In this study a gold electrode was used to indirect determination of melittin in nonaqueous medium by square wave stripping Voltammetric (SWSV) method. The method is based on measuring the decrease of oxidation peak current of ferrocene with addition melittin. The optimum conditions include an accumulation potential of 0 mV (versus Ag/Ag+), an accumulation time of 10 s. A calibration curve was obtained for melittin in a linear range of 40 to 240 nM with a low detection limit CLOD) of 12 nM (at a signal-to-noise ratio of 3). The validated SWSV method showed good linearity as well as satisfactory repeatability and immediate precision values, for both instrument and method. The method was successfully applied to the determination of melittin in honeybee venom sample. (C) 2015 Elsevier B.V. All rights reserved.