Evaluation of the antioxidant activity of extracts from buntan (Citrus grandis Osbeck) fruit tissues

The goal of the present work was to evaluate the antioxidant properties of buntan (Citrus grandis Osbeck) using various solvents, such as n-hexane, ethyl acetate (EtOAc), butanol and methanol. The antioxidant activities of crude extracts were evaluated by using the free radical scavenging β-carotene assay and total polyphenol. Ethyl acetate extracts of falvedo exhibited high antioxidative activities, followed by albedo and segment membrane extracts. Chromatography separation of EtOAc extract of flavedo using a silica gel column, yielded six fractions (A, B, C, D, E and F) using gradient elution with benzene and acetone (19:1, 14:1, 9:1, 5:1, 1:1 and 0:l). Among them, two fractions (C and D) showed strong antioxidant activities using the free radical scavenging activity (DPPH) antioxidant assay. These two fractions were further purified using silica gel column chromatography and preparative TLC. Their extracts could well be useful to prevent oxidation in fruit juices and essential oil food products as well as for health supplements. Identification of the responsible components is underway.     

Antioxidant and antidiabetic activity of Dangyuja (Citrus grandis Osbeck) extract treated with Aspergillus saitoi

Dangyuja is a citrus fruit of Cheju Island in Korea, which is known to have a high content of flavanone glycosides, such as naringin and neohesperidin. Flavanone glycosides of Dangyuja (Citrus grandis Osbeck) extract were converted into their aglycones by naringinase and hesperidinase, and into their hydroxylated forms by Aspergillus saitoi.Dangyuja extract treated with A. saitoi had significantly higher antioxidant and antidiabetic activity than both its glycosides and aglycones in oxygen radical absorption capacity (ORAC) and supercoiled DNA strand scission assay, rat intestinal α-glucosidase and porcine pancreatic α-amylase inhibition, suggesting that the fermentation of flavanone aglycones of Dangyuja extract with A. saitoi can increase antioxidant and antidiabetic activity, compared to their glycosides and aglycones. This result suggests that Dangyuja extract prepared with enzymatic and microbial treatments could be used for the development of pharmaceutical foods to control the blood glucose level of diabetic patients by inhibiting α-amylase and α-glucosidase in the intestinal tract.     

Induction of apoptosis in U937 human leukaemia cells by the hexane fraction of an extract of immature Citrus grandis Osbeck fruits

The antiproliferative effect of an immature Citrus grandis Osbeck fruit extract was investigated using U937 human leukaemia cells. Maximum cytotoxicity was observed using the hexane fraction (HF) of the extract. Cell death was dose-dependent (IC₅₀ = ca. 60 μg/ml) and was characterised by chromatin condensation, apoptotic body formation, and DNA fragmentation. The induction of apoptosis was confirmed by caspase-3 activity assays and by immunoblotting using antibodies against Bcl-2, Bax, poly(ADP-ribose) polymerase (PARP), caspase-9, and caspase-3. The molecular mechanism underlying HF-induced apoptosis in U937 cells may involve a mitochondria-mediated signalling pathway, as demonstrated by an increase in the Bax/Bcl-2 expression ratio. Analyses of the HF by gas chromatography (GC) and GC-mass spectrometry (MS) tentatively identified 19 compounds, including γ-sitosterol (17.5%), 7-methoxy-8-(2-oxo-3-methylbutyl) coumarin (6.8%), stigmasterol (3.8%), and campesterol (3.4%). Together, our results provide the first evidence that the HF of an immature C. grandis Osbeck fruit extract induces apoptosis in U937 cells.     

