Which polyphenolic compounds contribute to the total antioxidant activities of apple?

The antioxidant activities of eight apple cultivars were studied by using the ferric reducing/antioxidant power (FRAP), the beta-carotene-linoleic acid model system (beta-CLAMS), and the photochemiluminescent (PCL) assays. The antioxidant activity of apples is highly correlated to the total phenolic content (TPC) measured by the Folin-Ciocalteu method and the total polyphenolic index (TPI) obtained by HPLC. Extracts of the peel and flesh were analyzed and assayed separately. The FRAP activities of both peel and flesh extracts correlate well with the TPC (r = 0.95 and 0.99, respectively) and the TPI (r = 0.82 and 0.99, respectively). Similar results were found in the beta-CLAMS activities, showing correlation coefficients of r = 0.90 and 0.91 with the TPC for the peel and flesh extracts and of r = 0.90 and 0.84 with the TPI for the peel and flesh extracts, respectively. The antioxidant activity measured by the PCL assay was not correlated with TPC or TPI due to the lack of integratable lag phase in this method with the flavan-3-ols/procyanidins. Among the five major polyphenolic groups, flavan-3-ols/procyanidins had the highest positive correlation with the FRAP and beta-CLAMS activities: r = 0.84 and 0.88 for the peel extracts, respectively; and r = 0.98 and 0.87 for the flesh extracts, respectively. At individual compound level, epicatechin and procyanidin B2 were the major contributors to the antioxidant activity of apple. Hydroxycinnamic acids may have a significant role in the flesh.     

Polyphenolic profiles in eight apple cultivars using high-performance liquid chromatography (HPLC)

Polyphenolic compounds of apple may play an important role in physiologic functions related to human health. Different polyphenolics may have varied biological activities including antioxidant activity. The objective of this study was to investigate the profiles of polyphenolic compounds in different apple varieties and different parts of an apple. The total and individual polyphenolics differed significantly among the eight apple cultivars grown in Ontario, and the peels had higher concentrations than the flesh. Among the tested cultivars, Red Delicious and Northern Spy had the highest concentrations and Empire the lowest. Five major polyphenolic groups with a total of 16 identified individual compounds were found, among which the dihydroxycinnamic acid esters, phloretin glycosides, and flavan-3-ols were found in both flesh and peel, whereas quercetin glycosides were almost exclusively found in the peel. Cyanidin 3-galactoside was unique to and found only in red apple peels. In both apple peel and flesh, the predominant group of polyphenolics was the procyanidins, followed by quercetin glycosides in the peel and hydroxycinnamic acid esters in the flesh. 3-Hydroxyphloretin 2'-xyloglucoside was newly identified in apple. The results obtained in this study will further the understanding of the polyphenolic composition of apples and their roles in health-promoting physiological functions.     

Apple and pear peel and pulp and their influence on plasma lipids and antioxidant potentials in rats fed cholesterol-containing diets

The aim of this study was to assess the bioactive compounds of apple and pear peel and pulp in vitro and their influence on plasma lipids and antioxidant potentials in vivo. The antioxidant potentials measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH), beta-carotene bleaching (beta-carotene), and nitric oxide inhibition radical scavenging (NO) tests in apple peel and pulp were significantly higher than in pear peel and pulp, respectively. The ethanol extract of apple peels showed the strongest inhibition of lipid peroxidation as a function of its concentration and was comparable to the antioxidant activity of butylated hydroxyanisole. The pear pulp extract had the weakest antioxidant ability, whereas other extracts such as apple pulp and pear peel were nearly equal. The antioxidant activities comprised contributions from polyphenols, phenolic acids, and flavonoids and correlated well with polyphenols and flavonoids. The correlation coefficients between polyphenols and antioxidant activities by DPPH, beta-carotene, and NO were as follows: 0.9207, 0.9350, and 0.9453. Contrarily, the correlation coefficient between the content of dietary fiber and the antioxidant activities test was low. The content of all studied indices in apple and pear peel was significantly higher than in peeled fruits (p < 0.05). Diets supplemented with fruit peels exercised a significantly higher positive influence on plasma lipid levels and on plasma antioxidant capacity of rats than diets with fruit pulps.     

