Refractive index of D-Glucose solution in many areas such as scientific research and industry has very important applications. It′s very important to measure the refractive index accurately. The beam splitter prism interferometry is used to measure refractive index of D-Glucose solution at different temperatures with different concentrations. The basic principle of this method and apparatus is introduced. The refractive index of a sample of known calcium chloride solution is measured by the system and contrasted with literature values to verify its reliability. The accuracy is up to 10-4. In order to research the characteristics of D-Glucose solution, diversifications of D-Glucose solution′s refractive index at different temperatures measured and plotted into graphs. Error analysis is conducted as well. The results can provide help for medicine and biology.