Session: Parallel session 5 - Forensic and cultural heritage

Combine stable isotopes, carbon-14 dating and paleoproteomics in a single sample preparation protocol.

Louis DELEGUE¹, Delphine DECRUYENAERE^2,3, Stéphanie FLAMENT¹, Satine DALESSANDRO¹, Elise DUFOUR², Christian ROLANDO¹, Fabrice BRAY¹

¹UAR 3290 MSAP, Miniaturisation pour l'Analyse, la Synthèse et la Protéomique, Villeneuve d'ascq, France
²UMR 7209 MNHN, National Museum of Natural History, Paris, France
³Department of History and Cultural Heritage, Silk Road University of Tourism and Cultural Heritage, Samarkand, Uzbekistan

Archaeological teeth and bones contain a wealth of information such as the age of samples through carbon-14 dating or amino acid racemization, the diet of animals through stable isotope analysis, and the identification of taxa and proteomes through paleoproteomics. At present, there is no single preparation method for all these techniques. Unfortunately, it's not always possible to take an archaeological sample more than once, due to its rarity or size. The aim of this study is to demonstrate that it is possible to combine isotopic measurement, paleoproteomic and amino acid racemization analyses using a single bone collagen extraction protocol.

To this end, we have based ourselves on a collagen extraction method used for isotopic analysis and developed at MNHN. 8 samples from Uzbekistan dating from the 7th to the 11th century A.D. were studied. Paleoproteomic analysis was carried out on the demineralization, decontamination and bone powder fractions using ZooMS analysis on the MALDI FTICR. Amino acid racemization was performed on freeze-dried collagen after acid hydrolysis and derivatization with Marfey's reagent. The analysis was performed using nanoLC-MSMS orbitrap.

ZooMS analysis on demineralization, decontamination and bones fraction from the isotope protocol enables taxonomic identification of all our samples. The deamidation value, is higher in demineralization, decontamination fraction as HCl and NaOH utilization time is long and induces deamidation. For amino acid racemization, the derivatization protocol with Marfey's reagent works on standard amino acids and separates the enantiomer.

In conclusion, this study demonstrates for the first time that paleoproteomics and amino acid racemization can be combined with isotope analysis, and that the integration of is on the right track.