Antioxidant and Antiproliferative Activity of <Emphasis Type="Italic">Diospyros lotus</Emphasis> L. Extract and Isolated Compounds

The object of the study was to determine the chemical composition of Diospyros lotus L. extract and their antioxidant and antiproliferative properties. Eight compounds were isolated from D. lotus and identified as gallic acid, methylgallate, ellagic acid, kaempferol, quercetin, myricetin, myricetin 3-O-β-glucuronide, and myricetin-3-O-α-rhamnoside. D. lotus extract tested in different in vitro systems (DPPH, ABTS, FRAP, and Fe²⁺ chelating activity assay) showed significant antioxidant activity. The potential antiproliferative properties of D. lotus extract and isolated compounds against nine human cancer cell lines such as COR-L23, CaCo-2, C32, ACHN, A375, A549, Huh-7D12, MCF-7, and LNCaP were investigated in vitro by SRB assay. D. lotus extract demonstrated the highest inhibitory activity against COR-L23 with an IC₅₀ value of 12.2 μg/ml. Among identified hydrolysable tannins, ellagic acid evidenced strong antiproliferative activity against both C32 and A375 cells with IC₅₀ values of 0.8 and 4.1 μg/ml, respectively. Interesting results were observed, also, with gallic acid that showed the highest cytotoxic activity against CaCo-2 (IC₅₀ 2.6 μg/ml). Overall, the results of this study suggest that D. lotus displays a good antioxidant activity and has antiproliferative effects. Both activities are related to identified phenolic compounds.     

Antioxidative and Anticancer Activity of Extracts of Cherry (<Emphasis Type="Italic">Prunus serrulata</Emphasis> var. <Emphasis Type="Italic">spontanea</Emphasis>) Blossoms

Organic solvent (methanol, ethanol, and acetone) extracts and water extracts of cherry (Prunus serrulata var. spontanea) blossoms were prepared, and antioxidant activities of the extracts were evaluated. Methanolic CBE (100 μg/ml) showed the highest total phenol content (104.30 μM), radical scavenging activity (34.2%), and reducing power (0.391). The effect of CBE on DNA damage induced by H₂O₂ in human leukocytes was evaluated by Comet assay. All CBE was a potent dose dependent inhibitor of DNA damage induced by 200 μM of H₂O₂, methanolic CBE showed the most strong inhibition activity. The methanolic CBE of 500 μg/ml showed 38.8% inhibition against growth of human colon cancer cell line HT-29. These results indicated that cherry blossoms could provide valuable bioactive materials.     

Antioxidant Capacity and Phenolics Content of Apricot (<Emphasis Type="Italic">Prunus armeniaca</Emphasis> L.) Kernel as a Function of Genotype

Fourteen apricot genotypes grown under similar cultural practices in Trans-Himalayan Ladakh region were studied to find out the influence of genotype on antioxidant capacity and total phenolic content (TPC) of apricot kernel. The kernels were found to be rich in TPC ranging from 92.2 to 162.1 mg gallic acid equivalent/100 g. The free radical-scavenging activity in terms of inhibitory concentration (IC₅₀) ranged from 43.8 to 123.4 mg/ml and ferric reducing antioxidant potential (FRAP) from 154.1 to 243.6 FeSO₄.7H₂O μg/ml. A variation of 1–1.7 fold in total phenolic content, 1–2.8 fold in IC₅₀ by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay and 1–1.6 fold in ferric reducing antioxidant potential among the examined kernels underlines the important role played by genetic background for determining the phenolic content and antioxidant potential of apricot kernel. A positive significant correlation between TPC and FRAP (r = 0.671) was found. No significant correlation was found between TPC and IC₅₀; FRAP and IC₅₀; TPC and physical properties of kernel. Principal component analysis demonstrated that genotypic effect is more pronounced towards TPC and total antioxidant capacity (TAC) content in apricot kernel while the contribution of seed and kernel physical properties are not highly significant.     

