In this paper, an electrochemical sensor for theophylline (ThPh) was prepared by electropolymerizing o-phenylenediamine (o-PD) on a glassy carbon electrode in the presence of ThPh via cyclic voltammetry, followed by deposition of gold nanoparticles to enhance the sensitivity of the sensor. Therefore, the sensor showed a high sensitivity for ThPh determining. Peak current of Fe(CN)6]3?/Fe(CN)6]4? varied linearly with the concentration of ThPh in the range of 4.0×10-7~1.5×10-5 mol·L-1 and 2.4×10-4~3.4×10-3 mol·L-1, and the detection limit reached 1.0×10-7 mol·L-1. Compared to structurally related molecules, the sensor also has a high recognition capability for ThPh. With excellent regeneration property and stability, the present sensor maybe provides a new class of polymer modified electrodes for sensor applications.