Black surfaces on ancient leather tefillin cases and straps from the Judean Desert: Macroscopic, microscopic and spectroscopic analyses.

Tefillin are Jewish ritual artifacts consisting of leather cases, containing inscribed slips, which are affixed with leather straps to the body of the tefillin practitioner. According to current Jewish ritual law, the tefillin cases and straps are to be colored black. The present study examines seventeen ancient tefillin cases discovered among the Dead Sea Scrolls in caves in the Judean Desert. All seventeen cases display grain surfaces with a very dark, nearly black appearance. We start with a hypothesis that the cases were intentionally colored black in antiquity using either a carbon-based or iron-gall-based paint or dye. The aim of this study is to test this hypothesis by subjecting these tefillin cases to a battery of examinations to assess the presence of carbon and iron used as pigments, and of organic materials which may have been used as binding agents in a paint. The tests deployed are: (1) macroscopic and microscopic analyses; (2) multispectral imaging using infrared wavelengths; (3) Raman spectroscopy; (4) Fourier transform infrared spectroscopy (FTIR); and (5) scanning electron microscope (SEM) and energy dispersive X-ray (EDX) spectroscopy. The results of these tests found no traces of carbon-based or iron-gall-based pigments, nor of organic compounds which may have served as binders in a paint. These results suggest that our posited hypothesis is unlikely. Instead, results of the SEM examination suggest it more likely that the black color on the surfaces of the tefillin cases is the result of natural degradation of the leather through gelatinization. The Judean Desert tefillin likely represent tefillin practices prior to when the rabbinic prescription on blackening tefillin was widely practiced. Our study suggests that the kind of non-blackened tefillin which the later rabbis rejected in their own times may well have been quite common in earlier times.

Metal record of copper-based antifouling paints in sediment core following marina construction and operation.

Marinas are central hubs of global maritime leisure and transport, yet their operations can deteriorate the environmental quality of sediments. In response, this study investigated the metal contamination history associated with antifouling paint uses in a sediment core collected from Bracuhy marina (Southeast Brazil). Analysis target major and trace elements (Cu, Zn, Pb, Cd and Sn), rare earth elements (REEs), and Pb isotopes. The modification in Pb isotopic ratios and REEs pattern unequivocally revealed sediment provenance disruption following the marina construction. Metal distribution in the sediment core demonstrates that concentrations of Cu and Zn increased by up to 15 and 5 times, respectively, compared to the local background. This severe Cu and Zn contamination coincides with the onset of marina operations and can be attributed to the use of antifouling paints.

Palaeoproteomic identification of the original binder and modern contaminants in distemper paints from Uvdal stave church, Norway.

Two distemper paint samples taken from decorative boards in Uvdal stave church, Norway, were analysed using palaeoproteomics, with an aim of identifying their binder and possible contaminants. The results point at the use of calfskin to produce hide glue as the original paint binder, and are consistent with the instructions of binder production and resource allocation in the historical records of Norway. Although we did not observe any evidence of prior restoration treatments using protein-based materials, we found abundant traces of human saliva proteins, as well as a few oats and barley peptides, likely deposited together on the boards during their discovery in the 1970s. This work illustrates the need to fully consider contamination sources in palaeoproteomics and to inform those working with such objects about the potential for their contamination.

New insight from MALDI-TOF MS and multivariate data analysis on the botanical origin of polysaccharide-based paint binders in ancient Egypt.

Polysaccharide-based materials of plant origin are known to have been used as binding media in paint and ground layers of artifacts from ancient Egypt, including wall paintings, cartonnages and sarcophagi. The use of gums from Acacia, Astragalus and Prunus genera has been suggested in the literature on the basis of their qualitative or quantitative monosaccharide profile after complete chemical hydrolysis. The introduction of partial enzymatic digestion of the polysaccharide material, followed by analysis of the released oligosaccharides by matrix assisted laser desorption ionization-time-of-flight mass spectrometry, has proved effective in discriminating among gums from different genera, as well as among species within the Acacia genus. In this study, the previously built Acacia database was expanded, principal component analysis (PCA) was used to aid in grouping of the samples, and data interpretation was refined following a modified acacieae taxonomy. Application of the analytical strategy to investigate the paint binders in artworks from ancient Egypt allowed qualitative discrimination of gums at a species level, and provided new insights into the artists' material choices.

