Variation in antioxidant enzyme activities and nonenzyme components among cultivars of rabbiteye blueberries (Vaccinium ashei Reade) and V. ashei derivatives

Fruit from 42 blueberry (Vaccinium species) cultivars, including 36 rabbiteye cultivars (Vaccinium ashei Reade), 3 V. ashei hybrid derivatives and 3 northern highbush (V. corymbosum L.) standards, were evaluated for antioxidant activities (AA), levels of antioxidant enzymes [catalase (CAT), ascorbate peroxidase (AsA-POD), glutathione reductase (GR), glutathione peroxidase (GSH-POD), superoxide dismutase (SOD), dehydroascorbate reductase (DHAR), monodehydroascorbate reductase (MDAR) and guaiacol peroxidase (G-POD)], and antioxidant nonenzyme components [ascorbic acid (ASA) and glutathione (GSH)]. The results of this study showed that cultivars had varying levels of AA, a wide range of antioxidant enzyme activities and various amounts of nonenzyme components. The correlations between AA and all of the enzyme activities or nonenzyme components were positive and high in the 42 cultivars tested in this study. The correlation (R²) values between AA and CAT, SOD, DHAR, MDAR, GSH-POD, GR, G-POD and AsA-POD were 0.91, 0.91, 0.91, 0.91, 0.90, 0.90, 0.90, and 0.90, respectively. The correlations between AA and ASA and GSH were 0.94 and 0.92, respectively. Among all 42 cultivars, ‘Early May’ had the highest activities of the AA, AsA-POD, G-POD, DHAR and MDAR and the highest amounts of ASA and GSH, whereas ‘Elliott’ had the highest levels of GR, GSH and SOD.     

Free radical scavenging capacity and antioxidant enzyme activity in deerberry (Vaccinium stamineum L.)

Fruit from three genotypes (‘B-76’, ‘B-59’ and ‘SHF-3A’) of deerberry [Vaccinium stamineum L.] were evaluated for fruit quality, total anthocyanin and phenolic contents, antioxidants, antioxidant capacity, and antioxidant enzyme activity. The fruit soluble solids, titratable acids, total anthocyanins, and total phenolic contents varied with genotypes. Cyanidin 3-galactoside and cyanidin 3-arabinoside were the two predominant anthocyanins. Resveratrol was also found in deerberries. Among the three genotypes, ‘B-76’ had higher amount of anthocyanins, phenolic compounds and resveratrol than ‘B-59’ and ‘SHF-3A’. Deerberries contained potent free radical scavenging activities for 2,2-Di (4-tert-octylphenyl)-1-picrylhydrazyl (DPPH), 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS⁺), peroxyl radical (ROO), superoxide radicals (), hydrogen peroxide (H₂O₂), hydroxyl radicals (OH), and singlet oxygen (¹O₂) radicals and also had high activities of antioxidant enzymes including glutathione-peroxidase (GSH-POD), glutathione reductase (GR), superoxide dismutase (SOD), ascorbate peroxidase (AsA-POD), guaiacol peroxidase (G-POD), monodehydroascorbate radical reductase (MDAR), dehydroascorbate reductase (DHAR), glutathione reductase (GR)] and non-enzyme antioxidants [ascorbic acid (ASA) and reduced glutathione (GSH)]. Antioxidant capacities were highly correlated to antioxidant enzymes activities. Among the three genotypes, ‘B-76’ had the highest level of antioxidants and antioxidant enzyme activity.     

Phenolic profile and antioxidant activity of highbush blueberry (Vaccinium corymbosum L.) during fruit maturation and ripening

The phenolic profile and quantitative composition of blueberries as well as the corresponding antioxidant activity of blueberries is well documented. Unfortunately, little is reported on the development of phenolic compounds and antioxidant activity during fruit maturation and ripening. In the present study, the total phenolic content and main phenolic compounds of four highbush blueberry cultivars (Vaccinium corymbosum L.) were analyzed at five stages of maturation and ripening. Antioxidant activity was screened with electron spin resonance spectrometry and trolox equivalent antioxidant capacity (TEAC) assay. An adequate picture of phenolic compounds developed during maturation and ripening was determined using HPLC-DAD. Anthocyanins of all varieties increased during successive harvest stages; meanwhile flavonols and hydroxycinnamic acids decreased from unripe green to ripe blue stage of berry ripening. Blueberry antioxidant activity, as well as total phenolic content tended to decrease during ripening.     

Comparative study of primary and secondary metabolites in 11 cultivars of persimmon fruit (Diospyros kaki L.)

