Hostname: page-component-78c5997874-fbnjt Total loading time: 0 Render date: 2024-11-20T01:32:28.177Z Has data issue: false hasContentIssue false
  • English
  • Français

The Chemical and Enzymatic Hydrolysis of Archaeological Wood Cellulose and Monosaccharide Purification by High Ph Anion Exchange Chromatography for Compound-Specific Radiocarbon Dating

Published online by Cambridge University Press:  18 July 2016

G W L Hodgins ,
T D Butters ,
C Bronk Ramsey  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
T D Butters
Affiliation:
Glycobiology Institute, Department of Biochemistry, University of Oxford, South Parks Road, OX1 3QU
C Bronk Ramsey
Affiliation:
Oxford Radiocarbon Accelerator Unit, Research Laboratory for Archaeology and the History of Art, 6 Keble Road, Oxford, United Kingdom OX1 3QJ
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].
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.

Preliminary experiments were carried out on archaeological wood to investigate methods of cellulose hydrolysis and carbohydrate monomer purification for the purpose of compound-specific radiocarbon dating. The Chelford log, a known 14C dead source of wood cellulose, was selected for study in order to investigate the levels of contamination introduced during sample purification. Two methods of hydrolysis were examined, mineral acid hydrolysis and enzyme hydrolysis using cellulase from Penicillium funiculosum. Under the conditions described, enzymolysis was far superior to acid hydrolysis in terms of the glucose monomer yield. Glucose monomer purification was accomplished using high pH anion exchange chromatography with pulsed amperometric detection. This high performance liquid chromatography (HPLC) method does not require sample derivatization and the chromatography products can be collected in water. These characteristics make it potentially well suited to carbon dating applications. 14C dating of chromatographically purified glucose fractions revealed significant levels of contamination had accumulated during both protocols. Glucose contamination from the cellulase enzyme preparation was a major source of contamination within the enzymatically hydrolyzed samples. Ultrafiltration of the enzyme removed some but not all of this contamination. The contamination must be reduced 10-fold before the methodology could be viable for dating. This hydrolysis/HPLC method is also being investigated for 14C dating of other carbohydrate polymers such as chitin.

Type
I. Becoming Better
Information
Radiocarbon , Volume 43 , Issue 2A: Proceedings of the 17th International Radiocarbon Conference (Part 1 of 3) , 2001 , pp. 209 - 215
Copyright
Copyright © The Arizona Board of Regents on behalf of the University of Arizona 

References

Hardy, MR, Townsend, RR. 1988. Separation of positional isomers of oligosaccharides and glycopeptides by high-performance anion-exchange chromatography with pulsed amperometric detection. Proceedings of the National Acadamy of Science USA 85:3289–93.Google Scholar
Hardy, MR, Townsend, RR, Lee, YC. 1988. Monosaccharide analysis of glycoconjugates by anion exchange chromatography with pulsed amperometric detection. Analytical Biochemistry 170:54–62.CrossRefGoogle ScholarPubMed
Hedges, REM, Law, I, Bronk, CR, Housley, RA. 1989. The Oxford accelerator mass spectrometry facility: technical developments in routine dating. Archaeometry 31(2):99–114.Google Scholar
Ramsey, CB Humm, MJ. On-line combustion of samples for AMS and ionsource developments at ORAU. Proceedings of the 8th International Conference on Accelerator Mass Spectrometry, 6–10 September 1999, Vienna, Austria. In press.Google Scholar
Sahasrabudhe, NA, Lachke, AH, Ranjekar, PK. 1987. Characterisation of the purified multifunctional cellulase component of Penicillium funiculosum. Biotechnology Letters 9(12):881–6.Google Scholar
Schimmelmann, A, DeNiro, MJ. 1983. Determination of carbon isotope ratios in plant starch via selective enzymatic hydrolysis. Analytical Chemistry 55:814–6.Google Scholar
Schimmelmann, A, DeNiro, MJ. 1986. Stable isotopic studies on chitin. I. Measurements on chitin/chitosan isolates and D-glucosamine hydrochloride from chitin. In: Muzzarelli, RAA, Jeuniaux, C, Gooday, GW, editors. Chitin in nature and technology. New York: Plenum. p 357–64.Google Scholar
Stafford, TW, Duhamel, RC, Haynes, CV Jr, Brendel, K. 1982. Isolation of proline and hydroxyproline from fossil bone. Life Sciences 31:931–8.Google Scholar
van Klinken, GJ, Hedges, REM. 1992. Experiments on 14C dating of contaminated bone using peptides resulting from enzymatic cleavage of collagen. Radiocarbon 34(3):292–5.Google Scholar
Wood, TM, McCrae, SI, MacFarlane, CC. 1980. The isolation, purification and properties of the cellobiohydrolase component of Penicillium funiculosum cellulase. Biochemical Journal 189:51–6.CrossRefGoogle ScholarPubMed
Wood, TM, McCrae, SI. 1982. Purification and some properties of a (1→4)-β-D-glucan glucohydrolase associated with the cellulase from the fungus Penicillium funiculosum Carbohydrate Research 110:291–303.Google Scholar