Chromatography Forum

Good morning,

From what I learnt I understand that in order to analyse sugars and organic acids you should use separate columns. However, right now I understand that there are a new set of columns that can be use for the simultaneous analysis of these analytes (sugars and O. acids) using a RI detector.

I would like to know if anybody has any good suggestion about a nice column and/or methods (solvents and type of elution), which I should consider for setting up this analysis for wine samples.

In addition to this last question, does anyone know about any cool method for aminoacid analysis by HPLC

Thanks in advance for your help and support.

Best regards

Maurole

Good morning,

From what I learnt I understand that in order to analyse sugars and organic acids you should use separate columns. However, right now I understand that there are a new set of columns that can be use for the simultaneous analysis of these analytes (sugars and O. acids) using a RI detector.

Maybe an Amide HILIC column?

- Karen

Do you mean carbohydrate column?

On page 9 and 10, there are some applications on analysis of carbohydrate and Alcohol in Wine: http://www.sepax-tech.com/catalog/Catalog_Carbomix.pdf

One more analysis of organic acid in wine data here: http://www.sepax-tech.com/application_notes/CO1008.pdf FYI.

Maurole -

A really easy way to analyze amino acids is to separate them isocratically on a HILIC column (our PolyHYDROXYETHYL A 100-Å material or some equivalent) using 15 mm ammonium formate in 60% ACN. They elute isocratically in about 4 minutes. You can detect and quantify all 20 amino acids this way using MS/MS. Contact me offlist if you like and I'll send you Bob Croes' poster from ASMS 1999 on the subject. He was using it to measure the amino acids in corn and soybeans. The same method will work for any other small, reasonably polar metabolite as long as there are no other compounds present that have the same mass spec signature as the daughter ion you're trying to measure in MS/MS. An example of that would be the interference of pipecolic acid with lysine, so they have to be separated chromatographically prior to MS.
A newer approach is ion-trap MS, now with a HILIC capillary. Ref.: Li et al., Anal. Bioanal. Chem. online Sept. 29, 2011 (measurement of pyruvate, lactate, glutamine, glutamic acid, etc.).