Skip to main content
Log in

Developing FTIR Microspectroscopy for the Analysis of Animal-Tissue Residues on Stone Tools

  • Published:
Journal of Archaeological Method and Theory Aims and scope Submit manuscript

Abstract

The analysis of microscopic residues on stone tools provides one of the most direct ways to reconstruct the functions of such artifacts. However, new methods are needed to strengthen residue identifications based upon visible-light microscopy. In this work, we establish that reflectance Fourier-transform infrared microspectroscopy (FTIRM) can be used to document IR spectra of animal-tissue residues on experimental stone tools. First, we present a set of reflectance FTIRM standards for the most commonly identified animal-tissue residues on stone tools: skin, meat, fat, hair, blood, feather barbules, fish scales, and bone. We provide spectral peak assignments for each residue and demonstrate that high-quality reflectance FTIRM spectra can be generated under ideal circumstances. Second, we document the spectra for these residues when they are located on a stone substrate such as flint or obsidian. We discuss procedures for correcting spectra that are affected by specular reflection and explain the effects of spectral interference from the stone. Our results show that reflectance FTIRM is sensitive to small intra-sample differences in composition. This means that it will record the effects of decomposition in ancient residues. The methodological developments we present here will help lithic residue analysts incorporate in situ reflectance FTIRM into their analysis protocols to strengthen identifications.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11
Fig. 12
Fig. 13
Fig. 14
Fig. 15
Fig. 16
Fig. 17

Similar content being viewed by others

References

  • Acerbo, A. S., Carr, G. L., Judex, S., & Miller, L. M. (2012). Imaging the material properties of bone specimens using reflection-based infrared microspectroscopy. Analytical Chemistry, 84, 3607–3613.

    Article  Google Scholar 

  • Baddiel, C. B. (1968). Structure and reactions of human hair keratin: an analysis by infrared spectroscopy. Journal of Molecular Biology, 38, 181–199.

    Article  Google Scholar 

  • Barth, A. (2007). Infrared spectroscopy of proteins. Biochimica et Biophysica Acta, 1767, 1073–1101.

    Article  Google Scholar 

  • Calabrò, E., & Magazù, S. (2014). Non-thermal effects of microwave oven heating on ground beef meat studied in the mid-infrared region by Fourier transform infrared spectroscopy. Spectroscopy Letters, 47, 649–656.

    Article  Google Scholar 

  • Cesaro, S. N., & Lemorini, C. (2012). The function of prehistoric lithic tools: a combined study of use-wear analysis and FTIR microspectroscopy. Spectrochimica Acta Part A-Molecular and Biomolecular Spectroscopy, 86, 299–304.

    Article  Google Scholar 

  • Chapman, R. E. (1986). Chapter 17: hair, wool, quill, nail, claw, hoof, and horn, in part VI: the skin of mammals. In J. Bereiter-Hahn, A. G. Matoltsy, & K. Sylvia Richards (Eds.), Biology of the integument, part 2: vertebrates (pp. 293–317). Berlin: Springer-Verlag.

    Chapter  Google Scholar 

  • Croft, S., Monnier, G. F., Radini, A., Little, A., & Milner, N. (2016). Lithic residue survival and characterisation at Star Carr: a burial experiment. Internet Archaeology, 1–88.

  • Crowther, A. Haslam, M., Oakden, N., Walde, D., & Mercader, J. (2014). Documenting contamination in ancient starch laboratories. Journal of Archaeological Science, 49, 90–104.

  • Deleris, G., & Petibois, C. (2003). Applications of FT-IR spectrometry to plasma contents analysis and monitoring. Vibrational Spectroscopy, 32, 129–136.

    Article  Google Scholar 

  • Duarte, R., Simoes, M., & Sgarbieri, V. (1999). Bovine blood components: fractionation, composition, and nutritive value. Journal of Agricultural and Food Chemistry, 47, 231–236.

    Article  Google Scholar 

  • Feughelman, M. (1997). Mechanical properties and structure of alpha-keratin fibres: wool, human hair, and related fibres. Sydney: University of New South Wales Press.