Primary metabolites (sugars, organic acids) and secondary metabolites (phenolics and carotenoids) were quantified by HPLC in fully ripe fruit of 11 kaki cultivars: ‘Amankaki’, ‘Cal Fuyu’, ‘Fuji’, ‘Hana Fuyu’, ‘Jiro’, ‘O’Gosho’, ‘Tenjin O’Gosho’, ‘Thiene’, ‘Tipo’, ‘Tone Wase’ and ‘Triumph’. Amongst the analysed cultivars, ‘Tone Wase’ stands out as the richest in sugars, particularly glucose, and cultivars ‘Tipo’ and ‘Triumph’ contained the highest amounts of organic acids. Cultivars ‘O’Gosho’, ‘Cal Fuyu’ and ‘Hana Fuyu’ contained the least sugars and cultivar ‘Jiro’ the least organic acids. Amongst the individual phenolic compounds catechin and gallic acid were present in highest concentrations. The predominant carotenoid in both skin and pulp of ripe persimmon fruit was β-carotene, the highest content was measured in skin of cultivar ‘Hana Fuyu’, which also contained the highest level of total carotenoids. In persimmon pulp, much lower values for carotenoids were obtained, particularly in fruit of cultivar ‘Cal Fuyu’.     

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.     

Dracaena draco L. fruit: Phytochemical and antioxidant activity assessment

The present study reports for the first time the metabolite profile and antioxidant activity of aqueous extract obtained from Dracaena draco L. fruit. Volatiles profile was determined by HS-SPME/GC-IT-MS, with 9 compounds being identified, distributed by several distinct chemical classes: 1 alcohol, 3 aldehydes, 2 carotenoid derivatives, and 3 terpenic compounds. Aldehydes constituted the most abundant class in this exotic berry, representing 59% of total identified volatile compounds. Phenolics profile was determined by HPLC/DAD and 5 constituents were identified: 5-O-caffeoylquinic, 3,5-O-dicaffeoylquinic, ferulic and sinapic acids, and quercetin-3-O-rutinoside. The major phenolic compound is quercetin-3-O-rutinoside, comprising 42% of the total phenolic content. Organic acids composition was also characterized, by HPLC-UV, and oxalic, citric, l-ascorbic, malic, quinic and shikimic acids were determined. The most abundant is quinic acid, representing 39% of the total organic acid content. The antioxidant potential of this matrix was assessed by (i) reducing power of Fe³⁺/ferricyanide complex, (ii) scavenging effect on DPPH free radicals, and (iii) ability to inhibit the 2,2´-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative hemolysis in human erythrocytes. Strawberry (Fragaria × ananassa Duch. cv. Camarosa) extract was used for comparison purposes. All assay models showed remarkable concentration dependent antioxidant activity, reducing power and radical scavenging efficiency for D. draco fruit, being invariably higher than that of strawberry extract. This is the first report showing that D. draco fruit is a promising new antioxidant agent.     

Major flavonoids with antioxidant activity from Teucrium polium L.

Teucrium polium L. (Lamiaceae) aerial parts are used widely in the daily diet and for medicinal purposes. This plant is used also as a spice and refreshing beverage. Phytochemical and bioactivity studies of this plant have been carried out. Aerial parts of the plant were extracted with petroleum ether, chloroform, methanol and water successively. Fractionation of the methanol extract yielded four major flavonoids. The crude extracts and isolated compounds were screened for their antioxidant and free radical scavenging activities using DPPH radical-scavenging, beta-carotene/linoleic acid and ammonium thiocyanate methods. Methanol extract, rutin and apigenin were found to be the most active fractions as radical-scavengers with IC₅₀ values of 20.1 ± 1.7, 23.7 ± 1.9 and 30.3 ± 2.1 μg/ml, respectively. The samples with the highest inhibition of oxidation of beta-carotene and lipid peroxidation in ammonium thiocyanate methods were also found to be methanol extract, rutin and apigenin. Methoxylated flavonoids exhibited a lesser antioxidant activity.     

The antioxidant properties of alcoholic extracts from Sambucus nigra L. (antioxidant properties of extracts)

The antioxidant properties of alcoholic extracts from the leaves, berries and flowers of Sambucus nigra L. are estimated by means of DPPH and β-carotene/linoleic acid methods, and considered in relation to the extraction temperature (in the range 20-200 °C) and to the level of flavonoids most representative for this plant. The extracts of S. nigra act as antioxidants neutralizing the activities of free radicals and inhibiting the co-oxidation reactions of linoleic acid and β-carotene. There is no direct correlation between the level of flavonoids in the extracts and their antioxidant activity. The data presented show that the extraction temperature strongly influences the antioxidant properties of the extracts, especially in the case of leaves.     

