Abstract: In this article, reduced graphene oxide-carbon dots (RGO-CDs) complex were deposited on the surface of glassy carbon electrode (GCE). RGO-CDs and rutoside were facilely embedded into poly(pyrrole) through electrochemical polymerization. After elution of rutoside, a novel sensing system of molecularly imprinted polymers (MIPs) was developed and used for rutoside detection. The use of RGO-CDs markedly improved the electrochemical signal responses on electroactive rutoside, which hence facilitated sensitive electrochemical sensing of rutoside. The as-synthesized MIPs/RGO-CDs/GCE sensing system was characterized by scanning electron microscope (SEM), cyclic voltammetry (CV) and differential pulse voltammery (DPV). Electrochemical responses of the sensing system were studied and optimized. Under optimal conditions, MIPs/RGO-CDs/GCE had a nearly linear relationship between the change of peak current intensities and the rutoside concentration in the range of 0.01-6.5 μM (R2 = 0.9932), with a low limit of detection of 3 nM. Moreover, the sensing system exhibited highly selective and sensitive electrochemical response on rutoside, over potential interferences. In real human serum sample, the sensing system possessed superior performance for rutoside detection, accompanied by high detection stability and recovery. These results proved the significant potential of MIPs/RGO-CDs/GCE for efficient determination of rutoside in real samples