Phytochemicals and antioxidant activity of different parts of bambangan (Mangifera pajang) and tarap (Artocarpus odoratissimus)

Mangifera pajang (family: Anacardiaceae; local name: bambangan) and Artocarpus odoratissimus (familiy: Moraceae; local name: tarap) are popular edible fruits in Sabah, Malaysia. The flesh, kernel and peel from M. pajang; seed and flesh from A. odoratissimus were analysed for total antioxidant activity, total polyphenol, total flavonoid and total anthocyanins contents. M. pajang kernel extract displayed the highest free radical scavenging and ferric reducing activities. Total phenolic content of the samples were in the range of 5.96–103.3 mg gallic acid equivalent/g. M. pajang kernel and M. pajang flesh contained the highest and lowest total flavonoid content with the values of 10.98 and 0.07 mg rutin equivalent/g, respectively. The antioxidant activities of extracts were significantly correlated with the total phenolic and flavonoid content (but not the anthocyanins content). The phytochemicals and antioxidant properties of M. pajang and A. odoratissimus, especially their by-products (kernel/seed), indicate that they may impart health benefits when consumed and should be regarded as a valuable source of antioxidant-rich nutraceuticals.     

Antioxidant activity of microwave-assisted extract of Buddleia officinalis and its major active component

Buddleia officinalis Maxim, commonly used as rice dye for festivals, was extracted with ethanol using microwave-assisted extraction and Soxhlet extraction. The antioxidant activities of microwave-assisted extract of B. officialis (MEB) and Soxhlet extract of B. officianils (SEB) at the optimum extraction conditions were evaluated and compared with synthetic antioxidant butylated hydroxytoluene (BHT) employing DPPH free radical assay, ABTS assay, total antioxidant activity and reducing power. MEB and SEB had stronger antioxidant activities than BHT in all assays except reducing power, and the effects decreased as follows: MEB > SEB > BHT. The total phenolic contents of MEB and SEB reached 113.56 mg/g and 100.94 mg/g dry weight of extract, respectively, expressed as pyrocatechol equivalents, while the total flavonoids contents were 75.33 mg/g and 62.56 mg/g dry weight of extract, respectively, expressed as catechin equivalents (P < 0.05). Higher phenolic and flavonoids compounds may be major contributors to their higher antioxidant activities. Following activity-oriented separation, luteolin was isolated as an active principle, which exhibited excellent free radical scavenging activities with DPPH IC₅₀ 3.09 μg/ml and ABTS IC₅₀ 2.20 μg/ml.     

Juice of New citrus hybrids (Citrus clementina Hort. ex Tan.×C. sinensis L. Osbeck) as a source of natural antioxidants

Fruit juices of pigmented and non-pigmented new citrus hybrids obtained by crossing clementine cv. Oroval with different cultivars of blood oranges were analysed to determine parameters related to fruit quality (total soluble solids titratable acidity, pH), and the content of ascorbic acid, total phenolics, flavanones, anthocyanins and phenolic acids. Antioxidant properties were evaluated by oxygen radical absorbance capacity (ORAC), and inhibition of induced linoleic acid peroxidation (InLAP) assays. The results of this study show that some hybrids with high antioxidant activity owing to considerable polyphenol content may be considered rich sources of phytochemicals. The OTA 9 hybrid was shown to be richest in polyphenols, suggesting that consumption of OTA 9 fruit or juice could be useful in health promotion and a disease-preventing diet. Moreover, the juice of this hybrid could be used as raw material to produce antioxidant ingredients for dietary, pharmaceutical, or cosmetic purposes.     

