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Home / Equine Research Bank / J Sep Sci / Proteotypic peptides of hairs for the identification of comm...
Journal of separation science2023; 46(13); e2300064; doi: 10.1002/jssc.202300064

Proteotypic peptides of hairs for the identification of common European domestic and wild animal species revealed by in-sample protein digestion and mass spectrometry analysis.

Abstract: The aim of this work is to offer an alternative or complementary analytical tool to the time-consuming and expensive methods commonly used for the recognition of animal species according to their hair. The paper introduces a simple and fast way for species differentiation of animal hairs called in-sample digestion. A total of 10 European animal species, including cat, cow, common degu, dog, fallow deer, goat, horse, sika deer, rabbit, roe deer, and 17 different breeds of dogs were examined using specific tryptic cleavage directly in hair followed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry and liquid chromatography-electrospray ionization quadrupole time of flight. Principal component analysis was used for the subsequent mass spectrometric data evaluation. This novel approach demonstrates the ability to distinguish among individual animal species, which is supported by finding characteristic m/z values obtained by the mass spectrometry for each animal species. The approach was successfully tested on two "blind" samples. On the other hand, the attempt to distinguish among hairs of different dog breeds has not been successful due to the very similar protein composition and their amino acid sequences.
© 2023 Wiley-VCH GmbH.
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Publication Date: 2023-05-03 PubMed ID: 37084407DOI: 10.1002/jssc.202300064Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This study presents an innovative method for identifying the species of European domestic and wild animals based on their hair. Using a method known as in-sample digestion, the researchers studied hair samples from 10 specific species and 17 different dog breeds, differing them by using a combined technique of mass spectrometry and liquid chromatography.

Research Methodology

  • The objective of the study was to create a simpler and faster tool that could be used to identify different species based on their hair. The tool was designed as an alternative to the existing time-consuming and costly methods.
  • They used a process called ‘in-sample digestion’ for species differentiation of animal hairs. This involved using specific tryptic cleavage directly in the hair sample.
  • Researchers studied the hair of ten specific European animal species including cat, cow, common degu, dog, fallow deer, goat, horse, sika deer, rabbit, and roe deer, and 17 different breeds of dogs.
  • The samples were analyzed using a technique combining matrix-assisted laser desorption/ionization-time of flight mass spectrometry and liquid chromatography-electrospray ionization quadrupole time of flight.
  • Subsequently, the mass spectrometric data were studied using Principal Component Analysis.

Results

  • The approach was successful in discerning between individual animal species thanks to the characteristic mass-to-charge ratio (m/z) value obtained by mass spectrometry for each animal species.
  • The researchers were able to accurately identify the species from two ‘blind’ samples, demonstrating the method’s effectiveness.
  • However, the study was unable to differentiate between hairs from different dog breeds, attributed to the similar protein composition and their amino acid sequences.

Implications

  • This work offers a new potential tool for species identification based on hair samples. This may find various applications, notably in ecology, wildlife management, and forensic biology.
  • The study did not conclude positively for the differentiation of various dog breeds. This reveals the complexity in distinguishing among closely related breeds or species and suggests the need for further research to include a broad range of genetic variability.

Cite This Article

APA
Kuckova S, Smirnova TA, Straka D, Meledina A, Santrucek J, Humpolakova K, Hoskova M, Cejnar P, Hynek R. (2023). Proteotypic peptides of hairs for the identification of common European domestic and wild animal species revealed by in-sample protein digestion and mass spectrometry analysis. J Sep Sci, 46(13), e2300064. https://doi.org/10.1002/jssc.202300064

Publication

Journal of separation science
ISSN: 1615-9314
NlmUniqueID: 101088554
Country: Germany
Language: English
Volume: 46
Issue: 13
Pages: e2300064

Researcher Affiliations

Kuckova, Stepanka
  • Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Czech Republic.
  • Department of Chemistry and Chemistry Education, Charles University, Prague, Czech Republic.
Smirnova, Tatiana Anatolievna
  • Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Czech Republic.
  • Department of Chemistry and Chemistry Education, Charles University, Prague, Czech Republic.
Straka, David
  • Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Czech Republic.
Meledina, Alena
  • Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Czech Republic.
Santrucek, Jiri
  • Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Czech Republic.
Humpolakova, Karin
  • Department of Chemistry and Chemistry Education, Charles University, Prague, Czech Republic.
Hoskova, Martina
  • Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Czech Republic.
Cejnar, Pavel
  • Department of Mathematics, Informatics and Cybernetics, University of Chemistry and Technology, Prague, Czech Republic.
Hynek, Radovan
  • Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Czech Republic.

