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.     

Simple DPPH.‐Based Electrochemical Assay for the Evaluation of the Antioxidant Capacity: a Thorough Comparison with Spectrophotometric Assays and Evaluation with Real‐World Samples

An assay based on the electrochemical detection of 2,2‐diphenyl‐1‐picrylhydrazyl radical (DPPH^.) for the evaluation of the total antioxidant capacity (TAC) was optimized. The assay is interchangeable with the classic spectrophotometric tests for TAC based on the same radical. In addition, it can be used for the analysis of dilute samples with low antioxidant capacities. A good linear correlation (R²=0.97) was obtained between the results obtained with the proposed electrochemical assay and the Trolox Equivalent Antioxidant Capacity test based on ABTS radical. The assay was successfully used to evaluate the antioxidant capacity of two red wines obtained by six different maceration‐fermentation techniques.     

Classification of Wines Based on Combination of ^1H NMR Spectroscopy and Principal Component Analysis

A combination of ^1H nuclear magnetic resonance （NMR） spectroscopy and principal component analysis （PCA） has shown the potential for being a useful method for classification of type, production origin or geographic origin of wines. In this preliminary study, twenty-one bottled wines were classified/separated for their location of production in Shacheng, Changli and Yantai, and the types of the blended, medium dry, dry white and dry red wines, using the NMR-PCA method. The wines were produced by three subsidiary companies of an enterprise according to the same national standard. The separation was believed to be mainly due to the fermentation process for different wines and environmental variations, such as local climate, soil, underground water, sunlight and rainfall. The major chemicals associated with the separation were identified.     

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.     

Varietal discrimination of Australian wines by means of mid-infrared spectroscopy and multivariate analysis

This study outlines the use of mid-infrared (MIR) spectroscopy combined with principal component analysis (PCA) and linear discriminant analysis (LDA) for the varietal classification of commercial red and white table wines. Three red varieties (Cabernet Sauvignon, Shiraz and Merlot) and four white varieties (Chardonnay, Riesling, Sauvignon Blanc and Viognier) were sourced from different wine regions in Australia. Wine samples were scanned in transmission on a FOSS WineScan FT 120 from wave numbers 926 to 5012 cm⁻¹. All samples were sourced from the 2006 vintage and had not been blended with any other variety or wine from other regions. Spectral data were reduced to a small number of principal components (PCs) and LDA was then performed to successfully separate the wines into the different varieties. To test the robustness of the LDA models developed for the red wines, a set of red wines scanned in 2005 were used. Correct classification of over 95% was achieved for the validation set.     

Occurrence of ochratoxin A in Turkish wines

A total of 95 wine samples including 34 white, 10 rosé and 51 red wines originating from four different Turkish areas were analysed for ochratoxin A (OTA). An analytical method based on immunoaffinity column (IAC) for clean-up and high performance liquid chromatography with fluorescence detection (HPLC-FD) was used to determine OTA in wines. The limit of detection (LOD) was estimated as 0.006 ng ml^− 1 for white wine and 0.010 ng ml^− 1 for rosé and red wines. The limit of quantification (LOQ) was estimated as 0.020 ng ml^− 1 in white wine and 0.030 ng ml^− 1 in rosé and red wines. Recovery experiments were carried out with spiked samples in the range 0.1–1 ng ml^− 1 of OTA. The average OTA recoveries from spiked white wine samples varied from 79.43% to 85.07%; while the mean recoveries for rosé and red wine samples were in the range of 77.48–83.96% and 76.61–83.55%, respectively. OTA was detected in 82 (86%) wine samples at levels of < 0.006–0.815 ng ml^− 1, which were below the maximum allowable limit established by the European Community. The mean OTA concentration in red wines was slightly higher than in white and rosé wines. Furthermore, our data indicate that the geographic region of origin has strong influence on OTA level for white, rosé and red wines: wines originating from Thrace (n = 44, mean = 0.158 ng ml^− 1) and Aegean (n = 28, mean = 0.060 ng ml^− 1) regions of Turkey were more contaminated with OTA compared with wines originating from central (n = 15, mean = 0.027 ng ml⁻¹) and east Anatolia (n = 8, mean = 0.027 ng ml^− 1) areas. This study showed that the occurrence of OTA in Turkish wines is high, but at levels that probably leads to a non-significant human exposure to OTA by consumption of wines.     

