This paper describes a stripping method for the determination of acyclovir at the submicromolar concentration level. This method is based on controlled adsorptive accumulation of acyclovir at thin-film mercury electrode, followed by a linear cyclic scan voltammetry measurement of the surface species. Optimal experimental conditions include a NaOH solution of 2.0 × 10(-3) mol L(-1) (supporting electrolyte), an accumulation potential of -0.40 V, and a scan rate of 100 mV s(-1). The response of acyclovir is linear over the concentration range 0.02 to 0.12 ppm. For an accumulation time of 4 minutes, the detection limit was found to be 0.42 ppb (1.0 × 10(-9) mol L(-1)). More convenient methods to measure the acyclovir in presence of the didanosine, efavirenz, nevirapine, nelfinavir, lamivudine, and zidovudine were also investigated. The utility of this method is demonstrated by the presence of acyclovir together with Adenosine triphosphate (ATP) or DNA.