Biodeterioration of carbographic ribbon: Isolation, identification of causal agents and forensic implications.

This study is part of a comprehensive investigation that was performed in regard to a case of alterations on a carbographic ribbon used in a typewriter that was found and seized by inner security operations of the Arma dei Carabinieri, Italy. Thirty-six coded scripts possessing potentially and criminally liable content were present on the tape; however, only the 6th and 7th scripts exhibited alterations of an uncertain nature. The study included sampling that was performed under sterile conditions of a large surface area of carbographic ribbons. A protocol based on physico-chemical, microbiological, and biomolecular tools was established. Preliminary results revealed the presence of fungal contamination that was primarily located on the inner surface of the 6th and 7th scripts on the black carbographic ribbon. One fungal strain was isolated and identified by universal ITS-PCR primer and rDNA sequencing as Alternaria infectoria strain NIS4. Fungal growth was monitored for 3 weeks in the laboratory under different environmental conditions (temperature, open-closed system, and substrate). The A. infectoria NIS4 strain exhibited the best growth at 28°C under a closed system with RH near saturation. We also noted that the fungal growth was abundant at 15°C. Moreover, this fungus (a potential human pathogen) possessed the ability to colonize the surface of the new carbographic ribbon even when using mineral medium; however, this only occurred in a closed system environment and not in open systems due to rapid desiccation. Under our experimental conditions, the A. infectoria NIS4 strain could degrade gelatin as an organic matter present in trace amounts that are often used as a binder in a carbographic ribbon emulsions. The results revealed that the isolated microorganism was the major biological candidate capable of altering the investigated carbographic ribbon; however, these alterations could only occur under favourable environmental conditions. AIMS: Identify the cause of microbial alterations on a carbographic ribbon in a typewriter used in a hypogean Italian criminal house named "covo." METHODS AND RESULTS: The isolation and identification of biodeteriogens (Alternaria infectoria NIS4) were performed using both culture-dependent and-independent methods, including ITS regions-primed PCR and rDNA techniques. Environmental scanning electron microscopy (ESEM) and optical observations were also performed. Growth tests and biodeterioration simulation tests on carbographic ribbons at the lab scale were performed under different environmental conditions. The A. infectoria NIS4 strain exhibited biodeterioration activity on carbographic ribbons under environmental conditions that were extremely favourable for growth. A high ability to colonize carbographic ribbon surfaces with fast and abundant growth at both 15°C and 28°C under lab-scale conditions at RH near saturation was observed. CONCLUSIONS: In this forensic case study, the ability of the isolated micromycetes A. infectoria NIS4 strain to colonize and induce alterations and degradation in a carbographic ribbon stored under indoor environmental conditions was examined. When favourable conditions change over time, the risk of microbial colonization and the damage produced by the fungal biodeterioration processes on the synthetic material objects has been confirmed. SIGNIFICANCE AND IMPACT OF STUDY: The current study contributes to the knowledge of biodeterioration processes in carbographic ribbon and the responsible agents, and our study provides an example of how environmental microbiology can also aid in forensic studies.

Violin Varnishes: Microstructure and Nanomechanical Analysis.

The aim of the current work is twofold: to demonstrate the application of in situ non-invasive imaging by portable atomic force microscopy (AFM) on the surfaces of a violin and to integrate compositional and mechanical analysis at the nano scale level on model samples of varnished wood. These samples were prepared according to traditional recipes by an Italian lute-maker family well practised in the art. Samples of oil and spirit-based varnishes on maple wood, naturally and accelerated light aged, were studied. AFM was used to measure the nanomechanical properties of the model samples and established that the spirit-based varnish was stiffer than the oil-based. Synchrotron radiation micro- Fourier Transform Infra-red analysis of the layer structure revealed that stiffer spirit-based varnish showed less penetration into the wood than the oil-based. Further PeakForce Quantitative Nanomechanical Mapping (QNM) demonstrated a difference in adhesion values between the oil- and spirit-based samples.

Plastics in Heritage Science: Analytical Pyrolysis Techniques Applied to Objects of Design.

