The identification and measuring method of petroleum pollutant is proposed by three-dimensional fluorescence spectroscopy combined with parallel factor analysis. Different concentration solutions of 97# gasoline, 0# diesel and kerosene in CCl4 are as measuring samples. Every petroleum product as one component is considered as a whole and the specific components are not taken into account. By mixing gasoline and diesel with different concentrations and taking kerosene as interfering substance, the three-dimensional fluorescence spectra of samples are measured with FLS920 fluorescence spectrometer. Instrumental error and effect of scattering are removed and the true spectra are obtained by using excitation and emission correction and blank subtraction. The experiments use the second-order calibration algorithms based on the parallel factor to analyze the spectral data and the second-order advantage is adequately exploited. It is proved that the identification and measurement of different components in mixed sample are achieved accurately in existence of interfering substance and good recovery is obtained.