Two types of rapeseed oil-based polyols in the content of 20 wt% and 50 wt% were used for synthesis of ureaurethane elastomers (EPUU) with isocyanate index 1.07. The bio-polyols were synthesized by epoxidation and opening of oxirane rings using isopropanol (RO) and diethylene glycol (MK). The aim of this work was to evaluate the impact of bio-polyols on the thermal and mechanical properties of the manufactured EPUUs, produced with poly(ethylene adipate) (PEA). Differences in the structure of used bio-polyols caused changes in the number of hydrogen bonds and degree of phase separation in EPUUs. As the content of bio-polyols increases, the number of hydrogen bonds connecting the rigid segments of these materials is reduced. The consequence of these changes is variation in the thermal characteristics of these materials. Cross-linking density calculated from dynamical mechanical thermal analysis increases for materials with 20% of both types of rapeseed oil-based polyols and is the highest for material 80PEA/20MK. An introduction of the MK polyol also resulted in an increase in the elongation at break of the modified elastomers.