The first synthetic polymers were introduced as constituents of everyday life, design objects, and artworks at the end of the 19th century. Since then, the history of design has been strictly connected with the 20th century evolution of plastic materials. Objects of design from the 20th century are today a precious part of the cultural heritage. They raise specific conservation issues due to the degradation processes affecting synthetic polymer-based plastics. Museums and collections dealing with the conservation of design objects and modern materials need to base their conservation strategies on compositional data that reveal the formulations of historical plastics and their decay processes. Specific and specifically optimized analytical tools are thus needed. We employed flash analytical pyrolysis coupled with gas chromatography and mass spectrometry (Py-GC/MS) and evolved gas analysis coupled with mass spectrometry (EGA-MS) to characterize "historic polymeric materials" (HIPOMS) and heritage plastics at the molecular level with high chemical detail. This approach complements non-invasive spectroscopic diagnosis whenever it fails to obtain significant or complete information on the nature and the state of preservation of the materials under study. We determined the composition of several 20th century design objects (1954-1994) from the Triennale Design Museum of Milan (Triennale Milano - Museo del Design Italiano), which for different morphological, chemical, or physical reasons were unsuitable for characterization by non-invasive spectroscopy. EGA-MS proved capable for the study of the different fractions constituting heterogeneous micro-samples and for gaining an insight into their degradation processes from the contextual interpretation of thermal and mass-spectrometric data.

A Mass Spectrometric Study on Tannin Degradation within Dyed Woolen Yarns.

Natural tannins from various plants have been used throughout human history in textile dyeing, often as mordant dyes. The ageing behavior of these dyes is a challenge in conservation science, requiring a thorough knowledge of the textile-mordant-dye system. In this work, we analyzed reference wool yarns dyed with natural tannins from oak gallnuts, walnut (Juglans regia), and catechu (Acacia catechu), after artificial ageing. To gain insights on the composition of the dyestuffs and on how they aged, an analytical procedure based on extraction with Na2EDTA/DMF (ethylenediaminetetraacetic acid/dimethylformamide) and high-performance liquid chromatography (HPLC) analysis using high-resolution mass spectrometry detection was used. Since conventional reversed-phase (RP) columns usually show poor retention efficiency of highly polar compounds such as tannins, an RP-amide embedded polar group stationary phase was used to achieve optimal retention of the most polar compounds. Tannins from oak gallnuts showed little degradation after ageing, while a significant increase in the content of hydroxybenzoic acids was observed for tannins from walnut and catechu. Finally, the analytical procedure was applied to characterize the tannin dyes in historical tapestries from the 15th to 16th century, and the results were discussed in comparison with the reference yarns.

Recent Advances in Analytical Pyrolysis to Investigate Organic Materials in Heritage Science.

The molecular characterization of organic materials in samples from artworks and historical objects traditionally entailed qualitative and quantitative analyses by HPLC and GC. Today innovative approaches based on analytical pyrolysis enable samples to be analysed without any chemical pre-treatment. Pyrolysis, which is often considered as a screening technique, shows previously unexplored potential thanks to recent instrumental developments. Organic materials that are macromolecular in nature, or undergo polymerization upon curing and ageing can now be better investigated. Most constituents of paint layers and archaeological organic substances contain major insoluble and chemically non-hydrolysable fractions that are inaccessible to GC or HPLC. To date, molecular scientific investigations of the organic constituents of artworks and historical objects have mostly focused on the minor constituents of the sample. This review presents recent advances in the qualitative and semi-quantitative analyses of organic materials in heritage objects based on analytical pyrolysis coupled with mass spectrometry.

Chemical characterisation of the whole plant cell wall of archaeological wood: an integrated approach.

