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Analysis of Bone “Collagen” Extraction Products for Radiocarbon Dating

  • F Brock (a1), V Geoghegan (a2), B Thomas (a2), K Jurkschat (a3) and T F G Higham (a1)...


Archaeological bones are now routinely dated in many radiocarbon laboratories through the extraction of “collagen.” Methods for “collagen” extraction vary, and several laboratories now apply an ultrafiltration step after gelatinization to extract the higher molecular weight (usually >10 or 30kDa) fraction for dating, thereby removing low molecular weight contaminants. Ultrafiltration has been demonstrated to result in products that are easier to handle and have more acceptable C:N ratios, and in some instances can result in significantly improved (generally older) 14C dates when compared to non-ultrafiltered products from the same bone. Although it has been suggested that ultrafiltration removes potential contaminants such as short-chain degraded collagen and other peptides and amino acids, fulvic acids, and salts, there remains little published evidence to support this. This paper presents data from a pilot study investigating the most suitable techniques with which to study the products of the routine “collagen” extraction procedures employed at the Oxford Radiocarbon Accelerator Unit (ORAU) (modified Longin followed by ultrafiltration). The preliminary data demonstrates that the final product of “collagen” extraction at ORAU appears to be an aggregate consisting of a range of proteins of different molecular weights, including collagen, as well as some other organic matter and inorganic species. Ultrafiltration is removing some, but not all, of the <30kDa fraction from the samples. Further work to investigate the nature of this aggregate and how best to improve the efficiency of “collagen” extraction procedures is discussed.


