Eastern Michigan University DigitalCommons@EMU Senior Honors eses Honors College 2009 Qualitative THM-GC-MS Analysis of Proteins and Amino Acids in Rock Painting Binders Melissa Doolin Eastern Michigan University Follow this and additional works at: hp://commons.emich.edu/honors Part of the Chemistry Commons is Open Access Senior Honors esis is brought to you for free and open access by the Honors College at DigitalCommons@EMU. It has been accepted for inclusion in Senior Honors eses by an authorized administrator of DigitalCommons@EMU. For more information, please contact lib- [email protected]. Recommended Citation Doolin, Melissa, "Qualitative THM-GC-MS Analysis of Proteins and Amino Acids in Rock Painting Binders" (2009). Senior Honors eses. 213. hp://commons.emich.edu/honors/213
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Eastern Michigan UniversityDigitalCommons@EMU
Senior Honors Theses Honors College
2009
Qualitative THM-GC-MS Analysis of Proteins andAmino Acids in Rock Painting BindersMelissa DoolinEastern Michigan University
Follow this and additional works at: http://commons.emich.edu/honors
Part of the Chemistry Commons
This Open Access Senior Honors Thesis is brought to you for free and open access by the Honors College at DigitalCommons@EMU. It has beenaccepted for inclusion in Senior Honors Theses by an authorized administrator of DigitalCommons@EMU. For more information, please contact [email protected].
Recommended CitationDoolin, Melissa, "Qualitative THM-GC-MS Analysis of Proteins and Amino Acids in Rock Painting Binders" (2009). Senior HonorsTheses. 213.http://commons.emich.edu/honors/213
Qualitative THM-GC-MS Analysis of Proteins and Amino Acids in RockPainting Binders
AbstractA vital step to radiocarbon dating rock paintings involves the characterization of organic material in the paint.Using THM-GC-MS we have created a library of mass spectrometric data on possible organic binders, tocompare to authentic paint samples. To reduce interference from contaminants and the mineral background,we are developing a simple purification technique to extract proteins and carbohydrates from these samples.Separating the organics can greatly enhance our results for GC-MS identification and subsequent radiocarbonanalysis by accelerator mass spectroscopy.
Degree TypeOpen Access Senior Honors Thesis
DepartmentChemistry
Subject CategoriesChemistry
This open access senior honors thesis is available at DigitalCommons@EMU: http://commons.emich.edu/honors/213
QUALITATIVE THM-GC-MS ANALYSIS OF PROTEINS AND AMINO ACIDS IN ROCK PAINTING BINDERS
by
Melissa Doolin
A Senior Thesis Submitted to the
Eastern Michigan University
Honors Program
In Partial Fulfillment of the Requirements for Graduation
With Honors in Professional Biochemistry
Approved at Ypsilanti, Michigan on this date ________________________________
_________________________________ _________________________________ Supervising Instructor Department Head _________________________________ _________________________________ Honors Advisor Honors Director
However, the absence of measurable amounts of any of the amino acids analyzed
is a significant concern. The most obvious conclusion would be that proteins are not
present in the paint samples or have deteriorated below detectable quantities. In none of
the samples is protein the last remaining possibility: the blind study samples have not
been analyzed for fatty acids, and all the samples have yet to be tested for carbohydrates.
Still, there are a number of reasons why we should not be hasty to discount proteinaceous
media as the carbon source in the paints.
Firstly, the clean-up technique utilized in this study was developed successfully
for identification of protein binders in museum art samples, which have the advantage of
being more recent and being housed in an environment favorable for preservation.
Should any proteins be present in our samples, the pipette-tip method may not be suitable
for such a small amount of material. There may be similar concerns in regard to the
method of analysis, that GC-MS may not be sensitive to miniscule sample quantities.
Tandem mass spectrometry such as MS/MS provides greater sensitivity and is used
extensively in peptide sequencing. Mori et al. [21] found Asp, Ala, Leu and Val, as well
as trace amounts of Thr and Gly by using electrospray ionization tandem mass
spectrometry for pictograph samples from Central Sahara.
Yet even more powerful methods exist in the field of proteomics that have been
used for over a decade, specifically peptide-mass fingerprinting [22]. Proteins are not
hydrolyzed in acid, but cleaved using a digestive enzyme, such as trypsin or
chymotrypsin. Resulting peptide fragments can be separated using liquid
chromatography and subsequently analyzed using a matrix-assisted laser
desorption/ionization mass spectrometer with a tandem time-of-flight analyzer, or
MALDI-TOF-TOF. This procedure has been applied by E. Cappellini et al. [23] for the
successful sequencing and identification of synthesized paints of known composition that
also underwent rapid ageing under UV light. Primary proteins of milk, glue, and egg-
based binders were identified using Swissprot software, which could account for amino
acid alterations due to hydrolytic and oxidative degradation. This is particularly
important, as the amino acids in this study were specifically selected by mass; any
chemical alterations due to ageing would render them invisible in SIS mode. Using
proteomics with powerful software that can take into account certain chemical changes
has great potential for ancient art samples, which have not yet been attempted using the
abovementioned procedure. This may be where the future of paint binder analysis lies,
and the techniques developed for cell biology may have a place in our study of the past.
ACKNOWLEDGEMENTS
The author would like to thank Marvin Rowe and Sara Scott for providing paint
samples; Sigma Xi, The Scientific Research Society for Grant-In-Aid of Research; as
well as the Eastern Michigan University Honors College and Chemistry Department.
REFERENCES
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