Vegetable oils offer a reliable chemical platform to synthesize polyols for polyurethane production to replace fully or partially polyurethane precursors that are derived from petrochemical resources. Rapeseed, a renewable feedstock widely available in Northern Europe, was used as a bio-based feedstock. In the present study, a cradle-to-gate Life Cycle Assessment for a pilot scale bio-based polyol production was carried out. Life Cycle Inventories were built on up-to-date regionalized inventory for rapeseed production and experimental data for polyol synthesis were obtained from a pilot scale (50 L) reactor. Bio-based polyols were compared with the petrochemical counterpart. Life Cycle Assessment was performed using SimaPro 9.0 and impacts were evaluated with ReCiPe Endpoint and Midpoint methods and Cumulative Energy Demand. The two developed rapeseed oil based polyols were analyzed with three different modelling approaches for the bio-based feedstock stage – system expansion, mass allocation and economic allocation. The results indicate that, compared to petrochemical polyols in all cases of allocation bio-based rapeseed oil polyols have a better environmental performance in 8 out of the 18 ReCiPe midpoint impact categories and lower Cumulative Energy Demand. However, results also showed that the choice of allocation in the bio-based feedstock production stage has a significant impact on the results.