Three-dimensional excitation emission matrix fluorescence spectroscopy (3D-EEM) was applied to analyze the fluorescence characterization of dissolved organic matter in landfill leachate. The results showed that fulvic-like, tryptophan-like and humic acids-like matters were the main compounds in landfill leachate. A number of UV fulvic-like matters led to ineffective biodegradation of leachate. HOA contained more UV fulvic-like and less Vis fulvic-like, by contrary to HIA; HIN was mainly composed of UV fulvic-like and Vis fulvic-like matters; HOB, HIB and HIN had more intense fluorescence signal, whose fluorescence peaks occurred in the region of fulvic-like, tryptophan-like and humic acids-like. This was similar to the original leachate as well as HON which had relatively weak fluorescence signal. Moreover, different fractions had different fluorescence signal intensity in different regions. HOB and HIB had more intense signal in the region of UV fulvic-like. HIN had more intense signal in the region of UV fulvic-like and Vis fulvic-like. HON had moderately intense signal in the whole region. HOA and HIA had relatively weak fluorescence signal.