During the past century, many lacustrine environments have changed substantially at the ecosystem level as a result of anthropogenic activities. In this study, the distributions of n-alkane homologues, carbon isotopes (delta C-13(org)), organic carbon, and the C/N atomic ratio in two sediment cores from Fuxian Lake (Yunnan, southwest China) are used to elucidate the anthropogenic impacts on this deep, oligotrophic, freshwater lake. The carbon preference index (CPI) of long-chain components, average chain length (ACL), proportion of aquatic macrophytes (Paq), and terrigenous/aquatic ratios (TAR) show different temporal patterns that reflect variations in biological production. Notably, the n-alkane homologues are shown to be more sensitive to environmental changes than delta C-13(org) and the C/N ratio. Prior to the 1950s, minor variations in the sedimentary geochemical record were likely caused by climate changes, and they represent a natural stage of lake evolution.The onset of cultural eutrophication in Fuxian Lake occurred in the 1950s, when the n-alkane proxies collectively exhibited high-amplitude fluctuations but overall decreasing trends that coincided with population growth and related increases in land-use pressure. In the 21st century, Fuxian Lake has become even more eutrophic in response to human activities, as indicated by sharp increases in C/N ratio, Paq, delta C-13(org), ACL, CPI, and TAR. Our findings provide robust molecular sedimentary evidence confirming that the environmental evolution of lakes in the Yunnan-Guizhou Plateau over the past century was closely associated with enhanced anthropogenic activities.