In this dataset, the branched glycerol dialkyl glycerol tetraethers (brGDGTs) data in a creek catchment located in southwest England are included, with 78 soil samples along 14 soil transects from hilltop to downslope, including 31 arable land sites, 14 permanent grassland sites, 24 temporary grassland (ley) sites and 5 woodland sites, 7 riverbed sediments at three locations along each of the two tributaries (upstream, midstream and downstream), and one more at the joint outlet, and a 50 cm long sediment core in the lake, which is about 150 m away from the joint outlet.The soils or sediments were freeze dried and homogenized, and then they were extracted with dichloromethane (DCM):MeOH (9:1, v/v) with an accelerated solvent extractor to obtain the total lipid extracts (TLEs). After adding of a known amount of internal standard, the TLEs were dried under a nitrogen stream, and then separated into apolar and polar fractions by passing them over an activated aluminum oxide column (Al2O3). The polar fractions containing GDGTs were evaporated under a gentle nitrogen stream, and then the samples were re-dissolved in a hexane:isopropanol (99:1, v/v) and filtrated through a polytetrafluoroethylene (PTFE) filter for UHPLC analyzing.