    Google Scholar 

  • Finlayson, C., Brown, K., Blasco, R., Rosell, J., Jose Negro, J., Bortolotti, G. R., et al. (2012). Birds of a feather: Neanderthal exploitation of raptors and corvids. PloS One, 7, e45927.

    Article  Google Scholar 

  • Fowler, B. O. (1974). Infrared studies of apatites. I. Vibrational assignments for calcium, strontium, and barium hydroxyapatites utilizing isotopic substitution. Inorganic Chemistry, 13(1), 194–207.

    Article  Google Scholar 

  • Fraser, R. D. B., & Parry, D. A. D. (2008). Molecular packing in the feather keratin filament. Journal of Structural Biology, 162, 1–13.

    Article  Google Scholar 

  • Fronticelli, C., Sanna, M., Perez-Alvarado, G., Karavitis, M., Lu, A., & Brinigar, W. (1995). Allosteric modulation by tertiary structure in mammalian hemoglobins—introduction of the functional characteristics of bovine hemoglobin into human hemoglobin by five amino acid substitutions. Journal of Biological Chemistry, 270, 30588–30592.

    Article  Google Scholar 

  • Greenwold, M. J., & Sawyer, R. H. (2011). Linking the molecular evolution of avian beta (beta) keratins to the evolution of feathers. Journal of Experimental Zoology Part B-Molecular and Developmental Evolution, 316B, 609–616.

    Article  Google Scholar 

  • Gregg, K., & Rogers, G. E. (1986). Chapter 33: feather keratin: composition, structure and biogenesis, in part XIII: skin proteins. In J. Bereiter-Hahn, A. G. Matoltsy, & K. Sylvia Richards (Eds.), Biology of the integument, part 2: vertebrates (pp. 666–694). Berlin: Springer-Verlag.

    Chapter  Google Scholar 

  • Hardy, B. L. (2004). Neanderthal behaviour and stone tool function at the middle Palaeolithic site of La Quina, France. Antiquity, 78, 547–565.

    Article  Google Scholar 

  • Hardy, B. L., & Moncel, M. (2011). Neanderthal use of fish, mammals, birds, starchy plants and wood 125-250,000 years ago. PloS One, 6, e23768.

    Article  Google Scholar 

  • Hardy, B. L., Kay, M., Marks, A. E., & Monigal, K. (2001). Stone tool function at the Paleolithic sites of Starosele and Buran Kaya III, Crimea: behavioral implications. Proceedings of the National Academy of Sciences of the United States of America, 98, 10972–10977.

    Article  Google Scholar 

  • Hardy, B. L., Bolus, M., & Conard, N. J. (2008). Hammer or crescent wrench? Stone-tool form and function in the Aurignacian of southwest Germany. Journal of Human Evolution, 54, 648–662.

    Article  Google Scholar 

  • Hardy, B.L., Moncel, M.-., Daujeard, C., Fernandes, P., Béarez, P., Desclaux, E., et al., 2013. Impossible Neanderthals? Making string, throwing projectiles and catching small game during marine isotope stage 4 (Abri du Maras, France). Quaternary Science Reviews, 82, 23–40.

    Article  Google Scholar 

  • Hortolà, P. (2016). Human bloodstains on bone artefacts: an SEM intra- and inter-sample comparative study using ratite bird tibiotarsus. Micron, 90, 108–113.

    Article  Google Scholar 

  • Ikoma, T., Kobayashi, H., Tanaka, J., Walsh, D., & Mann, S. (2003). Microstructure, mechanical, and biomimetic properties of fish scales from Pagrus major. Journal of Structural Biology, 142, 327–333.

    Article  Google Scholar 

  • Jung, C. (2000). Insight into protein structure and protein-ligand recognition by Fourier transform infrared spectroscopy. Journal of Molecular Recognition, 13, 325–351.

    Article  Google Scholar 

  • Kolczyńska-Szafraniec, U., & Bilińska, B. (1993). Infrared studies of natural pheomelanins. Current Topics in Biophysics, 16(2), 77–80.