Microplastic-antifouling paint particle contamination alters microbial communities in surrounding marine sediment.

Paint used to coat surfaces in aquatic environments often contain biocides to prevent biofouling, and as these coatings degrade, antifouling paint particles (APPs) end up in aquatic, and especially marine, sediments. However, it is currently unclear what further influence APPs in the sediment have on biotic communities or processes. This study investigates how a variety of commercially-available APPs effect the marine microbial community by spiking different laboratory-manufactured APPs to sediment. Following exposure for 30 and 60 days, APPs caused a clear and consistent effect on the bacterial community composition as determined by 16S metabarcoding. This effect was strongest between 0 and 30 days, but continues to a lesser extent between 30 and 60 days. APPs appear to inhibit the highly diverse, but in general rarer, fraction of the community and/or select for specific community members to become more dominant. 71 antifouling-presence and 454 antifouling-absence indicator taxa were identified by indicator analysis. The difference in the level of classification in these two indicator groups was highly significant, with the antifouling-presence indicators having much higher percentage sequence identity to cultured taxa, while the antifouling-absence indicators appear to be made up of undescribed taxa, which may indicate that APPs act as a proxy for general anthropogenic influence or that APP contamination selects for taxa capable of being cultured. Given the clear and consistent effect APPs have on the surrounding sediment microbial community, further research into how APPs affect sediment functional processes and how such effects scale with concentration is recommended to better assess the wider consequences of these pollutants for marine biogeochemical cycles in the future. SYNOPSIS: Microplastic-paint particles are commonly found in marine sediment but little is known about how these, especially antifouling, paint particles affect sediment microbial communities. This study demonstrates that antifouling paint particles fundamentally alter sediment microbial communities.

Using exterior housing conditions to predict elevated pediatric blood lead levels.

Housing-based lead paint dust is the most common source of lead exposure for US-born children. Although year of housing construction is a critical indicator of the lead hazard to US children, not all housing of the same age poses the same risk to children. Additional information about housing condition is required to differentiate the housing-based lead risk at the parcel level. This study aimed to identify and assess a method for gathering and using observations of exterior housing conditions to identify active housing-based lead hazards at the parcel level. We used a dataset of pediatric blood lead observations (sample years 2000-2013, ages 6-72 months, n = 6,589) to assess associations between observations of exterior housing conditions and housing-based lead risk. We used graphical and Lasso regression methods to estimate the likelihood of an elevated blood lead observation (≥3.5 µg/dL). Our methods estimate a monotonic increase in the likelihood of an elevated blood lead observation as housing conditions deteriorate with the largest changes associated with homes in the greatest disrepair. Additionally we estimate that age of home construction works in consort with housing conditions to amplify risks among those houses built before 1952. Our analysis indicates that a survey of external housing conditions can be used in combination with age of housing in the identification process, at the parcel level, of homes that pose a housing-based lead hazard to children.

Lead in painted surfaces and dusts from rented urban properties (Plymouth, UK).

