Comparison of Three Accelerated Oxidation Tests Applied to Red Wines with Different Chemical Composition

Background: Three accelerated oxidation tests were proposed to simulate red wine oxidation thus providing information useful to correctly manage moderate oxygen exposure of wine during aging in regard to phenolic composition and wine color. Since the results of the tests have never been compared on wines with different initial composition, the aim of this study was to find a suitable method to simulate oxidation of any still red wine. Methods: Aglianico, Barbera, Gaglioppo, Magliocco, and Nerello wines were treated with (1) three cycles of air saturation, (2) the addition of hydrogen peroxide, and (3) the addition of acetaldehyde. Changes in chromatic characteristics and phenolic composition were determined by spectrophotometric and HPLC methods. Results: Important differences in the behavior of the different wines were detected: the highest formation of polymeric pigments was observed in Barbera and Aglianico wines. In contrast, Gaglioppo and Magliocco wines showed a lower variability before and after the oxidation probably due to the lower anthocyanin/tannin ratio. Among the accelerated oxidation tests applied, no significant differences in color parameters and phenolic composition were detected in samples treated with the addition of H₂O₂ and the air saturation method. Conclusion: The study demonstrated that H₂O₂ addition is a successful tool to predict the evolution of different phenolic compounds during the air saturation treatment of wines.     

Phenolic Composition and Total Antioxidant Capacity by Electrochemical,Spectrophotometric and HPLC‐EC Evaluation in Portuguese Red and White Wines

Phenolic composition and in vitro total antioxidant capacity have been evaluated, in red and white Portuguese wines, by electrochemical methods, spectrophotometric methods, and reverse‐phase high performance liquid chromatography with electrochemical detection (HPLC‐EC). The different phenolic compounds present in seventeen red and white wines, with different grape varieties, and from different geographical locations, were successfully identified and detected. The wines in vitro total antioxidant capacity, using the method of capture of diphenilpicrilhydrazil (DPPH^.) free radical “efficient concentration” (EC₅₀), and the electrochemical quantitative index (EI), was determined. The wine with the highest total antioxidant capacity exhibited the lowest antioxidant power, and the EI and EC₅₀ assays had a very good correlation.     

Structural Characterization of a Novel Antioxidant Pigment Produced by a Photochromogenic <Emphasis Type="Italic">Microbacterium oxydans</Emphasis> Strain

The Microbacteriaceae family, such as Microbacterium, is well known for its ability to produce carotenoid-type pigments, but little has been published on the structure of such pigments. Here, we isolated the yellow pigment that is responsible for the yellowish color of a Microbacterium oxydans strain isolated from a decomposing stump of a resinous tree. The pigment, which is synthesized when the bacterium is grown under light, was purified and characterized using several spectroscopic analyses, such as ultraviolet-visible spectroscopy (UV–Vis), Fourier transform infrared spectroscopy (FTIR), ¹H and ¹³C nuclear magnetic resonance (¹H NMR, ¹³C NMR), and high-resolution mass spectrometry (HRMS). From these analysis, a molecular formula (C₂₇H₄₂O₂) and a chemical structure (8-hydroxymethyl-2,4,12-trimethyl-14-(2,6,6-trimethyl-cyclohex-2-enyl)-teradeca-3,7,9,11,13-pentan-2-ol) were deduced. The chemical properties of the pigment, such as aqueous stability at different pH, stability in different organic solvents, and antioxidant capacity, are also reported. Together, these data and previous studies have resulted in the identification of a new antioxidant pigment produced by M. oxydans. To the best of our knowledge, this is the first thorough investigation of this carotenoid-like pigment in the Microbacterium genera.     

The antioxidant activity of wines determined by the ABTS(+) method: influence of sample dilution and time

The free radical scavenging activity of 42 Spanish commercial wines was determined using the 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation (ABTS⁺). The ABTS⁺ radical was generated enzymatically using a horseradish peroxidase and hydrogen peroxide. The presence of wine phenolic compounds caused the absorbance of the radical to decay at 414 nm. The measurement conditions were optimised. The total phenolic content of wines ranged from 1262 to 2389 mg l⁻¹ for red wines and 70 to 407 mg l⁻¹ for white wines, expressed as gallic acid equivalents. The phenolic content of Sherry wines was similar to that of white wines. Optimum dilutions for white and Sherry wines were set up as a function of their total phenolic content (for total phenol index, TPI<300 mg gallic acid per liter, dilution 2.5:10 to 5:10; for TPI>300 mg gallic acid per liter, dilution 1:10 to 3:10). Red wines absorb at the wavelength of measurement and dilutions between 0.35:10 and 0.1:10 are advisable. Reaction kinetics were also monitored and the antioxidant activity, expressed as Trolox Equivalent Antioxidant Capacity (TEAC), was determined at 2 and 15 min of reaction. The mean values for TEAC_2 min were 5.01±1.40 mM for red wines, 0.46±0.32 mM for white wines and 0.26±0.19 mM for Sherry wines. At 15 min, mean values were 6.93±2.41 mM for red wines, 0.67±0.47 mM for white wines and 0.26±0.19 mM for Sherry wines. The correlation coefficients were better at 2 min (r=0.9012) than at 15 min (r=0.8462) when compared with TPI. Hence, TEAC_2 min seems to be a more appropriate measure.     

