Many achievements have been made on the research of composite polyurethane foams to improve their structure and mechanical properties, and the composite foams have been widely utilized in building insulation and furniture. In this work, rigid polyurethane foams (RPUFs) with the addition of different fillers (nano-SiO2, peanut shell, pine bark) were prepared through the one-step method. The effects of inorganic nano-SiO2 and organic biomass on foam properties were evaluated by means of physical and chemical characterization. The characterization results indicate that the compressive strength values of prepared foams could fully meet the specification requirement for the building insulation materials. The inorganic and organic fillers have no effect on the hydrogen bonding states in composite RPUFs. Furthermore, compared to the biomass fillers, the addition of nano-SiO2 greatly influenced the final residual content of the fabricated foam. All composite foams exhibit closed-cell structure with smaller cell size in comparison with the parent foam. The prepared composite foams have the potential for utilization in building insulation.