Changes in ochratoxin A and type B trichothecenes contained in wheat flour during dough fermentation and bread-baking.

Ochratoxin A (OTA) and type B trichothecenes are mycotoxins that occur frequently in cereals and thus can be found in cereal by-products such as bread. The aim of this work was to study the variation of the levels of OTA, deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON) and nivalenol (NIV) during the bread-making process. This was done by using wheat flour spiked with different levels of toxins. Mycotoxin levels were controlled after fermentation of the dough with yeasts (Saccharomyces cerevisiae) and after further baking at different temperature-time combinations. Analysis of variance (ANOVA) of the results showed a significant reduction in OTA level (p < 0.05) during fermentation of the dough. The reduction ranged between 29.8% and 33.5%, depending on the initial concentration of toxin in the flour. During this period, the level of the other mycotoxins studied was not modified. By contrast, in the baking phase there were significant changes in the levels of the four mycotoxins, although the reduction was similar under all the baking conditions. Considering all the temperature-time conditions tested, it can be concluded that during the baking period the average reduction of OTA, NIV, 3-ADON, and DON was 32.9%, 76.9%, 65.6%, and 47.9%, respectively.

Characterization of commercial synthetic resins by pyrolysis-gas chromatography/mass spectrometry: application to modern art and conservation.

To characterize a set of synthetic resins, a methodology by pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS) has been developed. The studied reference materials were commercial versions of a wide range of synthetic resins. For each polymer, the pyrolytic and chromatographic conditions were optimized to adequately resolve the fragment mixture in a short time. The proposed analytical method does not require previous treatment of the sample, and due to its high sensitivity, only a small sample quantity in the microgram range can be used. The pyrolysis temperature was found to have little effect on the obtained pyrograms. The summarized data set for the individual polymer materials, especially the characteristic fragments with a structure close to the monomeric unit, was useful to identify commercial synthetic resins. These materials were used in the art and conservation field, as binding media, paint additives, painting varnishes, coatings, or consolidants. Two case studies are introduced where direct Py-GC/MS and thermally assisted hydrolysis and methylation GC/MS were applied on art objects: first, a modern gluing material of a medieval reverse glass painting, and the second example, the binding medium of a painting by Georg Baselitz ("Senta", 1992/1993) from the Sammlung Moderne Kunst at the Pinakothek der Moderne, Munich.

Characterization of the alcoholic fraction of vegetable oils by derivatization with diphenic anhydride followed by high-performance liquid chromatography with spectrophotometric and mass spectrometric detection.

Aliphatic and triterpene alcohols present in vegetable oils have been identified and determined by HPLC using UV-vis and MS detection after previous derivatization with diphenic anhydride. The alcoholic fraction was obtained by saponification, extraction and TLC (according to the European Union official procedure). Derivatization was performed in tetrahydrofuran in the presence of suspended grinded urea, which increases the reaction rate and yield. Derivatized extracts were chromatographed on a C8 column using gradient elution with acetonitrile/water mixtures containing 0.1% acetic acid, with UV-vis followed by negative-ion mode MS detection. Using linear discriminant analysis of the HPLC-MS data (extracted ion chromatograms), oil samples belonging to seven botanical origins (hazelnut, sunflower, corn, extra virgin olive, soybean, peanut and grapeseed) were correctly classified with excellent resolution among all the categories.

Analytical and mineralogical studies of ore and impurities from a chromite mineral using X-ray analysis, electrochemical and microscopy techniques.

A wide analytical study of South African chromite ore, material with high interest in ceramic industry, has been carried out. With this purpose, an accurate chemical identification and mineralogical characterization of the mineral and the gangue have been performed using X-ray fluorescence (XRF), voltammetry, X-ray diffraction (XRD), light microscopy (LM), and scanning electron microscopy (SEM/EDX). The elemental composition of the sample (ore and gangue) has been obtained by XRF. The voltammetric analysis has allowed to demonstrate that iron in the sample was as Fe(II). The main compound of the chromite ore was a spinel (magnesiochromite ferroan), identified by XRD from the sample, which constitutes the chromite ore. This technique has also been useful to characterize some silicates as impurities in the chromite ore sample. Light microscopy has allowed the detection of the spinel and the identification of a silicate impurity (chrome chlorite), by means of their colouration. On the other hand, the other silicate impurity was identified as labradorite by means of X-ray microscopy by SEM/EDX. Finally, a strategy was developed to calculate the composition of each mineral in the unknown sample. The obtained results were: chromite spinel 82.89%, chlorite 12.79% and labradorite 4.32%.