    Google Scholar 

  • Kreplak, L., Doucet, J., Dumas, P., & Briki, F. (2004). New aspects of the alpha-helix to beta-sheet transition in stretched hard alpha-keratin fibers. Biophysical Journal, 87, 640–647.

    Article  Google Scholar 

  • Langejans, G. (2010). Remains of the day—preservation of organic micro-residues on stone tools. Journal of Archaeological Science, 37, 971–985.

    Article  Google Scholar 

  • Langejans, G. (2011). Discerning use-related micro-residues on tools. Testing the multi-stranded approach for archaeological studies. Journal of Archaeological Science, 38, 985–1000.

    Article  Google Scholar 

  • Lieber, R. L. (2002). Skeletal muscle structure, function, and plasticity. In The physiological basis of rehabilitation (2nd ed.). Philadelphia: Lippincott, Williams, & Wilkins.

    Google Scholar 

  • Lombard, M. (2005). Evidence of hunting and hafting during the Middle Stone age at Sibidu Cave, KwaZulu-Natal, South Africa: a multianalytical approach. Journal of Human Evolution, 48, 279–300.

    Article  Google Scholar 

  • Lombard, M. (2008). Finding resolution for the Howiesons Poort through the microscope: micro-residue analysis of segments from Sibudu Cave, South Africa. Journal of Archaeological Science, 35, 26–41.

    Article  Google Scholar 

  • Lombard, M. (2011). Quartz-tipped arrows older than 60 ka: further use-trace evidence from Sibudu, KwaZulu-Natal, South Africa RID A-2444-2012. Journal of Archaeological Science, 38, 1918–1930.

    Article  Google Scholar 

  • Lombard, M. (2014). In situ presumptive test for blood residues applied to 62,000-year-old stone tools. South African Archaeological Bulletin, 69, 80–86.

    Google Scholar 

  • Loy, T., & Dixon, E. (1998). Blood residues on fluted points from eastern Beringia. American Antiquity, 63, 21–46.

    Article  Google Scholar 

  • Loy, T. H., & Hardy, B. L. (1992). Blood residue analysis of 90,000-year-old stone tools from Tabun Cave, Israel. Antiquity, 66, 24–35.

    Article  Google Scholar 

  • Matoltsy, A. G. (1986a). Chapter 14 in biology of the integument 2: vertebrates. In J. Bereiter-Hahn, A. G. Matoltsy, & K. Syvia Richards (Eds.), Structure and function of the mammalian epidermis (pp. 255–271). Berlin: Springer-Verlag.

    Google Scholar 

  • Matoltsy, A. G. (1986b). Chapter 15 in biology of the integument 2: vertebrates. In J. Bereiter-Hahn, A. G. Matoltsy, & K. Syvia Richards (Eds.), Dermis (pp. 272–277). Berlin: Springer-Verlag.

    Google Scholar 

  • Miyazawa, T., Shimanouchi, T., & Mizushima, S. (1958). Normal vibrations of N-methylacetamide. The Journal of Chemical Physics, 29(3), 611–616.

    Article  Google Scholar 

  • Monnier, G. F., Ladwig, J. L., & Porter, S. T. (2012). Swept under the rug: the problem of unacknowledged ambiguity in lithic residue identification. Journal of Archaeological Science, 39, 3284–3300.

    Article  Google Scholar 

  • Monnier, G. F., Hauck, T. C., Feinberg, J. M., Luo, B., Le Tensorer, J., & al Sakhel, H. (2013). A multi-analytical methodology of lithic residue analysis applied to Paleolithic tools from Hummal, Syria. Journal of Archaeological Science, 40, 3722–3739.

    Article  Google Scholar 

  • Monnier, G. F., Frahm, E., Luo, B., & Missal, K. (2017). Developing FTIR microspectroscopy for analysis of plant residues on stone tools. Journal of Archaeological Science, 78, 158–178.

    Article  Google Scholar 

  • Morin, E., & Laroulandie, V. (2012). Presumed symbolic use of diurnal raptors by Neanderthals. PloS One, 7, e32856.

