A simple, rapid and versatile method for the separation and determination of sugars via high performance liquid chromatography is described. Virtually any combination of monosaccharides, disaccharides and/or higher oligomers can be separated. Detection via differential refractometry simplifies quantitative measurements and permits collection of emerging sugars for identifying tests. As little as 20 μg of an individual sugar can be determined and preliminary data indicate excellent precision and accuracy. Aqueous solutions of sugars, including samples containing substantial salts or other impurities, can be analyzed directly without derivatization or pretreatment.