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

IF 1.8 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS Rapid Communications in Mass Spectrometry Pub Date : 2024-08-01 DOI:10.1002/rcm.9874

Kelly R. Bowen, Carolyn M. Kurle

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Abstract

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 (δ¹³C) and nitrogen (δ¹⁵N) 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 δ¹³C and δ¹⁵N 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 δ¹⁵N values were observed for any of the experimental protocols. However, the δ¹³C 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 δ¹⁵N values from killer whale bone are of interest for analysis, no pre-treatment seems necessary. If the δ¹³C 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.

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处理前、历史年龄和样本特征对虎鲸（Orcinus orca）骨骼稳定同位素分析的影响。

理由：通过对骨骼进行稳定同位素分析，可以深入了解动物的觅食情况，并随着时间的推移进行生态重建，但要分离胶原蛋白，需要进行预处理。预处理通常包括去矿物质以去除无机成分和/或提取脂质以去除脂肪，但这些方法会因涉及的化学物质、组织和/或物种的不同而对稳定碳（δ13C）和氮（δ15N）同位素值产生不同的影响。针对不同物种的方法为各项研究的可比性提供了标准，并加深了对从现代鲸类骨骼中分离胶原蛋白的理解：方法：使用元素分析仪和同位素比质谱仪测量虎鲸（Orcinus orca）骨骼粉末的 δ13C 和 δ15N 值，这些骨骼粉末既可以是完整的（对照组），也可以在三种实验条件中选择一种：去矿物质、脂质萃取以及去矿物质和脂质萃取。此外，还对 C:N 比率进行了评估，作为胶原蛋白纯度的替代指标。最后，研究了对照组 C:N 比率与历史年龄之间的相关性，以及对照组 C:N 比率与样品特征之间的相关性：任何实验方案的 δ15N 值均无明显差异。但是，δ13C 值在所有三种实验方案中都有显著增加：脱矿、脂质提取和两种处理方法的结合。影响最大的实验方案是脱矿和脂质提取。C:N比率的测量值也因去矿物质化和两种处理方法的结合而显著降低，表明处理后的材料更接近于纯胶原蛋白。通过 C:N 比值显示的胶原蛋白纯度与历史年龄或样本特征无关：结论：如果只对虎鲸骨骼的 δ15N 值感兴趣，则无需进行预处理。如果对 δ13C 值感兴趣，则应对样本进行脱矿和脂质提取。由于历史年代和样本特征与样本污染无关，因此可以对所有样本一视同仁。

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来源期刊

Rapid Communications in Mass Spectrometry 化学-分析化学

CiteScore

4.10

自引率

5.00%

发文量

219

审稿时长

2.6 months

期刊介绍： Rapid Communications in Mass Spectrometry is a journal whose aim is the rapid publication of original research results and ideas on all aspects of the science of gas-phase ions; it covers all the associated scientific disciplines. There is no formal limit on paper length ("rapid" is not synonymous with "brief"), but papers should be of a length that is commensurate with the importance and complexity of the results being reported. Contributions may be theoretical or practical in nature; they may deal with methods, techniques and applications, or with the interpretation of results; they may cover any area in science that depends directly on measurements made upon gaseous ions or that is associated with such measurements.