<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.     

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">In vitro</Emphasis> Antioxidant Activity of Aged Extracts of some Italian <Emphasis Type="Italic">Allium</Emphasis> Species

Antioxidant activity of fresh Allium sativum L. (garlic) is well known and is mainly due to unstable and irritating organosulphur compounds. Fresh garlic extracted over a prolonged period (up to 20 months) produces odourless aged garlic extract (AGE) containing stable and water soluble organosulphur compounds that prevent oxidative damage by scavenging free radicals. The aim of this study was to investigate the in vitro antioxidant activity of aged (up to 20 months) 15% hydroethanolic extracts of different parts (bulbs, bulblets, flower bulblets, flowers, and leaves) of three Allium spontaneous species which are endemic for Italian flora: Allium neapolitanum Cyr., Allium subhirsutum L., Allium roseum L. and to compare it with the in vitro antioxidant activity of aged 15% hydroethanolic extracts of bulbs and leaves of garlic. The antioxidant potential of aged extracts of all species has been evaluated using two different spectrophotometric assays: 2,2-diphenylpicrylhydrazyl (DPPH) test and the ferric reducing/antioxidant power (FRAP) assay. Furthermore the polyphenol content was determined. The aged extracts obtained from the leaves showed the best antioxidant activity, followed by flowers and then by bulbs in both used tests, while flower bulblets and bulblets exhibited lower results or no activity. The polyphenol content was generally directly correlated with antioxidant/antiradical activity. This study confirms the data obtained in previous researches, the wild-type species of Allium and in particular organs other than bulbs are more active and efficacious than garlic bulb. Surely leaves of these Allium spp. deserve special attention.     

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.     

Comparative Study on the Chemical Composition,Antioxidant Properties and Hypoglycaemic Activities of Two <Emphasis Type="Italic">Capsicum annuum</Emphasis> L. Cultivars (<Emphasis Type="Italic">Acuminatum</Emphasis> small and <Emphasis Type="Italic">Cerasiferum</Emphasis>)

The present study aimed to evaluate for the first time the phenols, flavonoids, carotenoids, capsaicin and dihydrocapsaicin content and the antioxidant and hypoglycemic properties of Capsicum annuum var. acuminatum small and C. annuum var. cerasiferum air-dried fruits. The ethanol extract of C. annuum var. acuminatum small, characterized by the major content of total poliphenols, flavonoids, carotenoids and capsaicinoids, showed the highest radical scavenging activity (IC₅₀ of 152.9 μg/ml). On the contrary, C. annuum var. cerasiferum showed a significant antioxidant activity evaluated by the β-carotene bleaching test (IC₅₀ of 3.1 μg/ml). The lipophilic fraction of both C. annuum var. acuminatum and C. annuum var. cerasiferum exhibited an interesting and selective inhibitory activity against α-amylase (IC₅₀ of 6.9 and 20.1 μg/ml, respectively).     

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.     

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.     

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.     

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.     

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.     

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.     

Bread Supplemented with Amaranth (<Emphasis Type="Italic">Amaranthus cruentus</Emphasis>): Effect of Phytates on <Emphasis Type="Italic">In Vitro</Emphasis> Iron Absorption

The objective of the present study was to evaluate the effect of the bread supplemented with whole amaranth flour (0, 20 and 40%) on iron bioavailability using Caco-2 cells model. The phytate and lower myo-inositol phosphates content in in vitro bread digests were measured by high pressure liquid chromatography. The breads made with amaranth showed significant increase of soluble phytates levels (up to 1.20 μmol/g in dry matter for the 40% of substitution) in comparison with controls, which have not detectable values. A negative correlation among phytate and Fe availability was found when increased levels of amaranth. Ferritin concentration was found 2.7- and 2.0-fold higher (P < 0.05) in cultures exposed to 20% and 40% of amaranth formulated bread samples, respectively, compared to control bread. The soluble phytate/Fe molar ratio explained the whole amaranth flour-mediated inhibitory effect associated to the limitation of available Fe; however, the use up to 20% of amaranth in bread formulation appears as a promising strategy to improve the nutritional value of bread, as indicated by the ferritin concentrations quantified in cell cultures. Higher proportion of amaranth flour increased Fe concentration although there was not detected any increase in Fe uptake.     

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.     

Chemical and structural characterization of lignins isolated from <Emphasis Type="Italic">Caragana sinica</Emphasis>

Successive extractions of the dewaxed Caragana sinica with 70 % ethanol, 70 % ethanol containing 1 % NaOH, 1 M KOH, 1 M NaOH, 3 M KOH, and 3 M NaOH at 75 °C for 3 h released 80.4 % of the original lignin. The physico-chemical properties and structural features of these lignin fractions were comprehensively characterized by alkaline nitrobenzene oxidation, GPC and two-dimensional NMR spectroscopy. The results showed that all the lignin fractions had relatively lower molecular average-weights ranging between 910 and 1630 g/mol. Moreover, 2D-NMR showed that the lignin fraction L₂ isolated with 70 % ethanol containing 1 % NaOH from C. sinica was similar to a hardwood lignin, with a high S/G ratio accounted for 1.2, a predominance of β-O-4′ aryl ether linkages (77 % of all side-chains), followed by β-β′ resinol-type linkages (8 %) and lower amounts of β-5′ phenylcoumaran substructures (7 %), β-1′ spirodienone-type linkages (3 %), and cinnamyl end groups (5 %). The high predominance of the S-lignin units, together with the high proportion of β-O-4′ aryl ether linkages could contribute to a relatively high reactivity of C. sinica lignin during alkaline pulping.     

