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Effects of demineralization on the stable isotope analysis of bone samples Public Deposited

https://ir.library.oregonstate.edu/concern/articles/ns064779c

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Abstract
  • Rationale: The sampling of sequential, annually formed bone growth layers for stable carbon (δ¹³C values) and nitrogen (δ¹⁵N values) isotope analysis (SIA) can provide a time series of foraging ecology data. To date, no standard protocol exists for the pre-SIA treatment of cortical samples taken from fresh, modern, bones. Methods: Based on the SIA of historical bone, it is assumed that fresh bone samples must be pre-treated with acid prior to SIA. Using an elemental analyzer coupled to an isotope ratio mass spectrometer to measure stable carbon and nitrogen ratios, we tested the need to acidify cortical bone powder with 0.25 M HCl prior to SIA to isolate bone collagen for the determination of δ¹³C and δ¹⁵ values. We also examined the need for lipid extraction to remove potential biases related to δ¹³C analysis, based on a C:N ratio threshold of 3.5. Results: It was found that acidification of micromilled cortical bone samples from marine turtles does not affect their δ¹⁵N values, and the small effect acidification has on δ¹³C values can be mathematically corrected for, thus eliminating the need for pre-SIA acidification of cortical bone. The lipid content of the cortical bone samples was low, as measured by their C:N ratios, indicating that lipid extracting cortical bone samples from modern marine turtles is unnecessary. Conclusions: We present a standard protocol for testing fresh, modern cortical bone samples prior to SIA, facilitating direct comparison of future studies. Based on the results obtained from marine turtle bones, pre-acidification and lipid removal of cortical bone are not recommended. This is especially useful as there is frequently not enough bone material removed via micromilling of sequential growth layers to accommodate both acid treatment and SIA.
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  • Tomaszewicz, T., Calandra, N., Seminoff, J. A., Ramirez, M. D., & Kurle, C. M. (2015). Effects of demineralization on the stable isotope analysis of bone samples. Rapid Communications in Mass Spectrometry, 29(20), 1879-1888. doi:10.1002/rcm.7295
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  • 29
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  • 20
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  • This work was supported by NOAA Fisheries, and C.T.T. was supported by the University of California San Diego (UCSD), NIH T32 GM007240 Cell and Molecular Genetics Training Program, Jeanne Messier Memorial Fellowship, ARCS Foundation Scholarship, and a Hearts de Vite Research Grant. M.R. was supported by the Living Marine Resources Cooperative Science Center as part of the NOAA Educational Partnership Program and the NSF Graduate Research Fellowship Program. Additional support was provided by the US Fish & Wildlife Service, NMFS-SWFSC, Western Pacific Fisheries Management Council (NOAA Grant FNA05NMF4411092), and the David and Lucile Packard Foundation. For samples originating in Mexico, all research activities and permits were authorized by the Mexican government through SEMARNAP and SEMARNAT permits 150496-213-03, 280597-213-03, 190698-213-03, 280499-213-03, 280700-213-03, SGPA/DGVS/002 4661, SGPA/DGVS/10358, SGPA/ DGVS/03501/06, SGPA/DGVS/03406/07, SGPA/DGVS/03481/09, SGPA/DGVS/04990/10, and SGPA/DGVS/04568/11. All exported and imported CITES regulated samples were authorized by the Mexican government through export permit numbers MX-58124 and MX-64301, and the United States government through import permit numbers 11US844694/9 and 12US844694/9. Atlantic samples all originated from the US and samples were obtained under USFWS Permit # TE-676379-5.
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  • description.provenance : Made available in DSpace on 2015-10-29T15:16:52Z (GMT). No. of bitstreams: 2 RamirezMatthewFishWildlifeEffectsDemineralizationStable.pdf: 722525 bytes, checksum: f3370f73373a456e905c7a90422de8dd (MD5) RamirezMatthewFishWildlifeEffectsDemineralizationStableSupplementaryMaterial.pdf: 241412 bytes, checksum: 82695f1817a4b810ff44e34fe9d954a1 (MD5) Previous issue date: 2015-10-30
  • description.provenance : Approved for entry into archive by Patricia Black(patricia.black@oregonstate.edu) on 2015-10-29T15:16:52Z (GMT) No. of bitstreams: 2 RamirezMatthewFishWildlifeEffectsDemineralizationStable.pdf: 722525 bytes, checksum: f3370f73373a456e905c7a90422de8dd (MD5) RamirezMatthewFishWildlifeEffectsDemineralizationStableSupplementaryMaterial.pdf: 241412 bytes, checksum: 82695f1817a4b810ff44e34fe9d954a1 (MD5)
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