An electroanalytical method was developed for the direct quantitative determination of Acyclovir (Acy) in spiked human urine base on its oxidation behavior. The electrochemical oxidation and determination of Acy were easily carried out on ultra trace graphite electrode (UTGE) and glassy carbon electrode (GCE) using a variety of voltammetric techniques. The electrochemical measurements were carried out on these electrodes in various buffer solutions in the pH range of 3.66 to 9.08 by cyclic voltammetry (CV) and differential pulse voltammetry (DPV) techniques. The best results for the quantitative determination of Acy were obtained by DPV technique in 0.2 M acetate buffer (pH=4.66). In this acidic medium, one irreversible anodic peak was observed. The anodic peak current and peak potential depend on pH and scan rate were studied. The diffusion controlled nature of the peak was established. Acy was determined in the concentration range from 4x10(-6) to 7x10(-5) molL(-1) for UTGE and 2.0x10(-6) to 1.0x10(-4) molL(-1) for GCE by the applied electroanalytical procedure.