The organic matter of lake sediment plays an important role in paleolimnological reconstruction. Here, we report a detailed study of organic matter components (C_org%, N%, δ¹³C, δ¹⁵N) in a dated sediment core of Hulun Lake in northern China. Multiple mixing models based on the stoichiometric ratios and stable isotopic compositions were applied to quantify the contributions of organic matter sources in lake sediment. The results show that the organic matter in the sediments from Hulun Lake mainly comes from terrestrial organic matter: the proportion of terrestrial organic matter is more than 80%. The results of the SIAR mixing model further reveal that the proportions of terrestrial C₃ plants-derived organic matter, soil organic matter, and lake plankton-derived organic matter were 76.0%, 13.9%, and 10.1%, respectively. The organic matter content of lake sediment from terrestrial sources began to increase significantly from 1980 onward, which is consistent with the growth in overgrazing in the Hulun Lake basin. The content of organic matter from endogenous lake-derived sources began to increase significantly after 2000 due to the nutrients gradually becoming concentrated in lake water, indicating that the reduction in rivers’ discharge and the downgrade of the lake water level were the immediate causes of the lake’s environmental deterioration during this period.