Major phytochemicals in apple cultivars: contribution to peroxyl radical trapping efficiency

Forty-one samples of apples (peel plus pulp), obtained from eight cultivars, were examined for concentration of some important phytochemicals and for antioxidant activity expressed as peroxyl radical trapping efficiency. Five major polyphenolic groups plus ascorbate were identified and quantified by HPLC in the apple varieties. Oligomeric and polymeric proanthocyanidins were found to be about two-thirds of total polyphenols. The antioxidant efficiency of the apple extracts and of representative pure compounds for each group of phytochemicals was measured in a micellar system mimicking lipid peroxidation in human plasma. Although the amount of polyphenols measured by HPLC is similar to that measured by standard methods, the antioxidant efficiency calculated on the basis of the contribution of the pure compounds was lower than the antioxidant efficiency of the apple extracts. The higher efficiency of apples appears to be strictly related to the overwhelming presence of oligomeric proanthocyanidins.     

Cultivar and growing region determine the antioxidant polyphenolic concentration and composition of apples grown in New Zealand

Evidence suggests that increasing consumption of fruit and vegetables contributes to improved health and well-being by providing protection from diseases including various cancers and cardiovascular disease. Although there is uncertainty about which components generate this effect, an attractive hypothesis is that the antioxidants are at least partly responsible. We measured the polyphenolic concentrations in 10 different apple cultivars grown commercially in New Zealand, each sourced from three different geographic regions. Our results showed that the concentration of polyphenolics varied among the apple cultivars, with Pacific Queen containing 2.7 times the amount of polyphenolics found in Cox's Orange. Furthermore, there were significant differences in polyphenolic concentrations in fruit from different regions for some cultivars but not for others. We also measured the polyphenolic concentrations in apple skin and flesh and found that on average 46% of the polyphenolics in whole apples were in the skin. Essentially all of the flavonols (quercetin derivatives) were present in the skin. To maximize the intake of apple polyphenols, it is necessary to consume apples of cultivars with high polyphenolic concentrations such as Pacific Queen and include the skin. Our results also showed that there is potential for promoting apple fruit from specific geographical regions because they contained elevated concentrations of antioxidant polyphenolic compounds.     

Isolation, characterization, and antioxidant activity of E- and Z-p-coumaryl fatty acid esters from cv. Annurca apple fruits

A total of 12 fatty acid esters of Z- and E-p-coumaryl alcohol were isolated from cv. Annurca apple fruit and characterized. This apple variety is widely cultivated in the south of Italy, and the fruits typically undergoe a reddening treatment after harvest. Structures of the p-coumaryl esters were elucidated by GC-MS and (1)H and (13)C NMR after purification of individual compounds by HPLC. It was found that the esters are localized in the fruit peel. During reddening of the fruit, there was a substantial increase in the amount of esters and particularly in molecular species with unsaturated fatty acids. The individual compounds were tested for antioxidant activity, and over half were shown to be at least as effective as alpha-tocopherol.     

Free-radical-scavenging and antioxidant activities of secondary metabolites from reddened cv. Annurca apple fruits

Forty-three secondary metabolites were isolated and characterized from cv. Annurca apple fruit, an apple variety cultivated in the south of Italy. This apple cultivar undergoes a typical reddening treatment after collection. All of the compounds were characterized on the basis of their spectroscopic data. The compounds were tested for their radical-scavenging and antioxidant activities by measuring their capacity to scavenge DPPH* (2,2'-diphenyl-1-picrylhydrazyl radical), H2O2, and NO (nitric oxide) and to inhibit the formation of methyl linoleate conjugated diene hydroperoxides or TBARS (thiobarbituric acid reactive species).     

Identification of the strong vasorelaxing substance scirpusin B, a dimer of piceatannol, from passion fruit (Passiflora edulis) seeds

Piceatannol is present in passion fruit (Passiflora edulis) seeds in high amounts. In this study, we isolated the second major polyphenolic compound of passion fruit seeds and identified it as scirpusin B, which is a dimer of piceatannol. We investigated the antioxidant activities and vasorelaxing effects of these polyphenols. Their antioxidant effects were measured using an in vitro 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, and their vasorelaxant effects were determined ex vivo in rat thoracic aorta. Both polyphenolic compounds exhibited potent antioxidant activities and significant vasorelaxant effects in endothelium-intact aortas. More specifically, scirpusin B exerted a greater antioxidant activity and vasorelaxant effect compared with that of piceatannol. Additionally, the vasorelaxation effects of the compounds were induced via the NO derived from the endothelium. This study provides the possibility that polyphenols in passion fruit seeds are effective against cardiovascular diseases (CVDs).     

