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Three-dimensional fluorescence parameters can reflect classification, properties and content change of pollutants in wastewater treatment. In the present paper, by using three-dimensional fluorescence characteristic analysis, comparative analysis of conventional organic pollutants such as COD, TN and TP, and three dimensional fluorescence spectrum analysis, the classification and content of dissolved organic pollutants were identified. We studied fluorescence spectra, fluorescence peak (R. U.), fluorescence index (FI), humification index (HIX) of DOM's four components in the entrance and effluent water and interstitial water, as well as the correlation between these four components and COD, TN and TP. The results showed that the position and intensity of the characteristic fluorescence peak center changed significantly before and after sewage treatment, indicating that the relative composition and content of the organic wastewater varied with wastewater treatment. Furthermore, the test results presented that humic-like composition was not degraded significantly, while protein-like composition was degraded significantly. And the protein-like component and COD, TN and TP presented significant positive correlation. This paper analyzed the fluorescence characteristics changes of dissolved organic matter in sewage treatment by using three-dimensional fluorescence spectrometry, and discussed the feasibility of three-dimensional fluorescence technique applied for description of dissolved organic pollutant degradation rule in the wastewater treatment process.