X-ray fluorescence (XRF) spectrometry has been used to measure lead (Pb) in painted surfaces and indoor dusts from 14 rented properties in the city of Plymouth, UK. Lead was detected in 78 out of 164 surfaces measured (and in 69 out of 150 interior surfaces) with an overall median concentration of 7100 mg kg-1 (range from 157 to 139,000 mg kg-1), and was detected in all but two properties that included one building constructed within the last 25 years. Selected measurements on an area basis (n = 48) returned Pb concentrations ranging from below 0.01 to 16.1 mg cm-2 that were significantly correlated with concentrations on a mass basis but with scatter that was attributed to the degree to which leaded paint had been overpainted. As potential measures of Pb exposure, mean concentrations in samples of lint (n = 8) were <15 mg kg-1 while mean concentrations in hoovered dusts (n = 14) were more variable; specifically, mean Pb dust concentrations ranged from ∼20 to 140 mg kg-1 in 13 samples but was ∼4500 mg kg-1 in a property that had recently undergone extensive renovation. Although mean concentrations of Pb in lint or dust were not related to median Pb concentrations in paint, a strong correlation between barium (Ba) and Pb in dusts suggests that paint is the main source of dust Pb. This study indicates that, in most cases, leaded paints historically applied to interior surfaces have been over-coated and pose little risk, provided that surface coatings are in good condition. However, inappropriate practices during renovation can result in significant contamination of dusts. Our findings are likely to be more broadly applicable to the urban rented sector in the UK and landlords and tenants should, therefore, be made aware of government guidance on the safe decoration or remodelling of older properties where leaded paint is pervasive.

Lead and chromium in European road paints.

Lead chromate was commonly employed as a pigment in coloured road markings until restrictions led to the development of safer alternatives. In this study, the presence and concentrations of Pb and Cr have been determined in 236 road paints of various colours sampled from streets, highways, footways and carparks from eleven European countries. According to energy-dispersive X-ray fluorescence spectrometry, Pb was detected (>10 mg kg-1) in 148 samples at concentrations up to 17.2% by weight, and above 1000 mg kg-1 yellow was the dominant paint colour. Lead concentrations on an area basis varied from 0.02 to 8.46 mg cm-2 and the metal was located at different depths amongst the samples, suggesting that formulations had been painted both recently and historically (and overpainted). Chromium was detected (>5 to 50 mg kg-1) in 81 samples at concentrations between 20 and 20,000 mg kg-1 and most often in yellow paints, and concentrations co-varied with those of Pb. These observations, and results of scanning electron microscopy coupled with energy-dispersive X-ray spectrometry, suggested that heterogeneously dispersed PbCrO4 was the dominant, but not the only, Pb-based pigment in the samples. Although there were significant international differences in frequency of Pb detection and median Pb concentrations, overall, and despite various, albeit complex, regulations, recent or extant road paint pigmented with Pb and Cr remains a pervasive environmental problem and a potential health risk in many European countries.

Comparison of ATR-FTIR and O-PTIR Imaging Techniques for the Characterisation of Zinc-Type Degradation Products in a Paint Cross-Section.

ATR-FTIR (attenuated total reflection-Fourier-transform infrared) microscopy with imaging is widely used in the heritage field to characterise complex compositions of paint cross-sections. However, some limitations include the need for ATR crystal contact with the sample and the inability to resolve particle size below the IR diffraction limit. Recently, a novel O-PTIR (optical-photothermal infrared) spectroscopy technique claimed to open a new avenue for non-invasive, efficient, and reliable analysis at sub-micron resolution. O-PTIR produces transmission-like FTIR spectra for interpretation, without the need to touch the sample, which are highly favourable attributes for analysing heritage samples. This paper reports the comparison of O-PTIR and ATR-FTIR techniques applied to a cross-section embedding a thin paint fragment that delaminated from a late 19th to early 20th-century oil portrait. The hazy paint fragment consisted of zinc soaps (both crystalline and amorphous), gordaite (NaZn4Cl(OH)6SO4·6H2O), and zinc lactate, that could not all be well-resolved with ATR-FTIR imaging. With O-PTIR analysis, the degradation compounds could be resolved at sub-micron resolution with an equivalent or better signal-to-noise ratio. This case study shows how the two techniques can be used to obtain comprehensive information at a broad level with ATR-FTIR and a detailed level with O-PTIR.

Lead levels of new solvent-based household paints in Zimbabwe and Botswana: A preliminary study.