Genetic variability in apple fruit polyphenol composition in Malus × domestica and Malus sieversii germplasm grown in New Zealand

Variations in the concentrations of flavan-3-ol, oligomeric procyanidin, chlorogenic acid, dihydrochalcone, flavonol, and anthocyanin polyphenol groups and total polyphenols were examined in the fruit peel and cortical flesh of 93 (80 Malus × domestica and 13 Malus sieversii) apple genotypes in at least 1 year between 2003 and 2005 grown at one site in New Zealand (NZ). Differences among genotypes accounted for 46-97% of the total variation in the concentrations of total polyphenols and each of the individual phenol groups in the flesh and peel in both species, whereas effects of year and genotype × year were minimal, except for peel flavonols in M. × domestica and flesh flavonols in both species. In these cases, differences among genotypes accounted for less than 30% of the total variation, which was less than the variation found for the interaction between genotype and year. Total polyphenol concentrations among genotypes were spread over a 7- and 9-fold range in the flesh and a 4- and 3-fold range in the peel of M. sieversii and M. × domestica, respectively, with the spread in concentrations of individual polyphenol groups in each tissue and within each species varying from a 2-fold to over a 500-fold range. Higher concentrations were generally found in M. sieversii. In M. × domestica, cultivars and breeding selections originating in NZ had lower average flesh and peel total polyphenols and chlorogenic acid than older cultivars previously imported into NZ from overseas countries.     

Determination of polyphenolic profiles of Basque cider apple varieties using accelerated solvent extraction

Polyphenols in the peel and pulp of 15 Basque cider apple varieties were determined by accelerated solvent extraction followed by reversed phase high-performance liquid chromatography with diode array detection. It was observed that the polyphenolic composition in apple peel depended on varieties, whereas the main classes of phenolic compounds in apple pulp were flavan-3-ols and hydroxycinnamic acids in all cases, representing both together between 86 and 95% of total polyphenols assayed.     

Polyphenolic profiles of Basque cider apple cultivars and their technological properties

The polyphenolic compositions of 31 Basque cider apple cultivars were determined in pulp, peel, and juice by high-performance liquid chromatography with diode array detection analysis of crude extracts and after thiolysis. Total polyphenols are distributed in a wide concentration range depending on the cultivar. Procyanidins are the class of polyphenols that present major concentrations in apple. Their average degrees of polymerization range from 4 to 8 depending on the cultivar. Apple cultivars were technologically classified into bitter and nonbitter categories using different classification systems obtained by applying several pattern recognition techniques, such as principal component analysis, K-nearest neighbors, soft independent modeling of class analogy, partial least-squares, and multilayer feed-forward-artificial neural networks, to apple pulp, peel, or juice data (individual polyphenol concentrations, total procyanidin content, and the average degree of polymerization of procyanidins). Bitter apple cultivars present higher contents of flavan-3-ols and/or dihydrochalcones than nonbitter cultivars. Detailed knowledge of the polyphenolic profile of each apple cultivar affords information about their susceptibility to oxidation, their sensory properties (bitterness, astringency), and their possible influence on the characteristics and quality of the final product (juice, cider) when apples are processed.     

Influence of variety and storage on the polyphenol composition of apple flesh

Apple is among the most consumed fruits worldwide. It is available on the market for the whole year being a major source of dietary polyphenols. Several studies suggested that apple polyphenols could play a role in prevention of degenarative diseases. The action of these compounds has been partially ascribed to their antioxidative ability, and fruit antioxidants profile is influenced by apple variety and by the postharvest storage. In this work, the polyphenols composition of the flesh of four apple varieties cultivated in southern Italy were investigated by HPLC, and a flow injection MS/MS procedure to quantify cholorogenic acid and catechins was set up. Phenolic composition and the radical scavenging activity were monitored during a postharvest storage of four months. The quantification by flow injection procedure gives results comparable to those obtained by HPLC, and the increase of the antioxidant activity during storage correlated with an increase of the concentration of catechin and phloridzin. This trend is particularly evident for the variety "Annurca" which is a typical product cultivated in the area around Naples. The genetic characteristics of the Annurca variety together with the anticipated harvest time and the peculiar postharvest conditions are likely responsible for this increase of the antioxidant activity.     

