Hostname: page-component-78c5997874-mlc7c Total loading time: 0 Render date: 2024-11-17T21:32:35.707Z Has data issue: false hasContentIssue false

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*
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: ghodgins@arches.uga.edu.
Rights & Permissions [Opens in a new window]

Abstract

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

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
Copyright
Copyright © The Arizona Board of Regents on behalf of the University of Arizona 

References

Aalbersberg, G, Litt, T. 1998. Multiproxy climate reconstructions for the Eemian and early Weichselian. Journal of Quaternary Science 13(5):367–90.3.0.CO;2-I>CrossRefGoogle Scholar
Atkinson, TC, Briffa, KR, Coope, GR. 1987. Seasonal temperatures in Britain during the past 22,000 years, reconstructed using beetle remains. Nature 325: 587–92.Google Scholar
Bishop, WW, Coope, GR. 1977. Stratigraphical and faunal evidence for lateglacial and early Flandrian environments in south west Scotland. In: Gray, JM, Lowe, JJ, editors. The Scottish Lateglacial Environment, Pergamon Press. Oxford. p. 6188.Google Scholar
Coope, GR, Lemdahl, G, Lowe, JJ, Walking, A. 1998. Temperature gradients in northern Europe during the last glacial-Holocene transition (14-9 14C kyr BP) interpreted from coleopteran assemblages. Journal of Quaternary Science 13(5):419–33.Google Scholar
DeNiro, MJ, Epstein, S. 1981. Influence of diet on the distribution of nitrogen isotopes in animals. Geochimica et Cosmochimica Acta. 45:341–51.Google Scholar
Elias, SA, Carrara, PE, Toolin, LJ, Jull, AJT. 1991. Revised age of deglaciation of Lake Emma based on new radiocarbon and macrofossil analysis. Quaternary Research 36:307–21.Google Scholar
Elias, SA, Toolin, LJ. 1990. Accelerator dating of mixed assemblage of late Pleistocene insect fossils from the Lamb Spring site, Colorado. Quaternary Research 33: 122–6.CrossRefGoogle Scholar
Hedges, REM, Law, I, Bronk, CR, Housley, RA. 1989. The Oxford accelerator mass spectrometry facility: technical developments in routine dating Archaeometry 31(2):99114.Google Scholar
Hodgins, GWL, Butters, TD, Bronk Ramsey, C, Hedges, REM. 2001. The chemical and enzymatic hydrolysis of archaeological wood cellulose and monosaccharide purification by high pH anion exchange chromatography for compound-specific radiocarbon dating. Radiocarbon. This issue.CrossRefGoogle Scholar
Miller, RF, Voss-Foucart, M-F, Toussaint, C, Jeuniaux, C. 1993. Chitin preservation in Quaternary Coleoptera: preliminary results. Palaeogeography, Palaeoclimatology, Palaeoecology 103:133–40.Google Scholar
Morris, MG. 1997. Broad-nosed weevils, Coleoptera: curculionidae (Entiminae). In: Dolling, WR, Askew, RR, editors. Handbook for the identification of British insects, Vol. 5, Part 17a. London: Royal Entomological Society, p 1422.Google Scholar
Peter, MG, Kegel, G, Keller, R. 1986. Structural studies on sclerotized insect cuticle. In: Muzarelli, RAA, Jeuniaux, C, Gooday, GW, editors. Chitin in nature and technology. New York: Plenum Publishing Corporation, p 21–8.Google Scholar
Pigman, WW. 1957. The carbohydrates. New York: Academic Press, p 60.Google Scholar
Richards, AG. 1978. The chemistry of insect cuticle. In: Rockstein, M, editor. Biochemistry of insects. New York: Academic Press, p 205–32.Google Scholar
Schimmelman, A, DeNiro, MJ. 1986a Stable isotopic studies on chitin I. Measurements on chitin/chitosan isolates and D-glucosamine hydrochloride from chitin. In: Muzarelli, RAA, Jeuniaux, C, Gooday, GW, editors. Chitin in nature and technology. New York: Plenum Publishing Corporation, p 357–64.Google Scholar
Schimmelman, A, DeNiro, MJ. 1986b Stable isotopic studies on chitin II. The 13C/12C and 15N/14N ratios in arthropod chitin. Contributions in Marine Science 29: 113–30.Google Scholar
Stankiewicz, BA, Briggs, DEG, Evershed, RP, Duncan, IJ. 1997a Chemical preservation of insect cuticle from the Pleistocene asphalt deposits of California, USA. Geochimica et Cosmochimica Acta 61(11):2247–52.Google Scholar
Stankiewicz, BA, Briggs, DEG, Evershed, RP, Flannery, MB, Wuttke, M. 1997b. Preservation of Chitin in 25-million-year-old fossils. Science 276:1541–43.Google Scholar
Walker, MJC, Bryant, C, Coope, GR, Harkness, DD, Lowe, JJ, Scott, EM. 2001. Towards a radiocarbon chronology for the late-glacial in Britain. Radiocarbon. This issue.Google Scholar
Webb, S. 1998. Stable Carbon and Nitrogen isotopes in Insects: the influence of diet. D.Phil Thesis. University of Oxford.Google Scholar