In this paper electropolymerization of a thin film of para-phenylenediamine (PPD) is studied at glassy carbon electrode (GCE) in sulfuric acid media by cyclic voltammetry. The results showed that this polymer was conducting and had a reproducible redox couple in the potential region from 0.0 to 0.4 V in phosphate buffer solution. This modified GCE (p-PPD-GCE) was applied for simultaneous determination of ascorbic acid (AA), dopamine (DA) and uric acid (UA) using differential pulse voltammetry (DPV). The p-PPD-GCE in 0.1 M phosphate buffer solution (pH 5.0) separated the DPV signals of AA, DA and UA with sufficient potential differences between AA-DA and DA-UA and also enhanced their oxidation peak currents. The oxidation currents were increased from 2.0 to 2000.0 mM for AA, 10.0 to 1250.0 mM for DA and 50.0 to 1600.0 mM for UA. The detection limits were evaluated as 0.4, 1.0 and 2.5 mM for AA, DA and UA, respectively (S/N = 3).