AbstractThe electrochemical reduction of lactofen (LCT) at the glassy carbon (GCE) and silver amalgam film electrode (AMFE) is investigatedin detail by the means of square wave voltammetry (SWV), square wave stripping voltammetry (SWSV) and cyclic voltammetry. The influence of various factors such as supporting electrolyte composition and SW parameters were studied. The AMFE electrode showed an excellent electrochemical activity toward the electro‐reduction of LCT, leading to a significant improvement in sensitivity as compared to the glassy carbon electrode.The SWSV detection limits for GCE and AMFE were 285.0 nM and 2.0 nM, respectively. The applicability of the developed voltammetric method for analysis of tap water and river water is illustrated with spiked samples analysis. Moreover, as lactofen is highly toxic to fish and other aquatic organisms, its interaction with dsDNA isolated from salmon sperm was tested. The intercalative mode of LCT binding to dsDNA was estimated. The heterogeneous rate constants were calculated for the free LCT and the LCT‐dsDNA complex. Moreover, LCT‐dsDNA complex binding ratio and equilibrium constant were determined. The decrease in the SWV peak current of LCT in the presence of dsDNA was used for the determination of dsDNA.