Direct LSC method development for bio-component quantification in fuels is presented. Optimal parameters and measurement conditions on LS counter Quantlus are discussed. Two simple calibration procedures for 14C determination in biofuels were carried out. Fuel blends 1–100% FAME were produced from two feedstock and two fossil matrices. Minimal detectable biogenic content assessed is (0.47–1.1)%m for 300 min of counting.