Effectiveness of Cyclic Voltammetry in Evaluation of the Synergistic Effect of Phenolic and Amino Acids Compounds on Antioxidant Activity: Optimization of Electrochemical Parameters.

Antioxidant activity can be evaluated using cyclic voltammetry (CV). The aim of this work is to verify the efficacy of CV in evaluating the synergistic effect of bioactive compounds, such as phenolic and amino acid compounds, on antioxidant activity. Therefore, three types of model solutions were prepared: individual model solution (phenol and amino acid), (b) binary model solutions (phenol-phenol and amino acid-amino acid) and (c) mixed phenol-amino acid solutions. Electrochemical measurement conditions were optimized for phenolic compounds (pH 3.0, 1.0 g/L and 100 mV/s) and for amino acids (pH 7.0, 2.0 g/L for amino acids and 100 mV/s), and, for each solution, the functional groups responsible of the anodic and cathodic peaks were established. The peak anodic potential (Epa) and the onset potential (Eon) were two parameters of great importance. The first one was used to classify the solutions according to their antioxidant potential. In general, all the binary and mixed solutions had lower values of Epa than the corresponding individual model solution, which indicates an improvement in the antioxidant potential. The second one was used to evaluate the synergistic effects of phenolic compounds and amino acids.

Assessment of in vitro bioaccessibility of carotenoids and phenolic compounds in a model milk-mandarine beverage.

Mandarine juice is one of the richest sources of ß-cryptoxanthin and flavonoids, which have been positively associated with bone mineral density. Carotenoids are lipophilic isoprenoid compounds with a complex absorption process that can be affected by different factors. In this study, we have evaluated the effect of the food matrix on the in vitro bioaccessibility of carotenoids and phenolic compounds in a model milk-mandarine beverage (MMB). MMBs were formulated with mandarine juice and different dairy products to achieve three fat levels (0.2%, 1.7% and 3.2%) and three calcium levels (120, 310 and 500 mg Ca2+ per 100 ml). The bioaccessibility was evaluated using a harmonised in vitro digestion method. The results showed that the content of milk fat increased the bioaccessibility in vitro of phenolic compounds (p < 0.05), while a moderate fat level (1.7%) resulted in the highest bioaccessibility for bioactive carotenoids. On the other hand, calcium fortification at the highest level (500 mg Ca2+ per 100 mL) decreased the bioaccessibility of bioactive carotenoids from 76% to 43% (66% for the major ß-cryptoxanthin) compared to the lower calcium fortification level (120 mg Ca2+ per 100 mL). The bioaccessibility of hesperidin, the main flavanone in mandarine juice, was significantly (p < 0.05) reduced in the MMB with the highest calcium level. The bioaccessibility of carotenoids and phenolic compounds is affected by fat and calcium levels. When formulating functional beverages, the impact of the formulation should be carefully considered to optimize the bioaccessibility of the bioactive compounds.

Screening for Innovative Sources of Carotenoids and Phenolic Antioxidants among Flowers.

Flowers have been used for centuries in decoration and traditional medicine, and as components of dishes. In this study, carotenoids and phenolics from 125 flowers were determined by liquid chromatography (RRLC and UHPLC). After comparing four different extractants, the carotenoids were extracted with acetone: methanol (2:1), which led to a recovery of 83%. The phenolic compounds were extracted with 0.1% acidified methanol. The petals of the edible flowers Renealmia alpinia and Lantana camara showed the highest values of theoretical vitamin A activity expressed as retinol activity equivalents (RAE), i.e., 19.1 and 4.1 RAE/g fresh weight, respectively. The sample with the highest total phenolic contents was Punica granatum orange (146.7 mg/g dry weight). It was concluded that in most cases, flowers with high carotenoid contents did not contain high phenolic content and vice versa. The results of this study can help to develop innovative concepts and products for the industry.

Chemical characteristics and colorimetric properties of non-centrifugal cane sugar ("panela") obtained via different processing technologies.

Non-centrifugal cane sugar (NCS) samples obtained by traditional moulding and granulation, and also via a novel spray-drying powdering process without additives, were assessed to characterise their sugar and phenolic profiles, flavonoid content, as well as colour parameters. As expected, sucrose was the predominant sugar (91.9-95.5%), followed by glucose (2.9-4.6%), and fructose (1.6-3.7%). Total phenolic content was between 0.4 and 0.6% and total flavonoid content into the range of 0.2-0.4%. Six phenolic acids were found in all NCS samples: protocatechuic acid (0.36-0.94 µg/100 g), vanillic acid (0.70-1.45 µg/100 g), chlorogenic acid (2.08-3.82 µg/100 g), syringic acid (1.08-2.80 µg/100 g), p-coumaric acid (0.69-1.35 µg/100 g), and ferulic acid (0.50-0.95 µg/100 g). The thermal treatment under high temperatures required in the production of granulated products was related with darker colours and changes in phenol and flavonoid contents. In contrast, spray drying generates clearer products, but with slightly less phenol and flavonoid contents.

