Effects of Pressurized Low Polarity Water Extraction Parameters on Antioxidant Properties and Composition of Cow Cockle Seed Extracts

Antioxidant activity of pressurized low polarity water (PLPW) extracts of cow cockle seed and extraction residues were determined using DPPH, ABTS, and FRAP assays. The effect of extraction conditions (temperature (125, 150 and 175 °C) and time) on the antioxidant activity and the relationship amongst the antioxidant activity and extract composition (total phenolics and saponin content) were determined. The antioxidant activity of PLPW extracts increased with extraction temperature. Increasing activity with time was also observed at 175 °C. PLPW extraction residues had the highest activity suggesting antioxidant compounds were not completely extracted by PLPW. Antioxidant activity correlated well with total phenolics content of samples (R ² ≥ 0.94), however no correlation was observed with the saponin content. A strong correlation was observed between the antioxidant activity values obtained using different methods (R ² ≥ 0.94). These results point to the potential of PLPW extraction as a method to modify the activity of biological materials for the production of customized extracts.     

Studies on Indian Green Leafy Vegetables for Their Antioxidant Activity

To identify the potential of green leafy vegetables (GLV) as antioxidants, methanolic extracts of Amaranthus sp., Centella asiatica, Murraya koenigii and Trigonella foenum graecum were studied for their antioxidant activity in different systems at multiple concentrations. Total antioxidant activity assessed by phosphomolybdenum method, free radical scavenging activity by 1,1-diphenly-2-picryl hydrazyl (DPPH), reducing power and ferrous ion chelating activity were determined. The GLV were analyzed for ascorbic acid, total and β-carotene and total polyphenol contents. The ascorbic acid, total carotene, β-carotene and total phenolic content (tannic acid equivalents) of the GLV ranged between 15.18–101.36, 34.78–64.51, 4.23–8.84 and 150.0–387.50 mg/100 g GLV, respectively. The extracts were found to have significantly different levels of antioxidant activities in the systems tested. The total antioxidant activity was highest in Murraya koenigii (2,691.78 μmol of ascorbic acid/g sample) and least in Centella asiatica (623.78 μmol of ascorbic acid/g sample). The extract concentration causing 50% inhibition of DPPH (IC₅₀) was determined (M. koenigii < C.asiatica < Amaranthus sp. < T. graecum). The maximum DPPH scavenging activity and reducing power was exhibited by Murraya koenigii. Multiple regression analysis showed that the relationship of total antioxidant activity, free radical scavenging activity, and reducing power with polyphenol and total and β-carotene was highly significant. Paper awarded the Young Scientist Award in Experimental Nutrition at the Young Scientist Award Session of the 39th Annual Meet of the Nutrition Society of India, Hyderabad, India on 15–16th Dec 2007.     

Hypocholesterolemic Activity from the Leaf Extracts of Cnidoscolus chayamansa

The aim of this study was to determine the hypocholesterolemic activity of Cnidoscolus chayamansa. In an in vivo model, high-cholesterol diet administered to mice Balb/c induced hypercholesterolemia. Three extracts from Cnidoscolus chayamansa (ethanol, methanol and an aqueous extract) were tested on hypercholesterolemic mice. Active extracts were assessed against the in vitro inhibitory activity of the same three extracts on the HMG-CoA reductase enzyme by using Vero cells. The specific chemical groups present in the phytochemical extracts were also determined. Only the aqueous extract (at either doses employed) showed a significant cholesterol reduction (27.9 and 31.1%, for 50 and 100 mg kg⁻¹, respectively P < 0.01). The extract did not inhibit the HMG-CoA reductase enzyme, suggesting that its compounds act at another level in cholesterol metabolism. Reactions to secondary metabolites indicate the presence of alkaloids in the aqueous and ethanol extracts and phenol hydroxyls in the ethanol and methanol extracts.     

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.     

Effect of the Temperature on the Spray Drying of Roselle Extracts (<Emphasis Type="Italic">Hibiscus sabdariffa</Emphasis> L.)

