Antioxidant and semicarbazide‐sensitive amine oxidase inhibitory activities of alginic acid hydroxamates

The commercial polysaccharides of alginic acid (medium (3500 cps, 2% solution) and low (250 cps, 2% solution) viscosities) were esterified with acidic methanol (1 mmol L^?1 HCl) at 4 °C with gentle stirring for 5 days to obtain methyl esters of medium‐viscosity alginic acid (ME‐MVA) and low‐viscosity alginic acid (ME‐LVA). These ME‐MVA and ME‐LVA were reacted with alkaline hydroxylamine to obtain medium‐viscosity alginic acid hydroxamates (MVA‐NHOH) and LVA‐NHOH. The percentages of hydroxamic acid content in MVA‐NHOH and LVA‐NHOH were calculated as 25% and 20%, respectively. The hydroxamate derivatives of alginic acid were used to test the antioxidant and semicarbazide‐sensitive amine oxidase (SSAO) inhibitory activities in comparison with original materials (MVA and LVA). The half‐inhibition concentrations, IC₅₀, of scavenging activity against 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) were 24.5 and 29.8 µg mL^?1 for MVA‐NHOH and LVA‐NHOH, respectively. However, few scavenging activities of the MVA and LVA were found at the same concentrations. The IC₅₀ of the positive control of butylated hydroxytoluene was 5 µg mL^?1. The scavenging activity of DPPH radical was pH‐dependent, and the optimal pH for both of MVA‐NHOH and LVA‐NHOH was the Tris‐HCl buffer (pH 7.9). Using electron spin resonance (ESR) to detect the activity of scavenging hydroxyl radicals, both alginic acid hydroxamates showed dose‐dependent scavenging activities, and the IC₅₀ was 90 and 92 µg mL^?1, respectively, for MVA‐NHOH and LVA‐NHOH. Both alginic acid hydroxamates also exhibited protection against hydroxyl radical‐mediated DNA damage. Both MVA‐NHOH and LVA‐NHOH showed dose‐dependent inhibitory activities against bovine SSAO (2.53 units); the IC₅₀ was 0.16 and 0.09 µg mL^?1, respectively, for MVA‐NHOH and LVA‐NHOH, compared with 3.81 µg mL^?1 of semicarbazide (positive controls). Amine oxidase activity staining also revealed that both MVA‐NHOH and LVA‐NHOH exhibited SSAO inhibitory activities. Both MVA‐NHOH and LVA‐NHOH showed mixed non‐competitive inhibition against bovine SSAO. It was found that the V′_max value was reduced and the K′_m value was either increased (added MVA‐NHOH, 0.05 µg mL^?1) or reduced (added LVA‐NHOH, 0.11 µg mL^?1) in the presence of alginic acid hydroxamate. Copyright © 2006 Society of Chemical Industry     

Identification of polyphenols in tobacco leaf and their antioxidant and antimicrobial activities

Crude polyphenols were extracted from tobacco leaf by 80% ethanol solution with ultrasonic treatment and then purified by a macroporous resin. The polyphenols from tobacco leaf (PTL) were subjected to analyses by reverse-phase high-performance liquid chromatography (RP-HPLC) and electrospray ionization mass spectrometry (ESI-MS). The dominant polyphenols in tobacco leaf were identified as chlorogenic acid and rutin. Furthermore, the antioxidant activities of PTL were investigated, including scavenging activities of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals (5.02 μg/ml IC₅₀ value), hydroxyl radicals (49.6 μg/ml IC50 value) and superoxide anion radicals (44.0 μg/ml IC₅₀ value), inhibition activity of lipid peroxidation (132 μg/ml IC₅₀ value) and reducing power. The proliferation inhibition activities on Escherichia coli, Staphylococcus aureus and Bacillus subtilis were also measured for evaluating the antimicrobial activity of PTL. The diameters of inhibition zones were 20.23 ± 0.42, 17.66 ± 0.86 and 12.89 ± 0.29 mm, respectively. The results showed that PTL had great potential as antioxidant and antimicrobial agent.     

Antioxidant properties and chemical composition of technical Cashew Nut Shell Liquid (tCNSL)

The present study analysed the antioxidant activity of the technical Cashew Nut Shell Liquid (tCNSL) using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging assay and the xanthine oxidase assay, as well as in vivo evaluation by Saccharomycescerevisiae assay. Also, the chemical composition of tCNSL was determined by gas chromatography–mass spectrometry (GC–MS), revealing the presence of cardanols (40.26%), cardols (29.95%), phytosterol (10.68%), triacontanes (4.66%) and anacardic acid (1.79%). The DPPH-based assay results showed that tCNSL 1000 μg/ml reduced the radical level by 88.9%, and the scavenging of hydroxyl radicals in the xanthine oxidase assay indicated significant antioxidant activity (IC₅₀ = 702 μg/ml). In addition, tCNSL exerts an important protective effect against oxidative stress in yeast when used in 100–500 μg/ml concentration range, when exposed to paraquat or H₂O₂, indicating an in vivo antioxidant effect.     