MeSH Terms

  • Animals
  • Dogs
  • Rabbits
  • Horses
  • Animals, Wild
  • Proteolysis
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods
  • Deer
  • Peptides / chemistry
  • Proteins / analysis
  • Hair / chemistry
  • Spectrometry, Mass, Electrospray Ionization / methods

References

This article includes 51 references
  1. Tarditi CR, Grahn RA, Evans JJ, Jeffrey J, Kurushima D, Lyons LA. Mitochondrial DNA sequencing of cat hair: an informative forensic tool.. J Forensic Sci 2011;56(1):S36-46.
  2. Sato I, Nakaki S, Murata K, Takeshita H, Mukai T. Forensic hair analysis to identify animal species on a case of pet animal abuse.. Int J Legal Med 2010;124:249-56.
  3. Tridico S. Examination, analysis, and application of hair in forensic science - animal hair.. Forensic Sci Rev 2005;17(1):17-28.
  4. Spangenberg JE, Ferrer M, Tschudin P, Volken M, Hafner A. Microstructural, chemical and isotopic evidence for the origin of late neolithic leather recovered from an ice field in the Swiss Alps.. J Archeol Sci 2010;37(8):1851-65.
  5. Hollemeyer K, Altmeyer W, Heinzle E, Pitra C. Species identification of Oetzi's clothing with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry based on peptide pattern similarities of hair digests.. Rapid Commun Mass Spectrom 2008;22(18):2751-67.
  6. Hollemeyer K, Altmeyer W, Heinzle E, Pitra C. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry combined with multidimensional scaling, binary hierarchical cluster tree and selected diagnostic masses improves species identification of Neolithic keratin sequences from furs of the Tyrolean iceman Oetzi.. Rapid Commun Mass Spectrom 2012;26(16):1735-45.
  7. Solazzo C. Follow-up on the characterization of peptidic markers in hair and fur for the identification of common North American species.. Rapid Commun. Mass Spectrom 2017;31(17):1375-84.
  8. . Regulation (EC) No 1523/2007 of the European Parliament and of the Council of 11 December 2007 banning the placing on the market and the import to, or export from, the Community of cat and dog fur, and products containing such fur.. .
  9. de Oliveira CD R, Yonamine M, de Moraes Moreau RL. Headspace solid-phase microextraction of cannabinoids in human head hair samples.. J Sep Sci 2007;30(1):128-34.
  10. Naddaf E, Ebrahimi M, Es'haghi Z, Bamoharram FF. Application of carbon nanotubes modified with a Keggin polyoxometalate as a new sorbent for the hollow-fiber micro-solid-phase extraction of trace naproxen in hair samples with fluorescence spectrophotometry using factorial experimental design.. J Sep Sci 2015;38(13):2348-56.
  11. Karačonji I B, Zimić L, Brajenović N, Skender L. Optimisation of a solid-phase microextraction method for the analysis of nicotine in hair.. J Sep Sci 2011;34(19):2726-31.
  12. Xiao P, Bao C, Jia Q, Su R, Zhou W, Jia J. Determination of nitroanilines in hair dye using polymer monolith microextraction coupled with HPLC.. J Sep Sci 2011;34(6):675-80.
  13. Bagheri H, Zavareh A F, Koruni M H. Graphene oxide assisted electromembrane extraction with gas chromatography for the determination of methamphetamine as a model analyte in hair and urine samples.. J Sep Sci 2016;39(6):1182-8.
  14. Yu W, Cai W, Shao X. Chemometric approach for fast analysis of prometryn in human hair by GC-MS.. J Sep Sci 2013;36(14):2277-82.
  15. Štefan J, Hladík J. Soudní lékařství a jeho moderní trendy.. 1st ed. Praha: Grada Publishing; 2012.
  16. Tridico SR, Houck MM, Kirkbride KP, Smith ME, Yates BC. Morphological identification of animal hairs: myths and misconceptions, possibilities and pitfalls.. Forensic Sci Int 2014;238:101-7.