BACKGROUND: Lead paint remains a major potential source of lead poisoning globally, but there has been no documentation on lead content in solvent paints available on the markets in Zimbabwe and Botswana. AIM: To determine the lead content of solvent-based paints available on the market in Zimbabwe and Botswana and identify a need for a larger study to inform policy. METHODS: This pilot study was conducted in Harare, Zimbabwe, and Gaborone, Botswana. Popular brands of solvent-based household paints were bought from hardware shops in Harare (10 samples) and Gaborone (19 samples). Samples were analysed for lead content using inductively coupled plasma-atomic emission spectrometry. RESULTS: Seventy percent of samples from Zimbabwe were found to contain lead above 90 parts per million (ppm), the recommended regulatory limit, with ranges from less than 60 ppm to 12 000 ppm. Twenty percent of Zimbabwean samples had lead levels above 10 000 ppm. No samples from Botswana had lead concentration above the detection limit, with all levels below 100 ppm. LESSON LEARNT: Data strongly suggest very high lead content in popular brands of solvent paints in Zimbabwe, indicating a need for a larger, well-designed study for policy direction.

A novel approach for forensic identification of automotive paints using optical coherence tomography and multivariate statistical methods.

Automotive paint is one of the most important evidence in solving vehicle-related criminal cases. It contains the critical information about the suspected vehicle, providing essential clues for the investigation. In this study, a novel approach based on optical coherence tomography combined with multivariate statistical methods was proposed to facilitate rapid, accurate and nondestructive identification of different brands of automotive paints. 164 automotive paint samples from 8 different manufacturers were analyzed by a spectral-domain optical coherence tomography system (SD-OCT). Two-dimensional cross-sectional OCT images and three-dimensional OCT reconstruction of vehicle paints of different paints were obtained to show the internal structural differences. Visual discrimination of A-scan data after registration and averaging processing was first used to distinguish different samples. An scanning electron microscope was utilized to obtain the cross-sectional image of the sample to evaluate the effectiveness of OCT technique. Then the original A-scan data, first derivative data and second derivative data of 136 paints with four layers from 7 different manufacturers were collected. Multivariate statistical methods, including principal component analysis (PCA), multi-layer perceptron (MLP), k-nearest neighbor (KNN) algorithm and Bayes discriminant analysis (BDA), were used to analyze different datasets. The results show the hybrid PCA and BDA model based on the first derivative OCT data achieved the best result of 100% accuracy on the testing dataset for identifying automotive paints. It is demonstrated that the OCT technique combined with multivariate statistics could be a promising method for identifying the automotive paints rapidly and accurately.

Characterizing paint technologies and recipes in Levantine and Schematic rock art: El Carche site as a case study (Jalance, Spain).

This paper contributes to current debates on the technologies and practices of prehistoric artists using the rock art site of el Carche (Jalance, Spain) as a case study. The site preserves both Levantine and Schematic paintings, yet poorly understood from an analytical point of view. In the past, it has even been argued how little differentiation there is between these two post-Paleolithic traditions in terms of paint composition. Our aim with this paper was to identify pigments, paint recipes and technologies and decipher the order of the superimpositions, both between Levantine motifs of different styles, and between these and the Schematic ones. To do so, we adopted a multi-stage and multi-technical analytical strategy, trying to find a balance between sound scientific investigation and impact on the art, considering the irreplaceable nature of this World Heritage rock art. As such, our approach begins with in situ non-invasive investigations using portable EDXRF, to then collect micro-samples for non-destructive analyses by means of Optical Microscopy, Scanning Electron Microscopy coupled with Energy Dispersive X-Ray Spectroscopy (SEM-EDX), micro-Raman Spectroscopy and Fourier Transform Infrared Spectroscopy (FTIR). One of the key highlights of these paper is the identification of up to four different paint compositions, produced with various hematite-based raw materials and different processing techniques. This variability had not been previously documented. Interestingly though, no direct correlations appear to exist between styles or sub-styles and recipes. Some of these paint mixtures were even shared by both traditions. These results are discussed in cultural terms, challenging previous interpretations suggesting a similar pigment composition between Levantine and Schematic art. Microstratigraphic analysis of the cross-sections only partially clarified the overlapping sequence unveiling the complexity of these analysis. They also revealed several degradation layers and external crusts related to rock alteration processes and biological formations. Their role in rock art conservation is also discussed.