Flavonoid constituents and their contribution to antioxidant activity in cultivars and hybrids of rabbiteye blueberry (Vaccinium ashei Reade)

Fruit from 42 blueberry (Vaccinium spp.) cultivars, including 36 rabbiteye cultivars (Vaccinium ashei Reade), three V. ashei hybrid derivatives, and three northern highbush (Vaccinium corymbosum L.) standards were evaluated for antioxidant capacity, individual flavonoid content, and the contribution of each identified phenolic compound to total antioxidant activity. Considerable variation was found in flavonoid content, antioxidant activity, and their contribution to total antioxidant activity among cultivars. Among 42 blueberry cultivars, the rabbiteye ‘Early May’ contained the highest amount of chlorogenic acid, myricetin 3-arabinoside, quercetin derivatives, and delphinidin-, cyanidin-, petunidin-, and malvidin-basis anthocyanins. ‘Early May’ cultivar also had the highest antioxidant activity (88.2 μmol TE/g fw). ‘Owen’, ‘Bluegem’, ‘Clara’, Climax’, and ‘Centurion’ were among the other rabbiteye cultivars that also had high levels of flavonoids and antioxidant activities. In contrast, the pink-fruited V. ashei hybrid, ‘Pink Lemonade’, had the lowest content of flavonoids and lowest antioxidant activity. The mean flavonoid content and antioxidant activity of rabbiteye cultivars was higher than those among northern highbush and V. ashei hybrids. The antioxidant activity of V. ashei hybrid derivatives was derived mainly from chlorogenic acid, myricetin, and quercetin, which contributed 62.5% of total antioxidant activity, whereas anthocyanins (malvidin, petunidin, delphinidin, and cyanidin) were the main contributors to the antioxidant activity of rabbiteye cultivars (76.2%) and northern highbush standards (76.8%). Blueberry cultivars identified to have high phenolic content and high antioxidant activity could be used as parents for future blueberry breeding programmes to develop new blueberry cultivars with higher antioxidant activity and further improve human health.     

The influence of light and maturity on fruit quality and flavonoid content of red raspberries

The effects of various fruit maturities and different light intensities on berry fruit quality, antioxidant capacity and phytonutrient levels in red raspberries (Rubus ideaus L.) were determined. At harvest, immature berries contained significantly lower levels of sugars and acids than ripe berries. When berries were harvested at 5% or 20% maturity, they never developed the levels of soluble solids content (SSC) and titratable acid (TA) values observed in ripe berries at harvest. However, fruit harvested at 50% or more advanced maturity had the capacity of attaining comparable levels of SSC, TA and sugars as those harvested at 100% maturity. When 5% and 20% berries were stored under light, higher level of SSC and lower levels of TA values were observed than those kept in the dark. However, light condition showed little effect in 50% and 80% maturity fruit after 4 days at 24/16 °C (day/night). Ripe raspberries (100%) had stronger antioxidant activities and higher total anthocyanin content when compared with the pink stage (50% maturity). Fruit harvested at greener stages (5% and 20%) also consistently showed higher antioxidant activities and total phenolics than those harvested at 50%. Cyanindin-based anthocyanins increased during postharvest period. On the other hand, other polyphenols such as ellagic acid, quercetin 3-glucoside, quercetin derivative, and kaempferol 3-glucuronide were initially present at high levels but decreased drastically during storage. Red raspberries harvested at different developmental stages continued their development during storage even under the dark conditions. The antioxidant activity of red raspberries was directly related to the total amount of phenolics and flavonoids. Results of this study indicate that red raspberries harvested at 50% or more advanced maturity could develop comparable quality and antioxidant levels as those harvested at full maturity.     

Flavonoids, sugars and fruit acids of alpine bearberry (Arctostaphylos alpina) from Finnish Lapland

Alpine bearberry (Arctostaphylos alpina L.) is a special circumpolar edible berry primarily used for household purposes but with greater potential in commercial applications than utilized thus far. In this study, the flavonoids of alpine bearberry were investigated with HPLC-DAD and HPLC-MS, and the sugars and fruit acids as trimethylsilyl derivates with GC-FID. The most abundant anthocyanin was cyanidin-3-O-galactoside accounting for over 95% of the anthocyanins. Eight other anthocyanins existed in trace amounts only, the richest of them being cyanidin-3-O-arabinoside and cyanidin-3-O-glucoside. Cyanidin accounted for over 99% of the anthocyanin aglycones. All the flavonols identified were glycosides of quercetin. Majority of the sugars were glucose and fructose while the most abundant fruit acid was quinic acid. The unique anthocyanin composition of alpine bearberry encourages advanced use of the berry as a food ingredient in private households and in industrial applications. In addition, it serves as a unique reference material for research purposes.     

