Recent work has disclosed the critical role played by enamel peptides in sex classification of old skeletal remains. In particular, protein AMELY (amelogenin isoform Y) is present in the enamel dental tissue of male individuals only, while AMELX (isoform X) can be found in both sexes. AMELY can be easily detected by LC-MS/MS in the ion extracted chromatograms of the SM_(ox)IRPPY peptide (monoisotopic [M + 2 H]⁺² mass = 440.2233 m/z). In this paper, we exploited the dimorphic features of the amelogenin protein to determine the sex of the so-called ‘Lovers of Modena’, two Late Antique individuals whose skeletons were intentionally buried hand-in-hand. Upon discovery, mass media had immediately assumed they were a male-female couple, even if bad preservation of the bones did not allow an effective sex classification. We were able to extract proteins from the dental enamel of both individuals (~1600 years old) and to confidently classify them as males. Results were compared to 14 modern and archaeological control samples, confirming the reliability of the ion chromatogram method for sex determination. Although we currently have no information on the actual relationship between the ‘Lovers of Modena’ (affective? Kin-based?), the discovery of two adult males intentionally buried hand-in-hand may have profound implications for our understanding of funerary practices in Late Antique Italy.

Homogeneity, mass fractions of about forty trace elements and Sr isotope composition of Ca carbonate reference materials (RMs) between original and nano‐powdered pellets are compared. Our results using nanosecond and femtosecond LA‐(MC)‐ICP‐MS show that the nano‐pellets of the RMs MACS‐3NP, JCp‐1NP and JCt‐1NP are about a factor of 2–3 more homogeneous than the original samples MACS‐3, JCp‐1 and JCt‐1, and are therefore much more suitable for microanalytical purposes. With the exception of Si, the mass fractions of the synthetic RM MACS‐3 agree with its fine‐grained analogue MACS‐3NP. Very small, but significant, differences between original and nano‐pellets are observed in the RMs JCp‐1 and JCt‐1 for some trace elements with very low contents, indicating the need for re‐certification. Strontium mass fractions in the analysed RMs are high (1500–7000 mg kg⁻¹), and their isotope compositions determined by LA‐MC‐ICP‐MS in the original and the nano‐pellets agree within uncertainty limits.

Significance: The extent to which Neanderthals differ from us is the focus of many studies in human evolution. There is debate about their pace of growth and early-life metabolic constraints, both of which are still poorly understood. Here we use chemical and isotopic patterns in tandem with enamel growth rates of three Neanderthal milk teeth from northeastern Italy to explore the early life of these individuals. Our study shows that these Neanderthals started to wean children at 5 to 6 months, akin to modern humans, implying similar energy demands during early infancy. Dental growth rates confirm this and follow trajectories comparable with modern humans. Contrary to previous evidence, we suggest that differences in weaning age did not contribute to Neanderthals’ demise.