Billions of road miles are driven every year by vehicles which generate millions of tyre wear particles (TWPs) due to the frictional interaction between the tyres and the road surfaces. The behaviour and impact of TWPs in the environment remains poorly understood due to the challenge in identifying them amongst other particulates in the samples. The aim of this research was to develop an analytical approach for the analysis and characterisation of TWPs in road dust samples given that road dust would be the primary deposition matrix. A cryomill method was developed to produce cryomilled tyre particles (CTPs) as an in-house reference material, which facilitated the development of techniques for comprehensive characterisation of size, surface area, pH at point of zero charge (pHpzc) and chemical composition. In the latter, thermogravimetric analysis (TGA) provided critical insights into the degradation of tyre particles which prompted the development of a novel technique, pyroTGA-GC-MS, to understand the evolved gases released during the thermal decomposition of tyre particles. The evolved gases would then serve as markers for TWPs in road dust and the technique revealed the prevalence of TWPs in road dusts collected from central London and two highway roads. Elemental analysis, using microwave plasma-atomic emission spectroscopy (MP-AES) for the first time, showed limited reliability for detecting TWPs in the environment because of the inherent natural abundance of Al, Fe and Mg in the environment. Zinc being abundant in CTPs was believed to be a suitable elemental marker for TWPs in the road dusts but lacked specificity without sample pretreatment, as it is also used in road signs and vehicle components. Hence novel oil extraction protocol (OEP) was developed to enhance the detection of TWPs in the environment which was proven via application of the pyroTGA-GC-MS on selected road dust samples before and after the OEP. Future studies should consider combining the techniques discussed in this thesis for a comprehensive understanding of TWPs from the point of deposition on the roads to their distribution into soils and receiving water bodies.