Determination of proline in wine using flow injection analysis with tris(2,2'-bipyridyl)ruthenium(II) chemiluminescence detection

Flow injection methodology is described for the determination of proline in red and white wines using tris(2,2′-bipyridyl)ruthenium(II) chemiluminescence detection. Selective conditions were achieved for proline at pH 10, while other amino acids and wine components did not interfere. The precision of the method was less than 1.00% (R.S.D.) for five replicates of a standard (4 × 10⁻⁶ M) and the detection limit was 1 × 10⁻⁸ M. The level of proline in white and sparkling wines using the developed methodology was equivalent to those achieved using HPLC-FMOC amino acid analysis. SPE removal of phenolic material was required for red wines to minimize Ru(bipy)₃³⁺ consumption and its associated effect on accuracy.     

The use of cyclic voltammetry for wine analysis: Determination of polyphenols and free sulfur dioxide

The use of cyclic voltammetry to characterize wines and wine polyphenols in a pH 3.3 model wine solution has been extended to take into account the effects of sulfur dioxide and polyphenol adsorption processes. A good correlation was obtained between a cyclic voltammetric measure, based upon the response produced before and after acetaldehyde additions, and the concentration of free sulfur dioxide in eight white wines (r² = 0.974). By the addition of acetaldehyde to the white wines, an important new step in the methodology, the area under the anodic scan in the potential range from −100 to 1200 mV (Ag/AgCl) closely matched the spectroscopic measure of total polyphenols (absorbance at 280 nm) for the white wines, when both were measured in terms of caffeic acid equivalents (r² = 0.949). The anodic peak area accounted for about 70% of the 280 nm total phenols measure, in catechin equivalents, for the red wines, and a good linear correlation was also obtained (r² = 0.942). The level of catechol and galloyl-containing polyphenols in the wines was calculated by measuring the size of the first anodic peak at around 450 mV after treatment of the wines with acetaldehyde; the peak current correlated well with the total caffeic acid derivatives in the white wines determined by HPLC (r² = 0.982). The concentration of flavonols was estimated by selective adsorption of these compounds onto the carbon electrode and determining the anodic peak current at 1120 mV, with good correlations obtained when compared to total flavonols as measured by HPLC (r² = 0.984 for the red wines, and r² = 0.987 for the white wines).     

Comparison of a Rapid Light-Induced and Forced Test to Study the Oxidative Stability of White Wines

The oxidation processes of white wines can occur during storage and commercialization due to several factors, and these can negatively affect the color, aroma, and quality of the wine. Wineries should have faster and simpler methods that provide valuable information on oxidation stability of wines and allow fast decision-making procedures, able to trigger suitable technological interventions. Using a portable prototype instrument for light irradiations at different wavelengths and times was considered and evaluated on sensorial, spectrophotometric, and colorimetric parameters of white wines. The sensorial analysis revealed that white and light blue were the most significant, after only 1 h of irradiation. The experimental results showed that hydrogen peroxide could enhance the effect of light treatment, allowing a contemporary evaluation of the oxidation stability of wine against light and chemical stresses. As expected, a good correlation (R² > 0.89) between optical density at 420 nm and b* parameter was highlighted. The synergic effect of light and H₂O₂ was also studied on the hydrolyzable and condensed tannins’ additions to white wine. The proposed methodology could be used to evaluate the oxidative stability of white wines, but also to evaluate the effect of some oenological adjuvants on wine stability.     