Comparative analysis of polyphenolic profiles and antioxidant and antimicrobial activities of tunisian pome fruit pulp and peel aqueous acetone extracts

Pome trees, apple, pear, and quince, are classified into the subfamily Pomoideae, belonging to the Rosaceae family. Their autumnal fruits are consumed worldwide in different forms, that is, fresh or transformed into jams, jelly, juices, etc. Their well-established beneficial properties to human health were found mainly related to their phenolic content. Pulp and peel aqueous acetone extracts obtained from Tunisian fruits at commercial maturity were comparatively evaluated for their phenolic profiles and antioxidant and antimicrobial potentials. The phenolic compounds present in the extracts were identified and quantified using RP-HPLC-DAD and ESI-MS techniques. Significant differences in the chromatographic profiles among these fruits, as well as between pulp and peel extracts of each fruit, were observed. Quince, followed by 'Red Delicious', peel extracts showed the highest phenolic content (160.33 and 110.90 mg/100 g of fresh weight). The stronger inhibitory effect on DPPH radicals corresponded to those obtained from peel materials. A comparative analysis of the antimicrobial potential against a range of microorganism strains was also carried out. Staphylococcus aureus, Pseudomonas aeruginosa, and Bacillus cereus were the most sensitive to the active extracts. Among the examined phenolic extracts, 'Red Delicious' and quince peels showed the highest effects for inhibiting bacteria growth. Minimum inhibitory and bactericide concentrations ranged from 10(2) to 10(4) microg of polyphenol/mL. Red skin apple and quince peels could be of great interest as important antioxidant and antimicrobial polyphenol sources.     

Antioxidant effectiveness of phenolic apple juice extracts and their gut fermentation products in the human colon carcinoma cell line caco-2

Apples represent a major dietary source of antioxidative polyphenols. Their metabolic conversion by the gut microflora might generate products that protect the intestine against oxidative damage. We studied the antioxidant effectiveness of supernatants of fermented apple juice extracts (F-AEs, 6 and 24 h fermentation) and of selected phenolic degradation products, identified by HPLC-DAD-ESI-MS. Cell free antioxidant capacity of unfermented apple juice extracts (AEs) was decreased after fermentation by 30-50%. In the human colon carcinoma cell line Caco-2, F-AEs (containing <0.5% of original AE-phenolics) decreased the reactive oxygen species (ROS) level more efficiently than the F-blank (fermented without AE) but were less effective than the respective AEs. Similarly, antioxidant effectiveness of individual degradation products was lower compared to respective AE constituents. Glutathione level was slightly increased and oxidative DNA damage slightly decreased by fermented AE03, rich in quercetin glycosides. In conclusion, F-AEs/degradation products exhibit antioxidant activity in colon cells but to a lesser extent than the respective unfermented AEs/constituents.     

Absorption and biological activity of phytochemical-rich extracts from açai (Euterpe oleracea Mart.) pulp and oil in vitro

Polyphenolic extracts from various fruits and vegetables have been shown to exert growth inhibitory effects in cell culture studies. Whereas individual polyphenolic compounds have been extensively evaluated, understanding of the biological activity of polyphenolic extracts from natural sources is limited and critical to the understanding of their potential effects on the human body. This study investigated the absorption and antiproliferative effects of phytochemical extracts from acai pulp and a polyphenolic-enriched acai oil obtained from the fruit pulp of the acai berry ( Euterpe oleracea Mart.). Chemical composition, antioxidant properties, and polyphenolic absorption of phytochemical fractions in a Caco-2 monolayer were determined, along with their cytotoxicity in HT-29 human colon adenocarcinoma cells. Standardized extracts were characterized by their predominance of hydroxybenzoic acids, monomeric flavan-3-ols, and procyanidin dimers and trimers. Polyphenolic mixtures (0-12 microg of gallic acid equiv/mL) from both acai pulp and acai oil extracts inhibited cell proliferation by up to 90.7%, which was accompanied by an increase of up to 2.1-fold in reactive oxygen species. Absorption experiments using a Caco-2 intestinal cell monolayer demonstrated that phenolic acids such as p-hydroxybenzoic, vanillic, syringic, and ferulic acids, in the presence of DMSO, were readily transported from the apical to the basolateral side along with monomeric flavanols such as (+)-catechin and (-)-epicatechin. Results from this study provide further evidence for the bioactive properties of acai polyphenolics and offer new insight on their composition and cellular absorption.     

