A new method was developed in this work to investigate the toxic Sb(III) and Sb(V) species distribution in soil and grass samples along two major traffic routes (Rankweil A14 and Knittelfeld S36) in Austria, since vehicle emissions are an important anthropogenic source of Sb in soil. The developed speciation analysis method (HPLC-ID-ICP-MS) depends in using isotope dilution techniques to quantify Sb species after a chromatographic separation using an anion exchange column (PRP-X100) and 10 mmol L-1 EDTA plus 1 mmol L-1 phthalic acid at pH 4.5 as a mobile phase. The optimized extraction for Sb species was carried out using 0.1 M citric acid at pH 2.08 applying an ultrasonic bath extraction for 45 min at room temperature. A combination of results of species unspecific and species specific isotope dilution was used to calculate for Sb species interconversion during analysis. Results of Sb and its inorganic species distribution in roadside soil and grass samples confirmed significant enrichment of Sb (exceeding the natural background values more than 10-fold) and its species predominantly Sb(V) near the edge of the traffic lane. Their concentrations decreased sharply to natural background levels within a few meters from the road edge. Magnetic susceptibility data of all soil samples show the same distribution pattern as Sb and Sb(V) concentrations with an excellent correlation. This is an evidence for an anthropogenic source of Sb such as abrasions of braking linings.