Wood artefacts undergo complex alteration and degradation during ageing, and gaining information on the chemical composition of wood in archaeological artefacts is fundamental to plan conservation strategies. In this work, an integrated analytical approach based on innovative NMR spectroscopy procedures, gel permeation chromatography and analytical pyrolysis coupled with gas chromatography/mass spectrometry (Py-GC-MS) was applied for the first time on archaeological wood from the Oseberg collection (Norway), in order to evaluate the chemical state of preservation of the wood components, without separating them. We adopted ionic liquids (ILs) as non-derivatising solvents, thus obtaining an efficient dissolution of the wood, allowing us to overcome the difficulty of dissolving wood in its native form in conventional molecular solvents. Highly substituted lignocellulosic esters were therefore obtained under mild conditions by reacting the solubilised wood with either acetyl chloride or benzoyl chloride. A phosphytilation reaction was also performed using 2-chloro-4,4,5,5-tetramethyl-1,3,2-dioxaphospholan. As a result, the functionalised wood developed an enhanced solubility in molecular solvents, thus enabling information about modifications of lignin, depolymerisation of cellulose and structure of lignin-carbohydrate complexes to be obtained by means of spectroscopic (2D-HSQC-NMR and 31P-NMR) and chromatographic (gel permeation chromatography) techniques. Py-GC-MS was used to investigate the degradation undergone by the lignocellulosic components on the basis of their pyrolysis products, without any pre-treatment of the samples. The application of all these combined techniques enabled a comprehensive characterisation of the whole cell wall of archaeological wood and the evaluation of its state of preservation. High depletion of carbohydrates and high extent of lignin oxidation were highlighted in the alum-treated objects, whereas a good preservation state was found for the untreated wood of the Oseberg ship. Graphical abstract ᅟ.

Terpenoid Oligomers of Dammar Resin.

Dammar is a triterpenoid resin containing a volatile fraction, a monomeric fraction, and a high-molecular weight fraction. Although the low-molecular-weight components comprising sesquiterpenoids and triterpenoids have been extensively studied, the nature of the macromolecular components is still not fully understood, and different and sometimes contradictory theories have been proposed. The aim of this paper is to clarify the nature of the macromolecular components of dammar resin. A multianalytical approach was adopted based on thermoanalytical-thermogravimetric analysis (TGA), and thermogravimetric analysis coupled with Fourier transform infrared spectroscopy (TGA/FTIR)--and mass spectrometric techniques-direct exposure mass spectrometry (DE/MS), pyrolysis coupled to gas chromatography and mass spectrometry (Py/GC/MS), flow injection analysis electrospray ionization mass spectrometry (FIA/ESI/MS), and gas chromatography/mass spectrometry (GC/MS). The data indicate that the oligomeric fraction comprises triterpenoids bound through ester bonds, and that these triterpenoids are the same as those found in the free terpenoid fraction. The oligomeric fraction also includes triterpenoids containing carbonyl moieties, such as formyl groups, thus suggesting that these are involved in the esters in their corresponding enolic form.

Ingredients of a 2,000-y-old medicine revealed by chemical, mineralogical, and botanical investigations.

In archaeology, the discovery of ancient medicines is very rare, as is knowledge of their chemical composition. In this paper we present results combining chemical, mineralogical, and botanical investigations on the well-preserved contents of a tin pyxis discovered onboard the Pozzino shipwreck (second century B.C.). The contents consist of six flat, gray, discoid tablets that represent direct evidence of an ancient medicinal preparation. The data revealed extraordinary information on the composition of the tablets and on their possible therapeutic use. Hydrozincite and smithsonite were by far the most abundant ingredients of the Pozzino tablets, along with starch, animal and plant lipids, and pine resin. The composition and the form of the Pozzino tablets seem to indicate that they were used for ophthalmic purposes: the Latin name collyrium (eyewash) comes from the Greek name κoλλυρα, which means "small round loaves." This study provided valuable information on ancient medical and pharmaceutical practices and on the development of pharmacology and medicine over the centuries. In addition, given the current focus on natural compounds, our data could lead to new investigations and research for therapeutic care.

Industrial alkyd resins: characterization of pentaerythritol and phthalic acid esters using integrated mass spectrometry.

