Preparation of a Ligand Exchange Column for Carbohydrate Separation and Glucose Determination in Cell Culture Medium

Abstract

The packing and testing of a ligand exchange column for the separation of carbohydrates was investigated. Resin with a 5 mm particle size and 8% crosslinking provided the best overall column performance. A range of column packing pressures were tested and revealed an optimum of 750 psi to provide good separation and low back pressure. Baseline separation of standard carbohydrates was achieved using an isocratic elution with pure water as the mobile phase and a column temperature of 85 C. The column was applied to the separation of a range of carbohydrates used commonly in the food industry, as well as the specific analysis of glucose in tissue culture medium successfully.