In this study, a new selective and sensitive voltammetric procedure for determination of acyclovir (ACV) was proposed using a disposable electrode, pencil graphite electrode (PGE). Cyclic and differential pulse voltammograms of ACV were recorded in Britton–Robinson buffer solution containing 0.10M KCl with pH of 4.0 at PGE. The PGE displayed a very good electrochemical behavior with significant enhancement of the peak current compared to a glassy carbon electrode (GCE). Under experimental conditions, the PGE had a linear response range from 1.0μM to 100.0μM ACV with a detection limit of 0.3μM (based on 3 Sb). Relative standard deviations of 4.8 and 3.6% were obtained for five successive determinations of 10.0 and 50.0μM ACV, respectively, which indicate acceptable repeatability. This voltammetric method was successfully applied to the direct determination of ACV in real pharmaceutical samples. The effect of various interfering compounds on the ACV peak current was studied.