Free radical scavenging activity and phenolic content in achenes and thalamus from Fragaria chiloensis ssp. chiloensis, F. vesca and F. x ananassa cv. Chandler

The total phenolic, flavonoid and anthocyanin content of achenes (true fruit) and thalamus (receptacle) from the native South American Fragaria chiloensis ssp. chiloensis (f. patagonica and f. chiloensis), Fragaria vesca and Fragaria x ananassa cv. Chandler was determined by spectrophotometric means. Highest phenolic content was found in F. vesca while lowest content was measured for white strawberry (F. chiloensis ssp. chiloensis, f. chiloensis). The total anthocyanin and total flavonoid contents in the samples investigated was lower for the white strawberry and higher in F. x ananassa cv. Chandler. Total flavonoid content showed a better correlation than total anthocyanins with the free radical scavenging effect of the extracts measured by means of the DPPH discoloration assay. In the superoxide anion assay all the acetone extracts of strawberries showed similar activity. The data presented in this study demonstrate that the amount of phenolic compounds differ significantly between species and subspecies and determine the free radical scavenging activity of fruits. On a w/w basis, higher total phenolics including flavonoids was found in achenes. The highest total anthocyanin content was found in the achenes of F. chiloensis and F. vesca, while F. ananassa presented higher antocyanin content in thalamus. The main anthocyanin in thalamus of F. ananassa (95%) were pelargonidin derivatives which were also present in F. chiloensis ssp. chiloensis f. patagonica (62.6%) but were not detected in F. vesca and F. chiloensis ssp. chiloensis f. chiloensis. In F. vesca and F. chiloensis ssp. chiloensis f. chiloensis only cyanidin derivatives were found both in thalamus and achenes.     

Identification of phenolic compounds in strawberries by liquid chromatography electrospray ionization mass spectroscopy

Strawberry (Fragaria x ananassa Duch.) fruits contain phenolic compounds that have antioxidant, anticancer, antiatherosclerotic and anti-neurodegenerative properties. Identification of food phenolics is necessary since their nature, size, solubility, degree and position of glycosylation and conjugation influence their absorption, distribution, metabolism and excretion in humans. Freeze-dried whole strawberry fruit powder and strawberry fruit extracts were analyzed by liquid chromatography electrospray ionization mass spectrometry (LC–ESI–MS) methods. Phenolics were identified as ellagic acid (EA), EA-glycosides, ellagitannins, gallotannins, anthocyanins, flavonols, flavanols and coumaroyl glycosides. The anthocyanidins were pelargonidin and cyanidin, found predominantly as their glucosides and rutinosides. The major flavonol aglycons were quercetin and kaempferol found as their glucuronides and glucosides. LC–ESI–MS/MS methods differentiated EA from quercetin conjugates since both aglycons have identical molecular weights (302 g/mol). The identification of strawberry phenolics is necessary to generate standardized materials for in vitro and in vivo studies and for the authentication of strawberry-based food products.     

Antioxidant compounds and antioxidant capacity of Peruvian camu camu (Myrciaria dubia (H.B.K.) McVaugh) fruit at different maturity stages

The antioxidant capacities of ascorbic acid and phenolic compounds present in camu camu fruit were screened during ripening. Ascorbic acid decreased, and anthocyanin, flavonol and flavanol contents, and DPPH antioxidant capacity increased during ripening. Antioxidant compounds from camu camu were fractionated in two fractions: an ascorbic acid-rich fraction (F-I) and a phenolics-rich fraction (F-II). F-I was the major contributor to the DPPH antioxidant capacity (67.5–79.3%) and F-II played a minor role (20.7–32.5%). A total of 30 different phenolic compounds were detected by HPLC-PAD. The presence of catechin, delphinidin 3-glucoside, cyanidin 3-glucoside, ellagic acid and rutin was elucidated. Other phenolic compounds, such as flavan-3-ol, flavonol, flavanone and ellagic acid derivatives, were also present. For the three ripening stages the flavan-3-ols and ellagic acid group were the most representative phenolic compounds in this fruit. Acid hydrolysis of F-II revealed the presence mainly of gallic and ellagic acids, suggesting that camu camu fruit possesses important quantities of hydrolysed tannins (gallo- and/or ellagitannins). These results confirm that camu camu fruit is a promising source of antioxidant phenolics.     