Apple peel polyphenols protect against gastrointestinal mucosa alterations induced by indomethacin in rats

The stability of an apple peel polyphenol extract (APPE) with powerful antioxidant activity was evaluated under acidic conditions in vitro, and its protective effect against gastrointestinal damage was investigated in rats treated with indomethacin. The antioxidant activity of APPE remained stable at pH 2.0 for 4 h. In rats treated with indomethacin (40 mg/kg ig), the previous administration of APPE protected the gastric, intestinal, and colonic mucosa from oxidative stress by preventing increased malondialdehyde concentrations and decreasing the GSH/GSSG ratio. APPE also displayed anti-inflammatory effects by preventing neutrophil infiltration in the mucosa, as evidenced by the lower myeloperoxidase activity. These protective effects of APPE resulted in the prevention of macro- and microscopic damage and of barrier dysfunction along the gastrointestinal tract of the indomethacin-treated animals. This study supports the concept that apple peel polyphenols may be useful in the prevention and/or treatment of nonsteroidal anti-inflammatory drug-associated side effects.     

Antioxidant activity of apple peels

Consumption of fruits and vegetables has been shown to be effective in the prevention of chronic diseases. These benefits are often attributed to the high antioxidant content of some plant foods. Apples are commonly eaten and are large contributors of phenolic compounds in European and North American diets. The peels of apples, in particular, are high in phenolics. During applesauce and canned apple manufacture, the antioxidant-rich peels of apples are discarded. To determine if a useful source of antioxidants is being wasted, the phytochemical content, antioxidant activity, and antiproliferative activity of the peels of four varieties of apples (Rome Beauty, Idared, Cortland, and Golden Delicious) commonly used in applesauce production in New York state were investigated. The values of the peels were compared to those of the flesh and flesh + peel components of the apples. Within each variety, the total phenolic and flavonoid contents were highest in the peels, followed by the flesh + peel and the flesh. Idared and Rome Beauty apple peels had the highest total phenolic contents (588.9 +/- 83.2 and 500.2 +/- 13.7 mg of gallic acid equivalents/100 g of peels, respectively). Rome Beauty and Idared peels were also highest in flavonoids (306.1 +/- 6.7 and 303.2 +/- 41.5 mg of catechin equivalents/100 g of peels, respectively). Of the four varieties, Idared apple peels had the most anthocyanins, with 26.8 +/- 6.5 mg of cyanidin 3-glucoside equivalents/100 g of peels. The peels all had significantly higher total antioxidant activities than the flesh + peel and flesh of the apple varieties examined. Idared peels had the greatest antioxidant activity (312.2 +/- 9.8 micromol of vitamin C equivalents/g of peels). Apple peels were also shown to more effectively inhibit the growth of HepG(2) human liver cancer cells than the other apple components. Rome Beauty apple peels showed the most bioactivity, inhibiting cell proliferation by 50% at the low concentration of 12.4 +/- 0.4 mg of peels/mL. The high content of phenolic compounds, antioxidant activity, and antiproliferative activity of apple peels indicate that they may impart health benefits when consumed and should be regarded as a valuable source of antioxidants.     