NMR spectroscopy evaluation of direct relationship between soils and molecular composition of red wines from Aglianico grapes

¹H NMR spectroscopy was employed to investigate the molecular quality of Aglianico red wines from the Campania region of Italy. The wines were obtained from three different Aglianico vineyards characterized by different microclimatic and pedological properties. In order to reach an objective evaluation of “terroir” influence on wine quality, grapes were subjected to the same winemaking procedures. The careful subtraction of water and ethanol signals from NMR spectra allowed to statistically recognize the metabolites to be employed in multivariate statistical methods: Principal Component Analysis (PCA), Discriminant Analysis (DA) and Hierarchical Clustering Analysis (HCA). The three wines were differentiated from each other by six metabolites: α-hydroxyisobutyrate, lactic acid, succinic acid, glycerol, α-fructose and β-d-glucuronic acid. All multivariate analyses confirmed that the differentiation among the wines were related to micro-climate, and carbonate, clay, and organic matter content of soils. Additionally, the wine discrimination ability of NMR spectroscopy combined with chemometric methods, was proved when commercial Aglianico wines, deriving from different soils, were shown to be statistically different from the studied wines. Our findings indicate that multivariate statistical elaboration of NMR spectra of wines is a fast and accurate method to evaluate the molecular quality of wines, underlining the objective relation with terroir.     

Hemisynthesis and structural characterization of flavanol‐(4,8)‐vitisins by mass spectrometry

Vitisins constitute the major group of pyranoanthocyanins naturally occurring in red wines. Regarding their chemical structure, only carboxypyranoanthocyanins have been detected and quantified in red wines, but no vitisin with substitutions in the carbons of the A ring has been identified. However, considering the chemical reactions that take place in red wine, the existence of flavan‐3‐ol‐(4,6/8)‐vitisins is expected. This paper reports for the first time the structural identification of catechin‐vitisin A and catechin‐vitisin B based on their mass data and fragmentation patterns. This work also provides some chromatographic and visible spectroscopic features of these pigments and documents the existence of both in red table wines. Moreover, it is also proposed that Cat‐Vit A pigments arise from the cycloaddition of pyruvic acid to an anthocyanin moiety of a flavanol‐anthocyanin adduct rather than by direct nucleophilic attack of a vitisin A on the carbocation C₄ of catechin. Copyright © 2010 John Wiley & Sons, Ltd.     

The Management of Dissolved Oxygen by a Polypropylene Hollow Fiber Membrane Contactor Affects Wine Aging

Background: Numerous oenological practices can cause an excess of dissolved oxygen in wine, thus determining sensory and chromatic defects in the short- to long-term. Hence, it is necessary to manage the excess of oxygen before bottling. Methods: In this study, the management of the dissolved oxygen content by a polypropylene hollow fiber membrane contactor apparatus was performed in two wines from different grape varieties (Aglianico and Falanghina). The wines were analyzed after an 11-month aging. Anthocyanins and acetaldehyde content were evaluated by HPLC. In addition, other phenolic compounds and chromatic characteristics were analyzed by spectrophotometric methods. NMR and HR ESIMS analyses were conducted to evaluate the amount of pyranoanthocyanins and polymeric pigments. Results: After 11 months of aging, in both wines a decrease of free and total SO₂ with respect to initial values was detected. In the wines with the highest dissolved oxygen levels, a more remarkable loss was observed. No significant differences in terms of color parameters were detected. In red wine with the highest oxygen content, a massive formation of polymeric pigments and BSA reactive tannins was observed, as opposed to wines with lower oxygen levels. Conclusion: The study demonstrated that the membrane contactor can prove a successful tool to manage dissolved oxygen in wines as to prevent their oxidative spoilage.     

