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Protocol Development for Purification and Characterization of Sub-Fossil Insect Chitin for Stable Isotopic Analysis and Radiocarbon Dating

Published online by Cambridge University Press:  18 July 2016

G W L Hodgins ,
J L Thorpe ,
G R Coope  and
R E M Hedges
G W L Hodgins*
Affiliation:
Oxford Radiocarbon Accelerator Unit, Research Laboratory for Archaeology and the History of Art, 6 Keble Road, Oxford, United Kingdom OX1 3QJ.
J L Thorpe
Affiliation:
School of Geography, University of Oxford, Mansfield Road, Oxford, UK, OX1 3TB
G R Coope
Affiliation:
Centre for Quaternary Research, Department of Geography, Royal Holloway, University of London, Egham, Surrey, United Kingdom TW20 0EX
R E M Hedges
Affiliation:
Oxford Radiocarbon Accelerator Unit, Research Laboratory for Archaeology and the History of Art, 6 Keble Road, Oxford, United Kingdom OX1 3QJ.
*
Corresponding author. Current address: CAIS, University of Georgia, 120 Riverbend Rd., Athens, Georgia 30602 USA. Email: [email protected].
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Abstract

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Reliable radiocarbon dating depends upon well-defined samples. We have been investigating whether or not reliable 14C dates can be obtained directly from sub-fossil insect cuticle or biochemical fractions derived from it. Initial carbon and nitrogen stable isotope measurements on sub-fossil insect chitin from species with known feeding behaviors found within a single site (St Bees, Cumbria) clustered in a manner reminiscent of trophic level effects seen in terrestrial ecosystems. Although this finding implied some chemical stability, the measurement of CN ratios from the same samples indicated compositional variability. In addition, 14C dates obtained from these same samples were different from dates obtained from plant macrofossils found at the same depth. We have experimented with protocols designed to biochemically reduce chitin to its principle carbohydrate component glucosamine with the aim of using this compound to generate reliable 14C dates. Solvent extractions of sub-fossil chitin were carried out to remove both endogenous and exogenous lipid-soluble materials. Base hydrolysis reactions designed to extract polypeptides retained surprisingly high levels of contaminating amino acids. Proteinase K enzyme treatment had little affect on the level of amino acid contamination. Strong acid hydrolysis reactions designed to depolymerize chitin to glucosamine yielded only 5% glucosamine. Clearly alternative methods of chitin depolymerization must be identified before the purification and 14C dating of glucosamine from sub-fossil chitin becomes practical.

Type
I. Becoming Better
Information
Radiocarbon , Volume 43 , Issue 2A: Proceedings of the 17th International Radiocarbon Conference (Part 1 of 3) , 2001 , pp. 199 - 208
Copyright
Copyright © The Arizona Board of Regents on behalf of the University of Arizona 

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