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How Clean is Ultrafiltration Cleaning of Bone Collagen?

  • Matthias C Hüls (a1), Pieter M Grootes (a1) and Marie-Josée Nadeau (a1)

Abstract

As part of our bone dating development, we have tested the ultrafiltration of bone gelatin using 2 different filters—Vivaspin 20™ (VS20), a polyethersulfone, and Vivaspin 15R™ (VS15R), a cellulose, both with a 30,000 molecular weight cutoff—and bone collagen from dated samples ranging in age from 1.5 to >50 kyr BP. A direct accelerator mass spectrometry (AMS) measurement yielded radiocarbon concentrations of ∼0.5 pMC (∼42 kyr) for the polyethersulfone, ∼14.4–17.5 pMC (∼15.6–14 kyr) for the cellulose, and ∼107.4 pMC for the glycerin. The filters were cleaned before use similar to the Oxford protocol (Bronk Ramsey et al. 2004), and a series of freeze-dried archaeological bone gelatin samples and a modern pig-skin gelatin were passed through VS20 and VS15R filters (Vivascience™). We recovered both the eluent (<30-kD fraction) and the liquid that stayed above the filter (>30 kD) in order to obtain a carbon mass and isotope balance. While the >30-kD collagen fraction that is usually selected for AMS analysis does not appear to be significantly contaminated, measurements show significant age differences between the eluent <30 kD and the unfiltered bone collagen, indicating that, despite cleaning, both glycerin and filter still give off contaminants in the eluent. Ultrafiltration with young collagen from pig skin generally confirms these results for the <30-kD fraction but also shows the possibility of small contaminations in the >30-kD fraction. Until a contamination with filter carbon of the >30-kD collagen fraction can be excluded, we would recommend caution in the use of ultrafiltration for cleaning bone collagen with VS20 or VS15R ultrafilters.

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Copyright

Corresponding author

Corresponding author. Email: mhuels@leibniz.uni-kiel.de

References

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Brock, F, Bronk Ramsey, C, Higham, TFG. 2007. Quality assurance of ultrafiltered bone dating. Radiocarbon , these proceedings.
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.
Grootes, PM, Nadeau, M-J, Rieck, A. 2004. 14C-AMS at the Leibniz-Labor: radiometric dating and isotope research. Nuclear Instruments and Methods in Physics Research B 223–224:5561.
Longin, R. 1970. Extraction du collagène des os fossiles pour leur datation par la méthode du carbone 14 [PhD dissertation]. Lyon: Université de Lyon. In French.
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.

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