This study investigated how the anions had the effect on the spectral characteristics of dissolved organic matter (DOM) of secondary effluent from municipal wastewater treatment by using three-dimensional fluorescence spectroscopy and UV spectrophotometry in order to provide the data foundation for DOM’s environmental behavior. The results showed that the addition of SO2-4 and Cl- could increase or decrease the fluorescence intensity of DOM’s fluorescence peaks. In addition, the fluorescence peak representing the visible humus-like was observed with the additional concentration of NO-3 with 0.005~0.1 mol·L-1. Moreover, the fluorescence intensity of each fluorescence peak decreased obviously with the increase of NO-3 concentration, and the excitation and emission wavelengths of the fluorescence peaks representing the humus-like were red shifted with the right side of UV absorption peak. The added NO-3 concentration was positively correlated with HIX with the determination coefficient of 0.993. This was similar to that with UV254(0.994), UV253/UV203(0.987), α300(0.998) and α250/α365(0.995). And it had the negative correlation with BIX as the determination coefficients was 0.949. However, it had no significant effect on α350 and α355. In conclusion, the addition of SO2-4 and Cl- had little effect on the fluorescence and UV characteristics of secondary effluent DOM except fluorescence intensity. When the concentrations of NO-3 in the samples were different, it had a certain effect on analyzing the source and the properties of the different DOMs by using fluorescenct parameters and UV parameters.