The effect of the drying temperature on the volatile components and sensory acceptance of the Roselle (Hibiscus sabdariffa) extract in powder was investigated. The Roselle extraction was carried out by maceration with 7 L of 30% ethanol (v/v), 560 g of fresh Roselle calyces for 168 h. The Roselle extracts were spray dried at different temperatures 150, 160, 170, 180, 190, 200 and 210 °C, giving different outlet values about yield and final moisture. The volatile compounds in Roselle extract and dried samples were performed using needles of solid phase microextraction (SPME) and gas chromatography/mass spectrometry (GC/MS HP-5890). Twenty volatile compounds were identified in the extracts among them terpenoids, esters, hydrocarbons and aldehydes. Fourteen volatile compounds were identified in the powder sample, but only ten were present in the Roselle extract. This indicates that some compounds were lost and some others were generated due to a degradation process. An acceptability sensory analysis showed that the best powder sample was the Roselle extract dehydrated using temperature between 190 °C and 200 °C (p < 0.05). There was not statistically significant difference in the pH of Roselle extracts ranging from 3.4 to 3.9. It was concluded that the spray drying temperature of the Roselle extracts has an effect on the volatile compounds losses.     

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.     

Determination of the Antimicrobial and Antioxidative Properties and Total Phenolics of Two “Endemic” Lamiaceae Species from Turkey: <Emphasis Type="Italic">Ballota rotundifolia</Emphasis> L. and <Emphasis Type="Italic">Teucrium chamaedrys</Emphasis> C. Koch

This study was designed to examine the in vitro antimicrobial and antioxidant activities and the amount of total phenolics of the methanol extracts of Ballota rotundifolia L. and Teucrium chamaedrys C. Koch. In the case of antimicrobial activity tests, polar sub-fractions of the methanol extracts of plant species exhibited weakest antimicrobial activity when compared with the non-polar ones. While, non-polar sub-fraction of B. rotundifolia showed moderate activity against A. lwoffii, C. perfringens and the yeasts, T. chamaedrys performed excellent activity pattern against all of the tested microorganisms. The sub-fractions were also screened for their possible antioxidant activities by two complementary tests, namely DPPH free radical-scavenging and β-carotene/linoleic acid assays. Non-polar extracts of the plant species remained inactive in both test systems. On the other hand polar extracts showed remarkable antioxidant activities. In DPPH system, free radical scavenging effect of T. chamaedrys was measured as 18.00 ± 1.42 μg.mg⁻¹. It is extremely important to point out that, polar sub-fraction of T. chamaedrys is found as effective as the positive control BHT. Non-polar sub-fraction of T. chamaedrys found to have the highest total phenolic amount (97.12 ± 1.28 μg/mg). Results obtained from this experiment confirm the relationship between the amount of phenolics and biological activities.     

Antioxidant Activities of Four Edible Seaweeds from the Southern Coast of Thailand

The aqueous extracts of four marine algae, Caulerpa racemosa var. macrophysa, Gracilaria tenuistipitata var. tenuistipitata, Sargassum sp., and Ulva lactuca, from the coastal areas in Southern Thailand, were prepared by boiling dried seaweed powder in water for 3 h, and by autoclaving each sample at 120 °C for 3 h. They were then freeze-dried and evaluated for their antioxidant activities using DPPH (1, 1-diphenyl-2-picrylhydrazyl), hydroxyl radical (OH^•) and superoxide anion (O₂^•−) scavenging assays. Boiling extracts of the seaweeds, except C. racemosa, were found to have higher total phenolic contents (TPC) than those obtained from the autoclave method. The antioxidant results also showed that O₂^•− scavenging activity existed only in the boiling extracts of C. racemosa, G. tenuistipitata, and U. lactuca. In DPPH and OH^• assays, however, almost all the boiling extracts were less active than the autoclave ones. Among the four alga species, Sargassum sp. was the most active. Both extracts of this seaweed had the highest TPC and also displayed the strongest DPPH^• and OH^• inhibitory activities. A strong positive-correlation between the antioxidant potency and TPC of the autoclave extracts was found, while for the boiling extracts such relation was very weak. This result thus reflected that in addition to the phenolic compounds, there might be some other active components present in these extracts involved in the antioxidant activity.     

