• A Fresh Water Diet-Derived 14C Reservoir Effect at the Stone Age Sites in the Iron Gates Gorge

      Cook, Gordon T.; Bonsall, C.; Hedges, Robert E. M.; McSweeney, K.; Boronean, V.; Pettitt, Paul B. (Department of Geosciences, The University of Arizona, 2001-01-01)
      Human bones from single inhumation burials and artifacts made from terrestrial mammal (ungulate) bone found in direct association with the skeletons were obtained from the Stone Age site of Schela Cladovei situated just below the Iron Gates Gorge of the River Danube. The results of stable isotope analyses of the human bone collagen are consistent with a heavy dependence on aquatic protein while radiocarbon dating of the samples reveals an offset of 300-500 years between the two sample types, indicating a freshwater reservoir effect in the human bone samples. Since protein consumption is by far the major source of nitrogen in the human diet we have assumed a linear relationship between delta-15N and the level of aquatic protein in each individual's diet and derived a calibration for 14C age offset versus delta-15N which has been applied to a series of results from the site at Lepenski Vir within the gorge. The corrected 14C ages (7310-6720 BP) are now consistent with the previous 14C age measurements made on charcoal from related contexts (7360-6560 BP). In addition, the data indicate a change from a primarily aquatic to a mixed terrestrial/aquatic diet around 7100 BP and this may be argued as supporting a shift from Mesolithic to Neolithic. This study also has wider implications for the accurate dating of human bone samples when the possibility exists of an aquatic component in the dietary protein and strongly implies that delta-15N analysis should be undertaken routinely when dating human bones.
    • 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.