RATIONALE: Alkyd resins are synthetic polyesters used as paints and coatings. Current approaches for their analysis do not allow the characterization of pentaerythritol and phthalic acid esters, whose detection is interesting to fully characterize the materials, e.g. for forensic or cultural heritage applications. METHODS: A combined analytical approach based on Gas Chromatography/Mass Spectrometry (GC/MS), High-Performance Liquid Chromatography (HPLC)/MS and flow injection analysis (FIA)/MS was adopted. GC/MS was used to characterize the fatty acid profile and the polybasic acids in extracts from industrial alkyd resins. HPLC/MS and FIA/MS were used for the characterization of the triglyceride profile of the oil used to manufacture the resin and for the identification of reaction products deriving from the synthesis process. RESULTS: The multi-analytical approach was applied on two different industrial alkyd resins produced from two different oils. The GC/MS analysis was successful in characterizing the fatty acid profile and the aromatic fraction of the resin. The HPLC/MS analysis allowed us to characterize the pentaerythritol and phthalic acid ester and the triglycerides residues from the synthesis process, by studying their high-resolution tandem mass spectra. CONCLUSIONS: The application of liquid chromatography coupled with high-resolution tandem mass spectrometry to the study of industrial alkyd resins allowed us to characterize for the first time the esters formed by the transesterification reactions involving pentaerythritol, phthalic acid and triglycerides.

Py-GC/MS applied to the analysis of synthetic organic pigments: characterization and identification in paint samples.

A collection of 76 synthetic organic pigments was analysed using pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS). The purpose of this work was to expand the knowledge on synthetic pigments and to assess characteristic pyrolysis products that could help in the identification of these pigments in paint samples. We analysed several classes of synthetic pigments not previously reported as being analysed by this technique: some metal complexes, ß-naphthol pigment lakes, BONA pigment lakes, disazopyrazolone, triarylcarbonium, dioxazine, anthraquinone, indanthrone, isoindoline and thioindigo classes. We also report for the first time the Py-GC/MS analysis of a number of naphthol AS, benzimidazolone, phthalocyanine and perylene pigments and other miscellaneous pigments including pigments with unpublished chemical structure. We successfully used the Py-GC/MS technique for the analysis of paints by artists Clyfford Still and Jackson Pollock to identify the synthetic organic pigments and the binding media.

Early evidence of San material culture represented by organic artifacts from Border Cave, South Africa.

Recent archaeological discoveries have revealed that pigment use, beads, engravings, and sophisticated stone and bone tools were already present in southern Africa 75,000 y ago. Many of these artifacts disappeared by 60,000 y ago, suggesting that modern behavior appeared in the past and was subsequently lost before becoming firmly established. Most archaeologists think that San hunter-gatherer cultural adaptation emerged 20,000 y ago. However, reanalysis of organic artifacts from Border Cave, South Africa, shows that the Early Later Stone Age inhabitants of this cave used notched bones for notational purposes, wooden digging sticks, bone awls, and bone points similar to those used by San as arrowheads. A point is decorated with a spiral groove filled with red ochre, which closely parallels similar marks that San make to identify their arrowheads when hunting. A mixture of beeswax, Euphorbia resin, and possibly egg, wrapped in vegetal fibers, dated to ∼40,000 BP, may have been used for hafting. Ornaments include marine shell beads and ostrich eggshell beads, directly dated to ∼42,000 BP. A digging stick, dated to ∼39,000 BP, is made of Flueggea virosa. A wooden poison applicator, dated to ∼24,000 BP, retains residues with ricinoleic acid, derived from poisonous castor beans. Reappraisal of radiocarbon age estimates through bayesian modeling, and the identification of key elements of San material culture at Border Cave, places the emergence of modern hunter-gatherer adaptation, as we know it, to ∼44,000 y ago.

Border Cave and the beginning of the Later Stone Age in South Africa.

