Three- dimensional fluorescence spectroscopy is one of the most important techniques to analyze substance. The adulterated edible oil is detected, and Rayleigh scattering, spectral intensity and position of characteristic peak are analyzed through three-dimensional fluorescence spectroscopy. It is found that the Rayleigh scattering, emission wavelength and spectral intensity vary with the increase of concentration of fried oil. In order to exactly analyze the impact of the fried oil on edible oil, the fluorescence peak of vitamin E is discussed. The experimental results show that the redshift of the fluorescence peak of vitamin E is observed and the fluorescence intensity declines with the dosage concentration increasing. The sensitivity and reliability of the three-dimensional fluorescence spectroscopy technique is demonstrated. As the results show, the detection of fried oil in edible oil based on three-dimensional fluorescence spectroscopy is feasible and effective.