Secret Recipe Revealed: Chemical Evaluation of Raw Colouring Mixtures from Early 19th Century Moravia.

An archaeological excavation in Prostejov (Czech Republic) revealed a workshop of a local potter with colourless, pink, and blue powders presumably used to produce faience/surface decoration. A comprehensive analytical study, which combined elemental and molecular analysis techniques, was performed to shed light on the chemical composition of these unique findings. Scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM EDX), inductively coupled-plasma mass spectrometry (ICP MS), flow injection analysis (FIA) with electrospray ionisation mass spectrometry (ESI MS), laser desorption ionisation mass spectrometry (LDI MS), and Raman spectroscopy were applied to reveal the elemental composition of the powders and identify the colouring agents in the pink and blue powders. The colouring agents in the pink powder were probably iron and the agent in the blue powder is Prussian blue. On top of that, it was also possible to determine the organic additives in these powders through pyrolysis gas chromatography with mass spectrometric detection (Py GC/MS), atmospheric solids analysis probe ion mobility mass spectrometry (ASAP IM MS), and LDI MS. The organic constituents were identified as plant resin, beeswax, and fats. These results point to the preparation of faience/pigment mixtures as oil paint.

Palaeoproteomics guidelines to identify proteinaceous binders in artworks following the study of a 15th-century painting by Sandro Botticelli's workshop.

Undertaking the conservation of artworks informed by the results of molecular analyses has gained growing importance over the last decades, and today it can take advantage of state-of-the-art analytical techniques, such as mass spectrometry-based proteomics. Protein-based binders are among the most common organic materials used in artworks, having been used in their production for centuries. However, the applications of proteomics to these materials are still limited. In this work, a palaeoproteomic workflow was successfully tested on paint reconstructions, and subsequently applied to micro-samples from a 15th-century panel painting, attributed to the workshop of Sandro Botticelli. This method allowed the confident identification of the protein-based binders and their biological origin, as well as the discrimination of the binder used in the ground and paint layers of the painting. These results show that the approach is accurate, highly sensitive, and broadly applicable in the cultural heritage field, due to the limited amount of starting material required. Accordingly, a set of guidelines are suggested, covering the main steps of the data analysis and interpretation of protein sequencing results, optimised for artworks.

Qualitative and quantitative determination of butanol in latex paint by fast gas chromatography proton transfer reaction mass spectrometry.

Butanol is a common organic solvent used in latex paint, and one of its isomers, tert-butanol, is toxic and can cause potential harm to the human body. Therefore, it is of great significance to develop a qualitative and quantitative detection method for butanol isomers. In this study, we combined the advantages of rapid detection of proton transfer reaction mass spectrometry (PTR-MS) with the separation and qualitative capabilities of gas chromatography-mass spectrometry (GC-MS) to achieve the detection of isomers, building a fast gas chromatography proton transfer reaction mass spectrometry (FastGC-PTR-MS) equipment. Firstly, the developed technology was optimized using standard samples of several common volatile organic compounds. The retention times of acetonitrile, acetone, and alcohols were less than 50 s, and the retention times of the benzene series were less than 110 s, on the premise that these isomers could be basically separated (resolution R > 1.0). Compared with a commercial GC-MS equipment, the detection times were shortened by 5-6 times and 2-4 times, respectively. Then the FastGC-PTR-MS was applied to detect the isomers of butanol in latex paint. The results showed that the headspace of brand D latex paint mainly contained five substances: tert-butanol, n-butanol, acetaldehyde, methanol, and acetone. Tert-butanol and n-butanol could be completely separated (R > 1.5). The concentration of tert-butanol was 4.41 ppmv, far below the 100 ppmv maximum allowable workplace concentration. The developed FastGC-PTR-MS can be used for rapid qualitative and quantitative detection of butanol isomers in latex paint. The new equipment has the potential to play an important role in indoor environmental safety applications.

