In this study, the injection processes and spray behavior of diesel fuel (DF), rapeseed oil (RO) and their fuel blends (RO25, RO50, RO75) have been qualitatively and quantitatively analyzed to identify the differences in the injection rate and the spray parameters. The volumetric and mass injection rates, the spray penetration, the spray cone angle and the spray area under non-evaporation conditions in a nitrogen-filled camber were analyzed. The results showed that rapeseed oil leads to a decrease in the peak injection rate due to its higher density and viscosity. Moreover, rapeseed oil and its blends with mineral diesel oil exhibited smoother rising slopes at the start of injection. The spray tip penetration was the longest for the rapeseed oil and the spray tip penetrations of the fuel blends RO25, RO50 and RO75 were arranged between the values of DF and RO. As the injection pressure increased, the differences in the spray tip penetrations diminished. Increasing the amount of rapeseed oil in fuel blends resulted in a smaller spray cone angle compared to diesel fuel. The spray area of all tested fuels increased significantly with increasing spray tip penetration, the spray area of RO was consistently lower than that of diesel fuel.