Extraction of Antioxidants from Winemaking Byproducts: Effect of the Solvent on Phenolic Composition, Antioxidant and Anti-Cholinesterase Activities, and Electrochemical Behaviour.

Extraction solvent is a very important factor in the recovery of antioxidants from natural matrices. In this study, the effect of three solvents (ethanol, ethanol/water and water) on the phenolic composition, antioxidant and anti-cholinesterase activities and electrochemical behaviour of four winemaking byproducts (seeds, skins, stems, and pomace) was evaluated. Phenolic composition was determined by the Folin-Ciocalteu method and ultra-high-performance liquid chromatography (UHPLC), antioxidant activity by the capacity to scavenge 2,2-diphenyl-1-picrylhydrazyl and hydroxyl radicals, anti-cholinesterase activity by the Ellman's method, and electrochemical behaviour by cyclic voltammetry. Eight phenolic compounds were quantified with higher content in water/ethanol extracts (e.g., epicatechin in pomace: 17 mg/100 g vs. 7 and 6 mg/100 g in ethanol and water extracts, respectively), although there were some exceptions (e.g., gallic acid in seeds was most abundant in water extracts). Moreover, the highest total phenolic content (TPC) and antioxidant activity were found in ethanol/water extracts (between 2 and 30-fold the values of the other extracts). Overall, the most active extracts in inhibiting both acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) enzymes were ethanol/water and ethanol extracts from seeds (between 31.11 and 53.90%). The electrochemical behaviour allowed for differentiating the extracts depending on the solvent and the byproduct. Our findings indicate that winemaking byproducts represent a source of phenolic compounds with antioxidant and anti-cholinesterase activities and suggest that cyclic voltammetry is a promising technique to evaluate the phenolic extraction process from these byproducts.

Potential of Cooperage Byproducts Rich in Ellagitannins to Improve the Antioxidant Activity and Color Expression of Red Wine Anthocyanins.

Cooperage byproducts are an important source of phenolic compounds that could be used for wine technology applications. The effects of the addition of two types of oak wood shavings (American, AOW, and Ukrainian, UOW) on the antioxidant activity and color of red wine anthocyanins, in a wine model solution, were evaluated by spectrophotometric and colorimetric analyses. Phenolic compounds from shavings, mainly ellagitannins, were determined by ultra-high-performance liquid chromatography/mass spectrometry (UHPLC/MS). Antioxidant and copigmentation effects varied depending on the type of shavings (AOW and UOW) and the phenolic concentration (100, 400, and 500 mg/L). Phenolic compounds from shavings improved the color characteristics (darker and more bluish color) and the copigmentation effect of red wine anthocyanins, being UOW a better source of copigments than AOW shavings. The best antioxidant activity was found for the 400 and 500 mg/L model solutions for both types of shavings. Results show a winemaking technological application based on the repurposing of cooperage byproducts, which could improve color and antioxidant characteristics of red wines.

Removal of phenolic, turbidity and color in sugarcane juice by electrocoagulation as a sulfur-free process.

This work analyzed the use of electrocoagulation as substitute for sugarcane clarification process using sulfitation. It was evaluated technological parameters (Icumsa color and turbidity), phenolic compounds content and CIELAB color parameters. Four kinetics of reduction color from sugarcane juice were carried out. The essays were divided according to the voltage applied: 35, 45, 55 and 65 V (also based on previous tests). Higher voltage treatments achieved greater reduction of Icumsa color, turbidity and total phenolic compounds. However, none of treatments impacted simple phenolic content analyzed in this work. Tristimulus analysis presented some pattern that went beyond technological analysis, including that 65 V essay changed the pigmentation of sugarcane juice and had an early stabilization on chroma. This kind of results could be useful for industry, once they could correlate quality with different color parameters and finally improve the clarification in general with finer settings of technique according to different situations.

Simultaneous determination of dietary isoprenoids (carotenoids, chlorophylls and tocopherols) in human faeces by Rapid Resolution Liquid Chromatography.

