In this work we used controlled microcosms to study the effect of temperature and pH on brGDGTs in lake water. We collected surface water from Kennedy Lake, Tucson, AZ, a shallow eutrophic artificial reservoir previously described by Martínez-Sosa & Tierney (2019). From the collected samples we set up a series of microcosms, consisting of 1L glass flasks filled with lake water, and manipulated single environmental factors including temperature and pH. For our temperature incubations, we selected four conditions (9C, 18C, 27C and 35C) and incubated 3 1L flasks under each condition for two periods of time (4 or 6 weeks). For the pH incubations, we used commercially available freshwater aquarium non-phosphate buffers (Proprietary composition, Seachem, Madison, GA, USA) to manipulate the pH of the microcosms. For these experiments we targeted four pH conditions (4, 5, 6 and 7), and included two control samples: one where we added enough buffer to maintain the initial pH (Control + Buffer), and another to which we added no buffer (Control - Buffer). GDGTs were analyzed on an Agilent 1260 Infinity HPLC coupled to an Agilent 6120 single quadrupole mass spectrometer using two BEH HILIC silica columns (2.1 x 150 mm, 1.7 um; Waters) and the methodology of Hopmans et al. (2016). We calculated peak areas using the MATLAB package software ORIGAmI (Fleming et al. 2016) and estimated the concentration of brGDGTs by comparing the obtained peaks with a C46 internal standard (Huguet et al. 2006) normalized to the volume of each sample.