Murta leaves (Ugni molinae Turcz) as a source of antioxidant polyphenols

Extracts from Murta leaves are used by Chilean natives for their benefits on health and cosmetic properties, which are mainly due to the presence of polyphenolic compounds. Extraction of such compounds is strongly influenced by several variables, the effects of which are studied in this work; the antioxidant power of the resulting extracts was measured by two different methods [2,2-diphenyl-1-picrylhydrazyl (DPPH) and thiobarbituric acid reactive substances (TBARS)]. On the whole, maximum values of polyphenolic yields and antiradical power (DPPH method) were attained at 50 degrees C (from 25 to 50 degrees C) and a solvent-to-solid ratio (v/w) of 15:1 (15:1-25:1). The solvents assayed were ethanol, methanol, and water. The highest polyphenolic yield values (2.6% expressed as gallic acid) were reached with methanol, whereas maximum EC50 was attained by the ethanol extract (0.121 mol gallic acid/mol DPPH). Contact time was shown to have only a slight influence in alcoholic extraction, while in water a remarkable effect of increasing contact times (30-90 min) was observed. Just water was the solvent that offered the best result when the antioxidant power was measured by the TBARS method. High-performance liquid chromatography-mass spectrometry analysis revealed the presence of polyphenols, basically flavonols and flavanols, sometimes glycosilated; myricetin and quercetin glycosides were detected in all extracts, whereas epicatechin was present in alcoholic extracts and gallic acid was only present in water.     

Polyphenol composition and antioxidant activity of Kei-apple (Dovyalis caffra) juice

The polyphenolic and ascorbate (ASC) components as well as the antioxidant capacity of Kei-apple (Dovyalis caffra) juice were analyzed and compared to three other fruit juices. The Kei-apple juice had significantly the highest total polyphenolic concentrations (1013 mg gallic acid equivalent/L), and solid phase (C(18)) fractionation identified the majority of these polyphenols to be phenolic acids. The Kei-apple juice also had significantly the highest ASC concentrations (658 mg/L), which showed exceptional heat stability with very little conversion to dehydroascorbate (DHA). Antioxidant capacities of both the unfractionated fruit juices and their solid phase-extracted fractions, as determined by oxygen radical absorbance capacity and ferric reducing antioxidant power analyses, correlated well to the polyphenol concentrations. Gas chromatography-mass spectrometry analyses showed caffeic acid as the most abundant polyphenol present (128.7 mg/L) in the Kei-apple juice; it contributed to 63% of the total antioxidant capacity (of all of the individual compounds identified). Other notable polyphenols identified in higher concentrations included p-coumaric acid, p-hydroxyphenylacetic acid, and protocatechuic acid. Our results therefore support the putative high antioxidant value linked to this fruit and better define this potential in terms of the major antioxidants that exist in the Kei-apple.     

Radical scavenging activities of peels and pulps from cv. Golden Delicious apples as related to their phenolic composition

The relationship between phenolic composition and radical scavenging activity of apple peel and pulp was investigated in fruit produced according to both organic and integrated agricultural methods. Apple tissue extracts were subjected to high-performance liquid chromatography separation, which showed that as compared with pulps, peels are richer in almost all of the quantified phenolics. Flavonols, flavanols, procyanidins, dihydrochalcones, and hydroxycinnamates were the identified phenolic classes in peel tissue, and the most abundant compounds were epicatechin, procyanidin B2, and phloridzin. Pulps were poorer in phytochemicals. Their major phenolics were procyanidins and hydroxycinnamates. Flavonols in amounts <20 mg kg(-1) fresh weight (fw) were also found. In both peels and pulps, integrated production samples were richer in polyphenols. Among the 14 compounds identified, only phloridzin had a tendency to appear higher in organic peels. The total antioxidant capacities (TAC) of extracts were evaluated using the 1,1-diphenyl-2-picrylhydrazyl radical assay and were expressed as Trolox equivalents. Integrated peels gave the highest TAC (18.56 mM kg(-1) fw), followed by organic peels (TAC = 14.96), integrated pulps (TAC = 7.12), and organic pulps (TAC = 6.28). In peels, the top contributors to the antioxidant activity were found to be flavonols, flavanols, and procyanidins, which accounted for about 90% of the total calculated activity whereas in pulps, the TAC was primarily derived from flavanols (monomers and polymers) together with hydroxycinnamates. A good correlation between the sum of polyphenols and the radical scavenging activities was found. Among the single classes of compounds, procyanidins (in peels and pulps) and flavonols (in peels) were statistically correlated to the TAC.