A speciation study of Pb and Mn in roadside dust along major roads in Ile-Ife, South Western Nigeria, was investigated. Pb and Mn values obtained by total digestion ranged from 22.23 ± 3.52 to 43.48 ± 3.05 μg/g and 35.93±0.15 to 83.76 ± 0.06 μg/g, respectively. The results of speciation analysis of Pb and Mn in the samples showed that the mean levels of these metals in the various fractions followed the order: organic matter>residual>Fe-Mn>carbonate>exchangeable and organic matter-bound>exchangeable>carbonate-bound>residual>Fe-Mn oxide-bound respectively. The speciation study therefore revealed that most of the Pb and Mn were associated with the organic matter fraction and that they were least available in the exchangeable and Fe-Mn oxide fractions, respectively. The apparent mobility and potential bioavailability for these metals in the road dust was Mn>Pb. There is a significant difference between the means of Pb and Mn in the road dust of the study area at p≤0.05, which strongly suggests that they may not have come from the same source; different sources may be responsible, which may be anthropogenic, such as tyre wear, vehicular emission, brake linings and natural.