Poster Presentation 28th Annual Lorne Proteomics Symposium 2023

Species identification of a collection of worked bone artefacts from Pyrmont, Australia using mass spectrometry of bone collagen (#150)

Dylan H. Multari 1 , Geraldine J Sullivan 1 , Mary Hartley 1 , Ronika K. Power 1 , Paul A. Haynes 1
  1. Macquarie University, NSW, Australia

Zooarchaeology by Mass Spectrometry (ZooMS) is an increasingly utilised peptide mass fingerprinting (PMF) technique in archaeological science that analyses collagen 1A1 and 1A2 (COL1A1 and COL1A2, respectively) marker peptides for the genus- or species-level identification of fragmentary bones in the archaeological record. Traditionally, this analysis is performed using matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-ToF-MS) to identify characteristic m/z values of known marker peptides however MALDI instrumentation is becoming increasingly niche in mainstream proteomic laboratories, thus limiting the availability of this analysis. Here we present data on the application of a modified paleoproteomics approach, using nanoflow liquid chromatography - tandem mass spectrometry proteomics, to the analysis of a collection of six early colonial Australian (early to mid-19th Century CE) worked bone artefacts, believed to be mostly knife handles, excavated from a site in Pyrmont, Sydney, Australia in 2017. The historical context suggested that the bones were likely to be from sheep, cattle, whale or perhaps Australian native species. These bone handles display no distinguishable morphological features that are typically used in osteoarchaeological analyses of species; thus, a molecular approach is required for their identification. Mass spectrometric analyses were performed on a Thermo Q Exactive Orbitrap coupled with a Thermo Easy-nLC1000 liquid chromatography system. Peptide-to-spectrum matching was performed against the total curated SwissProt database supplemented with additional mammalian collagen sequences from NCBI, using the X! Tandem algorithm operating under the GPM user interface, where high-confidence positive identifications for Bos taurus COL1A1 and COL1A2 peptides were returned. Pairwise statistical analysis of identified peptide sequences was performed using a Fitch Matrix against a database of common domesticated species and selected Australian native mammals. This confirmed high correlation to sequences from the Bovinae subfamily, while excluding other possibilities. Taken together, these data indicate that the bone artefacts were prepared from bones of domestic cattle.