Bio-efficacy of the Essential Oil of Oregano (<Emphasis Type="Italic">Origanum vulgare</Emphasis> Lamiaceae. Ssp. <Emphasis Type="Italic">Hirtum</Emphasis>)

The aim of this study was to investigate the bioactivity of the essential oil isolated from Origanum vulgare L. (EOv). We analyzed the in vivo anti-inflammatory properties in a mouse-airway inflammation model and the in vitro antimicrobial activity, genotoxicity over the anaphase-telophase with the Allium cepa strain and its cytotoxicity/viability in A549 culture cells. In vivo, EOv modified the levels of tumor necrosis factor -α and viable activated macrophages and was capable to mitigate the effects of degradation of conjugated dienes. In vitro, EOv reduced the viability of cultured A549 cells as well as the mitotic index and a number of chromosomal aberrations; however, it did not change the number of phases. We found that EOv presents antimicrobial activity against different Gram (?) and (+) strains, measured by disc-diffusion test and confirmed with a more accurate method, the AutoCad software. We postulate that EOv presents antibacterial, antioxidant and chemopreventive properties and could be play an important role as bioprotector agent.     

<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.     

Identification and Quantification of Steroidal Alkaloids from Wild Tuber-Bearing <Emphasis Type="Italic">Solanum</Emphasis> Species by HPLC and LC-ESI-MS

Wild Solanum species are characterized by several types of glycoalkaloids (GAs), which are usually not found in commercial potato cultivars. These alkaloids serve as defence compounds against herbivores and microorganisms, especially fungi. In this study, the GA composition of tuber and leaf material of 17 wild Solanum species was analysed qualitatively by liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) and quantitatively by high-performance liquid chromatography. The GAs identified were α-solanine, α-chaconine, β-chaconine, solasonine, solamargine, demissine, dehydrodemissine, α-tomatine, dehydrotomatine, commersonine, dehydrocommersonine, leptine I and leptine II. Most species accumulated the common potato GAs α-solanine and α-chaconine in tubers and solasonine and solamargine additionally in leaves. In some species, such as S. acaule ssp. acaule, S. demissum and S. polyadenium, substantial amounts of unusual alkaloids were detected. By using LC-ESI-MS, we detected several minor alkaloids such as dehydrogenated forms of α-tomatine, demissine and commersonine for the first time. Total GA content, expressed as the sum of the four main alkaloids α-solanine, α-chaconine, solamargine and solasonine differed from species to species. In general, GA contents in leaf tissue were higher and GA patterns were more complex than those of tubers.     

Variation in Contents of Total Phenolics and Flavonoids and Antioxidant Activities in the Leaves of 11 <Emphasis Type="Italic">Eriobotrya</Emphasis> Species

Eriobotrya plants are known to have significant amounts of phenolics and flavonoids, and exhibit a strong antioxidant activity. Experiments were conducted to examine variation in the contents of total phenolics and flavonoids, and antioxidant activities in the leaves of 11 Eriobotrya species (Tibet loquat, Daduhe loquat, Hengchun loquat, Taiwan loquat, Oak leaf loquat, Bengal loquat, Fragrant loquat, Guangxi loquat, Obovate loquat, Big flower loquat, and common loquat, the last species include two materials, one is a cultivar ‘Zaozhong 6’, another is a wild tree). In these species, ‘Zaozhong 6′ loquat is a cultivar. The leaf extracts of ‘Tibet’, ‘Obovate’, ‘Taiwan’, ‘Bengal’ and ‘Hengchun’ loquats exhibited significantly higher contents of total flavonoids and total phenolics, compared with those of other species. Of these 11 species, the highest contents of total phenolics and total flavonoids were observed in ‘Tibet’ and ‘Obovatae’ loquats, respectively. The significantly stronger antioxidant abilities assessed by the DPPH radical scavenging activity and reducing power were obtained in the leaf extracts of ‘Taiwan’, ‘Tibet’, ‘Bengal’, ‘Oak leaf’, ‘Hengchun’ and ‘Obovate’ loquats, compared with the other species. In addition, significant correlations were found between the contents of total phenolics or flavonoids and DPPH radical scavenging activity/reducing power. This work indicates that the leaf extracts of the wild Eriobotrya species, ‘Tibet’, ‘Obovatae’, ‘Taiwan’, ‘Bengal’, ‘Oak leaf’ and ‘Hengchun’ loquats, exhibited significantly higher levels of total phenolics and flavonoids, and significantly stronger antioxidant activities, compared with the cultivated species, ‘Zaozhong 6′ loquat, which suggests that these wild species have a better utilization value.