An analytical method was validated for the quantitative determination of isoprenoids compounds in faecal samples, based on liquid-liquid extraction from a small aliquot (0.3-0.5 g of sample) and subsequent analysis by Rapid Resolution Liquid Chromatography (RRLC) on a C30 column. An excellent linear response was observed over the range specified for all dietary isoprenoids, as confirmed by the correlation coefficient, which ranged from 0.9977 to 0.9999. LODs ranged from 0.002 µg to 0.036 µg for lutein and α-tocopherol, respectively. Depending on the compound, LOQs ranged from 0.001 µg (lutein) to 0.120 µg (α-tocopherol). For accuracy testing, spiking of faeces samples with trans-ß-apo-8'-carotenal, α-tocopherol and chlorophyll a were performed (three concentration levels). Excellent recoveries were obtained in all levels (>90%). The intra-day RSD% ranged from 0.86 to 9.78%. The inter-day RSD% was not higher than 10%, except to α-tocopherol (11.34%). In order to assess the applicability of the method faecal samples from a baby fed with different purees formulated from various vegetables were analysed during a six month period. α-carotene, ß-carotene, capsanthin, lycopene, lutein, phytoene, phytofluene, violaxanthin, zeaxanthin and ζ-carotene), and their isomers were identified and quantified using this method. Besides, 2 tocopherols and 9 chlorophylls and derivatives were identified and quantified in the faecal samples analysed. This method is suitable to determine dietary isoprenoids from complex matrices such as human faeces within 28 min.

Study of commercial quality parameters, sugars, phenolics, carotenoids and plastids in different tomato varieties.

The aim of this study was to assess commercial quality parameters, sugars, phenolics, carotenoids and plastid in diverse and little studied tomato varieties to gain insight into their commercial and functional quality and reveal possible noticeable differences. Five cherry tomato varieties and six common (i.e., non-cherry) tomatoes were evaluated. The highest levels of lycopene were detected in 'Tigerella' and 'Byelsa', and those of phytoene in 'Orange', those of phenolics in 'Green Zebra', all of them common tomatoes. The levels of sugars in both groups of tomatoes were comparable. Interesting differences in plastid carotenoid-accumulating sub-structures as a function of the carotenoid profile were observed. Given the importance of chromoplasts in the deposition of carotenoids in plants and their release during digestion, this information can be valuable in investigations on the regulation of the biosynthesis and the bioavailability of tomato carotenoids.

Effects of in vitro gastrointestinal digestion on phenolic compounds and antioxidant activity of different white winemaking byproducts extracts.

The effect of in vitro gastrointestinal digestion on phenolic composition and antioxidant activity of different white winemaking byproducts extracts (grape pomace and its parts: seeds, skins and stems) was evaluated. Fourteen individual phenolic compounds were evaluated by UHPLC. The antioxidant activity was measured by DPPH and ORAC assays. Differences on phenolic profile and antioxidant activity were observed depending on the digestion phase, the type of byproduct, the phenolic group and the antioxidant activity assay. In general, digestion had a reducing effect on TPC and antioxidant activity; however, ORAC values of seed and stem extracts increased after digestion and some recovery indexes of the phenolic groups were very high. Results indicate that extracts from white winemaking byproducts are a reliable source of bioaccessible antioxidant compounds, which could be used as functional food ingredients.

Antioxidants (carotenoids and phenolics) profile of cherry tomatoes as influenced by deficit irrigation, ripening and cluster.

The purpose of this study was to assess the relationship between the effect of regulated deficit irrigation, cluster, developmental stages and two seasons (autumn 2015 and spring 2016) on the commercial and functional quality (carotenoids and plenolics levels) in 'Lazarino' and 'Summerbrix' tomatoes. Autumn had a positive effect on the commercial quality, with larger fruits (22% in 'Summerbrix'; 26% in 'Lazarino') and higher soluble solids (16% in 'Summerbrix'; 12% in 'Lazarino'). Total carotenoids did not change significantly with irrigation and variety while total phenolics did with the cluster and season. In most cases, the main amounts of carotenoids and phenolic were found in the higher cluster and carotenoids in ripe fruit. Thus, irrigation of such varieties could be reduced drastically (ca. 80%) without affecting considerably the overall quality of their fruits (changes not greater than 30%).

Effect of the fruit position on the cluster on fruit quality, carotenoids, phenolics and sugars in cherry tomatoes (Solanum lycopersicum L.).