Emerging nuclear methods for historical painting authentication: AMS-14C dating, MeV-SIMS and O-PTIR imaging, global IBA, differential-PIXE and full-field PIXE mapping.

There is a considerable interest in developing new analytical tools to fight the illicit trafficking of heritage goods and particularly of easel paintings, whose high market values attract an ever-increasing volume of criminal activities. The objective is to combat the illicit traffic of smuggled or forged paintworks and to prevent the acquisition of fakes or looted artefacts in public collections. Authentication can be addressed using various investigation techniques, such as absolute dating, materials characterization, alteration phenomena, etc.; for paintings this remains a challenging task due to the complexity of the materials (paint layers, ground, varnish, canvas, etc.) and preferable use of non-destructive methods. This paper outlines results from concerted action on detecting forged works of art within the framework of a Coordinated Research Project of the International Atomic Energy Agency (IAEA) called Enhancing Nuclear Analytical Techniques to Meet the Needs of Forensic Sciences1. One of the main objectives is to foster the use of emerging Nuclear Analytical Techniques (NAT) using particle accelerators for authentication of paintings, with potential application to other forensics domains, by highlighting their ability to determine painting authenticity and to track restorations or anachronistic clues. The various materials comprising a test painting were investigated using an array of NAT. Binder, canvas and support were directly dated by 14C using Accelerator Mass Spectrometry (14C-AMS); binder and pigments' molecular composition was determined using Secondary Ion Mass Spectrometry with MeV ions (MeV-SIMS); paint layer composition and stratigraphy were accurately determined using Ion Beam Analysis (IBA) and differential Particle-Induced X-ray Emission (PIXE); and pigment spatial distributions were mapped using full-field PIXE. High resolution Optical Photothermal Infrared Spectroscopy (O-PTIR) molecular imaging was also exploited. Obtained results are presented and discussed. It is shown that the combination of the above-mentioned techniques allowed reconstructing the history of the test painting.

Concentrations and risk assessment of metals and microplastics from antifouling paint particles in the coastal sediment of a marina in Simon's Town, South Africa.

Maintenance of maritime vessels includes the removal of paint from hulls that are sources of metals, antifouling paint particles (APPs) and microplastics (MPs) that end up in the coastal environment. Simon's Town is a small urban town in False Bay, Cape Town, South Africa, where maritime activities take place (there is a naval harbour, marina and shipyard). The aim of this study was to measure metals, APPs and MPs in Simon's Town, to assess the impact of maritime activities and a storm water pipe in a sheltered marina. Sediment samples were collected from six sites during winter 2018. Sediment and extracted APPs were analysed for metal concentrations (Al, As, B, Ba, Cd, Co, Cr, Cu, Hg, Mo, Ni, Pb, Se, Sb, Sn, Sr, V and Zn) and MPs characterised based on type (shape and polymer), colour and size. Highest average metal concentrations in sediment for all sites were Fe (32228 ± SEM 4024), Al (12271 ± 1062) and Cu (1129 ± 407). Metals in paint particles were highest for Fe (80873 ± 19341), Cu (66762 ± 13082) and Zn (44910 ± 1400 µg/g). Metal and MP fragment concentrations were highest at the slipway of the shipyard, decreasing with increased distance from the slipway. MP filaments were highest close to the storm water outfall pipe. Our results suggest that shipyards are potential sources of metals and MP fragments (mainly APPs), with storm water pipes potential sources of MP filaments. Various indices applied to assess the potential impacts of metals and MPs suggest that these contaminants have the potential to adversely impact the intertidal ecosystem investigated.

Biocides in antifouling paint formulations currently registered for use.

