Earth's climate is prone to natural and episodic cycles. The most recent period of climate change, the only one to be caused by humans, is significantly affecting species composition and landscapes. Northern expansion of the Boreal Forest in Canada is one of the expected outcomes, and tree line migration northward is one of the anticipated changes. Previous studies have found that many human activities like agriculture, grazing, and pastoralism can significantly affect tree line movement. Improving our ability to examine past tree line dynamics can be achieved using archival records in lake sediments. In this study, we focused on three sediment biomarkers that may be related to the presence of trees in a lake's catchment: n-alkanes, lignin-derived phenols, and stable isotopes of carbon. We examined the composition of these markers in sediment from 19 lakes in Saskatchewan spanning 4 ecoregions, from Prairie Grassland to Boreal Plain, to determine the biomarker signature for lakes in each ecoregion and relating them to land cover (trees vs herbaceous plants) in the catchments of each lake. The results showed that n-alkane composition was significantly correlated to the proportion of trees to herbaceous plants in a lake's catchment, raising the possibility that these can be used to infer the presence of trees in sediment records. The C/N ratio and δ¹³C were not effective in distinguishing ecoregions or land cover composition, likely due to algal production in the lake and agricultural activities in the surrounding farmland, while lignin-derived phenols appeared to be affected by unknown factors.