Collection Citation: Samantha Brown, Sandra Hebestreit, Naihui Wang, Nicole Boivin, Katerina Douka, Kristine Korzow Richter 2020. Zooarchaeology by Mass Spectrometry (ZooMS)- Pretreatment protocols for bone material. protocols.io https://dx.doi.org/10.17504/protocols.io.bf5djq26
Manuscript citation:
Wang, N; Brown, S; Richter, K. K; Ditchfield, P; Hebsetreit, S; Kozilikin, M; Luu, S; Wedage, O; Grimaldi, S; Chazen, M; Horwitz, K. L; Spriggs, M; Summerhayes, G; Shunkov, M; Douka, K. (2020). Testing the efficacy and comparability of ZooMS protocols on archaeological bone. Under review.
License: This is an open access collection distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol in our group and it is working. It is a part of a collection of several ZooMS protocols.
Created: May 07, 2020
Last Modified: June 29, 2020
Collection Integer ID: 36741
Keywords: ZooMS, Zooarchaeology, Archaeology, mass spectrometry, MALDI, peptide mass fingerprinting, collagen, protein extraction, bone,
Abstract
This collection details some of the different established protocols for Zooarchaeology by Mass Spectrometry (ZooMS) for use on archaeological bone. ZooMS allows for taxonomic identification by the peptide mass fingerprinting of collagen type I. These protocols can be used individually or combined depending on the preservation, sample size, and ability to do destructive analysis. All protocols are optimized for bone as the starting material.
The AmBic protocol can be used on samples where distructive analysis cannot be undertaken. Samples are pretreated by soaking in ammonium bicarbonate at room temperature followed by a brief heating step to "melt" a small amount of collagen out of the bone. The bone can then be dried at room temperature.
Both acid based protocols are destructive as the samples are pretreated with hydrocholric acid to demineralize the bone. In the acid soluble protocol the acid is removed and the collagen is filtered from the acid. In the acid insoluble protocol the bone shaddow that remains after demineralization is washed to remove the acid and then heated in ammonium bicarbonate to gelitinize the collagen. Both acid based protocols can be done on the same sample in conjunction with each other and then either analyzed separately on the MALDI or combined before analysis.
In all protocols the extracted collagen is then digested with trypsin and the peptides are purified using C18 ZipTips.
If you are using any of the protocols please cite the DOI for the protocol, the following paper, and any papers in the additional individual protocols:
Buckley, M., Collins, M., Thomas-Oates, J., & Wilson, J. C. (2009). Species identification by analysis of bone collagen using matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. Rapid Communications in Mass Spectrometry: RCM, 23(23), 3843–3854. https://doi.org/10.1002/rcm.4316
Image created by Kristine Korzow Richter. Rat by Rebecca Groom from phylopic.org, cow by Alessandro Suraci and scissors by Lluisa Iborra from thenounproject.com, collagen from smart.servier.com.
