Chromatography Forum

Hi I have been looking into the phenomenex columns. Specially on the synergi columns ( maxRP, Hydro RP, and Fusion-RP) for the purification of this sulfated carbohydrates.

have anyone have any experience on which of those would be a better column or if any other brand has higher quality for these carbohydrates.

you should probably consider hilic, but frankly you have a very difficult analyte! retention on reverse phase columns, even so-called polar reverse phase columns, will be minimal (based on my limited experience of phosphorylated sugars).

ooh, and when I say Hilic, I don't necessarily mean Phenomenex's Hilic column. You can also do Hilic on amino phases, and a range of other column chemistries. Ask your column suppliers for more details...

thank you for your input

it depends what sort of sulphated carbohydrates you are taliking about (chain length?, degree of sulphation, etc). We were able to separate the disaccharide components of chondroitin sulphate fairly easily using reversed phase (Waters, Symmetry C18) and some co-workers did the same using a zorbax column. However you need to use an ion pairing reagent - we used TBAOH.

Also had excellent results with sulphated mono and disaccharides on the Dionex PA1 column. This uses sodium hydroxide as the mobile phase so you need a system designed for the separation. In some cases you may need to reduce the sugars to prevent degradation. You also need a gradient. Typically acetate is used in these applications but sodium trifluoroacetate works better.

The HILIC suggestion previously is also good - there are many publications describing suitable conditions on such columns (amino phase).

oh, yes! I never think of ion-pair reagents, being a general-purpose MS person...