The Glucose detection in human blood and urine is very vital and the unusual level of it in the body shows that the corporal biological system has been hampered. Therefore, in this study, the new nanocomposite with reduced Graphene oxide-polydopamin-Copper(I) oxide (RGO-PDA-Cu 2 O) structure was designed and deposited on the glassy carbon electrode (GCE) surface for this subject. The UV-Visible, fourier transform infrared (FT-IR), X-ray diffraction (XRD), Energy-dispersive X-ray (EDX), transmission electron microscopy (TEM) and field emission scanning electron microscopy (FESEM) techniques were applied to the characterization of as-fabricated nanocomposite electrode. The electroanalysis of the new electrode were investigate by the cyclic voltammetry (CV), differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS) methods. The detection limit of glucose (Glu) was also obtained and showed that the deposited GCE by RGO-PDA-Cu 2 O nanocomposite has a high potential for Glu detection. In addition, this electrode was applied to the Glu detection as biosensor in real samples in order to utilizing in commercial applications.