Multiple forms of polygalacturonase from mango (Mangifera indica L. cv Alphonso) fruit

Polygalacturonase (PG) from mango pulp revealed three isoforms (I, II, III) upon ion exchange and gel filtration chromatography, each having an abundance of 68%, 6% and 26%, and molecular weights (M_r) 40, 51 and 45 kDa, respectively. The pH optimum for the isoforms was between 3 and 4. PG-I was stable over a wide pH range (4–7.5) unlike PG II and III, which were stable at pH 4 and 5, respectively. The optimum temperature was around 40 °C for all the three isoforms. Their apparent K_m for pectic acid was in the range 0.22–0.25 mg ml⁻¹. The V_max for PG I, II and III was 5.7, 3.6 and 4.4 μmol GalA equivalent h⁻¹, respectively. Cd²⁺, Cu²⁺ and Fe²⁺ and EDTA inhibited whereas GalA, Gal, Fuc, Rha and Ara stimulated PG-I activity, in particular. The major endogenous substrates for mango PG were identified to be two rhamnogalacturonans varying in their sugar ratio. These results are discussed in the light of pectin dissolution in vivo in ripening mango.     

In vitro antioxidant activity and in vivo anti-fatigue effect of loach (Misgurnus anguillicaudatus) peptides prepared by papain digestion

The in vitro antioxidant activity and in vivo anti-fatigue activity of loach peptide (LP) were determined. Results showed that LP contained the amino acids, which were expected to contribute to its antioxidant and anti-fatigue activities. LP could scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) (IC₅₀ 17.0 ± 0.54 mg/ml) and hydroxyl radicals (IC₅₀ 2.64 ± 0.29 mg/ml). It could chelate cupric ion and inhibit the lipid peroxidation in a linoleic acid emulsion system. It also prolonged the swimming time to exhaustion of mice by 20–28% compared to the control. It increased the levels of blood glucose (28–42% increase) and liver glycogen (2.3–3.0-fold increase). It decreased the levels of lactic acid and blood urea nitrogen by 10.9–27.5% and 8.6–17.5%, respectively. It also improved the endogenous cellular antioxidant enzymes in mice by increasing the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px). Therefore, LP can increase an endurance capacity and facilitate recovery from fatigue.     

Effects of nicotinic acid derivatives on tyrosinase inhibitory and antioxidant activities

The tyrosinase inhibitory and antioxidant activities of four nicotinic acid derivatives, namely, nicotinamide, methyl nicotinate, nicotinic acid hydroxamate (NAH), N-methyl nicotinic acid hydroxamate (NAH-M), and kojic acid, were studied. The tyrosinase inhibitory activity was found to follow the trend: NAH > kojic acid > NAH-M > methyl nicotinate > nicotinamide. The concentrations of half-inhibition (IC₅₀) of NAH against monophenolase and diphenolase activity were 2 and 1 μM, respectively; the inhibition mode was a mixed-type in the former and an uncompetitive type in the latter. In the antioxidant activity assays, NAH and NAH-M, but not nicotinamide or methyl nicotinate, showed dose-dependent DPPH radical scavenging activity, and their corresponding IC₅₀ values were 65.81 and 125 μM, respectively. NAH and NAH-M also showed hydroxyl radical scavenging activity and anti-low-density-lipoprotein peroxidation. These results indicate that NAH and NAH-M have the potential to be useful in cosmetics or food processing, and therefore, they should be investigated further.     

Antioxidant potential of meso-zeaxanthin a semi synthetic carotenoid

Semi synthetic carotenoid meso-zeaxanthin was evaluated for its antioxidant potential in vitro and in vivo. Meso-zeaxanthin was found to scavenge superoxide radicals, hydroxyl radicals and inhibited in vitro lipid peroxidation. Concentrations needed for 50% inhibition (IC₅₀) were 27.0, 3.5 and 3.2 μg/ml, respectively. It scavenged 2,2-azobis-3-ethylbenzthiozoline-6-sulphonic acid and 2,2-diphenyl-1-picryl hydrazyl radicals and IC₅₀ were 46.5, 6.25 μg/ml, respectively. It also scavenged nitric oxide radicals and IC₅₀ was found to be 2.2 μg/ml. Oral administration of meso-zeaxanthin inhibited superoxide radicals generated in macrophages by 25.2%, 50.1% and 67.2% at doses of 50, 100 and 250 mg/kg b.wt., respectively. One month oral administration of meso-zeaxanthin to mice significantly increased catalase, superoxide dismutase, glutathione and glutathione reductase levels in blood and liver. Levels of glutathione peroxidase and glutathione-S-transferase were also found to be increased in the liver, in a dose dependent manner. These results showed that meso-zeaxanthin has significant antioxidant activity in vitro and in vivo.     