Use of linear discriminant analysis applied to vibrational spectroscopy data to characterize commercial varnishes employed for art purposes.

An improvement of methodologies for characterising synthetic resins used in varnishes employed for art purposes has been suggested. Several kinds of standard of the most common polymeric resins (acrylic, vinyl, poly(vinyl alcohol), alkyd, cellulose nitrate, latex, polyester, polyurethane, epoxy, organosilicic, and ketonic) were analyzed by Fourier transform infrared (FTIR) spectroscopy. Synthetic resins characterization is based on the mathematical treatment of their whole spectrum, dividing it in 13 sections, avoiding the one-by-one interpretation of the absorption bands. The mathematical model takes as variables the maximal absorbance of each section, and each synthetic standard resin as categories. Two exploratory analysis methods, Hierarchical Clustering and Principal Component Analysis (PCA), and a classificatory chemometric tool, linear discriminant analysis (LDA), are tested, validating the models by leave-one-out method. LDA is proved to be a powerful tool for grouping objects in categories, providing a satisfactory distinction of polymeric resin standards. The described analytical procedure has successfully been applied to characterization of synthetic resins contained in commercial varnishes.

Gas chromatography/mass spectrometry characterization of historical varnishes of ancient Italian lutes and violin.

The organic constituents of historical vanishes from two ancient Italian lutes and a Stradivari violin, kept in the Musée de la musique in Paris, have been characterized using gas chromatography-mass spectrometry. Results have been compared with the chromatograms and mass spectra of recent as well as old naturally aged reference materials. The three historical varnishes analyzed have been shown to be oil varnishes, probably mixtures of linseed oil with resins. Identification of diterpenoids and triterpenoids compounds, and of the resins that may have been ingredients of the varnishes, are discussed in this paper.

Infusion mass spectrometry as a fingerprint to characterize varnishes in oil pictorial artworks.

Mass spectrometry methodology to characterize drying oil used as binding media and varnishes in pictorial artworks, prior to conservation or restoration treatment, is proposed. The analytical treatment requires prior basic hydrolysis of the samples to release the fatty acids: caprylic, pelargonic, capric, sebacic, azelaic, suberic, eicosanoic, lauric, mirystic, palmitic, linolenic, linoleic, oleic and stearic, followed by separation from the matrix by a hexane/water extraction. After removing the solvent, the remaining solid is dissolved in potassium hydroxide, propanol and methanol. The mixture is directly infused into a mass spectrometer without any previous derivatization or separation steps. The detector is operated in electrospray negative ion mode and the [M-H](-) ions of the fatty acids enable identification of the acids. Obtained data for fatty acid ion abundances are analyzed by linear discriminant analysis. The drying oils studied (linseed, poppy seed and walnut) were satisfactorily distinguished. The analytical method shows adequate sensitivity, reproducibility, speed and ease. The proposed methodology has been successfully applied to samples from artistic samples belonging to the Cultural Heritage of Valencia (Spain).

Fumonisin production in rice cultures of Fusarium verticillioides under different incubation conditions using an optimized analytical method.

Fumonisin B1 (FB1) and fumonisin B2 (FB2) are the main members of a family of mycotoxins produced by various fungal species belonging to the Gibberella fujikuroi complex. The present work shows the results of a comparative study of various clean-up and derivatization procedures for analysis of fumonisins in rice cultures. Fumonisins were extracted from rice with acetonitrile/water (50/50, v/v). For clean-up, three solid-phase extraction procedures were assayed (C18 cartridge, SAX cartridge, and a combination of both). Two reagents (o-phthaldialdehyde and 4-fluoro-7-nitro-benzofurazan) were studied comparatively for formation of fluorescent derivatives. The separation was carried out by LC using a fluorescence detector. The best procedure for analysis of fumonisins in rice involved clean-up with C18 cartridge and derivatization with o-phthaldialdehyde. The limit of detection was 0.010 mg kg(-1) for both toxins. In the 10-500 mg kg(-1) spiking level range, the recovery rates for FB1 and FB2 in rice varied from 94.6% to 103.6% and from 96.3% to 101.9%, respectively. The optimized analytical method for determination of fumonisins in rice was applied to the study of FB1 and FB2 production by four isolates of the G. fujikuroi species complex in rice cultures carried out at different temperatures and water activities to establish the influence of strain and environmental conditions on fumonisin production in this cereal. In general, fumonisin production was the highest at 20 degrees C and lowest at 37 degrees C. Four of the five assayed water activity (aw) values (0.97, 0.98, 0.99, and 1.0) did not affect significantly fumonisin accumulation but fumonisins were not detected in cultures when aw was 0.96.