Classification of Wines Based on Different Antioxidant Responses to Spectrophotometric Analytical Methods

Six well established spectrophotometric assays (Folin-Ciocalteu, DPPH, ABTS^?+, FRAP, CUPRAC, and the o-diphenols method) have been complementary implemented in order to estimate the total phenolic and o-diphenolic content as well as the free radical scavenging and reducing antioxidant capacities of 40 white and 10 red wines of different varieties and geographic origin. In white wines, the results were weakly correlated; whereas, the results in red wines were strongly correlated, therefore, postulating their separate use in order to estimate efficiently the correlation between results of these methods. The results were subjected to the unsupervised PCA pattern recognition method to investigate the possible classification of white wines. PC analysis framed the wine samples in clear clusters, when the extracted PCA model was based on the results of all six spectrophotometric assays. The usefulness of implementing assays based on different antioxidant mechanisms is discussed. To our knowledge, this is the first time different responses of antioxidants to these spectrophotometric assays have been used to classify white wines according to their variety.     

Synthesis and characterization of environmentally benign calcium-doped Pr2Mo2O9 pigments: Applications in coloring of plastics

A new class of environmentally benign rare earth pigments of general formula Pr_2−xCa_xMo₂O_9−δ (x ranges from 0 to 1.0) displaying colors ranging from green to yellow were synthesized by traditional solid-state route, as alternatives to lead, cadmium and chromium colorants. The products were characterized by X-ray powder diffraction, UV-vis spectroscopy and CIE-L*a*b* 1976 color scales. The coloring mechanism is based on the strong absorptions of the pigments in the blue and red regions due to electronic transitions between 4f²→4f¹5d¹ states of Pr³⁺. The designed pigments consist of non-toxic elements and further found to be thermally and chemically stable. The yellow-green pigments were found to be interesting alternatives to existing toxic pigments for coloration of plastics.     

Bioactivity fingerprint analysis of cyclooxygenase-2 ligands from radix Aconiti by ultrafiltration–UPLC–MSn

A novel fingerprinting method, bioactivity fingerprint analysis, based on an ultrafiltration–ultraperformance liquid chromatography–multistage tandem mass spectrometry (UPLC–MS ⁿ ) method is proposed for the quality control of herbal medicines from the bioactivity viewpoint concerning the efficacy of herbal medicines. The bioactivity fingerprints reflecting the anti-inflammatory activities of radix Aconiti and radix Aconiti preparata were established. With use of ultrafiltration UPLC–MS ⁿ , 11 cyclooxygenase-2 ligands from radix Aconiti preparata and 14 cyclooxygenase-2 ligands from radix Aconiti were found after incubation with cyclooxygenase-2. Twelve of the cyclooxygenase-2 ligands were identified by the ultraperformance UPLC–MS ⁿ method. The enrichment factor of each peak in the bioactivity fingerprint was calculated and was demonstrated to be characteristic, which makes bioactivity fingerprint analysis for the quality control of herbal medicines possible from the viewpoint of their bioactivities.

A noninvasive complementary study of an Egyptian polychrome cartonnage pigments using SEM,EPMA, and Raman spectroscopy

Raman spectroscopy, optical microscopy, scanning electron microscopy (SEM), and electron probe microanalysis (EPMA) were used to study pigments on an Egyptian cartonnage from the Ptolemaic period (305–30 bc ). The surface morphology of each color region was examined using backscattering (BS) and secondary electron imaging. SEM X-ray energy dispersive spectrometry and EPMA wavelength dispersive spectroscopy provided semiquantitative chemical analysis of each pigment. Raman spectroscopy was used to identify the minerals associated with the pigments. This technique confirmed the presence of cinnabar (α-HgS) in the red part of the fragments. A mixture of orpiment (As₂S₃) and bonazziite (β-As₄S₄) and/or alacránite (As₈S₉) was detected in the yellow regions of the fragments. The orange pigment was confirmed to be a mixture of orpiment, uzonite (χ-As₄S₅), and pararealgar (As₄S₄). Egyptian blue (CaCuSi₄O₁₀) and Egyptian green ((Cu,Ca)SiO₃) pigments were detected from blue/green dark-colored regions of the fragments.     

NMR identification of ethyl-linked anthocyanin-flavanol pigments formed in model wine ferments

The ethyl-linked pigments produced by the reaction between either catechin or epicatechin and malvidin 3-O-glucoside with added acetaldehyde have been isolated and characterised by NMR spectroscopy. These pigments are generated in high concentrations in model fermentations containing added malvidin 3-O-glucoside and (epi)catechin when inoculated with Saccharomyces cerevisiae yeast. This confirms that these pigments are produced during fermentation from metabolically produced acetaldehyde and provides evidence that the formation of these pigments may be a significant contributor to the purple colouration of young red wines.     

