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

      Hodgins, Gregory W. L.; Thorpe, J. L.; Coope, G. R.; Hedges, Robert E. M. (Department of Geosciences, The University of Arizona, 2001-01-01)
      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.
    • Sample Preparation of Dissolved Organic Carbon in Groundwater for AMS 14C Analysis

      Burr, George S.; Thomas, J. M.; Reines, D.; Jeffrey, D.; Courtney, C.; Jull, A. J. Timothy; Lange, Todd (Department of Geosciences, The University of Arizona, 2001-01-01)
      This study describes a sample preparation technique used to isolate dissolved organic carbon (DOC) in groundwater for radiocarbon analysis using accelerator mass spectrometry (AMS). The goal of the work is to improve our ability to determine groundwater residence times based on 14C measurements of the DOC fraction in groundwater. Water samples were collected from carbonate and volcanic rock aquifers in southern Nevada. Multiple measurements of total dissolved organic carbon (TDOC) in groundwater from one site are used to demonstrate the reproducibility of the analytical procedure. The reproducibility of the method is about one percent (1sigma) for a 0.5 mg sample. The procedural blank for the same size sample contains about 1 percent modern carbon (pMC).
    • The Chemical and Enzymatic Hydrolysis of Archaeological Wood Cellulose and Monosaccharide Purification by High pH Anion Exchange Chromatography for Compound-Specific Radiocarbon Dating

      Hodgins, Gregory L.; Butters, T. D.; Bronk Ramsey, Christopher; Hedges, Robert E. M. (Department of Geosciences, The University of Arizona, 2001-01-01)
      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.