The authors of the article explain the necessity of using eco-friendly biofuel for motor vehicles. The possibilities and ways of using biofuel in motor vehicles on the basis of alcohols (methanol, ethanol) and methyl ester of rapeseed oil are analyzed. The physical and chemical properties of these biofuel types are presented. In order to develop, determine and optimize the composition of eco-friendly biofuel for motor diesel engines, the authors have tested them during their work in methanol, ethanol-fuel emulsions and methyl ester of rapeseed oil. It has been experimentally established that the use of eco-friendly fuel in transport diesel engines is possible in the form of alcohol-fuel emulsions (alcohol (methanol or ethanol) is 25.0%, detergent-dispersant additive succinimide C-5A is 0.5%, water is 7.0%, diesel fuel is 67.5%) and in the form of separate supply of methanol (88.0%) and methyl ester of rapeseed oil (12.0%). The experimental research of motor diesel engines working on eco-friendly biofuel types of the above mentioned compositions has been carried out and the perspective decision of improving their environmental performance has been explained. When the diesel engine is operating on a methanol-fuel emulsion, the reduction in the pollutant content in the exhaust gases takes place: nitrogen oxides reduces by 41.3%, carbon soot reduces by 85.5%, carbon dioxide reduces by 6.7%, carbon monoxide reduces by 45.0%. When working on ethanol-fuel emulsion there is a decrease in nitrogen oxides by 50.2%, in carbon soot by 80.7%, in carbon dioxide by 23.8%, in carbon monoxide by 25.0%. And when working on methanol and rapeseed oil methyl ester there is a decrease in nitrogen oxides by 47.4%, in carbon soot by 90.4%, in carbon monoxide by 44.8%.