H-point standard addition method applied to solid-state stripping voltammetry: quantitation of lead and tin in archaeological glazes.

A solid-state electrochemical application of the H-point standard addition method to the quantification of two depositable metals A and B, which produce strongly overlapped stripping peaks, is described. The method is based on the mechanical transference of mixtures of the solid sample plus a selected compound, of a reference depositable metal R, and of known amounts of a reference material containing A or B, to paraffin-impregnated graphite electrodes. After a reductive deposition step, voltammograms recorded for those modified electrodes immersed into a suitable electrolyte produce stripping peaks for the oxidation of all of the metals deposited. Measurement of the currents at selected potentials in overlapping peaks corresponding to the stripping of A and B permits the quantitation of these metals in the solid sample, while avoiding matrix effects. The method was applied to the simultaneous determination of Pb and Sn in archaeological glazes using PbCO(3) and SnO(2) as standards and ZnO as a reference material.

Characterization of waxes used in pictorial artworks according to their relative amount of fatty acids and hydrocarbons by gas chromatography.

A study attempted to characterize natural waxes used in pictorial works of art was carried out by means of gas chromatography. The analytical treatment requires prior hydrolysis of the waxes to release the fatty acids (FA) (myristic (myr), palmitic (pal), oleic (ole), stearic (ste), araquidic (ara), behenic (beh), lignoceric (lig), cerotic (cer)) from the main esters of the waxes. The formation of volatile derivatives of the fatty acids was carried out by derivatization with ethyl choroformate (ECF). This derivatization reagent was chosen due to the speed, safety and quantitativity of the reaction. The analyzed hydrocarbons were n-eicosane, n-heneicosane, n-docosane, n-tricosane, n-tetracosane, n-pentacosane, n-hexacosane, n-heptacosane, n-octacosane, n-nonacosane, n-tricontane n-hentriacontane, n-dotriacontane, n-tritriacontane, n-tetratriacontane, n-pentatriacontane, main constituents of the waxes. No derivatization is needed to analyze the hydrocarbons. Ethyl ester derivatives and hydrocarbons are adequately separated by gas chromatography, identified by flame ionization detection and confirmed by mass spectrometry. To characterize natural waxes, peak area ratios of each fatty acids with respect to the palmitic acid and peak area ratios of each hydrocarbons with respect to n-heptacosane were calculated. The proposed method provides a good characterization of different waxes most frequently used in artworks, such as beeswax, carnauba wax and ceresin, and has been successfully applied to real samples. This is the first report on the application of ECF to the analysis of fatty acids in wax.

Characterization of proteinaceous glues in old paintings by separation of the o-phtalaldehyde derivatives of their amino acids by liquid chromatography with fluorescence detection.

A HPLC-fluorescence method for characterization of proteinaceous glues from binding media used in pictorial works of art prior to conservation or restoration treatment is proposed. Fluorescence derivatization of amino acids released by acid hydrolysis of standard proteins is studied. The derivatization reagent was o-phtalaldehyde with 2-mercaptoethanol as catalyst. Mobile phase was a programmed gradient among two eluents (water buffered at pH 5.8 wit 5% THF, and methanol) and is able to satisfactorily resolve the amino acid derivatives in 45min. Peak area ratios among amino acid derivatives and the leucine derivative are useful to characterize the proteins. The method shows good sensitivity and adequate linearity between 2.0x10(-3) and 3.3mmol/l of each amino acid, with a limit of detection of 6.0x10(-4)mmol/l. The proposed method has been successfully applied to artistic samples from items of the cultural heritage of Valencia (Spain).

Direct infusion mass spectrometry as a fingerprint of protein-binding media used in works of art.

