Roadway particles (RP) that can be collected with on-vehicle system, consist of a mixture of Tire and road wear particles (TRWP) with other traffic-derived particles (exhaust or nonexhaust) and/or biogenic compounds and represent a significant source of xenobiotics, susceptible to reach the different environmental compartments. The study of the RP fate is thus a major challenge to tackle in order to understand their degradation and impact. They offer a variety of carbon sources potentially usable by microorganisms, ranging from the tire-derived plasticizers, vulcanizing agents, protective agents and their transformation products, to other traffic, road and environmental-derived contaminants. A multi-analytical approach was implemented to characterize RP and study their biodegradation. Kinetics of RP extractions were monitored during 21 days in water, methanol, acetone and chloroform to identify leaching and extractable compounds and monitor the particle composition. The results confirmed that hundreds of readily leachable chemicals can be extracted from RP directly into water according 2 to a dynamic process with time while additional poorly soluble compounds remain in the particles. Mass spectrometry (LC-HRMS and GC-MS) allowed us to propose 296 putative compounds using an extensive rubber database. The capacity of 6 bacterial strains, belonging to Rhodococcus, Pseudomonas and Streptomyces genera, to biodegrade RP was then evaluated over 14 days of incubation. The selected strains were able to grow on RP using various substrates. Elastomer monitoring by 1 H NMR revealed a significant 12% decrease of the extractable SBR fraction when the particles were incubated with Rhodococcus ruber. After incubation, the biodegradation of 171 compounds among leachable and extractable compounds was evaluated. Fatty acids and alkanes from rubber plasticizers and paraffin waxes were the most degraded putative compounds by the six strains tested, reaching 75% of biodegradation for some of them.