Electrochemical properties of disulfiram, representative of highly significant bioactive compounds, were studied with a cyclic renewable silver amalgam film electrode (Hg(Ag)FE) using square wave cathodic stripping voltammetry (SWCSV). The influence of various factors such as pH, buffer concentration, buffer composition, and SWCSV parameters on current response was investigated. The optimum results in terms of signal shape and intensity were recorded in Britton Robinson buffer (pH 7.5) at 0.5 V versus Ag/AgCl/3 mol L-1 KCl. An elaborated electroanalytical procedure was used to determine disulfiram at the Hg(Ag)FE in the concentration range from 0.05 to 5.00 mu M. Precision, repeatability, and accuracy of the method as well as the influence of possible interferences were ascertained. The detection and quantification limits were 11 nM and 37 nM, respectively. The applicability of the developed method was tested in the determination of disulfiram in the commercial formulation Anticol. Thin-layer chromatography with image processing software was used to validate the accuracy of the method.