Influence of strawberry tree (Arbutus unedo L.) fruit ripening stage on chemical composition and antioxidant activity

Arbutus unedo is a widespread shrub with economic importance, derived from the use of its berries in the production of alcoholic beverages and in folk medicine. This work intends to evaluate for the first time the effect of fruit ripening stage on antioxidant activity, total phenolic content, fatty acid profile and tocopherol composition. Ripe fruits shown higher extraction yield (45.04 ± 2.23%) when compared to other fruit ripening stages. By contrast, total phenol contents were higher in the unripe and intermediate stage of ripeness (108 and 111 mg GAES/g dry fruit, respectively, against 60 mg/g dry fruit when ripe). Ripe and intermediate fruits shown the lower EC₅₀ values on the DPPH radicals (0.25 ± 0.02 mg/mL) and reducing power assay (1.09 ± 0.05 mg/mL), respectively. A significant correlation was established between antioxidant activity and fruits ripening stage. Fatty acid profiles were very similar between the ripening stages, being alfa-linolenic, linoleic and oleic, the three major ones. Polyunsaturated fatty acids (PUFA) represent as much as 60% of the total fatty acids, with a highly favorable omega 3/omega 6 ratio. From the analysis of the vitamin E vitamers, the most important was ??-tocotrienol, with a clear reduction in the total free vitamin E content with ripening.These results highlight that the fruits of intermediate ripeness can be regarded important sources of biologically active compounds with a fatty acid profile rich in omega-3 PUFA, properly supplemented with high vitamin E amounts.     

European elderberry (Sambucus nigra L.) rich in sugars,organic acids,anthocyanins and selected polyphenols

Sugars and organic acids in the fruit of two cultivars and three selections of black elderberry (Sambucus nigra L.): ‘Haschberg’, ‘Rubini’, ‘Selection 13’, ‘Selection 14’ and ‘Selection 25’ were quantified. The anthocyanin as well as quercetin profiles of this plant material were also established by the use of HPLC/MS. Significant differences in the concentration of sugars and organic acids were detected between the widely spread cultivar ‘Haschberg’ and all other cultivars/selections; ‘Haschberg’ was the richest in organic acids (6.38 g kg⁻¹ FW), and it contained the least sugar (68.5 g kg⁻¹ FW). The following major cyanidin based anthocyanins were identified in the fruit of black elderberry: cyanidin 3-sambubioside-5-glucoside, cyanidin 3,5-diglucoside, cyanidin 3-sambubioside, cyanidin 3-glucoside and cyanidin 3-rutinoside. The most abundant anthocyanin in elderberry fruit was cyanidin 3-sambubioside, which accounted for more than half of all anthocyanins identified in the berries. The ‘Rubini’ cultivar had the highest amount of the anthocyanins identified (1265 mg/100 g FW) and the lowest amount was measured in berries of the ‘Selection 14’ (603 mg/100 g FW). The ‘Haschberg’ cultivar contained a relatively low amount of anthocyanins in ripe berries (737 mg/100 g FW). From the quercetin group, quercetin, quercetin 3-rutinoside and quercetin 3-glucoside were identified; the latter prevailing in black elderberry fruit. The cultivar with the highest amount of total quercetins was ‘Selection 25’ (73.4 mg/100 g FW), while the ‘Haschberg’ cultivar contained average amounts of quercetins (61.3 mg/100 g FW). The chemical composition of the ‘Haschberg’ cultivar, the most commonly planted, conforms to the standards for sugars, anthocyanins and quercetins and exceeds them in the content levels of organic acids, the most important parameter in fruit processing.     

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.     

Constituents of Sideritis syriaca. ssp. syriaca (Lamiaceae) and their antioxidant activity

