Analytical characterization of diterpenoid resins present in pictorial varnishes using pyrolysis-gas chromatography-mass spectrometry with on line trimethylsilylation

A procedure based on the technique of the pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS) has been applied, in this work, in order to determine the composition of diterpenoid resin employed in art works. The method is based on the on line derivatization of these resins using hexamethyldisilazane (HMDS). Results obtained were compared with those previously reported in literature from Venice turpentine, Strasbourg turpentine, colophony, sandarac and Manila copal using this same method and with those others from in situ thermally assisted hydrolysis and methylation with tetramethylammonium hydroxide (TMAH). Canada balsam, copper resinate and Copaiba balsam have been also analyzed extending the scope of this method in the field of the analysis of artwork materials. Several non-reported trimethylsilylated derivatives of compounds present in the diterpenoid resins have been identified. An improvement in sensitivity has been obtained by using HMDS as derivatizing reagent, together with a better resolution of the most representative peaks. Additionally, this method reduces the number of pyrolytic fragmentation, recombination, dehydration and isomerization products formed during the pyrolysis process and, in consequence, more simplified chromatograms are obtained. Finally, the reported procedure has been successfully used for characterizing two diterpenoid resin-based varnishes present in the canvas painting "The Betrothal of the Virgin" (Anonymous, 17th century) which is included in the pictorial collection of Saint Joseph Church in Taormina (Italy) and the Magdalena Tryptich (Master of Alzira, 16th century, Valencia, Spain).     

Identification of natural dyes used in works of art by pyrolysis–gas chromatography/mass spectrometry combined with in situ trimethylsilylation

Samples of four natural dyes from different organic families—natural madder (anthraquinonoid), curcuma (curcuminoid), saffron (carotenoid) and indigo (indigotic)—were analysed using a new procedure based on pyrolysis–gas chromatography/mass spectrometry (Py–GC/MS), which includes the on-line derivatisation of the natural dyes using hexamethyldisilazane (HMDS). In addition, a previous procedure involving the addition of a 10% H₂SO₄ aqueous solution to the dye and further separation with ethyl acetate has been tested. This procedure enhances the sensitivity of the method by extracting the colouring compounds from the rest of the compounds present in the natural dye. Two possible derivatising reagents—HMDS and tetramethylammonium hydroxide (TMAH)—were compared in order to assess their effectiveness in the proposed method. Characteristic peaks from trimethylsilyl derivatives of alizarin, quinizarin, xanthopurpurin and purpurin were obtained for madder; peaks from safranal, isophorone and trimethylsilyl derivative of crocetin for saffron; peaks from 4-(4-hydroxy-3-methoxy)phenyl-3-buten-2-one and 4-(4-hydroxy-3-methoxy)phenyl-2-butanone, which are primary pyrolysis products of curcuma, and peaks from indole, 2-methylindole and 2,3-dihydroindol-2-one, which are primary pyrolysis products of indigo, among others, were obtained. The reported procedure leads to the unambiguous identification of the four studied dyes from solid samples formed by individual dyes.Electronic Supplementary Material Supplementary material is available for this article at     

Characterisation of varnishes used in violins by pyrolysis-gas chromatography/mass spectrometry

The correct characterisation and a detailed knowledge of the materials originally used in violin varnishes, like natural resins, is crucial for the conservation in museums and for a suitable restoration technique. The study presented here reports on the potential of pyrolysis (Py) coupled with gas chromatography (GC) and mass spectrometry (MS) for the identification of chemical markers of each resin; this technique is very sensitive and selective, it needs a small quantity of sample and does not require chemical treatments. To improve the chromatographic behaviour of polar compounds the derivatising agent tetramethylammonium hydroxide (TMAH) in combination with pyrolysis has been used, in the so-called TMAH thermochemolysis or thermally assisted hydrolysis and methylation (THM), or more simply pyrolysis-methylation. The natural resins studied were colophony, sandarac, manila copal, elemi, amber and benzoin, mainly composed of terpenic compounds, with the exception of the latter, composed of aromatic compounds. Many compounds were identified; in particular, methyl esters of resinous acids that, individually or in a group, can be used as chemical markers. However, through this technique it was not possible to distinguish between the sandarac and manila copal resins because their chromatographic behaviour is very similar. Finally, the procedure applied has been employed in the characterisation of original varnish samples.     