A direct infusion mass spectrometry method for the characterization of proteinaceous glues from binding media used in pictorial works of art prior to conservation or restoration treatment is proposed. Amino acids are released by acid hydrolysis and dissolved in a mixture of acidic water and ethanol. This mixture is directly infused into a mass spectrometer without any derivatization. The mass spectrometer is operated in positive ion electrospray mode (ESI-MS) to yield [M+H](+) ions for the amino acids. Relative amounts of each amino acid are calculated for each protein (beef and porcine gelatines, albumin, casein and egg). The analyzed proteins were satisfactorily distinguished. The method is easy and fast, and shows good sensitivity and resolution. The proposed method has been successfully applied to artistic samples from items of the cultural heritage of Valencia (Spain).

Identification of lipid binders in old oil paintings by separation of 4-bromomethyl-7-methoxycoumarin derivatives of fatty acids by liquid chromatography with fluorescence detection.

A HPLC-fluorescence method for identification of drying oils from binding media or protective film used in pictorial works of art prior to conservation or restoration is proposed. Fluorescence derivatization of fatty acids released by hydrolysis of structural drying oils is studied. The derivatization reagent was 4-(bromomethyl)-7-methoxycoumarin with 18-crown-6 as catalyst. Mobile phase was programmed from methanol-water (90:10 v/v) to methanol-water (100:0 v/v) in 25 min. The excitation and emission wavelengths were 325 and 395 nm, respectively. Under these chromatographic conditions, coumarin derivatives of myristic, palmitic, oleic and stearic acids were satisfactorily resolved. The method shows good sensitivity, with a detection limit of 6.0 x 10(-8) mmol, and good linearity between 1.0 x 10(-7) and 1.8 x 10(-4) mmol of each analyte. Peak area ratios among fatty acids derivatives, especially the stearic acid/palmitic acid peak area ratio, are useful to identify the drying oils. The proposed method has been successfully applied to artistic samples from items of the cultural heritage of Valencia (Spain).

Identification of diterpenes in canvas painting varnishes by gas chromatography-mass spectrometry with combined derivatisation.

A derivatisation method that combines the formation of ethyl esters from the carboxylic groups and trimethylsilyl ethers from hydroxyl groups of the components of diterpenic resins is presented in this paper. This methodology involves two experimental steps: (1) formation of ethyl esters using ethyl chloroformate; and (2) the esterified compounds are lead to react with trimethylsilylimidazole to form the corresponding trimethylsilyl ethers. The main advantage of the proposed method is the possibility of performing simultaneously the analysis of amino acids from proteins, fatty acids from drying oils, and diterpenic compounds from natural resins usually found in works of art. This methodology is of considerable interest due to the requirements of minimum sampling that usually involves the analysis of works of art. A chemometric study has been developed to adjust the optimal working conditions of the proposed derivatisation method in which chromatographic peak areas of the larixyl acetate derivative and the abietic acid derivative referred to n-hexadecane as internal standard have been compared. Samples of Venetian turpentine naturally aged have been used in this study. Finally, the efficiency of the proposed derivatisation method has been tested on other diterpenic resins and pigments commonly used in fine arts such as Strasbourg turpentine, Canada balsam, colophony, copper resinate and a sample from a Renaissance Altarpiece.

Characterization of bile acids and fatty acids from ox bile in oil paintings by gas chromatography-mass spectrometry.

Characterization of ox bile, traditionally used in painting, is of interest in the fields of archaeometry and conservation and restoration of works of art. Bile acids, fatty acids (F), and cholesterol found in ox bile have been identified using a derivatization method that combines the formation of ethyl esters from the carboxylic groups and the trimethylsilyl ethers from hydroxyl groups. This method of analysis is consistent with these others proposed by the authors to analyze drying oils, proteins, and diterpenic resins usually used as binders and varnishes by the painters. Bile acids from binary samples such as animal glue/ox bile, casein/ox bile and Arabic gum/ox bile have been successfully analyzed using the proposed method. Finally, a method of analysis of mixtures of drying oil and ox bile has been also proposed attempting to quantitatively characterize samples in which ox bile was added to the drying oil for increasing the surfactant properties.

Suppression of pigment interference in the gas chromatographic analysis of proteinaceous binding media in paintings with EDTA.

A method to suppress the interference of pigments in the analysis of proteinaceous media used in paintings is presented in this paper. This method is based on the formation of metallic ion-ethylenediaminetetraacetic acid (EDTA) complexes previous to the derivatisation process, using ethyl chloroformate (ECF), to transform the amino acids in N(O,S)-ethoxycarbonyl (EOC) ethyl esters. Test specimens, containing different proteinaceous media such as albumin, porcine gelatine and casein mixed with lead white, chalk, verdigris and raw Sienna have been prepared for carrying out this study. Different pH conditions have been probed for the different pigments studied. Values of peak area ratio of amino acids relative to the alanine, obtained using the proposed method on a series of protein-pigment test specimens, have been compared to those from specimens of pure protein in which direct method of derivatisation was applied. Finally, the method has been successfully applied to the analysis of 18th century wall paintings in which animal glue was used as binding medium.

