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The electrodeposition of a film of reduced graphene oxide (ERGO) on a glassy carbon electrode (GCE) was achieved using pulse potential method (PPM) in the graphene oxide colloidal solution. For the first time, the electrodeposition of a film was applied to develop a high-sensitive electrochemical sensor for determination of matrine using linear sweep adsorptive stripping voltammetry (LSASV). Compared with bare GCE and ERGO film prepared by potentiostatic method (PM) modified electrode, the resulting electrodes (PP-ERGO/GCE) exhibited excellent response toward the oxidation of matrine by significantly enhancing the oxidation peak currents and decreasing the overpotential of matrine. Under the selected conditions, there exist the linear relation between the oxidation peak currents and matrine concentration in the range of 2.0?×?10^?6–1.2?×?10^?4?mol?L^?1, with detection limit of 5.0?×?10^?7?mol?L^?1. At the same time, the method can be successfully applied to the quantitative determination of matrine in injection and its result is satisfactory.