Antifouling paints incorporate biocides in their composition seeking to avoid or minimize the settlement and growing of undesirable fouling organisms. Therefore, biocides are released into the aquatic environments also affecting several nontarget organisms and, thus, compromising ecosystems. Despite global efforts to investigate the environmental occurrence and toxicity of biocides currently used in antifouling paints, the specific active ingredients that have been used in commercial products are poorly known. Thus, the present study assessed the frequencies of occurrence and relative concentrations of biocides in antifouling paint formulations registered for marketing worldwide. The main data were obtained from databases of governmental agencies, business associations, and safety data sheets from paint manufacturers around the world. The results pointed out for 25 active ingredients currently used as biocides, where up to six biocides have been simultaneously used in the examined formulations. Cuprous oxide, copper pyrithione, zinc pyrithione, zineb, DCOIT, and cuprous thiocyanate were the most frequent ones, with mean relative concentrations of 35.9 ± 12.8%, 2.9 ± 1.6%, 4.0 ± 5.3%, 5.4 ± 2.0%, 1.9 ± 1.9%, and 18.1 ± 8.0% (w/w) of respective biocide present in the antifouling paint formulations. Surprisingly, antifouling paints containing TBT as an active ingredient are still being registered for commercialization nowadays. These results can be applied as a proxy of biocides that are possibly being used by antifouling systems and, consequently, released into the aquatic environment, which can help to prioritize the active ingredients that should be addressed in future studies.

Targeted proteomics for the analysis of cultural heritage: application of broadband collision-induced dissociation mass spectrometry.

A broadband collision-induced dissociation (bbCID) fragmentation mode was proposed for liquid chromatography-mass spectrometric targeted analysis of tryptic peptides obtained from proteins in samples of decoration paint coating. In this approach, a mass spectrometric dataset contains the information on the parent and all fragment ions. This maintains a balance between the quantity of simultaneously acquired data and the sensitivity of the method, which is beneficial under coupling with analytical chromatography. In this study, characteristic peptides were selected for casein, ovalbumin, and collagen, which are the most commonly used binder proteins in the artworks. A simplified sample preparation protocol including only protein extraction and trypsinization was tested and successfully implemented. The combination of analytical chromatography with bbCID MS technique is a lower cost alternative to the use of high-end nano-LC-MS approaches in the investigation of cultural heritage objects of regional or local importance, e.g., prior to and/or during restoration works. It was demonstrated that, for the paint coating samples, the required level of sensitivity could be acquired through the data-independent MS/MS strategy. The proposed approach was tested on a sample obtained during the restoration work at the Gromov cottage in the Lopukhin Garden (middle of the XIX century). As a result, the main protein component, collagen, was identified using 6 characteristic peptides, which may indicate the use of gelatin-based glue. For instance, the identification of the peptide GVQGPPoxGPAGPR of the incoming collagen composition α-1 was undertaken by three parameters: m/z of the precursor ion of 553.2910, m/z of the fragment ion y9 of 821.4238, and retention time of 1.9 min.

Critical evaluation of in situ analyses for the characterisation of red pigments in rock paintings: A case study from El Castillo, Spain.

Paint technology, namely paint preparation and application procedures, is an important aspect of painting traditions. With the expansion of archaeometric studies and in situ non-destructive analytical methods, a renewal of technological studies is being observed in rock art. In situ analyses have several limitations that are widely discussed in the literature, however. It is not yet clear whether they provide accurate information on paint technology, except under certain conditions. Here, we evaluated digital microscopic and pXRF in situ analyses for the characterisation of a large set of red and yellow paintings from the El Castillo cave, Cantabria, Spain. We have set experiments and used statistical methods to identify differences between paint components and determine factors impacting pXRF measurements. We found that the compositional heterogeneity of the paintings' environment, especially variations in secondary deposits, was responsible for most of the differences observed between the pXRF signals recorded on the paintings. We concluded that the El Castillo cave environment is not suitable for non-destructive technological studies, but that more favourable contexts might exist. Following previous works and our own results, we advocate a combination of both in situ and laboratory invasive analyses for the study of paint composition and paint technology. Our research protocol, based on the comparison of rock paintings, their substrate, experimental paintings and Fe-normalisation of the signals can improve the reliability of pXRF results. We also propose to include more systematic characterisation of rock wall heterogeneity and the use of microscopic analyses in non-destructive approaches.