Cherry tomato varieties are widely consumed in many countries. They contain nutrients and health-promoting compounds like phenolics and carotenoids. The main purpose of this study was to determine how the fruit position on the cluster affected quality parameters of diverse nature (weight, soluble solids, firmness, colour, carotenoids, phenolic compounds and sugars). For this purpose the fourth cluster of two cherry varieties (Summerbrix and Lazarino) were studied. The results indicated that the fruit position on the cluster decreased the fruit size between 14 and 16% and weight 40%; firmness in 'Lazarino' increased 56%; C*ab increased 12% in 'Lazarino' and decreased 13% in 'Summerbrix'; hab increased 9% in 'Summerbrix'. Total carotenoid levels were not statistically different in two positions, and in the other position the contents were higher, with a difference of 36% between these two values. The highest total phenolic levels were observed in the PII position. However, total sugars increased 36%, fructose 36% and glucose 35% from the PI to PIII position in 'Lazarino', while, in 'Summerbrix' higher values were observed in the PIII position.

Effect of regulated deficit irrigation on quality parameters, carotenoids and phenolics of diverse tomato varieties (Solanum lycopersicum L.).

This study aims to evaluate the effects of regulated deficit irrigation (RDI) and of cluster position (CI: first and second cluster; CII: third and fourth cluster; CIII: fifth and sixth cluster) on fruit quality parameters, carotenoids and phenolics in tomatoes. Three common ('Tigerella', 'Palamós' and 'Byelsa') and two cherry varieties ('Lazarino' and 'Summerbrix') were studied. The results showed that the regulated deficit irrigation with reduction of 40 and 50% in the leaf water potential in common and cherry tomatoes did not affect greatly the organoleptic quality of common tomatoes and 'Summerbrix', while cherry varieties were significantly affected with the cluster position. In most case, significant changes in the levels of carotenoids were observed depending on the treatment and the cluster position in all varieties. Significant changes with the treatment and no change with the cluster position were observed in phenolic compounds. Thus, in general, increased total carotenoid levels and reduced the content of phenolic compounds were observed. Considering the significance of changes in the levels of these groups of compounds it was concluded that 'Lazarino' was more susceptible to water deficit, whereas 'Summerbrix' and 'Palamós' were more resistant. On the other hand, the organoleptic and functional quality changed with the variety.

Cyclic voltammetry to evaluate the antioxidant potential in winemaking by-products.

Grape pomace is composed of seeds, skins and stems that are an important source of phenolic substances, which have antioxidant properties and potential benefits to human health. Cyclic voltammetry (CV) has been used to measure the total antioxidant potential of different winemaking by-products. The electrochemical behavior of pomace, seeds, skins and stems was measured by CV and lipid peroxidation inhibition by thiobarbituric acid reactive substances (TBARS) method. Differences for the electrochemical parameter were found between the by-products, pomace and seeds, which presented the greatest voltammetric peak area. Furthermore, the by-products induced inhibition of lipid peroxidation in rat liver homogenates. Pomace and seeds showed higher capacity to inhibit lipid peroxidation than stems and skins, which could be because these by-products are richer in flavanols. Simple regression analyses showed that voltammetric parameters are highly correlated to the values obtained for lipid peroxidation inhibition. CV is a promising technique to estimate the total antioxidant potential of phenolic extract from winemaking by-products.

Foam mat drying of Tommy Atkins mango: Effects of air temperature and concentrations of soy lecithin and carboxymethylcellulose on phenolic composition, mangiferin, and antioxidant capacity.

In this study, foam mat drying was applied to Tommy Atkins mango. Using a multifactorial design, the effect of soy lecithin (L) and carboxymethylcellulose (CMC) used as foam stabilizers (0-1.50g/100g), as well as temperature (T) (53-87°C), on phenolic content and antioxidant capacity of mango were evaluated. Mango pulp contains antioxidant, such as mangiferin, that can be utilized in foods to enhance their functional properties. Our results indicated that L and T had negative effects (p<0.05) on the phenolic content and antioxidant capacity, whereas CMC had a positive effect (p<0.05). Increasing the total amount of phenolic compounds present in dried mango contributed to the higher antioxidant capacity after the drying process. This study concluded that a drying T of 80°C, and a concentration of 0.30g/100g of CMC and L are optimal for increased retention of phenolic compounds and antioxidant capacity.

The Use of Grape Seed Byproducts Rich in Flavonoids to Improve the Antioxidant Potential of Red Wines.

The influence of adding seeds from grape pomace during Syrah wine fermentation in a warm climate has been studied. Seeds of Pedro Ximenez variety were rich in phenolic compounds, mainly flavonoids such as catechin and procyanidins. Changes in total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activity of red wines were observed. These changes depended on the vinification stage and the amount of seeds (SW: 450 g or DW: 900 g seeds/150 kg grapes) applied. In general, antioxidant activity was greater when a simple dose (SW) was considered. Results indicate that seeds rich in flavonoids could be used as wine additives, which could improve the antioxidant potential of red wines in a warm climate.

