In the present study,completely hydrolyzed sugarcane juice containing total sugar content of 140 g /L was adjusted with an equivalent amount of fructose and glucose to give a total sugar concentration of 220 g / L. Then the juices were supplemented with nitrogen sources at various stages,and used as ethanol production medium. The fructose and glucose concentrations were determined by HPLC( high performance liquid chromatography). Area under the curve( AUC) of fermentation of fructose and glucose was obtained using GraphPad Prism 5 software to analyze the discrepancy in fructose and glucose consumption. Results indicated that S. cerevisiae GJ2008 always showed a slightly higher preference for glucose than fructose,and fructose consumption always lagged behind glucose consumption. Nitrogen addition could reduce the discrepancy in glucose and fructose utilization effectively when it was added in the early stages of the fermentation.