Sweet potato (Ipomoea batatas [L.] Lam ‘Tainong 57’) storage root mucilage with antioxidant activities in vitro

Sweet potato storage root mucilage was extracted and purified by SDS and heating treatments. Total antioxidant activity, DPPH (1,1-diphenyl-2-picrylhydrazyl) staining, reducing power method, metal ion-dependent hydroxyl radical, FTC (ferric thiocyanate) method, and protection of calf thymus DNA against hydroxyl radical-induced damage were studied. Half-inhibition concentrations, IC₅₀, were 0.08 mg/ml and IC₅₀ > 0.1 mg/ml, respectively, for the crude and purified mucilage in the total antioxidant activity test. In the DPPH staining, the crude and purified mucilage appeared as white spots when they were diluted to 50 and 100 μg per application, respectively. Like total antioxidant activity, reducing power, scavenging capacity against hydroxyl radical, FTC activity and protection of calf thymus DNA against hydroxyl radical-induced damage were found in the mucilage. It is suggested that the mucilage might contribute its antioxidant activities against both hydroxyl and peroxyl radicals.     

Sustainable production of pectin from lime peel by high hydrostatic pressure treatment

The application of high hydrostatic pressure technology for enzymatic extraction of pectin was evaluated. Cellulase and xylanase under five different combinations (cellulase/xylanase: 50/0, 50/25, 50/50, 25/50, and 0/50 U/g lime peel) at ambient pressure, 100 and 200 MPa were used to extract pectin from dried lime peel. Extraction yield, galacturonic acid (GalA) content, average molecular weight (M_w,ave), intrinsic viscosity [η]_w, and degree of esterification (DE) were compared to those parameters obtained for pectins extracted using acid and aqueous processes. Pressure level, type and concentration of enzyme significantly (p < 0.05) influenced yield and DE of pectin. Enzyme and high pressure extraction resulted in yields which were significantly (p < 0.05) higher than those using acid and aqueous extraction. Although pressure-induced enzymatic treatment improves pectin yield, it does not have any significant effect on M_w,ave and [η]_w of pectin extracts indicating the potential of high pressure treatment for enzymatic pectin production as a novel and sustainable process.     

Chitosan and mint mixture: A new preservative for meat and meat products

Meat is prone to both microbial and oxidative spoilage and therefore it is desirable to use a preservative with both antioxidant and antimicrobial properties. Mint extract alone had good antioxidant activity but poor antimicrobial activity, while chitosan alone showed poor antioxidant activity with excellent antimicrobial properties. Therefore, the potential of chitosan and mint mixture (CM), as a preservative for meat and meat products, was investigated. Addition of chitosan to mint extract did not interfere with the antioxidant activity of mint. In the case of 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay, the IC₅₀ value for CM (17.8 μg/ml) was significantly (p ? 0.05) lower than that for mint extract (23.6 μg/ml). CM efficiently scavenged superoxide and hydroxyl radicals. The antimicrobial activities of CM and chitosan were comparable against the common food spoilage and pathogenic bacteria, the minimum inhibitory concentration being 0.05%. CM was more effective against Gram-positive bacteria. The shelf life of pork cocktail salami, as determined by total bacterial count and oxidative rancidity, was enhanced in CM-treated samples stored at 0–3 °C.     