Phenolic concentrations and antioxidant properties of wines made from north american grapes grown in china

The characteristics of wine phenolics found in several North American and (for comparison) European grape cultivars grown in China were analyzed. This was done to find non-Vitis vinifera wines with prominent features in order to diversify the kinds of wines. The phenolic richness and antioxidant activity decreased in the order: red > rose > white wines. In the red wines, the American grape 'Cynthiana' had the highest total concentrations of phenols, anthocyanins, flavonols and phenolic acids, as well as antioxidant capacity, followed by the French hybrid 'Chambourcin', the lowest were detected in two European grape varieties, 'Merlot' and 'Cabernet Sauvignon', while the total flavon-3-ols levels were reversed among these red grape cultivars. The highest concentration of stilbenes out of all the wines analyzed was found in the 'Merlot' variety. There were significant differences among wine phenolic compositions between North American and European grape cultivars. The antioxidant activities were significantly related to the concentrations of total phenols (r2 = 0.996), anthocyanins (r2 = 0.984), flavonols (r2 = 0.850) and gallic acid (r2 = 0.797). The prominent features of wine aroma and nutrition could make the American grape wines attractive to consumers. It is therefore necessary to perform further research on cultural practices and wine making involving these grapes.     

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.     

Automatic sequential determination of the hydrogen peroxide scavenging activity and evaluation of the antioxidant potential by the 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation assay in wines by sequential injection analysis

An automatic system that performs two analytical procedures, allowing the evaluation of the relative antioxidant capacity of wine samples, was developed. Automation was carried out using a sequential injection analysis (SIA) system that allowed, thanks to its versatility, the development of two methodologies. One is based on the decolorization assay of the 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical, using a spectrophotometric detector. A second methodology allowed the evaluation of the hydrogen peroxide scavenging activity by measuring the oxidation of homovanylic acid (HVA) to its fluorescent dimer, using a fluorescent detector.The developed automatic methodologies were evaluated using trolox as standard and subsequently using other antioxidant substances as gallic acid, caffeic acid, ascorbic acid, catechin and taxifolin which are abundant in wine and whose antioxidant activities were compared to that shown by trolox. The spectrophotometric and fluorimetric assays showed linearity intervals between 0.001 and 0.01 mM, and 0.001 and 0.008 mM of trolox, respectively.The evaluation of the antioxidant power of 20 white and red wine samples, from different Portuguese wine producing regions, was carried out sequentially, in the automatic system. The results were expressed in trolox equivalent antioxidant capacity (TEAC) and presented, for the ABTS and hydrogen peroxide scavenging activity methodologies, detection limits of 8.4 × 10⁻⁷ and 1.4 × 10⁻⁴ mM and relative standard deviation (R.S.D. (%)) in the range 0.6-2.4 and 1-1.8, respectively.     

Phytochemical Compositions and Antioxidant Activities of Essential Oils Extracted from the Flowers of Paeonia delavayi Using Supercritical Carbon Dioxide Fluid

Essential oils were extracted from dark-purple, red and yellow petals of Paeonia delavayi using Supercritical Carbon Dioxide method. The compositions of essential oils were analyzed using gas chromatography-mass spectrometry (GC-MS). Antioxidant activity assays were carried out using DPPH, ABTS- and FRAP methods. Total polyphenols and total flavonoids were measured to evaluate the in vitro antioxidant activity in addition to the volatile compounds contained in the essential oils extracted from the flower petals of P. delavayi with the three flower colors. A total of 194 compounds were detected from essential oils of P. delavayi flowers, including 83 in dark-purple petals, 90 in red petals and 80 in yellow petals. These compounds mainly include alcohols, aldehydes, ketones, alkenes, alkanes, esters and polyphenols. The results showed that the volatile compounds accumulated differentially among the essential oils from the different colors of flower petals. Principal component analysis (PCA) indicated that essential oils derived from dark-purple and red petals were more closely clustered while the yellow petal essential oil was very different with both the purple-red and red. Antioxidant assays suggested that the radical scavenging activity and the iron reduction antioxidant activity in the essential oils were highly correlated with the flower petal colors. These results suggest P. delavayi flower petals are potentially good resources for high quality essential oils and natural antioxidants.     

