Non-standard diesel blends can be harmful to the environment and human health. In this context, a simple analytical method to estimate the biodiesel mixture ratio in diesel was developed based on the impedance spectroscopy (IS) associated with the interdigitated sensors. In this article, four different interdigitated sensors, variable comb spacing (G), were simulated using the COMSOL Multiphysics software. Based on finite element simulations, four interdigitated electrode architectures by manufactured and evaluated. According to the X-ray powder diffraction technique, the deposition of the conductive layer (Au°) over the surface of the dielectric substrate (SiO2) did not alter its phase composition. In the analysis of AFM and SEM, it was possible to observe irregular edges on the electrodes, possibly related to thin layers' manufacturing process and mechanical stability. Another characteristic observed in the AFM images was the height of the step of the gold layer of the sensor. Several cross-sections were obtained, and the mean step value is 225.71 ± 0.0032 nm. Although there are differences in the roughness, the whole sensor has nanometric roughness. Based on the finite element method simulation performed, it can be assumed that the geometric parameters more suitable for the manufacturing of the electrode are: W = 20 µm, L = 1000 µm, G = 50 µm e N = 40 digits. The electrical characterization performed by impedance spectroscopy showed that we could differentiate between biodiesel and diesel fuels and their pseudo-binary mixtures in the low-frequency region.