Isolation and Isotopic Analysis of Individual Amino Acids from Archaeological Bone Collagen: A New Method Using Rp-hplc

Authors: O'Connell, T. C.; Hedges, R. E. M.

Source: Archaeometry, Volume 43, Number 3, August 2001 , pp. 421-438(18)

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Abstract:

This paper presents a new method for the isolation and isotopic analysis of some individual amino acids from proteins. The technique and its constituent steps are discussed; then isotopic analyses of amino acids from several samples of bone collagen from the Late Roman site of Poundbury, Dorset, UK are presented.

The applications of the method are discussed, as well as some advantages of this technique relative to other methods. Although developed for use with archaeological bone collagen, the technique is equally applicable to other proteinaceous materials. The use of reversed-phase HPLC avoids problems of isotopic fractionation inherent in using ion-exchange HPLC. Amino acids are isolated preparatively, allowing both carbon and nitrogen isotopic values to be measured on a single sample using CF-IRMS. Since amino acids are isotopically analysed in an underivatized form (unlike GC-C-IRMS), the method also presents the possibility of collecting the CO₂ generated during CF-IRMS: this would allow the subsequent dating by ¹⁴C-AMS of individual amino acids isolated from archaeological samples.

Keywords: CARBON; NITROGEN; ISOTOPES; AMINO ACIDS; ARCHAEOLOGY; PALAEODIET

Document Type: Research article

DOI: http://dx.doi.org/10.1111/1475-4754.00025

Affiliations: 1: Research Laboratory for Archaeology and the History of Art, University of Oxford, 6 Keble Road, Oxford OX1 3QJ, UK

Publication date: 2001-08-01