The transition from the Middle Stone Age (MSA) to the Later Stone Age (LSA) in South Africa was not associated with the appearance of anatomically modern humans and the extinction of Neandertals, as in the Middle to Upper Paleolithic transition in Western Europe. It has therefore attracted less attention, yet it provides insights into patterns of technological evolution not associated with a new hominin. Data from Border Cave (KwaZulu-Natal) show a strong pattern of technological change at approximately 44-42 ka cal BP, marked by adoption of techniques and materials that were present but scarcely used in the previous MSA, and some novelties. The agent of change was neither a revolution nor the advent of a new species of human. Although most evident in personal ornaments and symbolic markings, the change from one way of living to another was not restricted to aesthetics. Our analysis shows that: (i) at Border Cave two assemblages, dated to 45-49 and >49 ka, show a gradual abandonment of the technology and tool types of the post-Howiesons Poort period and can be considered transitional industries; (ii) the 44-42 ka cal BP assemblages are based on an expedient technology dominated by bipolar knapping, with microliths hafted with pitch from Podocarpus bark, worked suid tusks, ostrich eggshell beads, bone arrowheads, engraved bones, bored stones, and digging sticks; (iii) these assemblages mark the beginning of the LSA in South Africa; (iv) the LSA emerged by internal evolution; and (v) the process of change began sometime after 56 ka.

Field-emission scanning electron microscopy and energy-dispersive x-ray analysis to understand the role of tannin-based dyes in the degradation of historical wool textiles.

An innovative approach, combining field-emission scanning electron microscopy (FESEM) with energy dispersive X-ray spectroscopy (EDX) analysis, is presented to investigate the degradation mechanisms affecting tannin-dyed wool. In fact, tannin-dyed textiles are more sensitive to degradation then those dyed with other dyestuffs, even in the same conservation conditions. FESEM-EDX was first used to study a set of 48 wool specimens (artificially aged) dyed with several raw materials and mordants, and prepared according to historical dyeing recipes. EDX analysis was performed on the surface of wool threads and on their cross-sections. In addition, in order to validate the model formulated by the analysis of reference materials, several samples collected from historical and archaeological textiles were subjected to FESEM-EDX analysis. FESEM-EDX investigations enabled us to reveal the correlation between elemental composition and morphological changes. In addition, aging processes were clarified by studying changes in the elemental composition of wool from the protective cuticle to the fiber core in cross-sections. Morphological and elemental analysis of wool specimens and of archaeological and historical textiles showed that the presence of tannins increases wool damage, primarily by causing a sulfur decrease and fiber oxidation.

The role of organic and inorganic indoor pollutants in museum environments in the degradation of dammar varnish.

This paper investigates the effects of inorganic (NO(2) and O(3)) and volatile organic acid (acetic acid) pollutants on the degradation of dammar varnish in museum environments. Model paint varnish samples based on dammar resin were investigated by Gas Chromatography Mass Spectrometry (GC-MS), Dynamic Mechanical Analysis (DMA) and Atomic Force Microscopy (AFM). Dammar is a natural triterpenoid resin, commonly used as a paint varnish. Samples were subjected to accelerated ageing by different levels of pollutants (NO(2) and O(3) and acetic acid) over a range of relative humidities (RH) and then analysed. The results revealed that as the dose of the pollutant was increased, so did the degree of oxidation and cross-linking of the resin. Most interestingly, it was shown for the first time that exposure to acetic acid vapour resulted in the production of an oxidised and cross-linked resin, comparable to the resin obtained under exposure to NO(2) and O(3). These conclusions were supported by the analyses of model varnishes exposed for about two years in selected museum environments, where the levels of pollutants had been previously measured. Exposures were performed both within and outside the selected microclimate frames for paintings. Results showed that varnishes placed within the microclimate frames were not always better preserved than those exposed outside the frames. For some sites, the results highlighted the protective effects of the frames from outdoor generated pollutants, such as NO(2) and O(3). For other sites, the results showed that the microclimate frames acted as traps for the volatile organic acids emitted by the wooden components of the mc-frames, which damaged the varnish.

The role of early synthetic materials degradation in the downfall of the Ansaldo A.1, an Italian World War I biplane fighter.

