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Identification of protein binders in works of art by high-performance liquid chromatography–diode array detector analysis of their tryptic digests

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Abstract

Proteins in works of art are generally determined by the relative amounts of amino acids. This method, however, implies a loss of information on the protein structure and its modifications. Consequently, we propose a method based on the analysis of trypsin digests using high-performance liquid chromatography (HPLC) UV diode array detection (DAD) for painting binder studies. All reaction steps are done in the same vial; no extraction methods or sample transfer is needed, reducing the risk of sample losses. A collection of pure binders (collagen, ovalbumin, yolk and casein) as well as homemade and historical paint samples have been investigated with this method. Chromatograms of unknowns at 214 nm and 280 nm are compared with those of the reference samples as a fingerprint. There is a good agreement between many peptides, but others seem to have been lost or their retention time shifted due to small compositional changes because of ageing and degradation of the paint. The results are comparable with the results of other techniques used for binder identification on the same samples, with the additional advantage of differentiation between egg yolk and glair.

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References

  1. Colombini MP, Modugno F (2004) J Sep Sci 27:147–160

    Article  CAS  Google Scholar 

  2. Schilling MR, Khanjian HP, Souza LAC (1996) J Am Inst Conserv 35:45–59

    Article  Google Scholar 

  3. Schilling MR, Khanjian HP (1996) J Am Inst Conserv 35:123–144

    Article  Google Scholar 

  4. Casoli A, Musini PC, Palla G (1996) J Chromatogr A 731:237–246

    Article  CAS  Google Scholar 

  5. Colombini MP, Modugno F, Giacomelli M, Francesconi S (1999) J Chromatogr A 846:113–124

    Article  CAS  Google Scholar 

  6. Colombini MP, Carmignani A, Modugno F, Frezzato F, Olchini A, Brecoulaki H, Vassilopoulou V, Karkanas P (2004) Talanta 63:839–848

    Article  CAS  Google Scholar 

  7. Andreotti A, Bonaduce I, Colombini MP, Gautier G, Modugno F, Ribechini E (2006) Anal Chem 78:4490–4500

    Article  CAS  Google Scholar 

  8. Scott DA, Khandekar N, Schilling MR, Turner N, Taniguchi Y, Khanjian H (2001) Stud Conserv 46:93–108

    Article  CAS  Google Scholar 

  9. Sutherland K, Price BA, Passeri I, Tucker M (2005) A study of the materials of Pontormo's “Portrait of Alessandro de’ Medici”. In: Vandiver PB, Mass JL, Murray A (eds) Materials Issues in art and archaeology VII, MRS proceedings volume 852. MRS, Warrendale

