Effects of pre-treatment, historical age, and sample characteristics on the stable isotope analyses of killer whale (Orcinus orca) bone.

Publisher: John Wiley And Sons Ltd Country of Publication: England NLM ID: 8802365 Publication Model: Print Cited Medium: Internet ISSN: 1097-0231 (Electronic) Linking ISSN: 09514198 NLM ISO Abbreviation: Rapid Commun Mass Spectrom Subsets: MEDLINE

Publication: Chichester : John Wiley And Sons Ltd
Original Publication: London, UK : Heyden, c1987-

Carbon Isotopes*/analysis ; Nitrogen Isotopes*/analysis ; Whale, Killer* ; Bone and Bones*/chemistry ; Mass Spectrometry*/methods ; Collagen*/analysis ; Collagen*/chemistry; Animals ; Lipids/analysis ; Lipids/chemistry

Rationale: Stable isotope analysis of bone provides insight into animal foraging and allows for ecological reconstructions over time, however pre-treatment is required to isolate collagen. Pre-treatments typically consist of demineralization to remove inorganic components and/or lipid extraction to remove fats, but these protocols can differentially affect stable carbon (δ ¹³ C) and nitrogen (δ ¹⁵ N) isotope values depending on the chemicals, tissues, and/or species involved. Species-specific methodologies create a standard for comparability across studies and enhance understanding of collagen isolation from modern cetacean bone.
Methods: Elemental analyzers coupled to isotope ratio mass spectrometers were used to measure the δ ¹³ C and δ ¹⁵ N values of powdered killer whale (Orcinus orca) bone that was intact (control) or subjected to one of three experimental conditions: demineralized, lipid-extracted, and both demineralized and lipid-extracted. Additionally, C:N ratios were evaluated as a proxy for collagen purity. Lastly, correlations were examined between control C:N ratios vs. historical age and control C:N ratios vs. sample characteristics.
Results: No significant differences in the δ ¹⁵ N values were observed for any of the experimental protocols. However, the δ ¹³ C values were significantly increased by all three experimental protocols: demineralization, lipid extraction, and both treatments combined. The most influential protocol was both demineralization and lipid extraction. Measures of the C:N ratios were also significantly lowered by demineralization and both treatments combined, indicating the material was closer to pure collagen after the treatments. Collagen purity as indicated via C:N ratio was not correlated with historical age nor sample characteristics.
Conclusions: If only the δ ¹⁵ N values from killer whale bone are of interest for analysis, no pre-treatment seems necessary. If the δ ¹³ C values are of interest, samples should be both demineralized and lipid-extracted. As historical age and specimen characteristics are not correlated with sample contamination, all samples can be treated equally.
(© 2024 The Author(s). Rapid Communications in Mass Spectrometry published by John Wiley & Sons Ltd.)

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University of California, San Diego (UCSD); NIH Cell & Molecular Genetics Graduate Training Program; PADI Foundation; Lerner-Gray Fund for Marine Research by the American Museum of Natural History; Jeanne Marie Messier Memorial Endowed Fund (UCSD); American Cetacean Society Los Angeles Chapter John E. Heyning Research Grant; Association for Women in Science San Diego Chapter

0 (Carbon Isotopes)
0 (Nitrogen Isotopes)
9007-34-5 (Collagen)
0 (Lipids)

Date Created: 20240801 Date Completed: 20240801 Latest Revision: 20240801

20240802

10.1002/rcm.9874

39089821