences with a method described here:
Article Analysis of carbohydrates by anion exchange chromatography a...
It works like a charm, and since 2005 a lot of things changed in carbohydrate analysis with HPAEC-IPAD... in conjunction with MS or MS/MS.
Biggest benefit: Stable column chemistry, different and matched selectivities (sugar alcohols, or mono- and disaccharides, oligosaccharides, and polysaccharides),being available for a long time. Lot's of applications, and stable not only in the hands of "specialists"...
Please have a look and let us know, if this helps.
Cheers,
Detlef.
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