Measuring the ethanol/water ratio in biofuel of high ethanol content, such as E85, is important when used in a flex-fuel engine. A capacitive probe is generally used for measuring the ethanol/gasoline ratio. However, the water content in E85 biofuel cannot be disregarded or considered constant and full composition measurement of biofuel is required. Electric impedance spectroscopy with a customized coaxial probe operating in the 10 kHz to 1 MHz frequency range was investigated. An in-depth investigation of the electrical impedance domain has led to the conclusion that additional information is required to unambiguously determine the composition of the ternary biofuel mixture. Among the different options of measurement domains and techniques, optical absorption spectroscopy in the UV spectral range between 230 and 300 nm was found to be the most appropriate. The typical absorbance in the UV range is highly dominated by gasoline, while ethanol and water are almost transparent. This approach is experimentally validated using actual fuels.