Dissolved organic matter (DOM) is the most active component in an environmental system. It can influence the chemical and structural characteristics of soil. In this work, three-dimensional excitation-emission matrix (3D-EEM) fluorescence spectroscopy, parallel factor analysis (PARAFAC), and two-dimensional correlation spectroscopy (2D-COS) integrated with synchronous fluorescence were used to explore the interaction between soil-derived DOM and Pb(II) during the soil sorption process. According to the data of batch sorption experiments, the adsorbing capacities of soil, soil + 5 mL DOM, and soil + 10 mL DOM were 16.96, 18.29, and 19.32 mg g, respectively, which indicated that DOM significantly enhanced the adsorption efficiency of Pb(II). The pseudo-second-order kinetic equation could well explain the adsorption process. The adsorbing data conformed to the isotherm of Langmuir adsorption. According to EEM-PARAFAC results, there are two major components from DOM. Protein-like substances were represented by component 1, and humic-like and fulvic-like substances were represented by component 2. Based on 3D-EEM, the results further showed that the intensities of component 1 and component 2 were obviously quenched with the increase of Pb(II) concentrations. The combined interpretations of the 2D-COS map for the DOM revealed that Pb(II) binding might occur sequentially in the order of humic-like fraction > protein-like fraction (346 > 282 nm). According to synchronous fluorescence spectra, static fluorescence quenching was the major process of quenching. Graphical abstract ᅟ.