Antioxidative and Antigenotoxic Effects of Garlic (<Emphasis Type="Italic">Allium sativum</Emphasis> L.) Prepared by Different Processing Methods

This study describes the antioxidant activities and antigenotoxic effects of garlic extracts prepared by different processing methods. Aged-garlic extract (AGE) showed a significantly higher total phenolic content (562.6 ± 1.92 mg/100 g garlic acid equivalents) than those of raw garlic extract (RGE) or heated garlic extract (HGE). The SC₅₀ for DPPH RSA in HGE was significantly the highest at 2.1 mg/ml. The SC₅₀ for SOD-like activity in garlic extracts was, in decreasing order, RGE (7.3 mg/ml) > AGE (8.5 mg/ml) > HGE (9.2 mg /ml). The ED₅₀ of AGE was the highest (19.3 μg/ml) regarding H₂O₂ induced DNA damage and its inhibition rate was 70.8%. The ED₅₀ of RGE for 4-hydroxynonenal (a lipid peroxidation product) induced DNA damage was 38.6 μg/ml, followed by AGE > HGE. Although the heat treatment of garlic tended to decrease the TPC and SOD-like activity and increased DPPH RSA, garlic, in general, has significant antioxidant activity and protective effects against oxidative DNA damage regardless of processing method.     

Hypolipidemic Effect of Avocado (<Emphasis Type="Italic">Persea americana</Emphasis> Mill<Emphasis Type="Italic">)</Emphasis> Seed in a Hypercholesterolemic Mouse Model

Avocado seed contains elevated levels of phenolic compounds and exhibits antioxidant properties. We investigated the effect of Avocado Seed Flour (ASF) on the lipid levels in mice on a hyperlipidemic diet. The concentration of phenols was determined by high-performance liquid chromatography, antioxidant activity was evaluated using the Trolox equivalent antioxidant capacity method, and dietary fiber was measured using the Association of Official Analytical Chemists (AOAC) method. The LD₅₀ of ASF was determined using Lorke’s method and hypolipidemic activity was evaluated in a hypercholesterolemic model in mice. Protocatechuic acid was the main phenolic compound found in ASF, followed by kaempferide and vanillic acid. The total phenolic content in the methanolic extract of ASF was 292.00 ± 9.81 mg gallic acid equivalents/g seed dry weight and the antioxidant activity resulted in 173.3 μmol Trolox equivalents/g DW. In addition, a high content of dietary fiber was found (34.8%). The oral LD₅₀ for ASF was 1767 mg/kg body weight, and treatment with ASF significantly reduced the levels of total cholesterol, LDL-C, and prediction of the atherogenic index. Therefore, the antioxidant activity of phenolic compounds and dietary fiber in ASF may be responsible for the hypocholesterolemic activity of ASF in a hyperlipidemic model of mice.     

Carotenoid Composition of Jackfruit (<Emphasis Type="Italic">Artocarpus heterophyllus</Emphasis>), Determined by HPLC-PDA-MS/MS

Carotenoids are pigments responsible for the yellow-reddish color of many foods and are related to important functions and physiological actions, preventing several chronic-degenerative diseases. The objective of this study was to confirm the carotenoid composition of jackfruit by high-performance liquid chromatography connected to photodiode array and mass spectrometry detectors (HPLC-PDA-MS/MS). The main carotenoids were all-trans-lutein (24–44%), all-trans-β-carotene (24–30%), all-trans-neoxanthin (4–19%), 9-cis-neoxanthin (4–9%) and 9-cis-violaxanthin (4–10%). Either qualitative or quantitative differences, mainly related to the lutein proportion, were found among three batches of jackfruit. Since the fruits from batch A showed significantly lower contents for almost all carotenoids, it also had the lowest total carotenoid content (34.1 μg/100 g) and provitamin A value, whereas the total carotenoid ranged from 129.0 to 150.3 μg/100 g in the other batches. The provitamin A values from batches B and C were 3.3 and 4.3 μg RAE/100 g, respectively. The carotenoid composition of jackfruit was successfully determined, where 14 of the 18 identified carotenoids were reported for first time. Differences among batches may be due to genetic and/or agricultural factors.     

