The biocomponent ratio in liquid fuels as well as the usage of renewable resources for fuel consumption in the transport sector needs to be increased as a result of EU directive 2003/30/EC. Based on radiocarbon (14C) measurements, it should be relatively simple and fast to measure the weight percentage of the fossil and biological sources by accelerator mass spectrometry (AMS) as recommended in the ASTM D 6866-12 and EN 16640 standards. In this study, a relatively easy and fast sample preparation and measurement method based on AMS measurements was developed at the Hertelendi Laboratory of Environmental Studies (HEKAL) using reference samples from the Hungarian MOL Nyrt. oil company. Considering the recent EU regulation for mixing rates of liquid fuels in the transport sector (0.7–2% biofuel content) and the projected higher rates (2–10% biofuel content), the method is applicable to determine fatty acid methyl ester (FAME) and/or hydrotreated vegetable oil (HVO) derived proportions of fuel blends with a 1σ uncertainty better than±0.3% m/m.