Chemical Profiling,Bioactivity Evaluation and the Discovery of a Novel Biopigment Produced by Penicillium purpurogenum CBS 113139

Biobased pigments are environmentally friendly alternatives to synthetic variants with an increased market demand. Production of pigments via fermentation is a promising process, yet optimization of the production yield and rate is crucial. Herein, we evaluated the potential of Penicillium purpurogenum to produce biobased pigments. Optimum sugar concentration was 30 g/L and optimum C:N ratio was 36:1 resulting in the production of 4.1–4.5 AU (namely Pigment Complex A). Supplementation with ammonium nitrate resulted in the production of 4.1–4.9 AU (namely Pigment Complex B). Pigments showed excellent pH stability. The major biopigments in Pigment Complex A were N-threonyl-rubropunctamin or the acid form of PP-R (red pigment), N-GABA-PP-V (violet pigment), PP-O (orange pigment) and monascorubrin. In Pigment Complex B, a novel biopigment annotated as N-GLA-PP-V was identified. Its basic structure contains a polyketide azaphilone with the same carboxyl-monascorubramine base structure as PP-V (violet pigment) and γ-carboxyglutamic acid (GLA). The pigments were not cytotoxic up to 250 μg/mL.     

Classifying wine according to geographical origin via quadrupole-based ICP–mass spectrometry measurements of boron isotope ratios

The potential of quadrupole-based ICP–MS as a tool for B-isotopic analysis of wines and its usefulness in provenance determinations were assessed. A precision of 0.1–0.25% RSD (corresponding to a relative standard deviation of the mean of three replicate measurements of 0.06–0.12%) was sufficient to establish small differences in the B isotope ratios in wines from different geographical origins. Each sample measurement was bracketed by measurements of a standard and mass bias drift correction made by interpolation. Sample preparation was kept to a minimum to avoid possible fractionation. Dilution of the wine samples by a factor of 100 with 0.65% HNO₃ was found to reduce matrix-induced mass discrimination substantially. Wines from three wine-producing regions, Stellenbosch, Robertson, and Swartland, in the Western Cape Province of South Africa, and wines from specific regions in France (Bergerac) and Italy (Valpolicella) were analyzed by ICP–QMS for their B-isotopic compositions. It was concluded that the ¹¹B/¹⁰B ratios can be used to characterize wines from different geographical origins. Average ¹¹B/¹⁰B ratios in red wines from South Africa (Stellenbosch), France (Bergerac), and Italy (Valpolicella) were found to differ by between 0.5 and 1.5%.     

Elucidating the Color of Rosé Wines Using Polyphenol-Targeted Metabolomics

The color of rosé wines is extremely diverse and a key element in their marketing. It is due to the presence of anthocyanins and of additional pigments derived from them and from other wine constituents. To explore the pigment composition and determine its links with color, 268 commercial rosé wines were analysed. The concentration of 125 polyphenolic compounds was determined by a targeted metabolomics approach using ultra high-performance liquid chromatography coupled to triple quadrupole mass spectrometry (UHPLC-QqQ-MS) analysis in the Multiple Reaction Monitoring (MRM) mode and the color characterised by spectrophotometry and CieLab parameters. Chemometrics analysis of the composition and color data showed that although color intensity is primarily determined by polyphenol extraction (especially anthocyanins and flavanols) from the grapes, different color styles correspond to different pigment compositions. The salmon shade of light rosé wines is mostly due to pyranoanthocyanin pigments, resulting from reactions of anthocyanins with phenolic acids and pyruvic acid, a yeast metabolite. Redness of intermediate color wines is related to anthocyanins and carboxypoyranoanthocyanins and that of dark rosé wines to products of anthocyanin reactions with flavanols while yellowness of these wines is associated to oxidation.     

Starter units of the biosynthesis of blepharismins: self-defense pigments of Blepharisma japonicum

The heterotrich ciliate Blepharisma japonicum produces red pigment blepharismins, which function as self-defense toxin against predators and as a photoreceptor for step-up photonegativity. The dibenzoperylenequinone moiety of blepharismins was shown to be biosynthesized via the polyketide pathway. In this paper, the starter units of the biosynthetic pathway of blepharismins were determined to be isovaleryl-CoA and butyryl-CoA by HPLC, LC/ESI-MS, and ¹H and ²H NMR analyses of the pigments obtained from feeding experiment of l-leucine or sodium butyrate in excess and deuterium-labeled l-leucine.     

