In this research, graphene quantum dots (GQDs) was synthesized via the facile pyrolysis method. The properties of the prepared GQDs were examined by high-resolution transmission electron microscopy (HRTEM), scanning electron microscopy (SEM), X-ray diffraction (XRD), dynamic light scattering (DLS), photoluminescence (PL), Fourier transform infrared spectroscopy (FT-IR), and ultraviolet–visible (UV–Vis) methods. Afterward, the graphite/chitosan/graphene quantum dots (Gr/CS/GQDs) nanocomposite was prepared and used as a cathode electrode in electro-Fenton (EF) process to study the tetracycline (TCL) removal from the aqueous solution. The effect of operational parameters in EF, including current density (mA/cm2), initial pH, initial TCL concentration (mg/L), and process time (min) on the TCL removal efficiency, was investigated via response surface methodology (RSM). The results showed that the optimal values for current density, pH, initial TCL concentration, and process time were 60 mA/cm2, 5, 30 mg/L, and 150 min, respectively. Maximum TCL removal efficiency was 92.98%. FT-IR, XRD, Raman spectroscopy, atomic force microscopy(AFM), SEM, mapping, energy dispersive X-Ray (EDX), Brunauer Emmett Teller (BET), and cyclic voltammetry (CV) analyses were performed to investigate the Gr/CS/GQDs cathode. Finally, gas chromatography–mass spectrometry (GC-MS), total organic carbon(TOC), and atomic absorption spectroscopy (AAS) analyses were performed and the reusability of the Gr/CS/GQDs cathode was investigated.