Ethnographic observations suggest that Indigenous peoples employed a distinct regime of frequent, low-intensity fires in the Australian landscape in the past. However, the timing of this behaviour and its ecological impact remain uncertain. Here, we present detailed analysis of charcoal, including a novel measure of fire severity using Fourier transform infrared (FTIR) spectroscopy, at a site in eastern Australia that spans the last two glacial/interglacial transitions between 135–104 ka and 18–0.5 ka BP (broadly equivalent to Marine Isotope Stage (MIS) 6-5 and 2-1, respectively). The accumulation of charcoal and vegetation composition was similar across both periods, correlating closely with Antarctic ice core records, and suggesting that climate is the main driver of fire regimes. Fire severity was lower over the past 18,000 years compared to the penultimate glacial/interglacial period and suggests increasing anthropogenic influence over the landscape during this time. Together with local archaeological records, our data therefore imply that Indigenous peoples have been undertaking cultural burning since the beginning of the Holocene, and potentially the end of the Last Glacial Maximum. We highlight the fact that this signal is not easily discernible in the other proxies examined, including widely used charcoal techniques, and propose that any anthropogenic signal will be subtle in the palaeo-environmental record. While early Indigenous people’s reasons for landscape burning were different from those today, our findings nonetheless suggest that the current land management directions are based on a substantive history and could result in a reduction in extreme fire events.