Preparation of bone powder for FTIR-ATR analysis: The particle size effect |
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| Affiliation: | 1. BioArCh, Department of Archaeology, University of York, United Kingdom;2. BioArCh, Department of Chemistry, University of York, United Kingdom;3. Natural History Museum, University of Copenhagen, Denmark;1. Center for Lasers and Applications, Nuclear and Energy Research Institute, IPEN – CNEN, 05508-000, Brazil;2. Center for Radiation Technology, Nuclear and Energy Research Institute, IPEN – CNEN, 05508-000, Brazil;1. Department of Historical Geology and Palaeontology, Faculty of Geology and Geoenvironment, National and Kapodistrian University of Athens, Panepistimiopolis, 15784, Zografou, Greece;2. Wiener Laboratory, American School of Classical Studies at Athens ASCSA, 54 Souidias st., 10676, Athens, Greece;1. Laboratory of Histo-Embryology and Cytogenetics, Medicine Faculty of Sfax University, 3029, Sfax, Tunisia;2. Univ Rennes, CNRS, ISCR-UMR 6226, F-35000, Rennes, France;3. Laboratory of Animal Physiology, Sciences Faculty of Sfax University, 3064, Sfax, Tunisia;4. Laboratory of General Biology, Department of Biology, University of Ha’il, 81451, Ha’il, Saudi Arabia;5. Laboratory of Histology - Cytology, Medicine Faculty of Tunis, University of Tunis El Manar, 1007, La Rabta-Tunis, Tunisia;1. Institut Français d’Archéologie Orientale (IFAO), 37 al-Cheikh Aly Youssef Street, B.P. Qasr el-Ayni, 11652, 11441, Cairo, Egypt;2. Muséum National d’Histoire Naturelle, CNRS, UPVD, Histoire Naturelle de l’Homme Préhistorique (HNHP), UMR 7194, Musée de l’Homme, 17 Place du Trocadéro, 75116, Paris, France;3. Sorbonne Université, CNRS, Interactions et Spectroscopies, MONARIS, UMR 8233, de la Molécule aux Nano-objets: Réactivité, 4 Place Jussieu, 75005, Paris, France;4. University of Basel, Department of Ancient Civilisations, Petersgraben 51, 4051, Basel, Switzerland;1. Department of Archaeology, Faculty of Arts, University of Ljubljana, Zavetiška 5, 1000 Ljubljana, Slovenia;2. Institute of Forensic Medicine, Faculty of Medicine, Korytkova 2, 1000 Ljubljana, Slovenia;3. University Medical Centre Ljubljana, Zaloška 7, 1000 Ljubljana, Slovenia;4. National Institute of Chemistry, Hajdrihova 19, 1000, Ljubljana, Slovenia;5. Department of Archaeology, Faculty of Arts, University of Ljubljana, Zavetiška 5, 1000 Ljubljana, Slovenia;1. Max Planck Institute for the Science of Human History, Jena, Germany;2. School of Archaeology, University of Oxford, Oxford, UK;3. Faculty of Arts, Masaryk University, Czech Republic;4. University of Colorado, Boulder, USA |
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| Abstract: | Fourier transform infrared (FTIR) spectroscopy using attenuated total reflection (ATR) is commonly used for the examination of bone. During sample preparation bone is commonly ground, changing the particle size distribution. Although previous studies have examined changes in crystallinity caused by the intensity of grinding using FTIR, the effect of sample preparation (i.e. particle size and bone tissue type) on the FTIR data is still unknown.This study reports on the bone powder particle size effects on mid-IR spectra and within sample variation (i.e. periosteal, mesosteal, trabecular) using FTIR-ATR. Twenty-four archaeological human and faunal bone samples (5 heated and 19 unheated) of different chronological age (Neolithic to post-Medieval) and origin (Belgium, Britain, Denmark, Greece) were ground using either (1) a ball-mill grinder, or (2) an agate pestle and mortar, and split into grain fractions (>500 μm, 250–500 μm, 125–250 μm, 63–125 μm, and 20–63 μm).Bone powder particle size has a strong but predictable effect on the infrared splitting factor (IRSF), carbonate/phosphate (C/P) ratio, and amide/phosphate (Am/P) values. The absorbance and positions of the main peaks, the 2nd derivative components of the phosphate and carbonate bands, as well as the full width at half maximum (FWHM) of the 1010 cm−1 phosphate peak are particle size dependent. This is likely to be because of the impact of the particle size on the short- and long-range crystal order, as well as the contact between the sample and the prism, and hence the penetration depth of the IR light. Variations can be also observed between periosteal, cortical and trabecular areas of bone. We therefore propose a standard preparation method for bone powder for FTIR-ATR analysis that significantly improves accuracy, consistency, reliability, replicability and comparability of the data, enabling systematic evaluation of bone in archaeological, anthropological, paleontological, forensic and biomedical studies. |
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| Keywords: | Bone FTIR-ATR Sample preparation Particle size Bioapatite Crystal order/disorder |
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