Kinetics of the Inhibition of Renin and Angiotensin I Converting Enzyme by Polar and Non-polar Polyphenolic Extracts of <Emphasis Type="Italic">Vernonia Amygdalina</Emphasis> and <Emphasis Type="Italic">Gongronema Latifolium</Emphasis> Leaves

The aim of this study was to determine the in vitro modulation of the renin-angiotensin system by polyphenolic extracts and fractions of two green leafy vegetables, Vernonia amygdalina (VA) and Gongronema latifolium (GL), that are used for food and medicinal purposes. An 80% acetone extract of each leaf was fractionated on silicic acid-packed column to give two main fractions: acetone eluate (flow-through) and ethanol eluate (column-bound), that consist mostly of chlorophyllic and non-chlorophyllic fractions, respectively. Column fractionation resulted in polyphenolic fractions that displayed higher potency against angiotensin converting enzyme (ACE) and renin than the crude acetone extracts; generally, the chlorophyllic fraction was more active than the non-chlorophyllic fraction. ACE-inhibitory activity was significantly higher (p < 0.05) for the chlorophyllic fraction of VA than GL, with IC₅₀ values of 0.207 and 0.413 mg/ml, respectively. Similarly, the chlorophyllic fraction of VA had significantly higher (p < 0.05) renin inhibition than GL, with IC₅₀ values of 0.172 and 0.513 mg/ml, respectively. Kinetics studies showed that the chlorophyllic fractions of VA and GL exhibited mostly mixed-type ACE and renin inhibitions. We concluded that the hydrophobic nature of the chlorophyllic fraction may have contributed to the increased interaction with enzyme protein and inhibition of activities of ACE and renin.     

Antioxidant in Mango (<Emphasis Type="Italic">Mangifera indica</Emphasis> L.) Pulp

This work was carried out to investigate the pulp composition of four mango cultivars (Haden, Tommy Atkins and Ubá) at the ripening stage in relation to three components with antioxidant potential (total phenolics, carotenoids and ascorbic acid). Total phenolic compound content was estimated by the Folin-Ciocalteu reagent and total carotenoid content by spectrophotometry at 450 nm. The contents of β-carotene and total vitamin C (ascorbic acid and dehydroascorbic acid) were quantified by high performance liquid chromatography. Differences were found among the four mango cultivars in all the components analyzed. The content of phenolic compounds ranged from 48.40 (Haden) to 208.70 mg/100 g (Ubá); total carotenoid from 1.91 (Haden) to 2.63 mg/100 g (Palmer); β-carotene from 661.27 (Palmer) to 2,220 μg/100 g (Ubá) and total ascorbic acid ranged from 9.79 (Tommy Atkins) to 77.71 mg/100 g (Ubá). These results corroborated previous information that mangoes are a good source of antioxidants in human diet.     

Comparative Study of Lipophilic and Hydrophilic Antioxidants from <Emphasis Type="Italic">In vivo</Emphasis> and <Emphasis Type="Italic">In vitro</Emphasis> Grown <Emphasis Type="Italic">Coriandrum sativum</Emphasis>

Coriander is commonly used for medicinal purposes, food applications, cosmetics and perfumes. Herein, the production of antioxidants in vegetative parts (leaves and stems) of in vivo and in vitro grown samples was compared. In vitro samples were clone A- with notorious purple pigmentation in stems and leaves and clone B- green. Seeds were also studied as they are used to obtain in vivo and in vitro vegetative parts. Lipophilic (tocopherols, carotenoids and chlorophylls) and hydrophilic (sugars, ascorbic acid, phenolics, flavonols and anthocyanins) compounds were quantified. The antioxidant activity was evaluated by radical scavenging activity, reducing power and lipid peroxidation inhibition. The in vivo sample showed the highest antioxidant activity mainly due to its highest levels of hydrophilic compounds. Otherwise, in vitro samples, mainly clone A, gave the highest concentration in lipophilic compounds but a different profile when compared to the in vivo sample. Clones A and B revealed a lack of β-carotene, β- and δ-tocopherols, a decrease in α-tocopherol, and an increase in γ-tocopherol and clorophylls in comparison to the in vivo sample. In vitro culture might be useful to explore the plants potentialities for industrial applications, controlling environmental conditions to produce higher amounts of some bioactive products.     

