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The impact of fatty acid methyl ester (FAME) blended diesel fuel on engine/vehicle systems was comprehensively investigated by vehicle, laboratory and engine tests. In this study, 20% FAME blended fuel (B20) was mainly used and soy bean oil methyl ester (SME) was primarily selected as the FAME. Vehicle testing with long-term fuel storage in vehicle fuel tanks was conducted, considering the most severe conditions in market use. Laboratory and engine tests were also conducted to better understand the vehicle test results. In the vehicle test, engine startability, idle roughness and fuel injection control were evaluated using nine vehicles with plastic or metal fuel tanks. All vehicles showed no problems up to 7 months. While five vehicles with plastic fuel tank did not show any problems throughout the test period up to 18 months, four vehicles with metal fuel tanks experienced malfunctions in engine start or fuel injection control following 8, 13, 13 and 18 months respectively.