The fluorescence components of dissolved organic matter (DOM) in the water bodies of a mining area were studied using three-dimensional fluorescence spectroscopy (EEMs) combined with parallel factor analysis (PARAFAC), and their types, distribution, and sources were analyzed and discussed. The results showed 5 natural components in the water samples and sediments of the study area. Components C1 (275/325(335) nm) and C2 (290/345 nm) in the water samples were identified as protein-like substances (tryptophan-like). In comparison, components C3 (320/405 nm) and C5 (285/505 nm) were identified as visible region humic acids, and component C4 (265/430 nm) was identified as ultraviolet region humic acid. Components C1 (270(300)/340 nm), C2 (290/340 nm), C3 (300/360 nm), C4 (285/320(360) nm), and C5 (270/435 nm) in the sediments were identified as protein-like substances (tryptophan-like), and component C5 was identified as ultraviolet region humic acid. The contents of protein-like substances in the water samples and sediments of the mining area were 302.23 and 1 976.83, respectively, and the contents of humic acids were 96.72 and 41.11, respectively, indicating that both the water samples and sediments were mainly composed of protein-like substances. Additionally, since sediments are one of the main sources of protein-like fluorescence, the content of fluorescent substances in the sediments was much higher than that in the water samples. The spatial distribution of fluorescent substances in the water samples was higher at the mining wastewater outlet and the Shuangji River and lower along the river and at the Hutuo Ditch and the Shuangji River confluence. On the other hand, the spatial distribution of fluorescent substances in the sediments was higher at the Shuangji River and lower at the mining wastewater outlet along the river and at the confluence of the Hutuo Ditch and the Shuangji River, which was the opposite of the distribution in the water samples. The fluorescence index (FI), biological index (BIX), and humification index (HIX) in the water samples and sediments indicated strong autochthonous characteristics and weak humification characteristics. The correlation analysis between the components and the fluorescence characteristic parameters further indicated that the protein-like substances in the sediments were mainly derived from autochthonous sources rather than exogenous sources. The research findings can better understand the organic matter composition in the wastewater discharged from mining areas and serve as a reference for treating mining wastewater discharge.