The role of Amelogenin peptide analyses in the interpretation of a 5th century BCE Messapian funerary context from Roca Vecchia (Italy)

Biological sex in anthropological studies is of the utmost importance to inform paleo-demographic reconstruction. Historically, it has proven difficult to determine the sex of individuals based on morphological analysis due to insufficient conservation of the bones or the immature age of the individuals studied. To overcome these problems, we have used proteomic analysis to identify the Amelogenin protein in tooth enamel. This protein exists in two different isoforms linked to the X and Y chromosomes, X being present in both male and female sexes and Y being present only in the male sex. This identification is possible using liquid chromatography combined with mass spectrometry (LC-MS). In this paper, Amelogenin peptide analysis was applied to a sample of five individuals, with different age and sex, from a 5 th century BCE collective burial in Roca Vecchia (Italy). The analysis demonstrated that the method produces excellent results in the identification of the biological sex with a high reliability even in the case of low instrument resolution and in shorter time frames than other previous methods. The technique, therefore, has a strong prospect within the field of gender archaeology and offers the potential to better address the longstanding debate over the relationship between sex and grave goods.