  17. Sahajpal V, Goyal SP. Identification of a forensic case using microscopy and forensically informative nucleotide sequencing (FINS): a case study of small Indian civet (Viverricula indica).. Sci Justice 2010;50:94-7.
  18. Mariacher A, Garofalo L, Fanelli R, Lorenzini R, Fico R. A combined morphological and molecular approach for hair identification to comply with the European ban on dog and cat fur trade.. PEERJ 2019;7:e7955.
  19. Wortmann FJ, Augustin P. Quantitative fiber mixture analysis by scanning electron microscopy: part VII: modeling the microscopic analysis of binary animal fiber blends.. Text Res J 2004;74(3):248-52.
  20. Wortmann FJ, Phan KH, Augustin P. Quantitative fiber mixture analysis by scanning electron microscopy: part VI: possibilities and limitations of the analysis of binary specialty fiber/wool blends in view of test method IWTO-58.. Text Res J 2003;73(9):781-6.
  21. Wortmann FJ, Phan KH, Augustin P. Quantitative fiber mixture analysis by scanning electron microscopy: part V: analyzing pure fiber samples and samples with small admixtures according to test method IWTO-58.. Text Res J 2003;73(8):727-32.
  22. Harizi T. Thermoanalytical characterisation of dromedary hair.. J Tex Inst 2010;101(7) 668-73.
  23. Kerkhoff K, Cescutti G, Kruse L, Müssig J. Development of a DNA-analytical method for the identification of animal hair fibers in textiles.. Text Res J 2009;79(1):69-75.
  24. Melton T, Holland C. Routine forensic use of the mitochondrial 12S ribosomal RNA gene for species identification.. J Forensic Sci 2007;52(6):1305-7.
  25. Houck M, Budowle B. Correlation of microscopic and mitochondrial DNA hair comparisons.. J Forensic Sci 2002;47(5):964-7.
  26. Elkins KM. Forensic DNA biology: a laboratory manual.. 1st ed. San Diego, USA: Elsevier Science & Technology; 2012.
  27. Kitano T, Umetsu K, Tian W, Osawa M. Two universal primer sets for species identification among vertebrates.. Int J Legal Med 2007;121(5):423-7.
  28. Mills LS, Pilgrim KL, Schwartz MK, McKelvey K. Identifying lynx and other North American felids based on mtDNA analysis.. Conserv Genet 2000;1:285-8.
  29. Pilli E, Casamassima R, Vai S, Virgili A, Barni F, D'Errico G. Pet fur or fake fur? A forensic approach.. Investig Genet 2014;5:1-18.
  30. Solazzo C, Wadsley M, Dyer JM, Clerens S, Collins MJ, Plowman J. Characterisation of novel α-keratin peptide markers for species identification in keratinous tissues using mass spectrometry.. Rapid Commun Mass Spectrom 2013;27:2685-98.
  31. Wang B, Yang W, McKittrick J, Meyers MA. Keratin: structure, mechanical properties, occurrence in biological organisms, and efforts at bioinspiration.. Prog Mater Sci 2016;76:229-318.
  32. Bai L, Wang J, Zhou H, Gong H, Tao J, Hickford JG. Identification of ovine KRTAP28-1 and its association with wool fibre diameter.. Animals 2019;9(4):142.
  33. Folin M, Contiero E. Electrophoretic analysis of non-human primates hair keratin.. Forensic Sci Int 1996;83(3):191-9.
  34. Khawar SL, Watson K, Jones GL. Peptide mapping of S-carboxymethylated hair and feather proteins using two-dimensional electrophoresis.. Int J Biochem Cell Biol 1996;28(10):1155-62.
  35. Hollemeyer K, Altmeyer W, Heinzle E. Identification of furs of domestic dog, racoon dog, rabbit and domestic cat by hair analysis using MALDI TOF mass spectrometry.. Spectrosc Eur 2007;19:8-15.
  36. Hollemeyer K, Altmeyer W, Heinzle E. Identification and quantification of feathers, down, and hair of avian and mammalian origin using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.. Anal Chem 2002;74(23):5960-68.