Compositional analysis of UV-cured acrylic ester resins by pyrolysis–gas chromatography in the presence of organic alkali

Compositional analysis of UV-cured resins consisting of multi-component acrylic esters was studied by pyrolysis–gas chromatography (Py–GC) in the presence of organic alkali, tetramethylammonium hydroxide (TMAH). The pyrograms of the UV-cured resins formed from ethylene oxide modified bisphenol A diacrylate (EBADA) contained specific products such as methyl acrylate (MA) and various dimethyl ethers of ethylene oxide modified bisphenol A reflecting the numbers of ethylene oxide units in the original EBADA. Meanwhile the pyrograms of the UV-cured resins comprised of acrylated polyfunctional aliphatic alcohols such as pentaerythritol triacrylate (PETA) and dipentaerythritol hexacrylate (DPEHA) contained methyl ethers reflecting the structure of the original alcohols. In addition, considerable amounts of pyrolyzates with non-methylated hydroxyl groups were also detected for aliphatic alcohol moieties. The compositions of the UV-cured resins containing multi-component acrylic esters were also analyzed based on the relative yields of the characteristic pyrolyzates of each acrylic ester. For calibration purpose, a series of UV-cured standard samples, which contained known amounts of the individual acrylic ester and neopentylglycol diacrylate (NPGDA) used as an internal standard, were measured. The compositions of the multi-component UV-cured resins determined using this approach showed good agreement with the theoretical values estimated from the feed composition.     

Characterisation of Siccative Oils, Resins and Pigments in Art Works by Thermochemolysis Coupled to Thermal Desorption and Pyrolysis GC and GC-MS

Thermal desorption in the presence of tetramethylammonium hydroxide (TMAH) has been applied to materials belonging to the artistic field to verify the effectiveness of the derivatisation reaction. Thermal desorption of siccative oils and abietane resins give consistent results with traditional pyrolysis in methylation conditions, while as regard antraquinonic dyes the milder thermal treatment leads to the formation of more diagnostic compounds than pyrolysis.     

In-situ pyrolysis and silylation for analysis of lipid materials used in paint layers

Summary Although pyrolysis coupled with gas chromatography and mass spectrometry (Py-GC-MS) is a useful technique for the rapid characterization of the organic materials used by artists, diagnostic pyrolysis products bearing polar groups, for example carboxylic acids, require derivatization (e. g. methylation) before GC separation. In this study we propose the use of hexamethyldisilazane (HMDS) as an effective on-line derivatizing reagent to prepare the trimethylsilyl (TMS) derivatives of fatty acids released from the pyrolysis of fats. Pyrolysis in combination with HMDS has been applied to the analysis of lipid materials employed as painting media, for example siccative oils and egg.     

裂解色谱法甲基化试剂碱性对多不饱和脂肪酸降解和异构化的影响

采用最佳比例裂解甲基化试剂,消除了强碱性甲基化试剂对油脂中多不饱和脂肪酸降解和异构化作用的影响。考察了不同比例甲基化试剂的碱性对多不饱和脂肪酸降解和异构化的程度。     

Identification of Indigo Dyes in Painting Layers by Pyrolysis Methylation and Silylation. A Case Study: “The Dinner of Emmaus” by G. Preti

Reactive pyrolysis, under methylating or silylating conditions, in combination with gas chromatography-mass spectrometry (GC-MS) was evaluated as an analytical method for the detection of indigo dyes in painting layers. Samples with the addition of tetramethylammonium hydroxide (TMAH) or hexamethyldisilazane (HMDS), for pyrolysis/methylation and pyrolysis/silylation experiments, respectively, were pyrolysed at 600 °C by means of a heated filament pyrolyser and the evolved products were analysed on-line by GC-MS. Methyl and silylated pyrolytic markers, related to 2-aminobenzoic acid and the indoxyl moiety, were established from the analysis of synthetic indigo, neat or as a pigment of artificial painting layers containing a siccative oil. The occurrence of these markers was investigated in a real sample, taken from the painting The dinner of Emmaus by G.Preti (XVII century), in order to identify the blu pigment. Positive identification of indigo was achieved by pyrolysis/silylation and confirmed by RAMAN spectroscopy.     