Thermal analysis of the Bi2O3-Er2O3-Zro2 pigments

The synthesis of new compounds based on Bi₂O₃ is investigated because they can be used as new ecological inorganic pigments. Chemical compounds of the Bi_2−xEr_x/2Zr_3x/8O₃ type were synthetized. The host lattice of these pigments is Bi₂O₃ that is doped by Er³⁺ and Zr⁴⁺ ions. The incorporation of doped ions provides interesting colours and contributes to a growth of the thermal stability of these compounds. The simultaneous TG-DTA measurements were used for determination of the temperature region of the pigment formation and thermal stability of pigments. This paper also contains the results of the pigment characterization by X-ray powder diffraction and their colour properties.     

Synthesis and study of mixed oxide inorganic pigment from Bi<Subscript>2</Subscript>O<Subscript>3</Subscript>–ZnO–CeO<Subscript>2</Subscript> system

Novel environment-friendly yellow mixed oxide inorganic pigment from Bi₂O₃–ZnO–CeO₂ system with the composition 23 mol% Bi₂O₃, 15 mol% ZnO and 62 mol% CeO₂ was successfully synthesized by a conventional solid-state reaction method. Comprehensive analyses were carried out to characterize the develop pigment powder including simultaneous TG–DTA thermal analysis, colour properties and particle size distribution. The results demonstrated that the optimum calcination for pigment synthesis was located at a range 800–950 °C. The colour of the studied mixed oxide pigment is connected with the calcination condition. The substitution of Zn²⁺ changes the colour from orange to yellow. The colour of the obtained samples was dependent on the calcination condition and the particle size distribution. The most saturated yellow hue was obtained at the calcination temperature of 950 °C for 2 h in a furnace of pure air and after its application into organic binder in mass tone. The value C of this sample was approx. 65. The mixed oxide pigments were also evaluated from the standpoint of their particle size distribution. Bi₂Ce₂O₇ is considered to be a non-toxic compound, and the other component (Zn²⁺ ions) is also the safe element. Therefore, the present mixed oxide could be an attractive candidate as a novel environment-friendly inorganic yellow pigment.     

Mixed-mode solid-phase extraction followed by acetylation and gas chromatography mass spectrometry for the reliable determination of trans-resveratrol in wine samples

This work presents an advantageous analytical procedure for the accurate determination of free trans-resveratrol in red and white wines. The proposed method involves solid-phase extraction (SPE), acetylation of the analyte in aqueous media and further determination by gas chromatography (GC) with mass spectrometry detection (MS). The use of a mixed-mode SPE sorbent provides an improvement in the selectivity of the extraction step; moreover, the presence of several intense ions in the electron impact mass spectra of its acetyl derivative guarantees the unambiguous identification of trans-resveratrol. Considering a sample intake of 10 mL, the method provides a limit of quantification (LOQ) of 0.8 ng mL⁻¹ and linear responses for concentrations up to 2.5 μg mL⁻¹, referred to wine samples. The average recovery, estimated with samples fortified at different concentrations in the above range, was 99.6% and the inter-day precision stayed below 8%. Trans-resveratrol levels in the analyzed wines varied from 3.4 to 1810 ng mL⁻¹. Cis-resveratrol was also found in all samples. In most cases, equal or higher responses were measured for this latter form than for the trans-isomer. The reduced form of resveratrol, dihydro-resveratrol, was systematically identified in red wines.     

The photochromic effect of bismuth vanadate pigments. Part I: Synthesis, characterization and lightfastness of pigment coatings

We report on investigations of the photochromic effect of BiVO₄ pigments. Emphasis is placed on an approach widely used in industrial color testing. By means of colorimetry ΔE_ab^*-values, which measure the perceived color difference, can be calculated from reflectance spectra of non-illuminated and illuminated pigment coatings. Pigments were prepared by either wet-chemical precipitation or solid-state reactions. Depending on the choice of starting compounds, lightfastness was found to vary significantly. Small amounts of impurity phases do not seem to affect photochromism. In contrast, impurities like Fe and Pb cause intense photochromism. The role of Fe is suggested by trace analyses, which (in case of pigments synthesized by precipitation reactions) reveal a correlation between concentration and ΔE_ab^*. Indications are found that other effects like pigment-lacquer interactions might also be of importance. Difference reflectance spectra turn out to vary in shape depending on the type and concentration of impurities or dopants. For BiVO₄ at least three different mechanisms of photochromism can be assumed.