As global air traffic is expected to increase significantly in the coming decades, reducing the associated climate impact requires scalable solutions. While alternative propulsion technologies such as electric and hybrid-electric systems might offer long-term potential, their current applicability remains limited due to low energy density, limited range and scalability, and system complexity. Consequently, thermodynamic propulsion systems – such as gas turbines and piston engines – are expected to remain dominant in the medium term. In this context, sustainable hydrocarbon-based aviation fuels represent a practical and necessary solution. Certified sustainable aviation fuel (SAF) pathways are currently approved exclusively for use in gas turbines, with certification standards tailored to turbine-specific requirements. Consequently, fuel properties such as cetane number and evaporation behavior are not included in existing specifications. However, when SAF-kerosene blends are used in compression ignition engines, the impact of these properties on ignition quality, combustion behavior, and emissions must be specifically evaluated. For this purpose, a flight test campaign was conducted using a fully instrumented Diamond DA42 aircraft, configured as a flying laboratory and equipped with serial-production piston engines. Two synthetic fuel variants were evaluated: one certified according to ASTM D7566-23a Annex A2 (HEFA SPK) and a second, Tall Oil derived fuel with a distinctly different molecular composition – characterized by an increased content of cycloparaffins and low aromatics content. The aircraft as a flying air lab was equipped with special engine measurement technology including high-pressure in-cylinder indication to analyze the impact of these differing fuel compositions on engine efficiency and combustion characteristics, including ignition delay and peak pressure. Furthermore, a mobile emission and particle number measurement system enabled the assessment of environmental performance under real flight conditions. Both fuels demonstrated significant reductions in thermal NOx formation due to their low aromatics content. However, no clear benefit was observed in total particle number (PN), likely due to a shift in the particle size distribution towards the nanoparticle regime.