The incorporation of plant biomass into soil usually leads to long-chain n-alkanes with a relative predominance of odd carbon numbered homologues. Contrastingly, an increase in short-chain even carbon numbered n-alkanes was found in charred biomass with progressing temperatures. We applied lipid analysis to buried ancient topsoils that contained charred organic matter and to corresponding control soils, which were characterized by a lighter color and lower contents of charred materials. Commonly, the proportion of lipid extracts was found to be lower in the ancient soil than in the control samples, which indicated an enhanced degradation of organic matter, e.g., by thermal degradation. All samples displayed a particular pattern of short-chain and even carbon numbered n-alkanes (maximum at n-C16 or n-C18). The ratios CPI (carbon preference index) and ACL (average chain length) for the investigated soil samples matched the ratios found for maize and rye straw charred at 400 °C or 500 °C, respectively. These molecular ratios indicate the presence of charred biomass. The predominance of short-chain and even carbon numbered n-alkanes was a result of thermal degradation of biomass. The degradation products were preserved in ancient soils and could be applied as molecular markers in archaeological or palaeoenvironmental research.