This study investigated the competitive sorption of black soil to adsorb Pb(II) and methylene blue (MB) from multi-contaminated soils. According to the experimental data, the process of adsorption can be clearly explained by pseudo-second-order kinetic equation. Both single and binary systems of the adsorption isotherms had a good fit with Langmuir models. The maximal adsorption abilities of Pb(II) and MB acquired from binary systems sorption were attenuated compared to those from the single system (Pb(II): 77.70 > 65.96 mg g−1; MB: 242.31 > 222.36 mg g−1). Pb(II) and MB can inhibit each other's sorption ability. A combination of three-dimensional excitation-emission matrix (3D-EEM), synchronous fluorescence spectra as well as two-dimensional correlation spectroscopy (2D-COS) were employed to determine the binding of dissolved organic matter (DOM) for Pb(II) and MB during soil sorption process. As a result, 3D-EEM implicated that the two main composes of DOM were humic acid-like substances and the fluorescence of DOM specimens were gradually diminished with increasing concentrations of Pb(II) and MB. According to synchronous fluorescence spectra, static quenching of Pb(II) and MB mainly led to fluorescence quenching. Specifically, fluorescence-2D-COS implicated that Pb(II) and MB bound to fluorescence in the following sequence: the earlier occurrence of the humic-like fraction compared to that of protein-like fraction. FTIR-2D-COS results concluded that the structural change sequence of DOM by Pb(II) binding followed the order: 1700>863>1332>1529>1200>1086 cm−1 and the sequence of the MB binding affinities followed the order: 1520>1399>1345>1152>1602>993>881 cm−1. These findings would be beneficial to understand the mechanism of adsorb multi-component systems and have the potential to contribute significance to the interaction mechanism of multi-component with soil DOM at the molecular level.