This study was based on the determination of metals in particulate matter emitted by a typical diesel engine used by busses and trucks in Brazil. Emissions were sampled using a cascade impactor, and the engine was operated using diesel with 5% (B5), 10% (B10), 15% (B15), and 20% (B20) of biodiesel. The particulate matter was stratified in different sizes, i.e., 18, 5.6, 3.2, 1.8, 1.0, 0.560, 0.320, 0.180, 0.100, and 0.056 μm. Cd, Co, Cr, Cu, Mn, Mo, Ni, Pb, V, and Zn with concentrations within 10 to 1000 ng m were determined. The results indicate a trend in the prevalence of lead, nickel, and chromium in coarse particles and nanoparticles in all blends of fuels. By comparing the results of B5, B10, B15, and B20 fuels, we can confirm that the addition of biodiesel to diesel promotes a reduction of emissions, and by comparing the behavior of the concentration of all elements analyzed, emissions by B10 and B15 fuels are similar, while B5 and B20 suffer significant changes during the process of combustion. Multivariate statistical analysis was used, and it indicates possible sources in three clusters, one for Ni, other for Cr-Mn, and the last one for other metals.