A Search for Hepatoprotective Activity of Fruit Extract of Mangifera indica L. Against Oxidative Stress Cytotoxicity

Mango (Mangifera indica L.) and their components are commonly used in folk medicine for many curative effects. The protective effects of different concentrations of aqueous extract of Mangifera indica L. fruit (Mango Extract) (20, 50 and 100 μg/ml) and also gallic acid (100 μM) as a pure compound in the extract were examined against oxidative stress toxicity induced by cumene hydroperoxide (CHP) in isolated rat hepatocytes. The extracts and gallic acid (100 μM) protected the hepatocyte against all oxidative stress markers including cell lysis, ROS generation, lipid peroxidation, glutathione depletion, mitochondrial membrane potential decrease, lysosomal membrane oxidative damage and cellular proteolysis. Mango Extracts (20, 50 and 100 μg/ml) were more effective than gallic acid (100 μM) in protecting hepatocytes against CHP induced lipid peroxidation. On the other hand gallic acid (100 μM) acted more effective than Mango Extracts (20, 50 and 100 μg/ml) at preventing lysosomal membrane damage. In addition H₂O₂ scavenging effect of all extracts were determined in hepatocytes and compared with gallic acid (100 μM). There were no significance differences (P<0.05) between all plant extracts and gallic acid (100 μM) in H₂O₂ scavenging activity. These results suggest a hepatoprotective role for Mango Extract against liver injury associated with oxidative stress.     

Dual Functionality of Triticale as a Novel Dietary Source of Prebiotics with Antioxidant Activity in Fermented Dairy Products

The objectives of this study were to: i) define the optimum concentration of triticale bran (TB) that can be incorporated in yogurt, ii) evaluate the prebiotic effects of TB on microbial viability, pH and total titratable acidity (TTA) in yogurt across 28 days of cold storage, and iii) measure the oxygen radical absorbance capacity (ORAC) of water-extractable polysaccharides (WEP) in TB. Lactobacillus bulgaricus and Streptococcus thermophilus were used as starter cultures. Lactobacillus acidophilus and Bifidobacterium lactis were used as probiotics. A concentration of 4% TB in yogurt was determined to be the maximum amount that could be added without causing synersis. By day 7, the number of bacteria greatly increased in yogurt samples containing TB and maintained higher viable bacteria counts at the end of the cold storage period, in comparison to controls (P ≤ 0.05). Confirming this data was the lower pH levels and higher TTA values of TB yogurt samples exhibited throughout 28 days (P ≤ 0.05). Polysaccharide extracts of TB exhibited strong antioxidant activity with an ORAC value of 33.86 ± 2.30 μmol trolox equivalents (TE)/g of bran. Results of this study suggest that TB may serve as a new prebiotic and antioxidant source for functional foods and nutraceutical applications.     

Sugars Profiles of Different Chestnut (Castanea sativa Mill.) and Almond (Prunus dulcis) Cultivars by HPLC-RI

