Chromatography Forum

Hi All,

I am trying the separations of the following sugars on a Biorad HPX-87H column using an RI detector. (flow rate- 0.6ml/min & mobile phase - 0.005M H2SO4).However, apart from glucose, all the sugars co-elute on this column. Could anyone suggest any other column for these separations? Would lead based columns help separate these sugars?
The sugars are- glucose, galactose, mannose, fructose & xylose

Thanks

Bio-Rad's literature for the PB form column mentions glucose, galactose, mannose, & fructose. I'm not sure about xylose, but you might contact them to confirm.

Aminex HPX-87P gives the best monosaccharide separations

When the system is setup correct with short capillaries, exact temperature and the column is well equilibrated the chromatogram should look like in the application book of the Aminex columns. Reference flow cell is well rinsed before injecting the sample? What about sample preparation?

Aminex HPX-87P gives the best monosaccharide separations

Hi Karen01,

Thanks for your response. Have you used the Pb column for any separations?

When the system is setup correct with short capillaries, exact temperature and the column is well equilibrated the chromatogram should look like in the application book of the Aminex columns. Reference flow cell is well rinsed before injecting the sample? What about sample preparation?

Hi Gerhard Kratz,

Thanks for getting back on this. About the Aminex columns, I am using them according to the conditions prescribed by Bio-rad. Yes, the flow cell is well rinsed before injections. The sample is prepared in water then subjected to acid hydrolysis with the aid of the catalyst I synthesize

Aminex HPX-87P gives the best monosaccharide separations

Hi Karen01,

Thanks for your response. Have you used the Pb column for any separations?

Every day!

Overall it separates more monoscaraides than any other column we tried including the Aminex H and C columns, or the Water's UPLC amide column.

Every day!

Overall it separates more monoscaraides than any other column we tried including the Aminex H and C columns, or the Water's UPLC amide column.[/quote]


I am not sure of the type of samples you use, but my samples are acidic even before injecting them on the column (pH 2-4) . Can the column handle these acidic samples or do you have any specific guards in conjunction with the column?

Also, like I mentioned earlier I am looking for a column that can specifically separate xylose, mannose, fructose and galactose. The Aminex H column is of no use at it.

annbal,
I would also check out the website of Showa Denko. They are manufacturing some special phases for sugars and acids. Check the application data base. Out of my experience if you find an application it will work the same way also with your samples. Otherwise ask their excellent technical support.
Other manufacturers have also a lot of applications published using different column chemistries like NH3 columns.
A HILIC column can also do the job. My recommendation is to ask different manufacturers and also please ask them to get the column free of charge to test. If they are confident that the recommended column will work they will do so. For sure if the column works you should be ready to pay for the column, that would be fair.

Gerhard,
Thanks for your response. I did inquire with Shodex (Showa Denko) and they recommended an amino column. I shall inquire with them if they can provide any free column for testing before purchase.

Annbal