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In the present paper, a new biofuel ethylene glycol monomethyl ether soyate has been developed. The biofuel was synthesized with a refined soybean oil and ethylene glycol monomethyl ether as reactants and sodium as catalyst under 90°C. The synthesized crude product was purified and structurally identified through Fourier Transform Infrared Spectrum (FT-IR), 1H Nuclear Magnetic Resonance Spectroscopy (1H NMR) and Gel Permeation Chromatography (GPC) analyses. The physicochemical properties of the biofuel and its addition effects on properties of diesel fuel were measured according to China national standard test methods. A single cylinder diesel engine was employed to evaluate the influences of the biofuel on engine fuel economy and engine-out emissions of CO, HC, NOx and smoke.