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).
Polymeric strong-base anion exchange packing with limited porosity for separation of proteins
Sample recovery is excellent with PRP-X500 columns. The limited permeability prevents proteins from entering the pores and unfolding (a cause of ghosting).
Controlling Sample Retention
Applications#159 and #161 illustrate gradient separations on a PRP-X500 column. Mobile phases were buffered with TRIS, tris(hydroxymethyl)aminomethane at pH 8 or 9, and the proteins were eluted with a sodium chloride gradient. Other mobile phases such as phosphate, borate and lithium or potassium halides are also suitable for use on PRP-X500 columns. When additional resolution is needed, the gradient rate and mobile phase pH can be altered. When the isoelectric point (pI) of the proteins differ significantly, a mobile phase pH adjustment is very effective. Proteins will elute very early when the pH of the mobile phase is lower than its pI and later when the pH of the mobile phase is higher. A reduction in the gradient rate and range also improves chromatographic resolution.