Abstract: The organic matter release from the sediments in Lake Baiyangdian is investigated, which plays an important role in maintaining the water quality of the lake in the autumn-winter alternate seasons. Twenty samples were collected from representative sites in five representative areas, including the nature reserve areas, the tourist areas, the densely populated areas, villages in the middle of the lake, and in the lake outlet areas. The spectroscopy characteristics of the organic matter in the samples were determined, and the dissolution-release behavior of nitrogen and phosphorus in the samples were investigated. The results showed that:(1) The humification and aromatization of the organic matter in the sediment were relatively high in the nature reserve areas, villages in the middle of the lake and the lake outlet areas. The organic matters mainly consisted of the terrestrial visible light humic acid component (C1), marine visible light humic acid component (C3), and the UV-light humic acid component (C4). In contrast, the humification and aromatization of the organic matter were relatively low in the tourist areas and the densely populated areas, which was principally dominated by the tryptophan-like components (C2). (2) A close relationship between UV adsorption and fluorescence spectra parameters was obtained, which could be used to identify the characteristics of the organic matter in the sediment. A significant positive correlation existed between the fluorescence intensity of C2 and the w(TN) and w(TP), presenting a homologous characteristic. (3) A high release volume of nitrogen and phosphorus presented in the samples dominated by the protein-like substances rather than in those dominated by the humic-like substances. The results indicate that the protein-like substance and the humic-like substances as the main components exist in the sediment of the lake in the autumn-winter alternate seasons. The dissolution-release behavior of the coexistence nitrogen and phosphorus is accelerated in the presence of the protein-like substances.