  37. Solazzo C, Wadsley M, Dyer JM, Clerens S, Collins MJ, Plowman J. Characterisation of novel α-keratin peptide markers for species identification in keratinous tissues using mass spectrometry.. Rapid Commun Mass Spectrom 2013;27(23):2685-98.
  38. Zeman A, Smid M, Havelcova M, Coufalova L, Kuckova S, Velcovska M. The structure and material composition of ossified aortic valves identified using a set of scientific methods.. J Asian Earth Sci 2013;77:311-7.
  39. Kuckova S, Zitkova K, Novotny O, Smirnova T. Verification of cheeses authenticity by mass spectrometry.. J Sep Sci 2019;42(22):3487-96.
  40. Hynek R, Kuckova S, Hradilova J, Kodicek M. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry as a tool for fast identification of protein binders in color layers of paintings.. Rapid Commun Mass Spectrom 2004;18(17):1896-900.
  41. Krizkova MC, Kuckova SH, Santrucek J, Hynek R. Peptide mass mapping as an effective tool for historical mortar analysis.. Constr Build Mat 2014;50:219-25.
  42. Kuckova SH, Schultz J, Veiga R, Murta E, Sandu ICA. Proteomics tools for the contemporary identification of proteinaceous binders in gilded samples.. Int J Conserv Sci 2015;6:507-18.
  43. Krizova I, Schultz J, Nemec I, Cabala R, Hynek R, Kuckova S. Comparison of analytical tools appropriate for identification of proteinaceous additives in historical mortars.. Anal Bioanal Chem 2018;410(1):189-200.
  44. Strohalm M, Hassman M, Kosata B, Kodicek M. mMass data miner: an open source alternative for mass spectrometric data analysis.. Rapid Commun Mass Spectrom 2008;22(6):905-8.
  45. Strohalm M, Kavan D, Novak P, Volny M, Havlicek V. mMass 3: a cross-platform software environment for precise analysis of mass spectrometric data.. Anal Chem 2010;82(11):4648-51.
  46. Kuckova S, Rambouskova G, Hynek R, Cejnar P, Oltrogge D, Fuchs R. Evaluation of mass spectrometric data using principal component analysis for determination of the effects of organic lakes on protein binder identification.. J Mass Spectrom 2015;50(11):1270-8.
  47. Cejnar P, Kuckova S, Prochazka A, Karamonova L, Svobodova B. Principal component analysis of normalized full spectrum mass spectrometry data in multiMS-toolbox: an effective tool to identify important factors for classification of different metabolic patterns and bacterial strains.. Rapid Commun Mass Spectrom 2018;15(11):871-81.
  48. Smirnova T A, Viskin A, Hoskova M, Habartova L, Setnicka V, Cejnar P. Comparison of proteomic approaches used for the detection of potential biomarkers of Alzheimer's disease in blood plasma.. J Sep Sci 2021;44(22):41324140.
  49. Cejnar P, Smirnova T A, Kuckova S, Prochazka A, Zak I, Harant K. Acute and chronic blood serum proteome changes in patients with methanol poisoning.. Sci Rep 2022;12(1):21379.
  50. Tyanova S, Temu T, Cox J. The MaxQuant computational platform for mass spectrometry-based shotgun proteomics.. Nat Protoc 2016;11:2301-19.
  51. Elias JE, Gygi SR. Target-decoy search strategy for mass spectrometry-based proteomics.. Methods Mol Biol 2010;604:55-71.

Citations

This article has been cited 1 times.
  1. Peterková L, Tesařová M, Sukupová A, Michalus I, Cejnar P, Fík Z, Šantrůček J, Kašička V, Hynek R. Direct In-Bone Protein Digestion With Subsequent LC Separation and Trap Ion Mobility MS Detection of Released Peptides as an Effective Tool for the Proteomic Characterization of Bone Tissues. J Sep Sci 2025 Sep;48(9):e70277.
    doi: 10.1002/jssc.70277pubmed: 40965984google scholar: lookup
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