<Emphasis Type="Italic">In vitro</Emphasis> and <Emphasis Type="Italic">In vivo</Emphasis> Antioxidant Properties of Extracts from <Emphasis Type="Italic">Coptis chinensis</Emphasis> Inflorescence

The inflorescence of cultivated Coptis chinensis has been valued for tea production for many years in China. The antioxidant activities of C. chinensis inflorescence extracts prepared by various solvents were investigated by using several established in vitro systems: 2,2′-azinobis(3-ethylbenzthiazoline-6-sulphonic acid (ABTS), α,α-diphenyl-β-picrylhydrazyl (DPPH) and superoxide radical scavenging assays, reducing power assay, and ferrothiocyanate (FTC) and thiobarbituric acid (TBA) assays. The results showed that the 70% ethanol extract (EE) had the strongest antioxidant activity in vitro among the various extracts. Based on the in vitro results, EE was used to evaluate the antioxidant activity of C. chinensis inflorescence in vivo. The liver and kidney of intoxicated animals showed a significant decrease in superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) levels, while the malondialdehyde (MDA) level showed a significant increase. These changes were significantly reversed after treatment with EE and the standard vitamin E. Thus, the C. chinensis inflorescence may be a valuable natural source that can be applicable to food industries.     

Phenolic Composition,Antioxidant Capacity and <Emphasis Type="Italic">In Vitro</Emphasis> Cancer Cell Cytotoxicity of Nine Prickly Pear (<Emphasis Type="Italic">Opuntia</Emphasis> spp.) Juices

Juices of nine prickly pears (Opuntia spp.) were characterized in terms of color, acidity, sugar content, phenolics, flavonoids, betalains and antioxidant activity and tested in vitro against four cancer cell lines. The juices had pH′s, acidities and sugar ranging from 4.27 to 5.46, 0.03 to 0.27% and 8 to 14.7°Brix, respectively. Juices also varied in color from white to purple and contained total phenolics, flavonoids, betaxanthins, betacyanins and antioxidant capacity ranging from 22 to 226 μg gallic acid eq/g, 95 to 374 μg quercetin eq/g, 3 to 189 μg/g, 1.6 to 300 μg/g and 17 to 25 micromoles Trolox eq./mL, respectively. Among the cancer lines tested, viability of prostate and colon cells were the most affected. Moradillo contained the highest flavonoids and diminished both prostate and colon cancer cell viability without affecting mammary or hepatic cancer cells. Rastrero reduced the growth of the four cancer cell lines without affecting normal fibroblast viability. The research shows intervarietal differences among prickly pears in terms of juice properties and phytochemicals that could prevent oxidative stress and cancer.     

The Antioxidant and Anti-proliferative Activity of the Lebanese <Emphasis Type="Italic">Olea europaea</Emphasis> Extract

It is becoming increasingly evident that certain phytochemicals possess cancer chemopreventive properties. In this study, the anti-proliferative activity of plant extracts from olive (Olea europaea L.) leaves was tested on human leukemic cell line (Jurkat). Cytotoxicity of various concentrations of plant extracts was examined and the IC₅₀ was determined. Olive leaf extracts showed concentration-dependent anti-proliferative effect as determined by the WST-1 proliferation kit and [³H]-thymidine incorporation method. To study whether cell death was due to apoptosis, cells were stained with Annexin V-FITC and PI and the expression of important regulatory proteins (Bcl-2, Bax, and p53) involved in apoptosis were examined by Western blot. The antioxidant activity of olive leaves (SC₅₀ = 0.1 mg dry weight) was studied using the DPPH scavenging method. Present findings suggest that olive leaves extracts exhibit anti-proliferative effect on leukemic cells by inducing apoptosis.     

