Biofuels and alternative fuels are increasingly being blended to conventional gasoline fuel to reduce the overall CO₂ emissions. The effect on NOx and soot formation is still unclear as the atomization and evaporation of gasoline with biocomponents differ depending on fuel specific physiochemical properties. This work focuses on describing the biofuel evaporation behavior of gasoline sprays at homogeneous charge (early injection timing) and stratified-charge conditions (late injection timing mode) used in modern direct injection spark ignition engines (DISI). A spray plume of a 6-hole solenoid injector is analyzed in terms of liquid spray propagation, and local droplet sizes studied in an injection chamber. Depending on the operating conditions, different physiochemical properties are found to dominate the atomization and evaporation processes: For low and moderate ambient temperature and pressure, high-boiling point components show a strong influence on the spray droplet size distribution. However, at elevated temperature and pressure, the evaporation behavior changes completely. Due to a high degree of evaporation, the evaporation cooling effect dominates the local droplet sizes. Fuel mixtures owing a larger heat of vaporization show larger droplet sizes—even if these fuels have a lower boiling point. Depending on the local evaporation behavior, the different remaining droplet momentum in the spray controls the air entrainment and the subsequent progress of evaporation and mixing. Overall, it can be stated that the heat of vaporization is a dominating physiochemical property for the droplet evaporation rate at high-level supercharged conditions.