Analysis of organic residues in pottery vessels has been successful in detecting a range of animal and plant products as indicators of food preparation and consumption in the past. Panicoideae subfamily synthesize miliacin as their sole PTME28. The presence of high abundances of miliacin in sediment profiles indicating the locations of past millet cultivation has already been demonstrated27. In this study of Alpine lake sediments, was deemed to be the only possible source of miliacin, given the context and since it was the only PTME detected. High abundances of miliacin have also been 1472795-20-2 supplier reported in Early Iron Age palaesols (eastern Ukraine) from pits comprising broomcorn millet crop processing waste29. In contrast, a study of pit fills from Neolithic Cishan, China identified miliacin in authentic samples of both and and suggested differences between the two species based on the presence of other PTMEs in lower abundance7. The finding of miliacin in is not supported by investigations of authentic samples28. Unlike previous studies we focus on the identification of millet in pottery using molecular and isotopic characterisation. Ceramic vessels are abundant on archaeological sites during the period of millet domestication and dispersal. Prehistoric Eurasian pottery has been studied by archaeologists for centuries and often benefits from well-established chronologies. This approach also provides a means of contextualizing millet within the spheres of culinary practice and artefact use, allowing further and more detailed insight into the cultural significance of the crop. Archaeological samples Ceramics from two prehistoric sites, one in East Asia and one in Europe, were selected for analysis in order to demonstrate the approach (Fig. 1). These sites were chosen as charred millet grains were identified amongst the botanical remains at both sites. Figure 1 Location of sites investigated in Europe and East Asia (Adobe Illustrator CS4 14.0.0. http://www.adobe.com/uk/). Bruszczewo, a settlement in Poland some 60?km south of Pozna (520047N, 163507E), has been subject to comprehensive investigations30,31,32,33. Situated on a spur and close to a lake, two main settlement periods each with several sub-phases can be distinguished: An Early Bronze Age phase (EBA, 2100C1650?cal BC) with fortification, houses and clear settlement activities, and after an apparent gap of several hundred years, occupation dated to the Late Bronze Age/Early Iron Age (LBA/EIA, 1100C800?cal 1472795-20-2 supplier BC). The excavations uncovered several EBA layers and houses preserved in a wet area below the spur at the shore of the lake that date to ca. 1800C1650 BC cal33. Constructed originally on dry ground at the lakeshore during the EBA, the water level rose in the Middle Bronze Age and preserved parts of the site under wet conditions34. Archaeobotanical analyses revealed a diachronic differentiation in the occurrence of broomcorn millet. Only single charred grains were recovered from the EBA contexts, but they are numerous in the LBA thus indicating its relevance as food crop in this phase. Furthermore, there are impressions of millet grain in some sherds, particularly on the interior surface of the lower portion of the vessel, perhaps to facilitate the drying process prior to firing. Sixty-one visible charred deposits or foodcrusts adhering to pottery vessel surfaces were sampled. The samples include those from clearly defined and dated EBA and LBA/EIA contexts as well as a mixed transgression layer from the lake. This layer includes significant EBA material and very few younger finds; hence this group is identified as mostly EBA. The Majeon-ri site (360989N, 1270837E) is a settlement with a cemetery and paddy field system with wooden structures belonging to the Late Bronze Age and dated to 800C500?cal BC, located in the western region of the Korean peninsula35. Fifteen pottery sherds excavated from the irrigation ditch and puddle features at this site were selected for analysis. Unlike the pottery sherds from Bruszczewo, no charred surface deposits were observed. Results Mobilisation and absorption of 1472795-20-2 supplier miliacin in pottery To test whether miliacin is mobilised from millet AKT2 during processing and absorbed into the ceramic matrix, an experiment was undertaken involving the prolonged cooking of four species of millet; broomcorn millet (grain following solvent extraction and column chromatography to isolate the neutral fraction28. Much lower.