Abstract: Dissolved organic matter (DOM) significantly influences the carbon cycle, nutrient supply, and pollutant migration in both natural environments and engineered systems. Its characterization techniques primarily include ultraviolet-visible spectroscopy (UV-Vis) and three-dimensional excitation-emission matrix fluorescence spectroscopy (3D-EEM). This paper reviews the application of these two spectroscopic techniques in DOM analysis and the advantages of their combination. The results show that UV-Vis characterizes DOM's aromaticity, molecular weight, and humification degree through parameters such as absorption coefficients and spectral slope ratios. 3D-EEM, combined with Parallel Factor Analysis (PARAFAC), can identify DOM fluorescence components (such as humic acid) and evaluate DOM sources and transformation characteristics using indices like Fluorescence Index (FI) and Humification Index (HIX). The combined use of UV-Vis and 3D-EEM achieves complementary advantages in structural information, component identification, and source apportionment, providing an efficient tool for studying the physicochemical properties and transformation mechanisms of DOM. The combined technique is widely applied in freshwater, marine, soil, and water treatment engineering to track DOM sources, migration and transformation, and its complexation behavior with heavy metals, thereby optimizing water treatment processes and predicting the risk of Disinfection Byproducts (DBPs) formation. However, due to limitations such as spectral overlap, quenching effects, and interference from environmental variables, the combined technique still requires improvements in molecular-level structural analysis and standardization.