Identification and quantification of phenolic compounds and their effects on antioxidant activity in pomegranate juices of eight Iranian cultivars

Important health-promoting compounds, including six types of anthocyanins, phytoestrogenic flavonoids and ellagic acid were determined individually in pomegranate juices (Punica granatum L.) of eight Iranian cultivars by high performance liquid chromatography coupled to UV–vis detector (HPLC–UV) using individual calculation from the peak area based on standard curves of each component. Total phenolics and antioxidant activities were determined by Folin–Ciocalteu and 2,2-diphenyl-1-picrylhydrazyl (DPPH) methods, respectively, and compared among the cultivars. The predominant anthocyanins were delphinidin 3,5-diglucoside (372–5301 mg/l) followed by cyanidin 3,5-diglucoside (242–2361 mg/l), delphinidin 3-glucoside (49–1042 mg/l) and pelargonidin 3,5-diglucoside (7–90 mg/l), respectively. The highest level of total tannins was found in Sweet Alak cultivar (3 mg/l). Saveh Black Leather showed the highest level of ellagic acid (160 mg/l). Antioxidant activity varied among the cultivars (18–42 Trolox equivalents antioxidant capacity) and was directly related to the total phenolics in each type of juice.     

Chemical and antioxidative assessment of dietary turnip (Brassica rapa var. rapa L.)

The phenolic compounds and organic acids of turnip (Brassica rapa var. rapa L.) edible parts (leaves and stems, flower buds and roots) were determined by HPLC–DAD and HPLC–UV, respectively. The results revealed a profile composed of 14 phenolics (3-p-coumaroylquinic, caffeic, ferulic and sinapic acids, kaempferol 3-O-sophoroside-7-O-glucoside, kaempferol 3-O-sophoroside-7-O-sophoroside, kaempferol 3-O-(feruloyl/caffeoyl)-sophoroside-7-O-glucoside, kaempferol 3,7-O-diglucoside, isorhamnetin 3,7-O-diglucoside, kaempferol 3-O-sophoroside, 1,2-disinapoylgentiobiose, 1,2′-disinapoyl-2-feruloylgentiobiose, kaempferol 3-O-glucoside and isorhamnetin 3-O-glucoside) and six organic acids (aconitic, citric, ketoglutaric, malic, shikimic and fumaric acids). The quantification of the identified compounds showed kaempferol 3-O-sophoroside-7-O-glucoside, kaempferol 3-O-(feruloyl/caffeoyl)-sophoroside-7-O-glucoside, isorhamnetin 3,7-O-diglucoside and isorhamnetin 3-O-glucoside as the main phenolics, and malic acid as the organic acid present in highest amounts. A screening of the antioxidative potential was also performed by means of the DPPH radical scavenging assay. Turnip flower buds exhibited the strongest antioxidant capacity.     

Changes in Phenolic Compounds During Storage of Pasteurized Strawberry

This study aimed to establish mathematical models to describe changes in phenolics of pasteurized strawberry (Fragaria?×?ananassa Duch.) during storage at 23 °C for 90 days. Freshly cut strawberries cubes were pasteurized for 5 min in a water bath at 90 °C following a heating time of 15 min. Antioxidant activity, total phenolics, total anthocyanins, and individual phenolic compounds were assessed immediately before or after pasteurization and at regular time intervals during storage. The results indicated that (1) pasteurization did not affect (P?<?0.05) the concentrations of total phenolics or total anthocyanins, but significantly reduced the concentrations of quercetin-3-rutinoside, kaempferol, and cyanidin-3-glucoside, and increased the concentrations of (+)-catechin, (?)-epicatechin, epigallocatechin gallate, quercetin-3-galactoside, and ellagic acid; (2) changes in antioxidant capacity, total anthocyanin, and individual compounds during storage were described by a pseudo-first-order model with the exception of total phenolic and specifically kaempferol and ellagic acid which followed zero-order kinetic models. Pelargonidin-3-glucoside degraded at the highest rate (k?=?0.07 day^?1), followed by ellagic acid (k?=?0.004 day^?1) and kaempferol (k?=?0.003 day^?1). The rate constants can be used to predict phytochemical changes in strawberry products during storage.     