Antioxidant and Antimutagenic Activities of <Emphasis Type="Italic">Randia echinocarpa</Emphasis> Fruit

We report for the first time the antioxidant and antimutagenic activities of fractions from Randia echinocarpa fruit, which is a Rubiaceae plant native to Sinaloa, Mexico. This fruit has been traditionally used in the prevention or treatment of cancer, among other diseases. The pulp of the fruit was sequentially extracted with solvents of different polarity (i.e. hexane, chloroform, methanol and water). A high extraction yield was obtained with methanol (72.17% d.w.). The aqueous extract showed the highest content of phenolics (2.27 mg/g as ferulic acid equivalents) and the highest antioxidant activity based on the β-carotene bleaching method (486.15). The commercial antioxidant BHT was used as control (835.05). Antimutagenic activity of the aqueous extract (0–500 μg/tube) was evaluated using the Salmonella microsuspension assay (YG1024 strain) and 1-NP as the mutagen (50 and 100 ng/tube). The aqueous extract was neither toxic nor mutagenic and the percentage of inhibition on 1-NP mutagenicity was 32 and 53% at doses of 50 and 100 ng/tube, respectively. The results of the double incubation assay suggest that the extract inhibited the mutagenicity of 1-NP by a combination of desmutagenic and bioantimutagenic effects.     

Volatile Compounds and Antioxidative Activity of <Emphasis Type="Italic">Porophyllum tagetoides</Emphasis> Extracts

Porophyllum tagetoides is an annual warm-weather herb that has an intense typical smell. Its leaves are commonly used in soup preparation and traditional medicine for treatment of inflammatory diseases. Its volatile compounds and antioxidant properties were evaluated in crude, aqueous and ethanol leaf extract and an oil emulsion using different antioxidant assays in vitro, such as: DPPH radical scavenging activity, redox potential, polyphenol content, reducing power and optical density. A high antioxidative activity was found when comparing leaves with stems. The crude extract from leaves showed a very high reducing power (2.88 ± 0.20 O.D.) and DPPH radical-scavenging activity (54.63 ± 4.80%), in concordance with a major concentration of vitamin C (23.97 ± 0.36 mg/100 g). Instead, the highest polyphenol content (264.54 ± 2.17 mg GAE/g of sample) and redox potential (561.23 ± 0.15 mV) were found by the ethanol and aqueous extract, respectively. Aldehydes and terpenes such as nonanal, decanal, trans-pineno, β-myrcene and D-limonene were the major volatiles found. This study suggests that Porophyllum tagetoides extracts could be used as antioxidants.     

<Emphasis Type="Italic">Prunus persica</Emphasis> var. <Emphasis Type="Italic">platycarpa</Emphasis> (Tabacchiera Peach): Bioactive Compounds and Antioxidant Activity of Pulp,Peel and Seed Ethanolic Extracts

A comparative analysis of ethanol extracts from peel, pulp and seed of Prunus persica var. platycarpa (Tabacchiera peach) was done. The total phenol, flavonoid and carotenoid content as well as the antioxidant properties by using different in vitro assays (DPPH, ABTS, FRAP, Fe-chelating, β-carotene bleaching test) were evaluated. Pulp extract was subjected to liquid chromatography-electrospray-tandem mass spectrometry (HPLC-ESI-MS/MS). Gallic acid, protocatechuic acid, protocatechualdehyde, chlorogenic acid, p-coumaric acid, and ferulic acid were identified as main constituents. Pulp extract was characterized by the highest total phytonutrients content and exhibited the highest antioxidant activity in all in vitro assays (IC₅₀ values of 2.2 μg/mL after 60 min of incubation by using β-carotene bleaching test and 2.9 μg/mL by using Fe-chelating assay). Overall, the obtained results suggest that P. persica var. platycarpa displays a good antioxidant activity and its consumption could be promoted.     