The in vitro antioxidative properties of the essential oils and methanol extracts of Satureja spicigera (K. Koch.) Boiss. and Satureja cuneifolia ten

This study was designed to examine the in vitro antioxidant activities of the essential oil and methanol extracts of Satureja spicigera and S. cuneifolia from Turkish flora. GC and GC/MS analysis of the essential oils resulted in the identification of 40 and 29 compounds, representing the 99.4% and 99.5% of the oils, respectively. Major constituents of the oils were carvacrol (42.5% and 67.1%), γ-terpinene (21.5% and 15.2%) and p-cymene (20.9% and 6.7%), respectively. Methanol extracts were also obtained from the aerial parts of the plants. The samples were subjected to a screening for their possible antioxidant activities by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and β-carotene–linoleic acid assays. In general, samples obtained from S. cuneifolia exerted greater antioxidant activities than did those obtained from S. spicigera. In the DPPH test system, free radical-scavenging activity of S. spicigera oil was determined to be 127 ± 1.63 μg/ml, whereas IC₅₀ value of S. cuneifolia was 89.1 ± 2.29 μg/ml. In the β-carotene–linoleic acid test system, antioxidant activities of the oil were 81.7 ± 1.14% and 93.7 ± 1.83%, respectively. Antioxidant activities of the synthetic antioxidant, BHT, ascorbic acid, curcumin and α-tocopherol were also determined in parallel experiments.     

Chemical and biological variability of hot pepper fruits (Capsicum annuum var. acuminatum L.) in relation to maturity stage

The aim of the present work was to evaluate the chemical composition and the radical-scavenging and antioxidant activities of hot pepper fruits (Capsicum annuum L. var. acuminatum) at three maturity stages (small green, green and red). GC–MS analysis of n-hexane and chloroform fractions showed a different composition between the three stages of ripening. The first stage of maturation (small green) showed the highest radical-scavenging activity (IC₅₀ of 129 μg/ml). Using the bovine brain peroxidation assay, the methanolic extract of green pepper showed significant antioxidant activity (IC₅₀ of 522 μg/ml). Addition of methanolic extract of red and green pepper inhibited oxidation of linoleic acid. Methanolic extract of red pepper showed greater antioxidative potency than the others (IC₅₀ of 3 μg/ml). The different composition of lipophilic compounds and the various amount of phenolics, showed in the three stage of ripening of C. annuum var. acuminatum fruits, modifies the antioxidant activity.     

Antioxidant activity of sugarcane molasses against 2,2′-azobis(2-amidinopropane) dihydrochloride-induced peroxyl radicals

Sugarcane molasses is a rich source of antioxidant materials with peroxyl radical scavenging effects. To explore the potent antioxidant activity of sugarcane molasses against 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced peroxyl radicals, 7 methanolic fractions of sugarcane molasses (F1–F7) were separated via bioassay-guided fractionation and evaluated by oxygen radical absorbance capacity (ORAC), cellular antioxidant activity (CAA), and oxidative DNA damage protective activity assays. The ORAC values of sugarcane molasses fractions ranged from 4399 to 6266 μmol TE/g, whilst the EC₅₀ values for CAA ranged from 3.7 to 5.9 μg/ml. Moreover, it was found that sugarcane molasses fractions, particularly F6 and F7, could protect against oxidative DNA damage caused by peroxyl radicals at an effective concentration of 100 μg/ml. Ten phenolic constituents were identified in the fractions, including known antioxidative compounds, viz., schaftoside, isoschaftoside, ferulic acid, p-coumaric acid, and p-hydroxybenzaldehyde.     

Iron-chelating ability and antioxidant properties of phycocyanin isolated from a protean extract of Spirulinaplatensis

The invitro scavenger activities of different reactive oxygen species (superoxide radical, hydroxyl radical, hydrogen peroxide, hypochlorous acid and peroxyl radical), the effects on lipid peroxidation and the iron-chelating ability of a Spirulinaplatensis protean extract and the biliprotein, phycocyanin, isolated from this microalga were studied. S. platensis protean extract inhibited the generation of hydroxyl radical (IC₅₀ = 537 μg/ml for the system with EDTA and 1500 μg/ml without EDTA), the production of peroxyl radical (IC₅₀ = 230 μg/ml), and the lipid peroxidation process (IC₅₀ = 2320 μg/ml for the enzymatic system and 2180 μg/ml for the non-enzymatic system). Besides, phycocyanin inhibited hydroxyl and peroxyl radicals and the lipid peroxidation process. The iron ions decreased the maximum fluorescence emission spectra of S. platensis protean extract and phycocyanin and it was an indicator of the metal-chelating activity. The antioxidant properties of S. platensis and phycocyanin may arise from both radical-scavenging and metal chelation. Our results suggest that S. platensis could be used as a dietary supplement to prevent some diseases where free radicals are involved.     

Chemical composition and antioxidant activity of the essential oil of Clinopodium vulgare L.