HPLC–MS analyses and bioactivities of novel chemicals in Devil’s club (Oplopanax horridus (Sm.) Miq.)

Devil’s club (Oplopanax horridus (Sm.) Miq.) has traditionally been used as a folk medicine by native North Americans for the treatment of various chronic diseases. A methanolic extract and its sub-fractions by liquid–liquid re-extraction, i.e., chloroform (CHCl₃), ethyl acetate (EtOAc), n-butanol (BuOH), and water fractions, were analysed by their anti-cancer activity with in vitro cell proliferative bioassays, and their antioxidant capacities in terms of the determination of total phenolic content (TPC), ORAC value, and DPPH free radical scavenging activity. The whole extract of the dried root contained high TPC and possessed strong ORAC and DPPH radical scavenging activities. In addition, the extract exhibited a strong anti-proliferative ability against HT-29 cancer cells, e.g., with an inhibitive rate of 56.5% with a 0.2-mg/mL extract. Further investigation by HPLC–UV–MS identified significant bioactive phenolic compounds in the chloroform fraction, including gallic acid, caffeic acid, 4-O-feruloylquinic acid, 5-O-feruloylquinic acid, ferulic acid, methyl feruloylquinate, methyl ferulate, and quercetin. These results suggested that the associated bioactivity of the plant might result from the contribution of phenolic compounds.     

Antioxidant activity versus cytotoxic and nuclear factor kappa B regulatory activities on HT-29 cells by natural fruit juices

The antioxidant activity (free radical scavenging ability and lipid oxidation inhibitory activity) and the activity in cell culture [influence on HT-29 human colon cancer cell viability and nuclear factor kappa B (NF-κB) activation] of natural fruit juices from red grape, strawberry, cherry and sour cherry were investigated in vitro. Analysis of polyphenolic composition of fruit juices showed the dominant presence of anthocyanins (cyanidin-3-glucoside, cyanidin-3-rutinoside, peonidin-3-rutinoside, pelargonidin-3-glucoside), anthocyanidins (procyanidins B1, B2 and B3), flavonoids (glucosides of myricetin, quercetin and kaempferol), and phenolic acids (neochlorogenic acid, chlorogenic acid, ellagic acid, p-coumaroylquinic acid) in juices. DPPH test and β-carotene bleaching method indicated that grape, strawberry, cherry and sour cherry natural juices (at the concentration of 2.5%) showed very strong free radical scavenging (75–92%) and antioxidant activity (71–94%). In addition, at the concentration of 0.01% they showed complete inhibition of copper-induced oxidation of human LDL (low density lipoprotein). In contrast, selected fruit juices did not show cytotoxic effects on HT-29 cells and NF-κB inhibitory activity (at the concentrations of 0.5 and 2%). Results of this study showed no correlation between the antioxidant activity of natural fruit juices from red grape, strawberry, cherry and sour cherry and their effect on HT-29 cell viability and NF-κB inhibitory activity.     

Chemical composition,anthocyanins, non-anthocyanin phenolics and antioxidant activity of wild bilberry (Vaccinium meridionale Swartz) from Colombia

Berries of Vaccinium meridionale Swartz native to Colombia were analysed for chemical composition, total phenolic content, anthocyanin content, and antioxidant activity. In addition, high-performance liquid chromatography with photodiode array detection (HPLC–DAD) and HPLC-electrospray ionisation tandem mass spectrometry (ESI–MS/MS) were used to determine anthocyanin and phenolic composition. Anthocyanin content was 329.0 ± 28.0 mg cyanidin 3-glucoside equivalents/100 g (fresh weight) FW and total phenolic content was 758.6 ± 62.3 mg gallic acid equivalent/100 g FW. Cyanidin 3-galactoside was the major anthocyanin while the most abundant non-anthocyanin phenolic was chlorogenic acid.     

Influence of alcoholic fermentation process on antioxidant activity and phenolic levels from mulberries (Morus nigra L.)

