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The successful determination of vitamin B12 (VB12) was achieved by developing an electrochemical sensor that was prepared by the electrophoretic deposition of Au–SnO2 nanoparticles on ITO substrate. Sol gel method was used to synthesize SnO2 and Au–SnO2 nanoparticles. The incorporation of Au with SnO2 was verified by various characterization studies that include XRD, FESEM, EDX, TEM, UV–Vis, FTIR, Raman and XPS. The comparison of the electrochemical performance of Au–SnO2/ITO electrode was done with SnO2/ITO and ITO by cyclic voltammetry (CV), differential pulse voltammetry (DPV) cyclic and electrochemical impedance spectroscopy (EIS). DPV technique was used to show the electrocatalytic behavior of Au–SnO2/ITO electrode towards VB12. Excellent performance was exhibited by the fabricated sensor in a linear range from 0 to 1500 pM with 71.48 pM and 216.6 pM as the LOD (limit of detection) and LOQ (limit of quantification) values. The synthesized sensor exhibited good storage stability, specificity and reproducibility with RSD 1.73%. Real sample analysis was also done with the fabricated electrode. A graphical user interface has also been designed and proposed for easy handling of data in the graphical form. The results obtained in the form of data sheets can be constructed and compared with ease on this designed user interface platform.