A comprehensive understanding of tire and road wear particles (TRWPs) and their detection and quantification in soils is still challenged by the lack of well-set standardized methods, inherent technological inconsistencies, and generalized protocols. Our protocol includes soil sampling, size separation, and organic matter removal by using hydrogen peroxide followed by density separation and analysis. In this context, roadside soil samples from different sites in Kansas and Ohio, USA, were collected and analyzed. Tire cryogrinds analogous to TRWPs were used to evaluate various density separation media, and collected particles more than 1 mm in size were then subjected to infrared spectroscopy (IR), thermogravimetric analysis (TGA), and scanning electron microscopy coupled with energy dispersive X-ray spectroscopy (SEM-EDX) to confirm TRWP presence. Particles smaller than 1 mm were Soxhlet extracted, followed by gas chromatography-mass spectrometry (GC-MS) to validate the presence of tire-related intermediates. SEM-EDX validated the presence of elemental combinations (S + Zn/Na) ñ (Al, Ca, Mg, K, Si) attributed to tires. Ketones, carboxylic acids, epoxies, cyclohexane, and benzothiazole sulfenamide (BTS) intermediates were the most probable tire-related intermediates observed in the roadside soil samples. Thus, this simple, widely applicable, cost-effective sample preparation protocol for TRWP analysis can assist TRWP research advancement in terrestrial environments.