A gold electrode modified with a polypyrrole⁻mesoporous silica molecular sieves (polypyrrole-MCM-48) nanostructure film was used for the electrochemical determination of small concentrations of dopamine (DA) by cyclic voltammetry and square-wave voltammetry techniques. This electrode showed good electrocatalytic activity for the oxidation of dopamine. The oxidation potential of dopamine was decreased significantly compared with that obtained at the bare gold electrode. The observed linear range for the determination of the dopamine concentration, without interferents through cyclic voltammetry measurements, was from 10 μM to 1.2 mM (R² = 0.9989) for the gold electrode modified with the polypyrrole-MCM-48 nanostructure, with a detection limit of 2.5 μM. In the case of square-wave voltammetry, the linear range was 2⁻250 μM, with a correlation coefficient of 0.9996, and the detection limit was estimated to be 0.7 μM. The effects of interferents, such as ascorbic acid (AA) and uric acid (UA), on the electrochemical detection of dopamine were also examined. The modified electrode can successfully separate the oxidation potentials for ascorbic acid and dopamine, shifting the oxidation peak potential of ascorbic acid to a more positive potential, and significantly decreasing the peak current. The presence of ascorbic acid increased the sensitivity of dopamine determination at the modified electrode, and the detection limit was estimated to be 0.5 μM with 0.1 mM ascorbic acid to imitate physiological solutions. Additionally, studies showed that the presence of uric acid does not affect the electrochemical detection of dopamine. The modified electrode can be successfully applied for the quantitative analysis of dopamine both with and without interferents.