LC/MS column to separate glucose-1 phosphate and glucose-6 phosphate

Record: 706

Category: Specifications
Product: NH2P
Sample:

Question
I am interested in a column that can separate glucose-1 phosphate and glucose-6 phosphate and through detection of LC-MS.

I know that Shodex Asahipak NH2P-50 4E (4.6mmID*250mm) has been capable of this separation, however, not under a LC-MS setting. I am wondering whether you can recommend a column with same capability and can be used in LC-MS setting.

Answer
Unfortunately, phosphoric acid is essential for this analysis and cannot be replaced.

What we could suggest instead is our multi-mode column called JJ-50 2D.
http://shodex.net/index.php?seitenid=1&reiter=r_prod&prodid=30

JJ-50 works under mixture of reversed-phase and anion exchange modes.
Although we have not yet developed an optimized condition for the separation of glucose-1 and 6 phsphate, the condition like 20 mM ammonium formate (pH3.8) + ACN has a high potential of separating those. Most likely it will require optimization of pH and ACN concentration.

 
 
Additional Comments
NH2P series are modified with polyamine functional group.
Under aqueous condition, the amino groups and phosphates make strong ionic bonds.
Phospholic acid in mobile phase (which works as a counter ion and compete with phosphates) will release this bonding.
In other words, with other acids, phosphates will be retained in the column and will not elute.

JJ-50 is modified with trace amount of quaternary ammonium group – just like regular anion exchange column, but also provides reversed-phase mode.
The weaker ion exchange capacity (sorry, we don’t have a number to show you) does not retain phosphates as strong as NH2P series column.
We have seen that ammonium formate/ACN mobile phase releases the binding.
I’m afraid, but we do not have a technical notebook for JJ-50 column.
The pH range for JJ-50 is 2-10, so I do not see any problem using higher pH as long as it separates two compounds.

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