This dataset includes faecal lipid biomarker data from a lake sediment core from central Alaska. The purpose of this study was to assess the sensitivity and validity of lacustrine faecal steroid reconstructions of human and animal presence within the landscape by combining analyses of modern faecal samples and a sediment archive from Chisholm Lake, Alaska, where historical and archaeological population changes are well documented. To achieve this, we analysed faecal steroids (stanols and bile acids), which are biomolecules that are deposited in the landscape from faeces of higher vertebrates. The lake sediment core was recovered from Chisholm Lake in the Tanana Valley of interior Alaska (64.302267 N, 146.687331 W) in September 2019 using a Bolivia corer. The age-depth model was determined using lead-210 (210Pb). We extracted lipid biomarkers from 6 core subsamples and, following saponification and column chromatography, faecal biomarkers were identified and quantified by GC-MS at the University of Bristol. Stanol and bile acid data are reported as flux (accounting for the bulk density of the sediment samples and age-depth model inferred changes in sediment accumulation rates) and dry matter concentrations.