The aim of the present study was to evaluate the profile of the phenolic constituents of Morus nigra fruits and their antioxidant activity (DPPH) and to compare their contents before and after fermentation. Antioxidant phenolics of black mulberry (M. nigra L.) samples grown in Galicia (NW Spain) were extracted with methanol/formic acid/water (MFW) and determined by high performance liquid chromatography (HPLC). Two major anthocyanins (cyanidin 3-glucoside and cyanidin 3-rutinoside) and two flavonols (quercetin 3-glucoside and rutin) were isolated, together with caffeic acid and other hydroxycinnamic and ellagic acid derivatives. Their chemical structures were identified by spectral analyses with diode array detection (DAD), but also with alkaline saponification and acid hydrolysis of the mulberry phenolics. Good correlations (r² = 0.6229) were observed among total flavonols contents and the IC₅₀ radical scavenging capacities of mulberry fruits. Anthocyanins are the major flavonoids present in mulberry. It would be expected that anthocyanins contribute significantly to their antioxidant activity; nevertheless, alcohol generated during fermentation may also contribute to antioxidant activity. Our results provide useful antioxidant nutritional information of fresh and fermented mulberry fruits.     

Antioxidant activity and phenolic composition of sumac (Rhus coriaria L.) extracts

Antioxidant activity and phenolic compounds of sumac extracts were investigated. Sumac was extracted in methanol and subjected to solvent–solvent partitioning to yield two fractions as ethyl acetate and aqueous. Methanol extract was further fractioned over Sephadex LH-20 column. Antioxidant activity of extracts and fractions were screened using ferric thiocyanate and DPPH radical scavenging methods. Phenolic composition of active fraction(s) was determined by HPLC–MS systems. Those fractions which exhibited strong antioxidant activity were rich in anthocyanins and hydrolysable tannins. While gallic acid was the main phenolic acid in the extracts, anthocyanin fraction contained cyanidin, peonidin, pelargonidin, petunidin, and delphinidin glucosides and coumarates. Pentagalloyl glucose was abundant in the hydrolysable tannin fraction. Effective scavenging concentration (EC₅₀) on DPPH radical was 0.70 μg/mL both in ethyl acetate and tannin fractions, and 5.33 μg/mL in anthocyanin rich fraction. Same extracts and fractions showed moderate lipid peroxidation inhibition effect compared with the synthetic antioxidants. The findings demonstrate that sumac can be used as a natural antioxidant.     

A Comparative Study of Phenolic Antioxidant Activity and Flavonoid Biosynthesis‐Related Gene Expression Between Summer and Winter Strawberry Cultivars

Strawberry (Fragaria ananassa Duch.) possesses good antioxidant properties. Phenolic compounds in strawberries, such as anthocyanins and ellagic acid, mainly act as antioxidants. This study aimed to compare the phenolic content and expression patterns of genes involved in flavonoid biosynthesis between summer and winter strawberry cultivars affected by seasonal variation, degree of ripeness, and genotype. Antioxidant activity and the total content of phenols and flavonoids decreased with fruit ripening. Most notably, summer strawberry cultivars showed higher antioxidant activity than winter cultivars. The expression patterns of flavonoid biosynthetic genes tested were cultivar‐dependent and were also affected by ripening. These results help us understand the nutritional and physiological characteristics of selected cultivars and provide a range of information for strawberry consumption.     

Characterisation of volatiles and polyphenols for quality assessment of alcoholic beverages prepared from Corsican Myrtus communis berries

In the present work, the essential oil compositions of Myrtus communis berries from ten Corsican localities were studied and no chemical variability was observed. HS–SPME, GC and GC/MS analysis were carried out for the characterisation of volatile fractions of M. communis berries and two derived commercial alcoholic beverages (liqueur and eau-de-vie). Quantitative variations due to the distillation process were observed between the two beverages. The volatile compositions of Corsican myrtle alcoholic products were characterised by high amounts of monoterpene hydrocarbons and oxygenated monoterpenes with α-pinene and 1,8-cineole as major components. The polyphenolic compositions of the berry extract and the liqueur were also established using HPLC–DAD and LC–MS–MS; high concentrations of flavonol glycosides, flavonols and flavanols were reported.     