Antioxidative and anti-inflammatory properties of Citrus sulcata extracts

Citrus sulcata was subjected to ultrasound, high-pressure, and Soxhlet extractions. The antioxidant and anti-inflammatory activities of the extracts were evaluated qualitatively and quantitatively. The antioxidant content of the peel extract was twice that of the fruit extract. The quantitative analysis showed that the narirutin and hesperidin contents in the peel extracts were 8.8 and 7.5 mg/100 g, respectively. These extracts had a total phenolic content of 112.22 ± 2.89 gallic acid equivalent (GAE) mg/100 g, a total flavonoid content of 54.09 ± 1.01 rutin equivalent (RE) mg/100 g, DPPH radical scavenging activity of 46%, and antioxidant activity of 213.25 ± 2.82 μM of Trolox equivalents (TEAC). C. sulcata extracts could prevent hydrogen peroxide (H₂O₂)-induced oxidative stress, reduce expression of the inflammatory markers nuclear Factor kappa B (NFκB) and phosphorylated IκBα (p-IκBα) in A549 human lung carcinoma cells, and inhibit Lipopolysaccharide (LPS)-induced THP-1 monocyte differentiation to an extent of 85%.     

Antimicrobial activity of acid-hydrolyzed Citrus unshiu peel extract in milk

Citrus fruit (Citrus unshiu) peels were extracted with hot water and then acid-hydrolyzed using hydrochloric acid. Antimicrobial activities of acid-hydrolyzed Citrus unshiu peel extract were evaluated against pathogenic bacteria, including Bacillus cereus, Staphylococcus aureus, and Listeria monocytogenes. Antilisterial effect was also determined by adding extracts at 1, 2, and 4% to whole, low-fat, and skim milk. The cell numbers of B. cereus, Staph. aureus, and L. monocytogenes cultures treated with acid-hydrolyzed extract for 12 h at 35°C were reduced from about 8 log cfu/mL to <1 log cfu/mL. Bacillus cereus was more sensitive to acid-hydrolyzed Citrus unshiu peel extract than were the other bacteria. The addition of 4% acid-hydrolyzed Citrus unshiu extracts to all types of milk inhibited the growth of L. monocytogenes within 1 d of storage at 4°C. The results indicated that Citrus unshiu peel extracts, after acid hydrolysis, effectively inhibited the growth of pathogenic bacteria. These findings indicate that acid hydrolysis of Citrus unshiu peel facilitates its use as a natural antimicrobial agent for food products.     

Inhibitory effect of sour orange (Citrus aurantium) juice on Salmonella Typhimurium and Listeria monocytogenes

The sour orange (Citrus aurantium) juice is commonly used as flavoring and acidifying agent for vegetable salads and appetizers in Turkey. It was aimed to determine the survival and growth pattern of Salmonella Typhimurium and Listeria monocytogenes in sour orange juice. Different concentrations of neutralized and un-neutralized juice samples were inoculated with each of the test microorganisms (∼6 log CFU/mL) separately and then incubated at 4 °C and 37 °C for seven days. It was detected both of the test microorganisms could survive and even grow in neutralized juice samples at 37 °C for two days. However, none of them could survive at the end of seventh day of incubation at 37 °C. Low incubation temperature (+4 °C) increased the survival of the tested microorganisms. Also, it was detected that L. monocytogenes were less resistant to the variable conditions than S. Typhimurium. It was concluded that the antimicrobial effect of sour orange juice mainly depends on the low pH value of the product. However, incubation time and temperature are also effective on the survival of the tested pathogens.     

Chemical and aroma profiles of yuzu (Citrus junos) peel oils of different cultivars

The essential oils of six different yuzu cultivars, Kumon (KUM), Nagano (NAG), Yasu (YAS), Jimoto (JIM), Komatsu Sadao (KOS) and Komatsu Koichi (KOK), were extracted by cold-pressing method. A total of 69 compounds of the six samples were identified. Application of GC-olfactometry and aroma extraction dilution analysis technique in three-fold stepwise dilution of the neat oil for all samples indicated eight odourants with the highest flavour dilution (FD) values. Those were limonene, α-pinene, α- and β-phellandrene, myrcene, γ-terpinene, (E)-β-farnesene and linalool. ‘KOS’ was differentiated from the other oil samples by showing the highest number of components having yuzu-like odour notes and also from the PCA analysis of the FD-factor values. This is the first time the aroma characteristics of yuzu essential oils of specified cultivars were investigated.     