Sugar profiles of different almond and chestnut cultivars were obtained by high-performance liquid chromatography (HPLC), by means of a refractive index (RI) detector. A solid-liquid extraction procedure was used in defatted and dried samples. The chromatographic separation was achieved using a Eurospher 100-5 NH₂ column using an isocratic elution with acetonitrile/water (70:30, v/v) at a flow rate of 1.0 ml/min. All the compounds were separated in 16 min. The method was optimized and proved to be reproducible and accurate. Generally, more than 95% of sugars were identified for both matrixes. Sugars profiles were quite homogeneous for almond cultivars; sucrose was the main sugar (11.46 ± 0.14 in Marcona to 22.23 ± 0.59 in Ferragnes g/100 g of dried weight), followed by raffinose (0.71 ± 0.05 in Ferraduel to 2.11 ± 0.29 in Duro Italiano), glucose (0.42 ± 0.12 in Pegarinhos two seeded to 1.47 ± 0.19 in Ferragnes) and fructose (0.11 ± 0.02 in Pegarinhos two seeded to 0.59 ± 0.05 in Gloriette). Commercial cultivars proved to have higher sucrose contents, except in the case of Marcona. Nevertheless, chestnut cultivars revealed a high heterogeneity. Sucrose was the main sugar in Aveleira (22.05 ± 1.48), Judia (23.30 ± 0.83) and Longal (9.56 ± 0.91), while glucose was slightly prevalent in Boa Ventura (6.63 ± 0.49). The observed variance could serve for inter-cultivar discrimination.     

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 Activity and Phenolic Content of Betalain Extracts from Intact Plants and Hairy Root Cultures of the Red Beetroot Beta vulgaris cv. Detroit Dark Red

Betalains are water-soluble plant pigments that are widely used as food colorants, and have a wide range of desirable biological activities, including antioxidant, anti-inflammatory, hepatoprotective, anti-cancer properties. They can be produced from various plants, notably beetroot, but betalain products obtained in this way also have some undesirable properties and are difficult to standardize. A potentially attractive alternative is to use hairy root cultures. In the study reported here, we found that betalain extracts obtained from hairy root cultures of the red beetroot B. vulgaris cv. Detroit Dark Red also had higher antioxidant activity than extracts obtained from mature beetroots: six-fold higher 2,2-dyphenyl-1-picrylhydrazyl radical scavenging ability (90.7% inhibition, EC₅₀ = 0.11 mg, vs 14.2% inhibition, EC₅₀ = 0.70 mg) and 3.28-fold higher oxygen radical absorbance capacity (4,100 μM TE/g dry extract, vs 1,250 μM TE/g dry extract). The high antioxidant activity of the hairy root extracts was associated with increased concentrations (more than 20-fold) of total phenolic concomitant compounds, which may have synergistic effects with betalains. The presence of 4-hydroxybenzoic acid, caffeic acid, catechin hydrate, and epicatechin were detected in both types of extract, but at different concentrations. Rutin was only present at high concentration (1.096 mg.g⁻¹ dry extract) in betalain extracts from the hairy root cultures, whereas chlorogenic acid was only detected at measurable concentrations in extracts from intact plants.     

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.     

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.     

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.     

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.     

Seasonal Variation of Phenolic Antioxidant-mediated α-glucosidase Inhibition of <Emphasis Type="Italic">Ascophyllum nodosum</Emphasis>

Ascophyllum nodosum is a brown seaweed that grows abundantly in the US Northeast coastal region. This study examined the seasonal variation of A. nodosum in phenolic contents and subsequent antioxidant, α-glucosidase and α-amylase inhibitory activities. A. nodosum was harvested monthly and extracted in hot water and the resulting extracts were spray-dried. The results indicate a clear seasonal variation in terms of phenolic content, with June and July being the highest (36.4 and 37 mg/g, respectively) and May the lowest (21.8 mg/g). The antioxidant activities, in terms of 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, correlated with the phenolic contents observed (r = 0.81), with the month of July being the highest (58%) and April the lowest (26%). Similarly in terms of Trolox equivalent, July had the highest activity (15.53 μM) and April and May the lowest (8.40 and 8.27 μM, respectively). α-glucosidase inhibitory activity exhibited a pattern similar to the phenolic contents observed with July having the highest inhibitory activity (IC₇₀ 2.23 μg) and April the lowest (IC₇₀ 26.13 μg), resulting in an inverse correlation between IC₇₀ values and total phenolic content (r = −0.89). Such seasonal variation is believed to be caused by temperature-related stress considering that A. nodosum is a cold water species.