Stable carbon and oxygen isotope analysis of human and animal tooth enamel carbonate has been used as a proxy for individual diet and environmental reconstruction. Here, we provide a detailed description and visual documentation of bulk and sequential tooth enamel sampling as well as pretreatment of archaeological and paleontological samples.
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Method Article
Sampling and Pretreatment of Tooth Enamel Carbonate for Stable Carbon and Oxygen Isotope Analysis
In This Article
Summary
Abstract
Stable carbon and oxygen isotope analysis of human and animal tooth enamel carbonate has been applied in paleodietary, paleoecological, and paleoenvironmental research from recent historical periods back to over 10 million years ago. Bulk approaches provide a representative sample for the period of enamel mineralization, while sequential samples within a tooth can track dietary and environmental changes during this period. While these methodologies have been widely applied and described in archaeology, ecology, and paleontology, there have been no explicit guidelines to aid in the selection of necessary lab equipment and to thoroughly describe detailed laboratory sampling and protocols. In this article, we document textually and visually, the entire process from sampling through pretreatment and diagenetic screening to make the methodology more widely available to researchers considering its application in a variety of laboratory settings.
Introduction
Stable carbon and oxygen isotope analyses of tooth enamel carbonate has been used to study past human dietary intake, weaning, and mobility, as well as faunal reliance on vegetation, the movement of animals, and livestock foddering. These applications have been comprehensively discussed and reviewed for a variety of environmental conditions indicating the effects of local aridity, temperature, water sources, and vegetation compositions1,2,3,4,5,6. The diversity of potentia....
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Protocol
The following protocol follows the guidelines of the Light Isotope Mass Spectrometry Laboratory at the Max Planck Institute for the Science of Human History. Appropriate ethics permissions from national and international committees should be sought for analyses involving endangered modern or historical faunal specimens, and for the use of archaeological and faunal material of interest to contemporary stakeholders. In this paper, the samples used were archaeological and fossil specimens. No living humans were used in this study and full ethical, institutional, and governmental permissions have been gained for any destructive analysis.
1. Bulk ....
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Results
Using the sampling procedure presented above, incremental enamel bioapatite samples were prepared. The analysis of bioapatite in enamel depends on the accuracy of sampling, whether bulk or incremental. In this case, we have chosen to present the results of archaeological samples (two sheep) from different climatic zones. Incremental samples were analyzed from sheep second molars and labelled starting at the ERJ (Figure 4). Incremental sample locations were nu.......
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Discussion
The challenges of successful sampling (bulk and incremental) of dentition relies on the access to knowledge regarding drilling techniques and sample preparation, alongside the investment in relatively inexpensive equipment. These challenges are easily surmountable when clear instructions are available concerning sampling and pretreatment approaches. In this article, we hope to have disseminated these in a clear, concise fashion for researchers new to these methods. Scholars applying these methods for the first time .......
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Disclosures
The authors declare no conflict of interest.
Acknowledgements
We would like to thank the Max Planck Society for funding this research as well as the recent setting up of a Stable Isotope laboratory at the Department Archaeology, Max Planck Institute for the Science of Human History.
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Materials
| Name | Company | Catalog Number | Comments |
|---|---|---|---|
| Dremel Micro | Dremel | https://www.dremel.com/en_US/products/-/show-product/tools/8050-micro | |
| Diamond-tipped drill bit | Dremel | https://www.dremel.com/en_US/products/-/show-product/accessories/7122-diamond-wheel-point | |
| 1.5 mL micro-centrifuge tube | Sigma Aldrich | https://www.sigmaaldrich.com/catalog/product/sigma/t2422?lang=de®ion=DE&gclid=EAIaIQ obChMI7pHRpauW2QIV77ftCh1p1 wjhEAAYASAAEgKzkvD_BwE | |
| Methanol | Linear Formula: CH3OH | ||
| Acetic Acid | Linear Formula: CH3CO2H | ||
| Dremel rig set-up (workstation) | Dremel | https://www.dremel.com/en_US/products/-/show-product/tools/220-01-workstation | |
| Microcentrifuge | Thermo Scientific | http://www.thermofisher.com/order/catalog/product/75002401 | |
| Mini-centrifuge | Sprout | http://www.heathrowscientific.com/sprout-mini-centrifuge-4 | |
| Freeze drier | Zirbus Technology | http://www.zirbus.com |
References
- Balasse, M. Reconstructing dietary and environmental history from enamel isotopic analysis: time resolution of intra-tooth sequential sampling. International Journal of Osteoarchaeology. 12 (3), 155-165 (2002).
- Balasse, M. Potential biases in sampling design and interpretation of intra-tooth isotope analysis. International Journal of Osteoarchaeology. 13 (1-2), 3-10 (2003).
- Lee-Thorp, J. A.
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