Determination of volatile elements in biological materials by isotopic dilution ETV-ICP-MS after dissolution with tetramethylammonium hydroxide or acid digestion

Isotopic dilution for the determination of Ag, Cd, Hg, Pb and Tl in biological materials by ETV-ICP-MS is proposed. The sample was simply dissolved with tetramethylammonium hydroxide (TMAH) or acid digested in a microwave furnace, with an on line matrix separation. When the dissolution was employed, Ir was used as a chemical modifier for Hg and Pb and Pd was used for Cd and Tl. No modifier was used for Ag. The pyrolysis temperatures were taken from pyrolysis temperature curves. The on line preconcentration was performed in a flow injection system with solenoid valves and was based on the analyte complexation with ammonium diethyldithiophosphate and sorption of the complexes on C(18) bonded to silica gel in a minicolumn. For the digested sample submitted to the analyte preconcentration procedure, a modifier, Ir, was only used for Hg. For the other analytes, since a low pyrolysis temperature, 300 degrees C, was employed, no modifier was added. The isotopic dilution calibration was applied to two certified materials, bovine liver and dog fish muscle, dissolved with TMAH or acid digested, and to another two certified materials, corn bran and rice flour, acid digested and submitted to analyte preconcentration. The obtained concentration values agree with the certified ones, showing that this calibration procedure leads to accurate results in the determination of low concentrations of volatile elements. Due to simplicity, the dissolution with TMAH is very attractive.     

Analytical pyrolysis with in situ thermally assisted derivatisation, Py(HMDS)-GC/MS, for the chemical characterization of archaeological birch bark tar

An analytical method based on pyrolysis in the presence of hexamethyldisilazane followed by gas chromatographic/mass spectrometric analysis, Py(HMDS)-GC/MS, was used for the first time to study birch bark tar, an adhesive that has been used since the Palaeolithic period. Py(HMDS)-GC/MS enabled us to obtain information simultaneously both on polymerised and solvent soluble fractions of birch bark tar. A very particular chromatographic pattern was obtained, which was mainly characterised by the presence of homologous series of linear saturated and unsaturated monocaboxylic fatty acids, and homologous series of straight-chain alkenes and alkanes. The pattern can be linked to suberin and suberan, which are known to be present in birch barks. In addition, lupane-type triterpenoids were also revealed. Py(HMDS)-GC/MS was initially applied to a reference material, and was then used to study the archaeological adhesives from a wooden spindle recovered in a Gallo-Roman burial (3rd century AD) and from the golden sheath of a Thracian dagger (8th-7th centuries BC) recovered in Belogradets (Bulgaria).     

Pyrolysis and THM reactions of melamine and its resins

Melamine resins and a melamine polyester copolymer have been investigated with pyrolysis-GC/MS and thermally assisted hydrolysis and methylation (THM). Conventional pyrolysis yields characteristic volatile products only at temperatures above 500 °C and hexamethylenetetramine is the main product. A series of methylated melamines and secondary pyrolysis products were also found. THM yields high amounts of methylated melamines and therefore provides a sensitive method for the detection of melamine in resins and copolymers. Deduced from this results a preparative approach for the synthesis of methylated melamines with aqueous tetramethylammonium hydroxide solution was successfully developed.     

Dimethyl carbonate as a novel methylating reagent for fatty acids in analytical pyrolysis

Dimethyl carbonate (DMC) was investigated as a mild, harmless and odorless reagent for pyrolytic methylation of fatty acids. Soybean oil was selected as test material for its high content of (poly)unsaturated fatty acids. Pyrolyses were performed at 500, 700 and 900 degrees C by means of a heated platinum filament pyrolyser on-line and off-line to the GC-MS apparatus. Methyl esters of palmitic, linoleic, oleic and stearic acid were formed as prominent products from off-line pyrolysis of soybean oil in the presence of DMC and zeolite 13X. Fatty acid methyl esters (FAMEs) were not observed at important levels in the absence of zeolite, while on-line Py-GC-MS experiments resulted principally in the formation of free fatty acids and hydrocarbons. The FAME profiles obtained from the DMC/zeolite off-line pyrolysis were compared to those resulting from tetramethylammonium hydroxide (TMAH) thermochemolysis and BF3-methanol procedure. The observed differences between pyrolysis and methanolysis methods were principally attributed to the thermal degradation of unsaturated fatty acids. The effectiveness of the DMC/zeolite pyrolytic methylation was further demonstrated by the analysis of tripalmitine and soybean seeds.     

