In this work a series of experiments have been performed to study and simulate the separation of glucose and fructose as the most applicable sugars in the food industry. The objective of this work is understanding and modeling the behavior of these components in a chromatography column when they are introduced to the column as simple components, binary solutions, and also as the major species in date syrup. The column's diameter and height are 3.5 cm and 15 cm respectively, and Purolite PCR642Ca, which is an acidic gel-type cation exchange resin (Ca2+), was used for the separation of sugars. Static method was applied to determine the equilibrium constants, and frontal (dynamic) analysis method was used to determine the mass transfer coefficients at two different temperatures of 30 and 60 degrees C for single and multi-component mixtures for a solution with a sugar concentration of 70 g/L The kinetic studies for the prediction of mass transfer coefficients were performed according to the breakthrough curves by a linear driving force approximation (LDF), while the adsorption isotherms were estimated by linear model. Adsorption equilibrium constants and mass transfer coefficients between fructose, glucose and solid resin were determine by optimization procedure. The results of the optimization and parameter estimation for synthetic sugar solutions were also applied for modeling the separation of sugars in date syrup. The predicted results of the real mixtures (date syrup) have a good agreement with the experimental data and show that both sugars can be separated successfully by this method. (C) 2011 Elsevier B.V. All rights reserved.