    Article  Google Scholar 

  • Pearson, J. F., & Slifkin, M. A. (1972). The infrared spectra of amino acids and dipeptides. Spectrochimica Acta, Vol., 28A, 2408–2417.

    Google Scholar 

  • Pedergnana, A., Asryan, L., Fernández-Marchena, J. L., &. Ollé, A. (2016). Modern contaminants affecting microscopic residue analysis on stone tools: A word of caution. Micron, 86, 1–21.

  • Powell, B. C., & Rogers, G. E. (1986). Chapter 34: hair keratin: composition, structure, and biogenesis, in part XIII: skin proteins. In J. Bereiter-Hahn, A. G. Matoltsy, & K. Sylvia Richards (Eds.), Biology of the integument, part 2: vertebrates (pp. 695–721). Berlin: Springer-Verlag.

    Chapter  Google Scholar 

  • Prinsloo, L. C., Wadley, L., & Lombard, M. (2014). Infrared reflectance spectroscopy as an analytical technique for the study of residues on stone tools: potential and challenges. Journal of Archaeological Science, 41, 732–739.

    Article  Google Scholar 

  • Rahmania, H., Sudjadi, & Rohman, A. (2015). The employment of FTIR spectroscopy in combination with chemometrics for analysis of rat meat in meatball formulation. Meat Science, 100, 301–305.

    Article  Google Scholar 

  • Rao, C. N. R., & Venkataraghavan, R. (1963). Contribution to the infrared spectra of five-membered N- and N,S-heterocyclic compounds. Canadian Journal of Chemistry, 42, 43–49.

    Article  Google Scholar 

  • Rehman, I., & Bonfield, W. (1997). Characterization of hydroxyapatite and carbonated apatite by photo acoustic FTIR spectroscopy. Journal of Materials Science-Materials in Medicine, 8, 1–4.

    Article  Google Scholar 

  • Robertson, G., Attenbrow, V., & Hiscock, P. (2009). Multiple uses for Australian backed artefacts. Antiquity, 83, 296–308.

    Article  Google Scholar 

  • Sobolik, K. (1996). Lithic organic residue analysis: an example from the southwestern archaic. Journal of Field Archaeology, 23, 461–469.

    Google Scholar 

  • Solodenko, N., Zupancich, A., Cesaro, S. N., Marder, O., Lemorini, C., & Barkai, R. (2015). Fat residue and use-wear found on Acheulian Biface and scraper associated with butchered elephant remains at the site of Revadim, Israel. PloS One, 10,(3): e0118572. doi:10.1371/journal.pone.0118572.

  • Stephenson, B. (2015). A modified Picro-Sirius Red (PSR) staining procedure with polarization microscopy for identifying collagen in archaeological residues. Journal of Archaeological Science, 61, 235–243.

    Article  Google Scholar 

  • Stettenheim, P. (2000). The integumentary morphology of modern birds—an overview. American Zoologist, 40, 461–477.

    Google Scholar 

  • Stiner, M., Weiner, S., Bar-Yosef, O., & Kuhn, S. L. (1995). Differential burning, fragmentation, and preservation of archaeological bone. Journal of Archaeological Science, 22, 223–237.

    Article  Google Scholar 

  • Stiner, M., Kuhn, S., Surovell, T., Goldberg, P., Meignen, L., Weiner, S., et al. (2001). Bone preservation in Hayonim Cave (Israel): a macroscopic and mineralogical study. Journal of Archaeological Science, 28, 643–659.

    Article  Google Scholar 

  • Suzuki, S., Ohshima, T., Tamiya, N., Fukushima, K., Shimanouchi, T., & Mizushima, S. (1959). Infrared spectra of deuterated α-amino acids NH3 +CDRCOO-. Assignment of the absorption bands of α-alanine. Spectrochimica Acta, 11, 969–976.

    Article  Google Scholar 

  • Uitto, J. (1986). Chapter 40 in biology of the integument 2: vertebrates. In J. Bereiter-Hahn, A. G. Matoltsy, & K. Syvia Richards (Eds.), Interstitial collagens (pp. 800–809). Berlin: Springer-Verlag.

