General, chemical type or category of compound to be separated.
The type of resin that is packed inside the column tube.
United States Pharmacopeia HPLC column designation.
Size (in micrometers) of the polymer or silica bead packed inside the column tube.
Inside diameter (in millimeters) of the column tube.
Length of the column tube (in millimeters).
HC-40 and HC-75 Gel-Type Cation Exchange ColumnsThe HC-40 and HC-75 gel-type cation exchange columns are especially useful for determining ethanol in fermentation broths and for separating sugars and sugar alcohols in food samples like com syrup, chewing gum sweeteners, and fruit juices.
HC-40 and HC-75 columns separate compounds through size exclusion and ligand exchange. The 4% cross-linked HC-40 uses size exclusion as the primary mechanism of separation, while ligand exchange dominates in the more highly cross-linked HC-75.
The higher carbohydrate oligomers elute first; the smaller di- and monosaccharides elute later.
Column Selection and Applications
HC-40 – Separates oligo saccharides up to DP8
The HC-75 column, Application #126, provides a slightly faster (14-minute) separation up to DP 5, while the HC-40 column, Application #91, provides a much better separation of the oligomers up to DP 8 in 16 minutes. The HC-75 Hydrogen form column separates acids and alcohols (Application #209), while the HC-75 Calcium form column separates mono and disaccharides. The HC-75 Lead form column (Application #116) resolves sugar alcohols.
Water is the recommended mobile phase for gel-type columns. If a buffer is used, the counter cation must be the same as that on the column packing material. If not, counter ions on the column packing material will be displaced and replaced with ions present in the buffer, causing shrinkage or swelling of the polymer beads. This shrinkage and swelling disturbs the packing of the resin and destroys the column's performance. See Technical Specifications, including exchange capacity, temperature limits, particle size, mobile phase limits, buffer strength, maximum pressure, and pore size.
Because carbohydrates do not contain a chromophore, UV detection cannot be used without derivatization. The recommended detection method is refractive index. The control of carbohydrate retention lies in the selection of the correct column.
Mobile Phase Preparation
Use degassed and deionized water for mobile phase preparation.
Polymeric cross-linked soft-gel columns for cation, ligand exchange separation of carbohydrates
-Separate Mono and Disaccharides
-9 µm particle size