Phenolic content and antioxidant activity of cantaloupe (cucumis melo) methanolic extracts

The objectives of this study were to determine phenolic content and antioxidant activity of methanolic extracts from different parts of cantaloupe (leaf, stem, skin, seed and flesh). The flesh extract afforded the highest yield (89.6 ± 0.3%) whilst the lowest yield was obtained from the seed (13.7 ± 0.5%) (p < 0.05). The leaf extract showed the highest total phenolic content (26.4 ± 0.3 mg GAE/g extract) and total flavonoid content (69.7 ± 3.37 μg RE/g extract) accompanied with best antioxidant activity through all antioxidant assays (p < 0.05). In addition, the stem extract also exhibited good antioxidant activity. Thus, these results suggest that methanolic extracts of cantaloupe leaf and stem may serve as a potential source of natural antioxidant for food and nutraceutical application.     

Chemical composition and antimicrobial activity of the essential oil of Lippia grandis Schauer (Verbenaceae) from the western Amazon

Lippia grandis Schauer is an aromatic plant that has been used as a spice in Brazilian culinary and in traditional medicine to treat liver disease, disorders of the stomach and throat infections. We determined the chemical composition of the essential oil of L. grandis and evaluated its potential for the treatment of clinically-important pathogenic micro-organisms. The essential oil was obtained by hydrodistillation and analyzed by gas chromatography-mass spectrometry (GC-MS), giving carvacrol (37.12%), ρ-cymene (11.64%), and thymol (7.83%) as the main components. The agar disk diffusion method of the essential oil was effective against 75% of the micro-organisms analyzed, in particular, Staphylococcus aureus, Enterococcus faecalis, and Escherichia coli. The minimum inhibitory concentration was 0.57 mg/ml for E. faecalis and 1.15 mg/ml for all the other strains. The results indicate that the essential oil of L. grandis contains chemical compounds with good potential for the treatment of infections.     

Antioxidant activity of ethanolic extract of Cortex fraxini and use in peanut oil

Cortex fraxini was extracted with 95% ethanol to obtain a crude antioxidant extract. The antioxidant activity was evaluated using the linoleic acid peroxidation method and the free radical scavenging assays, namely 2,2′-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl radicals. Cortex fraxini extract (CFE) showed high inhibition of peroxidation of linoleic acid when compared with butylated hydroxytoluene (BHT). CFE also exhibited excellent scavenging activity on DPPH and hydroxyl radicals. Total antioxidant activity was measured by the reduction of Mo(VI) to Mo(V) by the extract, and subsequent formation of a green phosphate/Mo(V) complex at acid pH. CFE had significant total antioxidant activity and the effects were increased with increasing reaction time. The total phenolic content of the sample, analyzed by using Folin–Ciocalteu’s reagent, was 91.33 mg/g dry weight expressed as pyrocatechol equivalents. Then the suitability of CFE as an antioxidant was determined in peanut oil, and the decrease of lipid oxidation was monitored using thiobarbituric acid-reactive substances (TBARS) assay. CFE treatment significantly (P < 0.05) reduced lipid oxidation in peanut oil compared to the control. No significant differences (P = 0.05) in lipid oxidation were detected between CFE antioxidant and BHT antioxidant samples.     

Antioxidant capacity,total phenolic and ascorbate content as a function of the genetic diversity of leek (Allium ampeloprasum var. porrum)

Extracts of the white shaft and green leaves of 30 leek cultivars were investigated for their antioxidant properties, total phenolic (TP) and l-ascorbic acid (AA) content. The measured antioxidant properties included free radical scavenging activities against peroxyl (ORAC) and 2,2-diphenyl-1-picrylhydrazyl radicals (DPPH) and their Fe³⁺ reducing capacity (FRAP). The results from this study suggest that the green leek leaves generally have significantly stronger antioxidant properties than the white shaft. Correlation analysis between the TP and the AA content and the antioxidant activity showed that phenolics and ascorbic acid contribute significantly to the antioxidant activity of leek. The three antioxidant activity assays were all correlated for the extracts of the white shaft of the 30 leek cultivars. Principal component analysis (PCA) elucidated the influence of part and type of cultivar on the antioxidant capacity, TP, and l-ascorbic acid content, whilst the breeding strategy and seed company had no influence.     

