A label-free quantification method for assessing sex from modern and ancient bovine tooth enamel

Identification of the sex of modern, fossil and archaeological animal remains offers many insights into their demography, mortality profiles and domestication pathways. However, due to many-factors, sex determination of osteological remains is often problematic. To overcome this, we have developed an innovative protocol to determine an animal’s sex from tooth enamel, by applying label-free quantification (LFQ) of two unique AmelY peptides ‘LRYPYP’ (AmelY;[M+2] \(^{2+}\) $_{}^{2+}$ 404.7212 m/z) and ‘LRYPYPSY’ (AmelY;[M+2] \(^{2+}\) $_{}^{2+}$ 529.7689 m/z) that are only present in the enamel of males. We applied this method to eight modern cattle (Bos taurus) of known sex, and correctly assigned them to sex. We then applied the same protocol to twelve archaeological Bos teeth from the Neolithic site of Beisamoun, Israel (8-th–7-th millennium BC) and determined the sex of the archaeological samples. Since teeth are usually better preserved than bones, this innovative protocol has potential to facilitate sex determination in ancient and modern bovine remains that currently cannot be sexed.