Comparison of two methods of extracting bone collagen for stable carbon and nitrogen isotope analysis: comparing whole bone demineralization with gelatinization and ultrafiltration

Comparison of two methods of extracting bone collagen for stable carbon and nitrogen isotope analysis: comparing whole bone demineralization with gelatinization and ultrafiltration
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两种提取骨胶原进行稳定碳氮同位素分析的方法比较:全骨脱矿与糊化和超滤的比较

DOI:
10.1016/j.jas.2014.04.011
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发表时间:
2014
影响因子:
2.8
通讯作者:
Nehlich O
Nehlich O
中科院分区:
地球科学2区
文献类型:
--
作者:
Sealy J;Johnson M;Richards MP;Nehlich O

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我们比较了两种分离骨胶原用于稳定碳和氮同位素分析的方法。较旧的方法(如开普敦大学所采用的)对骨“块”进行脱矿,而较新的方法(如莱比锡马克斯·普朗克进化人类学研究所所采用的)涉及脱矿、糊化和超滤,以仅选择较高分子量的蛋白质片段进行同位素分析。后一种方法是针对有问题(即保存不良)的样品而开发的,虽然它更严格,但它也更加昂贵且更加劳动密集。我们的目的是找出使用这两种方法从保存相对完好的骨骼中分离出的骨胶原的 δ13C 和 δ15N 是否存在差异。我们的样本集包含来自南非南部和西部地区的 5 块现代动物和 47 块考古动物和人类骨骼。考古标本的年龄从几百年到大约六千年不等。提取胶原蛋白,使用 %C、%N 和 C:N 评估其质量,并在两个实验室独立测量 δ13C 和 δ15N 值。两个实验室的 δ13C 和 δ15N 值组之间没有统计学上的显着差异。对于保存相对完好的骨骼,胶原蛋白制备的“块”方法仍然是用于 C 和 N 同位素分析的更复杂的胶原蛋白提取方案的可接受的替代方案。
We compare two methods of isolating bone collagen for stable carbon and nitrogen isotope analysis. The older method (as practised at the University of Cape Town) demineralizes bone ‘chunks’, while the newer method (as practised at the Max Planck Institute for Evolutionary Anthropology in Leipzig) involves demineralization, gelatinization and ultra-filtration to select only higher molecular weight protein fragments for isotopic analysis. The latter method was developed for problematic (i.e. poorly-preserved) samples and while it is more rigorous, it is also significantly more expensive and more labor-intensive. Our aim is to find out whether there is any difference between the δ13C and δ15N of bone collagen isolated from relatively well-preserved bones using the two methods. Our sample set consists of 5 modern and 47 archaeological animal and human bones from the southern and western parts of South Africa. Archaeological specimens range in age from a few hundred to approximately six thousand years old. Collagen was extracted, its quality assessed using %C, %N and C:N, and δ13C and δ15N values measured independently in both laboratories. There are no statistically significant differences between the sets of δ13C and δ15N values from the two laboratories. For relatively well-preserved bones, the ‘chunk’ method of collagen preparation continues to be an acceptable alternative to more sophisticated collagen extraction protocols for C and N isotope analysis.
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