    Google Scholar 

  10. Prikryl P, Havlíková L, Pacáková V, Hradilová J, Tulík K, Hofta P (2006) J Sep Sci 29:2653–2663

    Article  CAS  Google Scholar 

  11. Gautier G, Colombini MP (2007) Talanta 73:95–102

    Article  CAS  Google Scholar 

  12. Wouters J, Van Bos M, Lamens K (2000) Stud Conserv 45:106–116

    Article  CAS  Google Scholar 

  13. Peris-Vicente J, Gimeno Adelantado JV, Doménech Carbó MT, Mateo Castro R, Bosch Reig F ((2006) Talanta 68:1648–1654

    Article  Google Scholar 

  14. Musumarra G, Fichera M (1998) Chemom Intell Lab Syst 44:363–372

    Article  CAS  Google Scholar 

  15. Messinger JM (1992) J Am Inst Conserv 31:267–274

    Article  Google Scholar 

  16. Carbo MTD, Reig FB, Adelantado JVG, Martinez VP (1996) Anal Chim Acta 330:207–215

    Article  CAS  Google Scholar 

  17. Jurado-López A, de Castro MD Luque (2004) Anal Bioanal Chem 380:706–711

    Article  Google Scholar 

  18. Vandenabeele P, Wehling B, Moens L, Edwards H, De Reu M, Van Hooydonk G (2000) Anal Chim Acta 407:261–274

    Article  CAS  Google Scholar 

  19. Nevin A, Osticioli I, Anglos D, Burnstock A, Cather S, Castellucci E (2007) Anal Chem 79:6143–6151

    Article  CAS  Google Scholar 

  20. Nevin A, Osticioli I, Anglos D, Burnstock A, Cather S, Castellucci E (2008) J Raman Spectrosc 39:993–1000

    Article  CAS  Google Scholar 

  21. Kockaert L, Gausset P, Dubi-Rucquoy M (1989) Stud Conserv 34:183–188

    Article  CAS  Google Scholar 

  22. Ramirez-Barat B, de la Viña S (2001) Stud Conserv 46:282–288

    Article  CAS  Google Scholar 

  23. Carbini M, Stevanato R, Rovea M, Traldi P, Favretto D (1996) Rapid Commun Mass Spectrom 10:1240–1242

    Article  CAS  Google Scholar 

  24. Chiavari G, Gandini N, Russo P, Fabbri D (2001) Chromatographia 53:311–314

    Article  CAS  Google Scholar 

  25. Chiavari G, Fabbri D, Prati S (2001) J Chromatogr A 922:235–241

    Article  CAS  Google Scholar 

  26. Ferreira ESB, van der Horst J, Boon JJ (2005) Chemical aspects of the binding media of the Oranjezaal ensemble: an insight into 17th century Netherlandish materials and methods. In: 14th Triennial Meeting the Hague Preprints, ICOM Committee for Conservation, vol II, pp 774–782

  27. Chiavari G, Fabbri D, Galletti GC, Prati S, Scianna N (2006) J Cult Heritage 7:67–70

    Article  Google Scholar 

  28. Tokarski C, Martin E, Rolando C, Cren-Olivé C (2006) Anal Chem 78:1494–1502

    Article  CAS  Google Scholar 

  29. Hynek R, Kuckova S, Hradilova J, Kodicek M (2004) Rapid Commun Mass Spectrom 18:1896–1900

    Article  CAS  Google Scholar 

  30. Kuckova S, Nemec I, Hynek R, Hradilova J, Grygar T (2005) Anal Bioanal Chem 382:275–282

    Article  CAS  Google Scholar 

  31. Kuckova S, Hynek R, Kodicek M (2007) Anal Bioanal Chem 388:201–206

    Article  CAS  Google Scholar 

  32. Kuckova S, Hynek R, Kodicek M (2008) Analysis of art objects using mass spectrometry. In: Proceedings of the 9th International Conference ART2008, Jerusalem, 25-30 May 2008

  33. Fremout W, Saverwyns S, Peters F, Deneffe D (2006) J Raman Spectrosc 37:1035–1045

    Article  CAS  Google Scholar 

  34. Deneffe D, Peters F, Fremout W, Sanyova J, Saverwyns S, Currie C, Depuydt-Elbaum L, Fraiture P (2009) Pre-Eyckian Panel Painting in the Low Countries, 1 Catalogue (Contributions to Fifteenth-Century Painting in the Southern Netherlands and the Principality of Liège, 9), edited by Stroo C. Brepols, Brussels (in press)

  35. Dresner E, Schubert M (1955) J Histochem Cytochem 3:360–368

    CAS  Google Scholar 

  36. Mills JS, White R (1987) The organic chemistry of the museum objects. Butterworths, London

    Google Scholar 

Download references

Acknowledgements

This investigation was funded by the Belgian Science Policy as a part of the doctoral research project of Wim Fremout. Support of the Interuniversity Attraction Poles Program—Belgian Science Policy is greatly acknowledged. The authors wish to thank Ing Ina Vanden Berghe and Marie-Christine Maquoi for the amino acid analysis with HPLC and Mathieu Douineau and Marie Poellaer for helping us prepare the homemade samples inspired by ancient recipes.

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Authors and Affiliations

  1. Royal Institute for Cultural Heritage (KIK/IRPA), Jubelpark 1, 1000, Brussels, Belgium

    Wim Fremout, Jana Sanyova & Steven Saverwyns

  2. Department of Analytical Chemistry, Ghent University, INW, Proeftuinstraat 86, 9000, Ghent, Belgium

    Wim Fremout & Luc Moens

  3. Department of Archaeology & Ancient History of Europe, Ghent University, Blandijnberg 2, 9000, Ghent, Belgium

    Peter Vandenabeele

Authors
  1. Wim Fremout
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  2. Jana Sanyova
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  3. Steven Saverwyns
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  4. Peter Vandenabeele
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  5. Luc Moens
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Correspondence to Wim Fremout.

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Fremout, W., Sanyova, J., Saverwyns, S. et al. Identification of protein binders in works of art by high-performance liquid chromatography–diode array detector analysis of their tryptic digests. Anal Bioanal Chem 393, 1991–1999 (2009). https://doi.org/10.1007/s00216-009-2686-z

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  • DOI: https://doi.org/10.1007/s00216-009-2686-z

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