The blending ratio of biodiesel in petroleum diesel has become one of the most important parameters to ensure the quality of diesel/biodiesel blend. This paper presents a fast and simple method based on electrochemical impedance spectroscopy to determine the biodiesel content in diesel fuel. Different from the method reported in the literature, using a pair of two identical screen printed carbon paste electrodes, in the present work we used two electrodes made from 304 stainless steel with dimensions of 0.8×0.3 cm for the EIS measurements. Improved results were obtained in terms of sensitivity, stability of measurements, electrode reuse, and cost. In this procedure the charge transfer resistance is proportionally related to the biodiesel content, which is used to construct a calibration curve for the analysis of biodiesel content. The procedure was validated by an official method, using two samples, being one of them, certified through an official interlaboratory program of the Brazilian government (Interlaboratory Program for Biodiesel of National Agency of Petroleum, Natural Gas and Biofuels (PIB)/ANP). Good results were obtained in terms of recovery (102.6 %-102.8 %), precision (coefficient of variation lower than 2.3 %), limit of detection (0.24 %) and limit of quantification (0.80 %). These results indicate that this method is sufficiently suitable as an alternative method to the official method for determining biodiesel content in commercial diesel fuel.