Effects induced by the nodulation with Bradyrhizobium japonicum on Glycine max (soybean) metabolism and antioxidant potential

This study was conducted in order to assess the effect of the inoculation of Glycine max (soybean) with Bradyrhizobium japonicum on the metabolite profile and antioxidant potential of its aerial parts. Extracts containing phenolic compounds and organic acids were analysed by HPLC–DAD and HPLC–UV, respectively. Volatiles profile was determined by HS-SPME/GC–IT-MS. The antioxidant activity of acidic and methanolic extracts was assessed against DPPH. Nodulation caused by B. japonicum led to an increase in the contents of phenolic compounds and organic acids. The same was observed for volatile compounds, with compounds like linalyl acetate, menthyl acetate and α-farnesene being detected only in inoculated plants. The phenolic extracts showed stronger antioxidant capacity than the organic acid extracts. In addition, extracts from plants nodulated with B. japonicum exhibited significantly higher antioxidant activity than control samples. These findings suggest that the inoculation with nodulating B. japonicum strains may be employed to manipulate the content of interesting metabolites in G. max aerial parts.     

Phenolics and antioxidant properties of bayberry (Myrica rubra Sieb. et Zucc.) pomace

Five cultivars of Myrica rubra, Biqi, Wandao, Dongkui, Dingao, and Zaodamei, were collected to analyze the phenolic compounds and evaluate the antioxidant properties of bayberry pomaces. The main anthocyanin was cyanidin-3-o-glucoside (3073.3–6219.2 mg/kg dry weight (DW)) and the main flavonol was quercetin-3-o-glucoside (296.2–907.9 mg/kg dry weight). Quercetin and myricetin were also found in the bayberry pomaces, and quercetin deoxyhexoside and myricetin deoxyhexoside were tentatively identified. The dominant phenolic acids were gallic acid (102.9–241.7 mg/kg dry weight) and protocatechuic acid (29.5–57.2 mg/kg dry weight). Other phenolic acids such as p-hydroxybenzoic, vanillic, caffeic, p-coumaric, and ferulic acids were also present in the bayberry pomaces, whereas, chlorogenic acid was only detected in Dongkui (1.58 mg/kg dry weight). The antioxidant activity of Wandao was the strongest of the five cultivars, whereas the activity of Dongkui was the weakest, and a significant positive relationship was observed between antioxidant activity and total phenolic content or total anthocyanins.     

A new flavone from antioxidant extracts of Pistacia terebinthus

Acetone and methanol extracts of the fruits of Pistacia terebinthus L. subsp. terebinthus L. were studied for their antioxidant activity by investigating their total phenolic and flavonoid contents, β-carotene bleaching potential, DPPH radical scavenging effect, scavenging activity on superoxide anion radical, reducing power, and metal chelating effect on ferrous ion. Both extracts showed very similar chemical profile by checking on TLC plates, and exhibited high scavenging activity on superoxide anion radical and DPPH radical. Due to these similarities they were combined and fractionated on a silica gel column for their constituents, and the most active three fractions in DPPH assay were purified to afford a new flavone 6′-hydroxyhypolaetin 3′-methyl ether (1) besides a group of known flavonoids apigenin, luteolin, luteolin 7-O-glucoside, quercetin, quercetagetin 3-methyl ether 7-O-glucoside, isoscutellarein 8-O-glucoside. Their structures were established by UV, UV shift reagents, and ¹H NMR spectroscopic techniques. Antioxidant activity of the new flavone was investigated by β-carotene bleaching and DPPH radical scavenging activity methods, and it showed a high activity in the first system, but not so good in the latter.     

Analysis of phenolic compounds in cork from Quercus suber L. by HPLC–DAD/ESI–MS

The aim of the present work was to identify the extractable phenolic compounds present in cork from Quercus suber L. The structures of thirty three compounds were tentatively identified by liquid chromatography coupled to electrospray ionisation mass spectrometry (HPLC–DAD/ESI–MS). The majority of those compounds were gallic acid derivatives, in the form of either galloyl esters of glucose (gallotannins), combinations of galloyl and hexahydroxydiphenoyl esters of glucose (ellagitannins), dehydrated tergallic-C-glucosides or ellagic acid derivatives. Others were found to correspond to low molecular weight phenolic compounds, like acids and aldehydes. Mongolicain, a flavanoellagitannin in which hydrolysable tannin and flavan-3-ol moieties are connected through a carbon–carbon linkage, was also detected in cork from Q. suber L. The results illustrate the rich array of phenolic compounds present in cork.     