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Ambrose, SH. 1990. Preparation and characterization of bone and tooth collagen for isotopic analysis. Journal of Archaeological Science 17(4):431–51.
Arslanov, KhA, Svezhentsev, YuS. 1993. An improved method for radiocarbon dating fossil bones. Radiocarbon 35(3):387–91.
Beaumont, W, Beverly, R, Southon, J, Taylor, RE. 2010. Bone preparation at the KCCAMS laboratory. Nuclear Instruments and Methods in Physics Research B 268(7–8):906–9.
Brock, F, Bronk Ramsey, C, Higham, T. 2007. Quality assurance of ultrafiltered bone dating. Radiocarbon 49(2):187–92.
Brock, F, Higham, T, Ditchfield, P, Bronk Ramsey, C. 2010. Current pretreatment methods for AMS radiocarbon dating at the Oxford Radiocarbon Accelerator Unit (ORAU). Radiocarbon 52(1):103–12.
Bronk Ramsey, C, Pettitt, PB, Hedges, REM, Hodgins, GWL, Owen, DC. 2000. Radiocarbon dates from the Oxford AMS system: Archaeometry dateiist 30. Archaeometry 42(2):459–79.
Bronk Ramsey, C, Higham, T, Bowles, A, Hedges, R. 2004. Improvements to the pretreatment of bone at Oxford. Radiocarbon 46(1):155–63.
Brown, TA, Nelson, DE, Vogel, JS, Southon, JR. 1988. Improved collagen extraction by modified Longin method. Radiocarbon 30(2):171–7.
Burba, P, Shkinev, V, Spivakov, BY. 1995. On-line fractionation and characterization of aquatic humic substances by means of sequential-stage ultrafiltration. Fresenius' Journal of Analytical Chemistry 351(1):7482.
Burns, DB, Zydney, AL. 1999. Effect of solution pH on protein transport through ultrafiltration membranes. Biotechnology and Bioengineering 64(1):2737.
Collins, MJ, Riley, MS, Child, AM, Turner-Walker, G. 1995. A basic mathematical simulation of the chemical degradation of ancient collagen. Journal of Archaeological Science 22(2):175–83.
Collins, MJ, Nielsen-Marsh, CM, Hiller, J, Smith, CI, Roberts, JP, Progodich, RV, Wess, TJ, Csapò, J, Millard, AR, Turner-Walker, G. 2002. The survival of organic matter in bone: a review. Archaeometry 44(3):383–94.
DeNiro, MJ, Weiner, S. 1988. Chemical, enzymatic and spectroscopic characterization of “collagen” and other organic fractions from prehistoric bones. Geochimica et Cosmochimica Acta 52(9):2197–206.
Di Lullo, GA, Sweeney, SM, Körkkö, J, Ala-Kokko, L, San Antonio, JD. 2002. Mapping the ligand-binding sites and disease-associated mutations on the most abundant protein in the human, type I collagen. The Journal of Biological Chemistry 277(6):4223–31.
Dobberstein, RC, Collins, MJ, Craig, OE, Taylor, G, Penkman, KEH, Ritz-Timme, S. 2009. Archaeological collagen: Why worry about collagen diagenesis? Archaeological and Anthropological Sciences 1(1):3142.
Fernández, J, Soria, J, Viseras, C. 1996. Stratigraphic architecture of the Neogene basins in the central sector of the Betic Cordillera (Spain): tectonic control and base-level changes. In: Friend, PF, Dabrio, CJ, editors. Tertiary Basins of Spain: The Stratigraphic Record of Crustal Kinematics. Cambridge: Cambridge University Press. p 353–65.
Fülöp, RH, Rethmeyer, J, Heinze, S, König, S. 2013. Why ultrafiltration is neither the problem nor the solution. Radiocarbon, these proceedings, doi:10.2458/azu_js_rc.55.16296.
Furthmayr, H, Timpl, R. 1971. Characterization of collagen peptides by sodium dodecyl sulfate-polyacrylamide electrophoresis. Analytical Biochemistry 41:510–6.
Ghabbour, EA, Davies, G. 2009. Spectrophotometric analysis of fulvic acid solutions – a second look. Annals of Environmental Science 3:131–8.
Gillespie, R. 1989. Fundamentals of bone degradation chemistry: collagen is not “the way.” Radiocarbon 31(3):239–46.
Havers, N, Burba, P, Lambert, J, Klockow, D. 1998. Spectroscopic characterization of humic-like substances in airborne particulate matter. Journal of Atmospheric Chemistry 29:4554.
Hedges, REM, Law, IA. 1989. The radiocarbon dating of bone. Applied Geochemistry 4:249–53.
Hedges, REM, van Klinken, GJ. 1992. A review of current approaches in the pretreatment of bone for radiocarbon dating by AMS. Radiocarbon 34(3):279–91.
Higham, TFG, Jacobi, RM, Bronk Ramsey, C. 2006. AMS radiocarbon dating of ancient bone using ultrafiltration. Radiocarbon 48(2):179–95.
Hüls, CM, Grootes, PM, Nadeau, M-J. 2009. Ultrafiltration: boon or bane? Radiocarbon 51(2):613–25.
Jacobi, RM, Higham, TFG, Bronk Ramsey, C. 2006. AMS radiocarbon dating of Middle and Upper Palaeolithic bone in the British Isles: improved reliability using ultrafiltration. Journal of Quaternary Science 21(5):557–73.
Jørkov, MLS, Heinemeier, J, Lynnerup, N. 2007 Evaluating bone collagen extraction methods for stable isotope analysis in dietary studies. Journal of Archaeological Science 34(11):1824–9.
Longin, R. 1971. New method of collagen extraction for radiocarbon dating. Nature 230(5291):241–2.
MacCarthy, P, Rice, JA. 1985. Spectroscopic methods (other than NMR) for determining functionality in humic substances. In: Aiken, GR, MacCarthy, P, McKnight, DS, Wershaw, RL, editors. Humic Substances in Soil, Sediment, and Water. Chichester: Wiley. p 457–76.
Marshall, PD, Meadows, J, Bayliss, A, Sparks, R, Bronk Ramsey, C, Beavan Athfield, N. 2010. Scientific dating. In: Ferris, I, editor. The Beautiful Rooms are Empty: excavations at Binchester Roman Fort, County Durham, 1976–1981 and 1986–1991. p 527–38. Durham: Durham County Council.
Maspero, F, Sala, S, Fedi, ME, Martini, M, Papagni, A. 2011. A new procedure for extraction of collagen from modern and archaeological bones for 14C dating. Analytical and Bioanalytical Chemistry 401(6):2019–23.
Piez, KA. 1984. Extracellular Matrix Biochemistry. New York: Elsevier.
Prigodich, RV, Vesely, MR. 1997. Characterization of the complex between bovine osteocalcin and type I collagen. Archives of Biochemistry and Biophysics 345:339–41.
Semal, P, Orban, R. 1995. Collagen extraction from recent and fossil bones: quantitative and qualitative aspects. Journal of Archaeological Science 22(4):463–7.
Termine, JD. 1984. The tissue-specific proteins of the bone matrix. In: Butler, WT, editor. The Chemistry and Biology of Mineralized Tissues. Birmingham: Ebsco Media Inc. p 94–7.
van Klinken, GJ. 1999. Bone collagen quality indicators for palaeodietary and radiocarbon measurements. Journal of Archaeological Science 26(6):687–95.
van Klinken, GJ, Mook, WG. 1990. Preparative high-performance liquid chromatographic separation of individual amino acids derived from fossil bone collagen. Radiocarbon 32(2):155–64.
Wilson, J, van Doorn, NL, Collins, MJ. 2012. Assessing the extent of bone degradation using glutamine deamidation in collagen. Analytical Chemistry 84(21):9041–8.
Wood, RE, Barroso-Ruíz, C, Caparrós, M, Jordá Prado, J, Galván Santos, B, Davidson, I, Higham, TFG. 2013. Radiocarbon dating casts doubt on the late chronology of the Middle to Upper Palaeolithic transition in southern Iberia. Proceedings of the National Academy of Sciences of the USA 110(8):2781–6.
Yuan, S, Wu, X, Gao, S, Wang, J, Cai, L, Liu, K, Li, K, Ma, H. 2000. Comparison of different bone pretreatment methods for AMS 14C dating. Nuclear Instruments and Methods in Physics Research B 172(1–4):424–7.
Yuan, S, Wu, X, Liu, K, Guo, Z, Cheng, X, Pan, Y, Wang, J. 2007. Removal of contaminants from oracle bones during sample pretreatment. Radiocarbon 49(2):211–6.
Zhang, Z, Li, G, Shi, B. 2006. Physicochemical properties of collagen, gelatin and collagen hydrolysate derived from bovine limed split wastes. Journal of the Society of Leather Technologists and Chemists 90(1):23–8.

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Analysis of Bone “Collagen” Extraction Products for Radiocarbon Dating

  • F Brock (a1), V Geoghegan (a2), B Thomas (a2), K Jurkschat (a3) and T F G Higham (a1)...


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