A new method of solubilizing, sensitizing and stabilizing petroleum substances by using sodium dodecyl sulfate (SDS) micellar solution as solvent was proposed. The variation relationship between fluorescence intensity of petroleum substances and SDS micellar solution concentration was studied, and the optimum concentration of SDS micelle solution as solvent was determined with 0.1 mol·L-1. Gasoline, diesel and kerosene SDS micellar solution of different diluted concentration were measured using FLS920 fluorescence spectrometer and fluorescence excitation-emission matrixes (EEMs) were obtained. After analyzing Rayleigh scattering, Raman scattering and instrument characteristics influence on measured spectrum, three-dimensional fluorescence spectra of three kinds of oil samples with excitation wavelength ranging from 250 to 400 nm and emission wavelength from 260 to 500 nm were established by spectral correction. The linear relationship between fluorescence intensity and concentration in certain concentration range was determined. By comparison with the spectra of gasoline, diesel, kerosene aqueous solution with the various concentrations under the same conditions and preparation method, the solubility and fluorescence intensity of petroleum pollutant in water was increased, and better stability was obtained. The measurement of petroleum substances without relying on some toxic solvent extraction was not only achieved, but its low solubility in water and difficult quantitative problems were solved.