A novel and non-destructive approach for ZooMS analysis: ammonium bicarbonate buffer extraction
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ORIGINAL PAPER
A novel and non-destructive approach for ZooMS analysis: ammonium bicarbonate buffer extraction Nienke Laura van Doorn & Hege Hollund & Matthew J. Collins
Received: 11 January 2011 / Accepted: 26 April 2011 / Published online: 17 May 2011 # Springer-Verlag 2011
Abstract Bone collagen is found throughout most of the archaeological record. Under experimental conditions, collagen is apparently preserved as an intact molecule, with amino acid compositions and isotopic profiles only changing when almost all of the protein is lost. The ubiquity of collagen in archaeological bone has lead to the development of the use of collagen peptide mass fingerprints for the identification of bone fragments— Zooarchaeology by Mass Spectrometry (ZooMS). We report a novel, but a simple method for the partial extraction of collagen for ZooMS that uses ammonium bicarbonate buffer but avoids demineralisation. We compared conventional acid demineralisation with ammonium bicarbonate buffer extraction to test ZooMS in a range of modern and archaeological bone samples. The sensitivity of the current generation of mass spectrometers is high enough for the non-destructive buffer method to extract sufficient collagen for ZooMS. We envisage that a particular advantage of this method is that it leaves worked bone artefacts effectively undamaged post-treatment, suitable for subsequent analysis or museum storage or display. Furthermore, it may have potential as a screening tool to aid curators in the selection of material for more advanced molecular analysis—such as DNA sequencing.
N. L. van Doorn (*) : M. J. Collins BioArCh, Biology (S-Block), University of York, York YO10 5YW, United Kingdom e-mail: [email protected] H. Hollund Department of Geoarchaeology, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The Netherlands
Keywords Bone . Collagen . Zooarchaeology . Non-destructive analysis . MALDI-TOF-MS
Introduction Collagen survival Collagen, the most abundant protein in bone and in dentin, is found widespread in the archaeological record (Higham et al. 2006; Tuross et al. 1980) and can persist in fossils up to at least 600 ka (Buckley et al. 2011), perhaps considerably longer (Schweitzer et al. 2009). Type I (bone) collagen is a triple helix composed of two alpha 1 (COL1A1) and one alpha 2 (COL1A2) chain. COL1A1 is more highly conserved between genera than COL1A2 (Buckley et al. 2009; Buckley et al. 2010). A third alpha chain, COL1A3, has been reported exclusively in fish (Piez 1965). Unlike DNA, which is typically fragmented to below 70 bp when recovered from bone (e.g. Krause et al. 2010), collagen is sometimes treated as if it remains an intact protein. Radiocarbon and stable isotope preparation methods typically isolate an acid insoluble fraction, gelatinise that fraction by heating in weak acid and subsequently retain a high molecular weight >30 kDa fraction. Covington et al (2008) argue that collagen is stabilised by physical compression of the collagen fibril by mineral or chrome tanning agents, in a m
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