In recent years, people have been beginning to pay attention to the issue of carbon emissions due to global warming. To reduce the dependence on petrochemical resources, thus, in this study, biomass polyol trimethylene ether glycol was used to synthesize waterborne polyurethane with different NCO/OH molar ratios.According to the results of particle size analysis, the particle size decreases with the increase of the content of hydrophilic groups. It will affect the viscosity and stability of the dispersion. When the molar ratios increase, the tensile strength increases. The thermogravimetric analysis found that materials with high soft segments have heat resistance. In the end, the results show that a molar ratio of 1.2 is the best tensile strength and heat resistance.