Rapid HPLC Methods for the Separation and Quantitation of a Mono-, Di-, and Tri-Saccharides Mixture and Applications

Abstract

A simple, rapid method has been developed for the separation and quantitation of mono-, di-, and tri-saccharides. The method utilizes a 30cmx 3.9mm i.d. Microbondapak NH₂ column, refractive index detection and water-acetonitrile elution. Two chromatographic systems are described. The isocratic mode was necessary to develop a procedure. 20 Carbohydrate's retention times were evaluated. To optimize the separation of nine water-soluble sugars, a gradient mode flow-programming was used. Separation was achieved within 28 minutes. The low detection limit (4 micrograms) of the above chromatographic procedure and its different possibilities could be of great interest to the analyst. The method has been successfully applied to quantify the major carbohydrates found in two types of commercial honey.