Fast ultrasonic-based extraction of peptides from teeth for forensics and archaeological purposes

This study presents a rapid, ultrasound-assisted solid-liquid extraction protocol for recovering peptides from the tooth surface, while preserving the macroscopically visible integrity of the dental morphology. The methodology described herein enables the concurrent use of a single peptide extraction for multiple analytical objectives, encompassing: (i) biological sex determination via detection of sex-dimorphic amelogenin isoforms; (ii) dietary reconstruction based on the identification of food-derived protein residues; (iii) oral microbiome profiling to infer historical health status and disease burden; and (iv) an assessment of age-at-death (AAD) estimation through quantification of asparagine and glutamine deamidation kinetics. Peptide extraction is achieved through an ultrasound-assisted solid–liquid interface, facilitating the release of biomolecules embedded within the dental tissues' matrix without producing macroscopically detectable damage to the sample. The resulting peptide fraction undergoes purification using hydrophilic-lipophilic balanced (HLB) cartridges and is subsequently analysed via high-resolution mass spectrometry. The protocol offers notable advantages, including speed, reproducibility, high throughput, and alignment with the principles of analytical minimalism. Collectively, this strategy represents a substantial advancement in non-invasive dental peptidomics, providing a robust and minimally destructive analytical framework for forensic and bioarchaeological investigations.