Zooarchaeology by Mass Spectrometry (ZooMS) for bone material - Acid insoluble protocol v1

Abstract

This collection details 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 the protocols are optimized for bone as the starting material. In the acid insoluble protocol bone is pretreated with hydrocholoric acid. The resulting collagen bone shaddow is washed to remove the acid and then heated in ammonium bicarbonate to gelitinize the collagen. The extracted collagen is then digested with trypsin and the peptides are purified using C18 ZipTips. This protocol can be used in conjuntion with the acid soluble protocol which analyzes the acid used to pretreat the samples. This protocol is suitable for a wide range of preservation conditions from very good to poor preservation. The ideal starting volume is 10-20mg of bone as a powerder or a bone chip. However, smaller sample sizes have yieled high quality results depending on preservation condidtions. This protocol can be modifed for high throughput in 96 well plates. If you are using this protocol, please cite the DOI for the protocol and the following two papers on which it is based: 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 Welker, F., Soressi, M., Rendu, W., Hublin, J.-J., & Collins, M. (2015). Using ZooMS to identify fragmentary bone from the Late Middle/Early Upper Palaeolithic sequence of Les Cottés, France. Journal of Archaeological Science, 54, 279–286. https://doi.org/10.1016/j.jas.2014.12.010