Antioxidant activity and rosmarinic acid changes in salicylic acid-treated Thymus membranaceus shoots

In order to characterise the antioxidant activity of Thymus membranaceus Boiss. subsp. membranaceus, an endemic species in Southeast Spain, five different analytical methods were used. Water, methanol and hexane extracts obtained from 60-day-old in vitro-grown shoots were assayed for their antioxidative properties using DPPH (1,1-diphenyl-2-picrylhydrazyl radical), ABTS (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonate), FRAP (ferric reducing antioxidant power), reducing power and conjugated diene assays. Total soluble phenol content, as well as rosmarinic acid, in the different extracts were also determined. Methanolic extracts exhibited the best antioxidant activity and the highest amounts of total soluble phenolics. A single application of salicylic acid (10 μM) on culture media resulted in an increase in rosmarinic acid and phenolic levels, which in turn improved the extracts’ antioxidant properties. These current findings open new opportunities for obtaining valuable natural antioxidants for commercial exploitation by using tissue culture systems.     

Effect of air-drying temperature on physico-chemical properties of dietary fibre and antioxidant capacity of orange (Citrus aurantium v. Canoneta) by-products

Dehydration promoted important modifications affecting both the physico-chemical properties of dietary fibre (DF) and the antioxidant capacity of orange by-products (peel and pulp remaining after juice extraction). The significance of such changes was largely dependent on the air-drying temperature used (from 30 °C to 90 °C). The major modifications on the DF components were observed when either extended drying periods, i.e. at lower temperatures, or elevated drying temperatures were applied. Dehydration around 50–60 °C apparently promoted the minor disruption of cell wall polymers, in particular of pectic substances. Pulp samples exhibited higher values of swelling (SW) and fat adsorption capacity (FAC) than those derived from orange peel. Although, significant decreases in water retention capacity (WRC), FAC and solubility values were detected for both by-products as the air-drying temperature increased. The antioxidant capacity associated to dehydrated citrus by-products was significantly higher for orange peel than for pulp samples. In general, the by-products studied proved to be quite resistant to the different heat treatments applied within the range of 40–70 °C. In overall, the study shows that, in order to preserve either the DF quality and/or the antioxidant capacity, air-drying temperature should be controlled since both types of compounds, DF components and antioxidants, might be degraded or modified either when extended drying periods and/or high drying temperatures are applied.     

Enhancement of antioxidant activity,total phenolic and flavonoid content of black soybeans by solid state fermentation with Bacillus subtilis BCRC 14715

In the present study, a solid state fermentation of black soybeans with Bacillus subtilis BCRC 14715 was performed. The effect of fermentation on the changes of total phenolic and flavonoid content and antioxidant activities including 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging effect, and Fe²⁺-chelating ability exerted by various solvent (water, 80% methanol, 80% ethanol, 80% acetone) extracts of black soybeans was examined. It was found that fermentation enhanced the total phenolic and flavonoid content as well as antioxidant activity of the black soybean extract. Among the various extracts examined, the acetone extract of fermented black soybeans showed the highest total phenolic and flavonoid content. The acetone extract and the methanol extract of fermented black soybeans showed the highest DPPH free radical-scavenging effect and Fe²⁺-chelating ability, respectively. Analysis of extraction yields showed that the active principle associated with the DPPH radical-scavenging effect was most efficiently extracted from black soybeans using water, regardless of fermentation. Water and methanol effectively extract the Fe²⁺-chelating principles from non-fermented and fermented black soybeans, respectively.