Road dust (RD) resuspension is one of the main sources of particulate matter in cities with adverse impacts on air quality, health, and climate. Studies on the variability of the deposited PM10 fraction of RD (RD10) have been limited in Latin America, whereby our understanding of the central factors that control this pollutant remains incomplete. In this study, forty-one RD10 samples were collected in two Andean cities (Bogotá and Manizales) and analyzed for ions, minerals, and trace elements. RD10 levels varied between 1.8–45.7 mg/m2, with an average of 11.8 mg/m2, in Bogotá and between 0.8–26.7 mg/m2, with an average of 5.7 mg/m2, in Manizales. Minerals were the most abundant species in both cities, with a fraction significantly larger in Manizales (38%) than Bogotá (9%). The difference could be explained mainly by the complex topography and the composition of soil derived from volcanic ash in Manizales. The volcanic activity was also associated with SO4⁻2 and Clˉ. Enrichment factors and principal component analysis were conducted to explore potential factors associated to sources of RD10. Elements such as Cu, Pb, Cr, Ni, V, Sb, and Mo were mainly associated with exhaust and non-exhaust traffic emissions.