From the Pioneer Era of the aviation to World War I the evolution of aircraft technology and chemical synthesis enabled a unique coexistence of traditional craftsmanship, artistic decoration practices, and technological advancements. The study of the materials used in these early years of aviation is still an uncharted territory: a vast portion of remaining planes has been partially or completely repaired and restored, usually by total replacement of the fabric. The Italian biplane Ansaldo A.1 (1918) is a fighter aircraft and is one of the few planes in the world that still preserves its own original materials. In the last years, the fabric sections of the airplane have started to become brittle and loose cohesion, severely compromising the integrity of the aircraft, and resulting in a general alteration of the pictorial layers of the painted sections. A chemical investigation was undertaken to unveil the materials, and to elucidate the causes of the degradation. This study presents one of the first steps into the study of early historical aircrafts, defining the background for the conservation plans to preserve these objects for future generations.

Physico-chemical characterization of protein-pigment interactions in tempera paint reconstructions: casein/cinnabar and albumin/cinnabar.

In this work, we characterized paint reconstructions using ovalbumin and casein as binders, and cinnabar (HgS) as a pigment, before and after artificial ageing. Egg and casein are common paint binders that were used historically in the technique of tempera painting. Despite extensive research on the identification of proteinaceous binders in paintings, there is a substantial lack of knowledge regarding the ageing pathway of their protein content, and their chemical interaction with inorganic pigments. Thermogravimetric analysis, infrared spectroscopy and size-exclusion chromatography (SEC) were used to reveal the physico-chemical processes involved in the ageing of proteins in paintings. Taken together, the three techniques highlighted that proteins are subject to both cross-linking and hydrolysis upon ageing, and to a lesser extent, to oxidation of the side chains. Mercury-protein interactions were also revealed using a cold vapour generation atomic fluorescence spectrometer mercury-specific detector coupled to SEC. The study clearly showed that HgS forms stable complexes with proteins and acts as a sensitizer in cross-linking, hydrolysis and oxidation.

The Cultural Heritage of "Black Stones" (Lapis Aequipondus/Martyrum) of Leopardi's Child Home (Recanati, Italy).

A macroscopic lithological study and physical (hardness, size, weight) investigations, coupled with laser-induced breakdown spectroscopy (LIBS) and X-ray fluorescence (XRF) chemical analyses of three egg- and one pear-shaped polished black stones, exposed in the library of the child home of the famous poet Giacomo Leopardi, at Recanati (Italy), were carried out. They are characterized by different sizes: two with the same weight of 16.9 kg and the two smaller ones of 5.6 kg each, corresponding to multiples of standard roman weights (drachma and scrupulum). These features and the presence of some grooves on the rock artefacts, probably for grappling hooks, suggest an original use as counterweight for the four black stones herein classified as amphibole-bearing serpentinites whose lithologies are far away from Recanati (probably coming from geological outcrops in Tuscany). The four serpentinite stones closely match with the so-called Lapis Aequipondus used in antiquity by the Romans as counterweights. Due to the presence of lead rings or iron hooks in these stones, Lapis Aequipondus were also used for martyrdoms during the persecution of Christians in the Roman period, attached to the necks of martyrs that were then thrown in the wells or attached to the ankles of hanging bodies. This is the reason why these stones are also known as Lapis Martyrum, venerated with the relative martyrs, in several churches of Rome. The four black stones investigated probably arrived at Recanati from Rome after the middle of the 19th century. In the past, Christians also called Lapis Martyrum the "devil's stones" (Lapis Diaboli). This could also be the reason for the popular belief that black stones cannot be touched by people, except those of the Leopardi dynasty. This work contributes to the cultural heritage of Leopardi's child home, as the four black stones had never been investigated.

Deterioration and discoloration of historical protective treatments on marble.

