This work describes an electroanalytical investigation of dopamine using cyclic voltammetry (CV) and the graphite–polyurethane composite electrode (GPU). In CV studies, well-defined redox peaks characterize the oxidation process at the GPU electrode, which is indicative of electrocatalytic effects associated with active sites on the GPU electrode surface. A new analytical methodology was developed using the GPU electrode and square wave voltammetry (SWV) in BR buffer solution (0.1 mol L−1; pH 7.4). Analytical curves were constructed under optimized conditions (f=60s−1, ΔE a=50 mV, ΔE I=2 mV) and detection and quantification limits of 6.4×10−8 mol L−1 (12.1 μg L−1) and 5.2×10−6 mol L−1 (0.9 mg L−1), respectively, were achieved. The precision of the method was checked by performing ten successive measurements for a 9.9×10−6 mol L−1 dopamine solution. For intra-assay and inter-assay precisions, the relative standard deviations were 1.9 and 2.3%, respectively. In order to evaluate the developed methodology, the determination of dopamine was performed with good sensitivity and selectivity, without the interference of ascorbic acid in synthetic cerebrospinal fluid, which indicates that the new methodology enables reliable analysis of dopamine.