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MALDI-TOF-MS methodologies for identification of paint binders


New protocols for MALDI-MS of lipids and proteins contained in a single small specimen of paint were developed.
The first protocol is based on Bligh–Dyer (BD) extraction followed by MALDI-TOF-MS analysis. For the first time it was demonstrated that the BD method is effective for the simultaneous extraction of lipids and proteins from complex, and atypical matrices, such as pigmented paint layers. The protocol makes use of an alternative denaturing anionic detergent (RapiGestTM) in order to improve efficiency of protein digestion and purification step. Detection of various lipid classes, such as TAGs and PLs and their oxidation by-products, was accomplished, whereas proteins could be identified by peptide mass fingerprinting. The effect of pigments on ageing of lipids and proteins was also investigated.
The second protocol contributes a step forward in the simplification of such a protocol as no conventional solvent extraction is required. The paint fragment (ca. 100 μg) is fixed on the MALDI target plate, and the wet chemistry steps required for protein digestion are performed directly on the plate that, after matrix addition, was analyzed by MALDI-MS for the simultaneous identification of proteinaceous and lipid markers of the most commonly used binders. Colloidal graphite is used to assure the adhesion of the painting fragment (as such or after grinding) on a conventional stainless steel target plate.
The potential of both protocols was demonstrated analyzing samples taken from different paintings.


MALDI-MS investigation of a 15th century panel painting by A. Vivarini
(Pinacoteca provinciale di Bari, Italy)


Published papers
• Calvano C.D., van der Werf I.D., Palmisano F., Sabbatini L., "Identification of lipid- and protein-based binders in paintings by direct on-plate wet chemistry and matrix-assisted laser desorption ionization mass spectrometry", Analytical and Bioanalytical Chemistry, 407 (2015) 1015–1022.
• Cosima Damiana Calvano, Inez Dorothé van der Werf, Luigia Sabbatini, Francesco Palmisano, On plate graphite supported sample processing for simultaneous lipid and protein identification by matrix assisted laser desorption ionization mass spectrometry, Talanta, DOI: 10.1016/j.talanta.2015.01.030
• I.D. van der Werf, A. Monno, C.D. Calvano, R. Laviano, L. Sabbatini, Il polittico di Antonio Vivarini: uno sguardo scientifico alla materia pittorica, in “Antonio Vivarini in Puglia” (a cura di C. Gelao), Marsilio editore, Venezia, 2014 (ISBN: 978-88-317-1879-0) pp. 77-89.
• V. Sajeva, Sviluppo di protocolli innovativi per l’identificazione di leganti pittorici mediante tecniche di spettrometria di massa MALDI-TOF, MSc thesis, 2013-2014, University of Bari Aldo Moro


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