A mixture design was used to optimize the mobile phase composition for separation of carbohydrates in honey by High Performance Liquid Chromatography. Nine ternary acetronitrile, water and ethyl acetate mobile phases corresponding to a simplex centroid design with axial points were test to separate those carbohydrates most frequently encountered in honey samples. The results suggest that a special cubic model accurately describes changes in the proportions of these solvents in the mobile phase close to the region of optimal peak separation. The addition of a third solvent, ethyl acetate, to binary mobile phase of water and acetonitrile permitted a significant improvement in carbohydrate separation that can be applied to the quantitative analysis of these compounds in honey.
mobile phase optimization; HPLC; carbohydrates; mixture design; honey