Analytical characterization of binding medium used in ancient Chinese artworks by pyrolysis–gas chromatography/mass spectrometry

In the present work, we attempted to characterize the natural organic binding medium mostly used in ancient Chinese artworks by using the pyrolysis–gas chromatography/mass spectrometry (Py–GC/MS) technique. Chinese lacquer, tung oil, deer glue, colophony and peach gum were broadly used as binding media in Chinese paintings or painted pottery, and have been selected as the study materials in this paper. Artificially aged samples have been analyzed by Py–GC/MS in order to establish a useful reference as convenient and rapid analyses of real samples. The method is based on the on-line derivatization in the presence of excess tetramethylammonium hydroxide (TMAH) (25% aqueous solution) as methylating agent. Analyses results revealed that this simultaneous pyrolysis with methylation derivation technique (SPM) is very useful to the characterization of Chinese lacquer, tung oil and colophony but no obvious improvement to deer glue and peach gum. The fingerprint ions of each studied binding medium were established.     

Characterization of polyvinyl resins used as binding media in paintings by pyrolysis–silylation–gas chromatography–mass spectrometry

Polyvinyl resins, in particular poly(vinyl acetate) emulsions (PVA), are widely used in contemporary paintings as binding media, because of the optimum mechanical and optical properties these polymers confer on the paint layers. A study has been carried out to chemically characterize samples of PVA resins prepared as coating films from commercial products currently available from fine arts suppliers. For this purpose, a new method has been based on "on-line" silylation-pyrolysis using hexamethyldisilazane as derivatisation reagent in pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS). This proposed procedure leads to unambiguous identification of this type of binder and improves conventional direct Py-GC-MS. PVA media used in three contemporary paintings from a private collection and from the Museum of Fine Arts of Málaga (Spain) have been successfully identified with this procedure. As a second step of this work, a study devoted to the characterization of changes in the chemical composition of the PVA commercial products studied has been carried out. Effects induced in the specimens by three different types of artificial accelerated ageing process--thermal, UV light, and in an SO(2)-polluted chamber--were compared.     

Mass spectrometric methodology for the analysis of highly oxidized diterpenoid acids in Old Master paintings

Diterpenoid resins from larch and pine trees and the corresponding fractions in a >100-year-old wax-resin adhesive and varnish and a 200-year-old resin/oil paint sample were analysed with by gas chromatography/mass spectrometry (GC/MS) using several off-line and on-line derivatization methods. The main resin compounds were highly oxidized abietic acids. Important products found are hydroxydehydroabietic acids (OH-DHAs), 7-oxoDHA, di-OH-DHAs and 15-OH-7-oxoDHA. The last two compounds have not been reported to occur in artworks before. Larixyl acetate, an important marker from larch resins, was found to be still present in high amounts in the adhesive. A large number of mass spectra of the different oxidation products and larixol and larixyl acetate are presented and their fragmentation behaviour under electron impact conditions is discussed. An index for the degree of oxidation (IDOX) of the abietic acids is presented as an indicator of the degree of oxidation of the matrix in which the resin is present. The IDOX was 0.10, 0.67, 0.81 and 0.76 for the fresh resins, the dark-aged adhesive, the aged varnish and the resin/oil paint, respectively (measured with pyrolysis (Py)-tetramethylammonium hydroxide (TMAH)-GC/MS). Py-TMAH-GC/MS and direct temperature-resolved mass spectrometry are reliable, valuable and fast techniques for the assessment of the presence and degree of oxidation of diterpenoid resins. Copyright 2000 John Wiley & Sons, Ltd.     

Study of ageing of ketone resins used as picture varnishes by pyrolysis–silylation–gas chromatography–mass spectrometry

This contribution presents a research aimed at the study of the commercial products Laropal K80, Keton N and MS2A, ascribed to ketone resins prepared as thin films, in an attempt to reproduce the pictorial layers and protective finishes that are commonplace in contemporary paintings. For this purpose, a new method based on “online” silylation–pyrolysis using hexamethyldisilazane as a derivatisation reagent in pyrolysis–gas chromatography–mass spectrometry (Py–GC–MS) has been proposed. This procedure leads to the unambiguous identification of such varnishes and improves direct Py–GC–MS. Chemical changes due to the degradation effect of environmental agents have been especially considered. To that end, three different accelerated ageing processes were applied to a series of specimens prepared from the studied commercial products: thermal, UV light and ageing in an SO₂-polluted chamber. Chemical changes due to UV light ageing of Keton N resin are in good agreement with those previously reported in the literature.     

