Signature pigments of photosynthetic green sulfur bacteria (GSB) were found in ancient sediments collected from an abandoned clay quarry located in the Banyoles lacustrine area (Spain). The sediments belong to the Interglacial Waalian of the lower Pleistocene (0.7–1.5 millions years old) and were deposited after a marshy event occurring during that geologic period. Reverse-phase high performance liquid chromatography (RP-HPLC) analyses of acetone:methanol sediment extracts revealed that the main pigments found were carotenoids of both eukaryotic and prokaryotic photosynthetic organisms. In particular, isorenieratene (Isr) and β-isorenieratene (β-Isr) constituted the larger bacterial fraction (35–40% of the total carotenoid content), whereas okenone, a signature pigment of purple sulfur bacteria, accounted for less than 2%. Xanthophylls from oxygenic photosynthetic organisms accounted for the remaining carotenoids. Likewise, traces of degradation products of both bacteriochlorophyll (BChl) and chlorophyll (Chl) were also detected. The low concentration of Chl degradation products made proper identification of these compounds impossible. In contrast, degradation products of BChl-e such as bacteriochlorophyllide-e and bacteriopheophorbide-e were identified in the HPLC analyses, suggesting that chemical degradation of naturally occurring BChl aggregates of GSB was slower in the clay quarry sediments. The presence of signature pigments of brown-colored species of GSB (Isr, β-Isr and degradation products of BChl-e) in the sediments suggests an ancient aquatic environment where GSB were present and where episodes of sulfide-rich anoxia reaching the photic zone could be envisaged. Similarly, the large percentage of algal xanthophylls supports that algae could be an important part of the microbial photosynthetic community in this ancient ecosystem.EU. TMR Program, Contract No. FMRXCT96-0081.Peer reviewed