Sex determination of mummies through multi-elemental analysis of head hair using electrothermal vaporization coupled to inductively coupled plasma optical emission spectrometry

Sex determination of human remains is of great archaeological significance, as it provides a more complete picture of social and familial structures within ancient societies. Typically performed through examination of bones in the pelvic region, accurate sex determination can be exceedingly challenging in the absence of a sufficiently preserved skeleton. Here, a method for sex determination in living humans, involving measurement of magnesium, strontium, sulfur, and zinc in head hair along with multivariate statistics, was applied for the first time to hair collected from 500 year-old mummies originating from Peru. Hair samples were washed in doubly deionized water, dried, and ground prior to analysis via electrothermal vaporization coupled to inductively coupled plasma optical emission spectrometry; only 2 mg of hair is required for analysis. Point-by-point internal standardization was performed with Ar i 430.010 nm to compensate for sample loading effects on the plasma. Peak areas were integrated and mass corrected before being used in combination with multivariate analysis. Although principal component analysis provided insufficient separation between the sexes, linear discriminant analysis (LDA) was highly effective for sex determination. Using mummy hair as the LDA model enabled accurate sex prediction of the mummies, showing that, despite the age of the hair, the samples still contain the necessary elemental information for sex determination. For accurate sex determination of mummies using hair collected from living humans, magnesium had to be replaced by sodium due to significant differences in dietary habits. With this simple modification, hair from living humans in North America could be used to successfully predict the sex of individuals who lived more than 500 years ago in Peru. This work paves the way for broader use of non-skeletal sex determination methods within the field of archaeology, filling a significant gap.