Siloxanes as sources of silanones

Pyrolysis of hexamethyldisiloxane (HMDS) and its copyrolysis with chlorotrimethylsilane and tetrachlorosilane were studied. Based on the data of GLC analysis and on the mass spectrum of the condensate obtained after the pyrolysis of HMDS, it was concluded that HMDS acts as a source of dimethylsilanone. The results of the copyrolysis of HMDS with chlorotrimethylsilane used as a trapping reagent indicate that the dimethylsilanone generated from HMDS can be inserted into the Si−Cl and Si−O bonds. In the copyrolysis of HMDS with tetrachlorosilane serving as a trapping reagent for dimethylsilanone, both dimethylsilanone and dichlorosilanone are generated. Translated fromIzvestiya Akademii Nauk. Seriya Khimicheskaya, No. 9, pp. 1663–1666, September, 1997.     

Behaviour of phospholipids in analytical reactive pyrolysis

Checking for the presence of egg in a painting layer allows to decide whether or not it is a tempera. Several already assessed analytical techniques may be used to perform the chemical analysis for the detection of egg in paintings. As an advantageous and alternative methodology for the determination of egg, a new application of analytical pyrolysis, hyphenated with gas chromatography–mass spectrometry (GC–MS) system, in presence of hexamethyldisilazane (HMDS) and tetramethylammonium-hydroxide (TMAH), is reported here. The innovation lays mainly in the choice of new markers for the presence of egg. It is here demonstrated that in art diagnostic tris-TMS-ester and methyl ester of phosphoric acid, generated by the pyrolysis of standard phospholipids and synthetic painting layers containing egg as binding medium, may be used as new markers for identification of egg in tempera layers. The adoption of these new markers in analytical pyrolysis allows to obtain higher analytical performance with respect to classical markers (fatty acids), especially in terms of yield and, as a consequence, in terms of limit of detection.     

Analytical pyrolysis vs. classical wet chemical analysis to assess the decay of archaeological waterlogged wood

The macromolecular complexity of wood limits the possibility of obtaining complete chemical information on its alteration in archaeological objects. This paper compares the results obtained in the characterisation of the components of archaeological wood by a classical wet chemical method and by an instrumental method based on pyrolysis in presence of hexamethyldisilazane coupled with gas chromatography/mass spectrometry, Py(HMDS)–GC/MS. We compare the results obtained with the two methods quantitatively. This enables us to evaluate the efficiency of Py(HMDS)–GC/MS in assessing the chemical composition and the state of conservation of degraded wood. The material analysed consisted of reference sound wood and waterlogged wood from the ?ó?te historical site, located on a small island on Lake Zarańskie in Poland. The samples are from the remains of settlements dating to a period between the 9th and the 12th centuries AD.     

Fast and selective extraction of nicotine from human plasma based on magnetic strong cation exchange resin followed by liquid chromatography-tandem mass spectrometry

In the study, a fast and selective method based on magnetic separation has been developed for the extraction of nicotine from human plasma using magnetic strong cation exchange (MSCX) resins as adsorbent. MSCX resins were prepared using hydrophobic Fe₃O₄ magnetite as magnetically susceptible component, styrene and acrylic acid as polymeric matrix components, and acetyl sulfonate as the sulfonation agent. The extraction procedure was carried out in a single step by stirring the mixture of diluted plasma sample and MSCX resins in the vortex for 5 min. Then, the resins with adsorbed nicotine were separated from the sample matrix by applying an appropriate magnetic field. Main factors affecting the extraction of nicotine such as the amount of MSCX resins, pH value of the extraction solvent, extraction time, and washing and eluting conditions were optimized. The nicotine eluted from the resins was determined by liquid chromatography–tandem mass spectrometry. The calibration curve obtained by analyzing matrix-matched standards shows excellent linear relationship (r ² = 0.9998) in the concentration range of 10–2,500 ng mL⁻¹. The limit of detection and quantification obtained are 2.9 and 9.7 ng mL⁻¹, respectively. The relative standard deviations of intra- and inter-day obtained are in the range of 1.9–6.9% and 2.5–7.8% with the recoveries ranging from 78.7% to 99.1%. The proposed method was successfully applied to determine nicotine in human plasma phlebotomized from ten male smokers. Nicotine was detectable with the contents ranging from 44.4 to 221.9 ng mL⁻¹ in five samples.