Electroanalytical Approach to Evaluate Antioxidant Capacity in Honeys: Proposal of an Antioxidant Index

A novel electrochemical route to estimate the antioxidant capacity in honey samples is proposed just using flow injection analysis. The analytical strategy involved the selective oxidation of polyphenolic compounds using two different target potentials, +0.8 and +0.5 V, at two different pHs. An oxidation current obtained at the fixed potential was used as an analytical guide of the antioxidant activity of the target honeys. Chemometrics (correlation and principal component analysis, PCA) demonstrated the significance of the electrochemical protocol versus the traditional spectrophotometric ones in the evaluation of antioxidant capacity and revealed the role of detection potential as a screening variable. The proposed protocol is very simple and fast. However, the most relevant merit of the electrochemical procedure is its inherent versatility which allows the evaluation of the antioxidant activity under predesigned controlled oxidation conditions. In addition, since intercept was statistically zero, its corresponding antioxidant content using just a calibration factor is proposed thus simplifying the calibration‐analysis process. As a result, an electrochemical antioxidant index (EAI) is proposed.     

Characterization of Wines by Trans-Resveratrol Concentration: A Case Study of Romanian Varieties

The goal of this study was to determine the trans-resveratrol concentration in wine. A set of 70 wine samples from Romania, produced according to the appellation of origin system, were analyzed by reverse-phase high performance liquid chromatography. White wines were found to have markedly lower concentrations of trans-resveratrol in comparison with red wines. The values were between 0.025 to 10.23 mg/L for red wine and from 0.074 to 2.572 mg/L for white wine. Red wine is a good dietary source of resveratrol.     

A microcalorimetric sensor for food and cosmetic analyses: l-Malic acid determination

Enzymatic microcalorimetry has been successfully employed in the reliable determination of the l-malic acid concentration in some foods and cosmetic products. The l-malic acid concentration during the wine-making process is particularly useful in order to control the progress of the malo-lactic fermentation. Total acidity, taste and flavour characteristics of wine depend on the l-malic acid quantity still present.To point out the analytical methodology the dehydration process of l-malic acid, in the presence of Fumarase enzyme, has been used. The new method has been compared with a common spectrophotometric one.By the proposed calorimetric method the l-malic acid concentration in different types of food (white and red wines, fruits and soft beverages) has been determined. In some cosmetic products too the l-malic acid was quantified.The method outlined resulted simple, direct and reliable (good accuracy and precision), in particular it does not require any pre-treatment or clean up of the samples, save the dilution in buffer.     

Determination of Flavonoids and Resveratrol in Wine by Turbulent-Flow Chromatography-LC-MS

Turbulent-flow chromatography (TFC) on-line coupled to liquid chromatography mass spectrometry (LC-MS) is used to determine flavonoids and resveratrol in different types of wines. A fully automated system was developed in which 10 mL of sample (diluted wine) was passed over a TFC column, after which the retained analytes were separated by reversed-phase LC and detected by negative ion mode atmospheric-pressure chemical ionization (APCI) MS. The method proved to be fast, non-laborious, robust and sensitive. The feasibility of the method was tested on several red, white and rose wines. Quantitation of resveratrol was possible using the standard addition procedure. Red wine showed the highest amount of resveratrol (4 mg L⁻¹), while rose and white wine contained concentrations which were about ten fold lower.

     

Phenolic profiles of Serbian autochthonous variety ‘Prokupac’ and monovarietal international wines from the Central Serbia wine region

Phenolic profiles of young monovarietal white and red wines from the Central Serbia were investigated. Quantification of phenolics was done using ultra-high-performance liquid chromatography (UHPLC) coupled with a diode array detector and a triple-quadrupole mass spectrometer. Generally, red wines were characterised with higher phenolics content when compared to the white wines. This applies also for the total phenolics content (TPC) and radical scavenging activity (RSA) values. Among all the samples, the red wine from autochthonous Serbian variety ‘Prokupac’ stood out with unique composition, showing the highest TPC, RSA and the highest concentration of the individual phenolics. Finally, based on a specific phenolic pattern, principal component analysis (PCA) indicated clustering of red and white wines, with the ‘Prokupac’ as a prominent outlier.     

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.