The aerial parts of Sideritis syriaca ssp. syriaca (Lamiaceae) were extracted, after defatting, with diethyl ether, ethyl acetate and n-butanol. The antioxidant activities of the extracts were evaluated through in vitro model systems, such as 1,1-diphenyl-2-picryl hydrazyl (DPPH) and Co(II) EDTA-induced luminol chemiluminescence. In both model systems the ethyl acetate extract was the most effective. Phytochemical analysis of ethyl acetate extract showed the presence of two new isomeric compounds (1 and 1′), identified as 1-rhamnosyl, 1-coumaroyl, dihydrocaffeoyl, protocatechuic tetraester of quinic acid, as well as chlorogenic acid (2), apigenin 7-O-glucoside (3), apigenin (4), 4′-O-methylisoscutellarein 7-O-[6′′′-O-acetyl-β-D-allopyranosyl-(1 → 2)-β-d-glucopyranoside] (5), isoscutellarein 7-O-[6′′′-O-acetyl-β-D-allopyranosyl-(1 → 2)-β-d-glucopyranoside] (6), 4′-O-methylisoscutellarein 7-O[β-d-allopyranosyl-(1 → 2)-β-d-glucopyranoside] (7) and 4′-O-methylisoscutellarein 7-O-[β-d-allopyranosyl-(1 → 2)-6′′-O-acetyl-β-d-glucopyranoside] (8). The above compounds were identified by spectroscopic methods.     

Antioxidant and cytotoxic flavonoids from the flowers of Melastoma malabathricum L.

Phytochemical and bioactivity studies of the flowers of Melastoma malabathricum L. (Melastomataceae) have been carried out. The ethyl acetate extract yielded three compounds, identified as naringenin, kaempferol and kaempferol-3-O-d-glucoside, and methanol extract gave kaempferol-3-O-(2″,6″-di-O-p-trans-coumaroyl)glucoside and kaempferol-3-O-d-glucoside. The crude extracts and isolated compounds were screened for their antioxidant and cytotoxic activities. The antioxidant assay was carried out by the DPPH radical-scavenging electron spin resonance (ESR) spectroscopic method. The cytotoxicity was measured by the MTT assay against a MCF7 cell line. Naringenin, kaempferol, kaempferol-3-O-d-glucoside, kaempferol-3-O-(2″,6″-di-O-p-trans-coumaroyl) glucoside, ethyl acetate and methanol extracts were found to be active as radical-scavengers with IC₅₀ values of 0.52 mM, 81.5 μM, 1.07 mM, 35.8 μM, 7.21 μg/ml and 6.59 μg/ml, respectively. Naringenin and kaempferol-3-O-(2″,6″-di-O-p-trans-coumaroyl)glucoside were also found to be active in inhibiting cell proliferation of MCF7 with IC₅₀ values of 0.28 μM and 1.3 μM, respectively.     

Comparative chemical composition,antioxidant and hypoglycaemic activities of Juniperus oxycedrus ssp. oxycedrus L. berry and wood oils from Lebanon

Juniper (Juniperus oxycedrus) is used in European cuisine for its distinguishing flavour. J. oxycedrus ssp. oxycedrus berry and wood essential oils were tentatively identified by GC and GC/MS. Fifty compounds were identified in the berry oil and 23 compounds were identified in the wood oil. The J. oxycedrus ssp. oxycedrus berry oil was characterised by high contents of α-pinene (27.4%) and β-myrcene (18.9%). Other important compounds were α-phellandrene (7.1%), limonene (6.7%), epi-bicyclosesquiphellandrene (2.3%) and δ-cadinene (2.2%) while, in the wood oil, δ-cadinene (14.5%) is a major main component, together with cis-thujopsene (9.2%) and α-muurolene (4.9%). In vitro evaluation of antioxidant activity by the DPPH method showed a significant activity for both oils with IC₅₀ values of 1.45 μl/ml for wood and 7.42 μl/ml for berries. Hypoglycaemic activity was investigated through the inhibition of α-amylase. The results revealed that oil obtained by hydrodistillation from J. oxycedrus ssp. oxycedrus wood exhibits an interesting activity with IC₅₀ of 3.49 μl/ml.     

Phenolic acid analysis and antioxidant activity assessment of oil palm (E. guineensis) fruit extracts

Phenolic compounds in oil palm fruit (E. guineensis) were extracted in soluble free (SFP), insoluble-bound (ISBP) and esterified (EFP) forms. The total phenolic content (TPC) of the oil palm fruit extracts was determined using the Folin–Ciocalteu method and found to range from 5.03 to 9.04 g/L per g of dried weight (DW). The antioxidant activities of oil palm phenolic extracts were analysed using free radical scavenging assays and results showed that oil palm phenolic extracts contained antioxidant activities in the order of ISBP > EFP > SFP. Eight different phenolic acids were identified and quantified using a simple reversed-phase high performance liquid chromatography (HPLC) with a diode array detector (DAD) and liquid chromatography/tandem mass spectrometry (LC/MS/MS). Ferulic, p-hydroxybenzoic and p-coumaric acid were the dominant phenolic acids found in oil palm fruit extracts and ranged from 55 to 376 μg/g of DW.