Complex Carbohydrate Analysis Columns
Complex Carbohydrate Analysis Columns
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PolyGLYCOPLEX™, PolyWAX HipH™, & PolySAX™ HipH for Carbohydrate HPLC Using HILIC Conditions
PolyGLYCOPLEX columns permit the separation of complex carbohydrates in volatile mobile phases; typically, acetonitrile and water, and due to their higher polarity surface, they require less acetonitrile than the PolyHYDROXYETHYL Aspartamide HILIC columns. Direct flow to Mass Spectrometry is convenient. Compared to reversed-phase HPLC, selectivity is generally superior. Selectivity is quite good for both native glycans and derivatives such as those with the 2-amino pyridine (PA-) fluorophore. The complete library of complex carbohydrates from a particular glycoprotein can often be resolved with isocratic elution, although gradients are recommended for especially varied samples.
Some advantages over basic HPLC materials (e.g., amino-silica) are:
1. Sialylated and asialo-glycans can be resolved using the same running conditions.
2. It is not necessary to operate at pH 7.3 or above with PA-derivatives or other reductively-aminated derivatives of carbohydrates.
For a discussion of the application of this chemistry, see Publications or Mfg. Instruction Sheet
The PolyWAX HipH and PolySAX HipH columns also separate small glycans and complex carbohydrates in volatile mobile phases, but without the separation of the individual anomers, due to their ability to operate at pH 9, which also confers the advantage of operating in negative mode MS for detecting sulfated saccharides.
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Last Updated: 08/20/21