Display omitted •Electrochemical properties of iron ions investigated by cyclic voltammetry using GCE.•Influence of pH, concentration and scan rate on electrochemical behavior.•Focus on the interaction of FeSO4 and K4Fe(CN)6 with dithizone at 293.15 K.•Assessment of scan rate, stability constants and kinetic parameters of iron-dithizone interaction.•Practical applications in detection of iron ions in biological samples and molecular docking studies. This study aims to investigate the electrochemical characteristics and complex formation of iron ions with dithizone in various solutions including human blood at 293.15 K, using a glassy carbon working electrode. Cyclic voltammetry was employed to analyze the oxidation and reduction processes of iron ions, specifically FeSO4 and K4Fe(CN)6, and determine their stability constants. The study revealed that the electrochemical behavior of FeSO4 and K4Fe(CN)6 are complex and influenced by various factors such as pH and concentration. The (pH 4) acidic medium acts as an electrochemical catalyst, enhancing redox peak at approximately −0.2 V and showing the strongest response with maximum sensitivity. An increase in the iron ion concentration causes an increase in Ip,a/Ip,c, highlighting kinetic and adsorption challenges for analytical examination. The study also determined the scan rate and kinetic parameters of the interaction between iron and dithizone, and calculated the stability constants of the complexes formed. Additionally, the practical applications of this research in detecting iron ions in biological samples and potential molecular docking studies against viral leukemia protein will be considered.