Polyphenolic profiles detected in the ripe berries of Vitis vinifera germplasm

Polyphenolic profiles in the berry samples of 344 European grape (Vitis vinifera) cultivars were evaluated for two consecutive years. These cultivars represent a diverse collection of V. vinifera germplasm maintained at the USDA-Agricultural Research Service Vitis Clonal Repository in Davis of California, USA. A total of 36 polyphenolic compounds, including 16 anthocyanins, 6 flavonols, 6 flavanols, 6 hydroxycinnamic acids and 2 hydroxybenzoic acids, were identified via HPLC–MS and quantified by HPLC–DAD. The mean contents for anthocyanins, flavanols, flavonols, hydroxycinnamic acids and hydroxybenzoic acids were 0.946 (coloured cultivars), 0.147, 0.043, 0.195 and 0.016 mg g⁻¹ FW, respectively. On average, wine grapes had higher concentrations than had table grapes for all of these compounds except hydroxycinnamic acids. Berry colours affected the total contents of anthocyanins, but not others. Positive correlations (0.151–0.535) were found among these groups of compounds. As expected, these groups of compounds were all negatively correlated with berry weight.     

Chemical characterization and antioxidant evaluation of muscadine grape pomace extract

Concentrated muscadine pomace extract was chromatographically analyzed for its individual phenolic, flavonoid, and anthocyanidin compounds. This extract was also characterized regarding its total phenolic content (TPC), total flavonoid content (TFC), antioxidative activities in terms of scavenging DPPH free radicals, reducing ferric, and chelating Fe²⁺. The TPC of this product was 34.1 ± 1.8 mg of gallic acid equivalents(GAE)/g of extract, and TFC was 3.0 ± 0.3 mg of quercetin equivalents/g of extract. Some phenolic compounds including ellagic acid, gallic acid, (−)-epicatechin, (−)-epigallocatechin, catechin, myricetin, quercetin, and kaempferol were identified. Some anthocyanidins including delphinidin, cyanidin, petunidin, peonidin, and malvidin were also identified in the extract by using a combination of retention and spectral properties on a reverse-phase HPLC–PDA. In addition, 3,3′,4,4′5,5′-hexahydroxystilbene-a resveratrol analogue present in the extract was identified for the first time by LC–MS. The results from this study demonstrate that the muscadine pomace extract is rich of natural antioxidants such as phenolics, flavonoids and anthocyanins, and possesses strong antioxidant properties. Besides, the developed methods can be used for routine quality control of the muscadine products for manufacturing efficiency and consistency.     

Non-toxic Salvia sclareoides Brot. extracts as a source of functional food ingredients: Phenolic profile,antioxidant activity and prion binding properties

Salvia sclareoides is an aromatic herb native to Portugal, of which phenolic content (Folin–Ciocalteau method), chemical profile (HPLC/DAD), antioxidant activity (DPPH, β-carotene/linoleic acid assays), acute toxicity (MTT method, adapted for non-adherent cells), genotoxicity (short-term chromosomal aberration assay) and prion binding properties were evaluated in the acetone, water, ethanol, methanol and n-butanol extracts. The latter presented the highest phenolic content and antioxidant activity (DPPH assay), and was the single one with the flavonoids (+)-catechin, kaempferol O-glucoside and quercetin. Vanillic acid was the major component of all extracts but gallic, gentisic, caffeic, syringic, coumaric and ferulic acids were also found in some extracts. Only the n-butanol extract had components binding to the cellular form of human prion protein detected by NMR which showed specificity for two regions of the folded domain and for the unstructured N-terminal region. Extracts were not cytotoxic nor genotoxic, reinforcing the potential of S. sclareoides for nutraceutical purposes.