Identification of drying oils used in pictorial works of art by liquid chromatography of the 2-nitrophenylhydrazides derivatives of fatty acids.

A new HPLC-UV-Vis method for identification of drying oils from binding media or protective film used in pictorial works of art prior to conservation or restoration is proposed. Chromophore derivatization of fatty acids released by hydrolysis of structural drying oils is studied. The derivatization reagent selected was 2-nitrophenylhydrazine with 1-ethyl-3-(3-dimethyl animopropyl)carbodiimide hydrochloride/pyridine as catalyst. This reaction was carried out using microwave heating. Mobile phase was methanol/water/n-propanol/acetic acid (80:14:5:1) running in isocratic mode. Absorbance was measured at 400nm. In these conditions, hydrazides of myristic, palmitic, oleic, and stearic acids were satisfactorily resolved. Method shows good sensitivity, with a detection limit of 15mumoll(-1), and good linearity between 0.03 and 3mmoll(-1). Peak area ratios among fatty acids derivatives allows identification of the drying oils. The stearic/palmitic ratio is the most important, because it allows to differentiate among the different drying oils. The proposed method has been successfully applied to real samples from items of the cultural heritage of Valencia (Spain).

Application of SEM/EDX and metallographic techniques to the diachronic study (6th-18th century) of metallurgical materials found in archaeological excavations on the island of Ibiza (Spain).

This work is the archaeometric study of the different types of materials recovered in the same excavation or archaeological dig (which makes it possible to accurately date the remains found) on the island of Ibiza (Spain). The samples found belong only to the phase of iron forging and span a very wide historical period from the 6th to the 18th century, including the Islamic period. Scanning Electron Microscopy (SEM) is proposed to observe the images obtained of the surfaces of the samples, in order to study the topographical, morphological and microstructural characteristics providing information on the materials contained in the samples. It also permits chemical analysis of the elements in the sample using X-ray Microanalysis (SEM/EDX), which provides both qualitative and semi-quantitative information on the elements in the sample. The archaeometric results suggest that the sole use of the area studied was as a forge workshop over a long period of time. The slag studied show that no reduction or refining activities took place in the area. However, in this forge, pieces of other metals were also produced, especially bronze alloys, polymetallism being a common characteristic throughout history.

Electrochemical identification of anthraquinone-based dyes in solid microsamples by square wave voltammetry using graphite/polyester composite electrodes.

An electrochemical method for identifying anthraquinone-type dyes in microsamples from works of art, based in the voltammetry of microparticles approach, is reported. Upon attachment onto graphite/polyester composite electrodes, natural pigments aloe, henna, cochineal red, madder lake, kermes, shellac, and alizarin and purpurin taken as reference materials can be identified from their square wave voltammetric profiles in MeCN (0.10 mol L(-1) Bu(4)NPF(6)) and aqueous (0.25 mol L(-1) acetic acid+0.25 mol L(-1) sodium acetate) electrolytes.

Analytical study by SEM/EDX and metallographic techniques of materials used in the iron production process during the iberian period.

This work is the archaeometric study of different types of materials used in iron production in the Iberian Period (6th-2nd century b.c.). The materials were recovered in several archaeological digs (which makes it possible to date the archaeological remains) on different sites in the Levante area of the Iberian Peninsula. The samples selected for this study belong to different materials used in the ancient iron production process such as ores, slags, finished objects, etc. The use of scanning electron microscopy (SEM) is proposed to observe surface images of the samples to determine their morphological, microstructural and topographic characteristics to obtain valuable information on the materials studied. The study also proposes chemical analysis of the elements in the sample by X-ray microanalysis (SEM/EDX) which provides both qualitative and quantitative information. The archaeometric study suggests that the iron obtained was very good quality, but the process was not economically efficient when viewed from today's perspective. Furthermore, the slag produced in the furnace can be related with the slag or impurities contained in the iron. It has also been possible to relate the ores and slag, some furnace conditions, the use of fluxes and also to differentiate types of slag.