    Google Scholar 

  • Vlachos, N., Skopelitis, Y., Psaroudaki, M., Konstantinidou, V., Chatzilazarou, A., & Tegou, E. (2006). Applications of Fourier transform-infrared spectroscopy to edible oils. Analytica Chimica Acta, 573, 459–465.

    Article  Google Scholar 

  • Warriss, P. D. (2010). Meat science, 2nd edition. In An introductory text. Oxfordshire, U.K.: CABI.

    Google Scholar 

  • Weiner, S. (2010). Microarchaeology: beyond the visible archaeological record. Cambridge, U.K.: Cambridge University Press.

    Book  Google Scholar 

  • Williamson, B. (2004). Middle stone age tool function from residue analysis at Sibudu Cave. South African Journal of Science, 100, 174–178.

    Google Scholar 

  • Zylberberg, L., Bereiter-Hahn, J., & Sire, J.-Y. (1988). Cytoskeletal organization and collagen orientation in the fish scales. Cell and Tissue Research, 253, 597–607.

    Article  Google Scholar 

Download references

Acknowledgements

This work was funded by NSF grant # BCS-1420702. It was carried out at the University of Minnesota in the Evolutionary Anthropology Laboratories and in the Characterization Facility, which receives partial support from the NSF through the MRSEC program. Many thanks to Matt Edling, Greg Haugstad, Keith Manthie, Colin McFadden, Marjorie Schalles, Nora Last, Kara Kersteter, and Gil Tostevin. Thanks also to the three anonymous reviewers whose comments and suggestions helped improve the final manuscript.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Gilliane Monnier.

Electronic supplementary material

Figure S1

FTIRM spectra of feather calamus residues obtained in reflectance mode (on a mirrored slide) and in transmission mode (on a NaCl plate), compared with an FTIR standard for keratin (prepared using the KBr pellet method) from the Kimmel Center for Archeological Science, Weizmann Institute of Science. Peak assignments are in Table 4. The reflectance spectrum (red line) is similar to the transmission spectrum (green line); both clearly exhibit the protein peaks seen in the keratin standard (blue line). The upside-down peaks on the reflectance spectrum (red line) at ~2350 cm−1 are due to atmospheric CO2 and should be ignored. Note: all spectra are graphed in calculated absorbance mode (log(1/R)). (PDF 264 kb)

Figure S2

The FTIRM spectrum for skin residue on obsidian with the Kramers-Kronig transform applied (blue line), compared with the skin reflectance standard (purple line, graphed in calculated absorbance mode [log(1/R)]). Above 1250 cm−1, the residue on obsidian spectrum exhibits peaks at the same locations as on the standard, although relative peak heights and the baseline are distorted. Peaks in the range of 1750–1200 cm−1 are also shifted to lower wavenumbers. Below 1250 cm−1, (to the right of the dashed line), however, the spectrum exhibits severe derivative peaks. Peak assignments are in Table 1. (PDF 217 kb)

Figure S3

The FTIRM spectra for feather calamus residue on English flint and obsidian compared with the feather calamus reflectance standard. Between 4000 and 1350 cm−1, the residue on flint is consistent with the calamus reference spectrum, although the latter exhibits a small carbonyl peak (7) and more absorbance in the CH3 stretching regions (peaks 4 and 6), which indicates compositional differences between the two. Below 1350 cm−1, as with almost all other spectra of animal residues on stone, the peaks no longer match, and derivative features are seen. The spectrum of the residue on obsidian differs because it exhibits very low reflectance, thereby resulting in low resolution. Finally, for both flint and obsidian spectra, many of the peaks are shifted to higher wavenumbers, relative to the standard, an important effect which needs to be taken into account (see peak assignments in Table 4). Note: all spectra are graphed in calculated absorbance mode (log(1/R)). (PDF 324 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Monnier, G., Frahm, E., Luo, B. et al. Developing FTIR Microspectroscopy for the Analysis of Animal-Tissue Residues on Stone Tools. J Archaeol Method Theory 25, 1–44 (2018). https://doi.org/10.1007/s10816-017-9325-3

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10816-017-9325-3

Keywords

Navigation