Antioxidant,Antimicrobial and Genotoxicity Screening of Hydro-alcoholic Extracts of Five Serbian <Emphasis Type="Italic">Equisetum</Emphasis> Species

The hydro-alcoholic extracts of five Equisetum species, E. arvense L., E. sylvaticum L., E. fluviatile L., E. palustre L. and E. telmateia Ehrh., growing-wild in Serbia were evaluated for their genotoxicity, antimicrobial activity, antioxidant capacity and the results related to the total phenol content and HPLC flavonoid profiles. The total phenol content was 92–349 μmol expressed as equivalents of chlorogenic acid per g of dried plant material. Main identified compounds were kaempferol-, quercetin- glycosides and caffeic acid derivatives. E. telmateia extract showed the greatest antioxidant capacity. Almost all tested microorganisms demonstrated some degree of sensitivity to the examined extracts. All tested extracts at 62.5 μg/ml showed higher incidence of micronucleus formation than in the control sample. The obtained data allowed mutual comparison of examined species and their assessment as possible sources of antioxidants, antimicrobials and/or genotoxic substances.     

Analysis of Flavonoids from Leaves of Cultivated <Emphasis Type="Italic">Lycium barbarum</Emphasis> L.

Leaves of Lycium barbarum are widely used as medicine vegetables and functional tea in China. The main flavonoids present in the leaves were separated and identified by high performance liquid chromatography (HPLC), liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (LC-(APCI) MS) and ultraviolet-visible spectra with shift additives. The predominant flavonoid was identified as rutin. Leaves are the rutin-rich parts (16.03–16.33 mg/g). In the wild and cultivated L. barbarum fruits, contents of rutin were determined very low (0.09–1.38 mg/g). The contents of total flavonoids (21.25 mg/g) of cultivated L. barbarum leaves were much higher than those in the wild L. barbarum leaves (17.86 mg/g), so cultivated barbarum leaves are a suitable source for medicine vegetables and functional tea.     

White Tea (<Emphasis Type="Italic">Camellia sinensis</Emphasis> Kuntze) Exerts Neuroprotection against Hydrogen Peroxide-Induced Toxicity in PC12 Cells

Tea is a popular beverage whose consumption is associated with prevention of certain disorders. The objective of the study was to investigate the potential neuroprotective effect of white tea extract (WTE) on hydrogen peroxide induced toxicity in PC12 cells. Cells were treated with various doses of WTE (10–250 μg/ml) before exposition to 250 μM hydrogen peroxide and cell survival was determined through the MTT and LDH assays. Oxidative stress was quantified in the cells after treatments as intracellular reactive oxygen species (ROS) production and the antioxidant activity of the extract was assessed in a cell free system in terms of free radical scavenging capacity. Results showed that WTE has a significant protective effect in the PC12 cell line against hydrogen peroxide as cell survival was significantly superior in WTE-treated cells compared to hydrogen peroxide-treated cells. A reduction on intracellular oxidative stress as well as radical scavenging properties were produced by WTE. Results suggest that WTE protects PC12 cells against H₂O₂-induced toxicity, and that an antioxidant mechanism through ROS scavenging may be in part responsible for cells neuroprotection.     

Antioxidant Capacity and Antimutagenic Activity of Natural Oleoresin from Greenhouse Grown Tomatoes (<Emphasis Type="Italic">Lycopersicon esculentum</Emphasis>)

Natural oleoresins rich in lycopene were obtained from two varieties of tomato (Zedona and Gironda) and their nutraceutical potential (antioxidant and antimutagenic capacity) was evaluated. Both oleoresins had a high content of lycopene, 58.33 ± 1.67 mg/g (Zedona) and 63.97 ± 0.80 mg/g (Gironda). The antioxidant activity (AA) of the oleoresins by β-carotene method were 56.4–74.5% (Zedona) and 51–72.8% (Gironda), while when using the free radical stable 2,2-diphenyl-picryl-hydrazyl (DPPH) method, the antiradical activity (ARA) was determined to be 18.2–32.7% (Zedona) and 16.6–26.7% (Gironda) for the concentrations tested that of 200–400 μM equivalents of lycopene. The antimutagenic activity of the oleoresins was tested against aflatoxin B₁ (AFB₁) using the microsuspension assay, both varieties had a very high antimutagenic potential against AFB₁ (60–66%).These results suggest the NCRT can be taken advantage to obtaining rich oleoresin in lycopene with a nutraceutical value.