Bioethanol is produced by bio-chemical process that converts sugar or biomass feedstock into ethanol. After bio-chemical process, the solution is distilled under controlled conditions of pressure and temperature, in order to obtain an ethanol-water solution. However, the ethanol concentration analysis is generally performed off-line and, sometimes, a re-distillation process becomes necessary. In this research, an optical apparatus based on Fresnel reflection has been used in combination with artificial neural networks for determination of bioethanol concentration in hydro-alcoholic solution at any temperature. The volumetric concentration and temperature effect was investigated. This intelligent system can effectively detect and update in real-time the correction of distillation parameters to reduce losses of bioethanol and also to improve the quality in a production plant.