Comparative Study of Phenolic Profile, Antioxidant Capacity, and Color-composition Relation of Roselle Cultivars with Contrasting Pigmentation.

Roselle is a plant that accumulates anthocyanins significantly, hence its importance as food coloring and as a source of antioxidant compounds for human health. This study was aimed to determine phenolic composition and antioxidant capacity of methanolic extracts, and beverages obtained from native roselle cultivars in Mexico (Negra, Sudan, Rosa and Blanca) with different degrees of pigmentation, and to establish the color-composition relationship. Chromatographic methods were used to determine phenolic compounds: flavanols, flavonols, benzoic, hibiscus and phenolic acids as well as two main anthocyanins (cyanidin 3-sambubioside and delphinidin 3-sambubioside). The antioxidant capacity was evaluated by ABTS and FRAP assays. Tristimulus colorimetry showed to be a useful technique to determine the color-composition relationship, leading to equations that allowed to predict anthocyanin content of roselle (R > 0.84). Also, a stepwise linear discriminant analysis (SLDA) was developed in order to classify roselle cultivars. The obtained mathematical model could be an important tool to be used in colorimetric characterization of functional compounds used in food processing.

Assessment of white grape pomace from winemaking as source of bioactive compounds, and its antiproliferative activity.

The antiproliferative effects of a purified white grape pomace extract (PWGPE), as well as of some phenolic standards on colon cancer cells were examined. The phenolic composition of the PWGPE was determined by rapid resolution liquid chromatography/mass spectrometry (RRLC/MS). The PWGPE had 92.6, 43.3 and 6.01mg/g of flavanols, flavonols and phenolic acids, respectively and, along with pure catechin, epicatechin, quercetin and gallic acid, they were all found capable of inhibiting cellular proliferation. PWGPE (100µg/ml) inhibited the proliferation of cells by 52.1% at 48h, whilst catechin, epicatechin, quercetin and gallic acid (60µg/ml) inhibited growth by 65.2%, 62.2%, 81.0% and 71.0%, respectively, at 72h. The PWGPE is an interesting source of phenolic compounds with antiproliferative properties, that could be of interest in the food and pharmaceutical industries.

Development and validation of a rapid resolution liquid chromatography method for the screening of dietary plant isoprenoids: carotenoids, tocopherols and chlorophylls.

A rapid resolution liquid chromatography (RRLC) method was developed and validated for the simultaneous determination of nine carotenoids compounds (violaxanthin, lutein, zeaxanthin, ß-cryptoxanthin, α-carotene, ß-carotene, lycopene, phytoene, phytofluene), four tocopherols and four chlorophylls and derivates (chlorophylls and pheophytins). The methodology consisted in a micro-extraction procedure with or without saponification and subsequent analysis by RRLC. The limits of detection were <0.07 µg for carotenoids and tocopherols and <0.08 µg for chlorophylls and derivatives. The overall precision values (intra- and inter-day) were lower than 12% when samples were not saponified and <27.6%, when the saponification step was performed. The recovery of the method without the saponification step ranged from 92% to 107%, whilst that when saponification was carried out ranged from 60% for α-tocopherol to 82% for ß-carotene. Finally, the applicability of the method was demonstrated by the identification and quantification of isoprenoids in different samples. The methodology is appropriate for the high-throughput screening of dietary isoprenoids in fruits and vegetables.

Detailed phenolic composition of white grape by-products by RRLC/MS and measurement of the antioxidant activity.

The development and validation of a rapid method of RRLC has been carried out to determine the phenolic composition of winemaking by-products (pomaces, seeds, skins and stems). Thirty-one phenolic compounds belonging to three groups (flavanols, flavonols and phenolic acids) have been identified by use of standards and mass spectrometric detection, and quantified by using the corresponding external standard calibration plot, in a 16-min run. The validation was realized calculating the repeatability, the reproducibility and the limits of detection (LOD) and quantification (LOQ), from standards solutions. The limits of detection and quantification were in the range of 0.16-1.09 and 0.52-3.63 mg/L, respectively, and good repeatability (R.S.D. values <1.5%) and reproducibility (R.S.D. values <5.5%) were found. Results confirmed that the method is effective and suitable for determination of phenolic compounds in winemaking by-products. Seeds, skins, stems and pomaces exhibited a different qualitative and quantitative phenolic profile and different antioxidant activities.