This study integrates the complex research conducted on the sources of brown discolorations that occur on marble statues (fifteenth century) of the Church of Orsanmichele in Florence (Italy). They underwent conservative interventions in the past and the brownish discolorations on their surfaces strongly altered the clear tone of the marble. In this study, Carrara marble model specimens were treated with organic and inorganic substances (non-pasteurised milk; linseed oil; walnut oil; ammonium oxalate; microcrystalline wax; beeswax; milk + linseed oil; and milk + ammonium oxalate + linseed oil) to simulate their effects on the stone. Some of the substances were commonly used in the past (as on the Orsanmichele statues) but most of them are still used in many countries. The treated specimens were exposed to natural and artificial ageing. The main results of the research were (i) the specimens treated with linseed oil, milk + linseed oil, and milk + linseed oil + ammonium oxalate showed a severe change of colour after either artificial or natural ageing; (ii) an extensive polymerisation of the organic substances occurred; (iii) calcium oxalate and several oxidised diacylglycerols (DAGs) and triacylglycerols (TAGs) were the last chemical products of the oxidation processes induced by ageing; (iv) Maillard reaction, producing brownish coloration, likely occurred in specimens containing milk as a result of the interaction between sugars and proteins.

Deamidation at asparagine and glutamine as a major modification upon deterioration/aging of proteinaceous binders in mural paintings.

Proteomic strategies are herein proved to be a complementary approach to the well established amino acid composition analysis for the characterization of the aging and deterioration phenomena occurring to proteinaceous materials in works-of-art. Amino acid analyses on several samples demonstrated that proteins in the frescoes from the Camposanto Monumentale in Pisa are deteriorated as revealed by the decrease in Met, Lys, and Tyr content and by the presence in all the samples of amino malonic acid as a result of Ser, Phe, and Cys oxidation. Proteomic analysis identified deamidation at Asn and Gln as a further major event occurred. This work paves the way to the exploitation of proteomic strategies for the investigation of the molecular effects of aging and deterioration in historical objects. Results show that proteomic searches for deamidation by liquid chromatography-tandem mass spectrometry (LC-MS/MS) could constitute a routine analysis for paintings or any artistic and historic objects where proteins are present. Peptides that can be used as molecular markers when casein is present were identified.

Analytical strategies for characterizing organic paint media using gas chromatography/mass spectrometry.

Throughout history, artists have experimented with a variety of organic-based natural materials, using them as paint binders, varnishes, and ingredients for mordants in gildings. Artists often use many layers of paint to produce particular effects. How we see a painting is thus the final result of how this complex, highly heterogeneous, multimaterial, and multilayered structure interacts with light. The chemical characterization of the organic substances in paint materials is of great importance for artwork conservation because the organic components of the paint layers are particularly subject to degradation. In addition, understanding the organic content and makeup of paint materials allows us to differentiate between the painting techniques that have been used over history. Applying gas chromatography/mass spectrometry (GC/MS) analysis to microsamples of paint layers is widely recognized as the best approach for identifying organic materials, such as proteins, drying oils, waxes, terpenic resins, and polysaccharide gums. The method provides essential information for reconstructing artistic techniques, assessing the best conditions for long-term preservation, and planning restoration. In this Account, we summarize the more common approaches adopted in the study of the organic components of paint materials. Our progress in developing GC/MS analytical procedures in the field of cultural heritage is presented, focusing on problems that arise from (i) the presence of mixtures of many chemically complex and degraded materials, (ii) the interference of inorganic species, (iii) the small size of the samples, and (iv) the risk of contamination. We outline some critical aspects of the analytical strategy, such as the need to optimize specific wet-chemical sample pretreatments in order to separate the various components, hydrolyze macromolecular analytes, clean-up inorganic ions, and derivatize polar molecules for subsequent GC/MS separation and identification. We also discuss how to interpret the chromatographic data so as to be able to identify the materials. This identification is based on the presence of specific biomarkers (chemotaxonomy), on the evaluation of the overall chromatographic profile, or on the quantitative analysis of significant compounds. GC/MS-based analytical procedures have for 20 years provided important contributions to conservation science, but challenges and opportunities still coexist in the field of organic-based paint materials. We give selected examples and provide case studies showing how a better understanding of the chemical composition of organic paint materials and of their degradation pathways contribute to a better knowledge our cultural heritage, and to its preservation for future generations.