This study was designed to examine the chemical composition and in vitro antioxidant activity of the essential oil of Clinopodium vulgare. GC–MS analysis of the oil resulted in the identification of 40 compounds, representing 99.4% of the oil; thymol (38.9%), γ-terpinene (29.6%) and p-cymene (9.1%) were the main components. The samples were subjected to a screening for their possible antioxidant activity by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and β-carotene-linoleic acid assays. In the first case, IC₅₀ value of the C. vulgare essential oil was determined as 63.0 ± 2.71 μg/ml. IC₅₀ value of thymol and γ-terpinene, the major compounds of the oil, was determined as 161 ± 1.3 μg/ml and 122 ± 2.5 μg/ml, respectively, whereas p-cymene did not show antioxidant activity. In β-carotene-linoleic acid system, C. vulgare essential oil exhibited 52.3 ± 1.19% inhibition against linoleic acid oxidation. In both systems, antioxidant capacities of BHT, curcumine and ascorbic acid were also determined in parallel experiments.     

Rosmarinic acid,scutellarein 4′-methyl ether 7-O-glucuronide and (16S)-coleon E are the main compounds responsible for the antiacetylcholinesterase and antioxidant activity in herbal tea of Plectranthus barbatus (“falso boldo”)

Plectranthus barbatus, known as “falso boldo” in Brazil, is used in herbal tea or cooked as a vegetable. Infusions and decoctions of leaves from P. barbatus were analysed for their inhibition of acetylcholinesterase and their antioxidant activity. The decoction showed high inhibition activity (31% inhibition with 0.5 mg of extract/ml) and also high antioxidant activity (IC₅₀ = 45.8 ± 0.5 μg of dry extract/ml in the DPPH test; IC₅₀ = 69.8 ± 3.1 μg of dry extract/ml in the β-carotene–linoleic acid test). Rosmarinic acid, scutellarein 4′-methyl ether 7-O-glucuronide and (16S)-coleon E were the main constituents identified. These compounds have antiacetylcholinesterase activity. Rosmarinic acid and the scutellarein derivative have IC₅₀ = 440 μg/ml and 1 mg/ml, respectively. One milligram per millilitre of (16S)-coleon E showed 61% inhibition of the enzyme. Other Plectranthus species, P. ecklonii, P. fructicosus, P. lanuginosus and P. verticillatus, were also analysed and the results obtained correlated with the content in rosmarinic acid.     

Chemical composition,antioxidative activity and cell viability effects of a Siberian pine (Pinus sibirica Du Tour) extract

Siberian pine (Pinus sibirica Du Tour) seeds, commonly known as cedar nuts, are ascribed a number of medicinal properties. In this study, we report the qualitative–quantitative composition, antioxidant activity and cell viability-related properties of a defatted aqueous-acetone-soluble P. sibirica seed extract. The total phenolic and total tannin contents were estimated at 266 ± 3.9 mg gallic acid/g and 115 ± 7.8 mg tannic acid/g, respectively. Reverse-phase chromatographic analysis of the crude extract indicated the presence of a chromatographic hump indicative of the presence of proanthocyanidins. After acid hydrolysis, the presence of hydroxylated benzoic and cinnamic acids, flavanones and flavan-3-ols was confirmed. After thiolysis, (+)-catechin was identified as more abundant than (−)-epicatechin, suggesting that this molecule was the main terminal unit of the proanthocyanidins within this extract. The extract demonstrated iron(III)-reductive (AscAE = 650 ± 5.10 μmol ascorbic acid/g) and iron(II) chelating (EC₅₀ = 20.1 ± 2.1) activities and the ability to scavenge 1,1-diphenyl-2-picrylhydrazyl (IC₅₀ = 257 ± 2.36 μg/ml) and hydroxyl (IC₅₀ = 338 ± 6.49 μg/ml) free radicals. When the effects of P. sibirica extract were assessed in a tumourigenic SH-SY5Y neuroblastoma cell line, it was found that the cell viability was diminished in the presence of P. sibirica extract (0.2–1.0 mg/ml), as indicated by decreased membrane integrity (LDH assay) and mitochondrial metabolic activity (MTT assay), but the level of p53 protein was not changed (Western blot).     

Antibacterial,anti-inflammatory and anti-allergic activities of standardised pomegranate rind extract

Antibacterial, anti-inflammatory and anti-allergic activities of standardised pomegranate rind extract (SPRE) containing 13% w/w ellagic acid were studied in vitro. The antibacterial activity of SPRE was determined using the disc diffusion and broth microdilution methods. SPRE exhibited a potent bacteriostatic effect against Propionibacterium acnes, a Gram-positive anaerobe, with a MIC of 15.6 μg/ml, and Gram-positive facultative anaerobic bacteria, Staphylococcus aureus and Staphylococcus epidermidis, with MICs of 7.8–15.6 μg/ml. Anti-inflammatory activity of SPRE was evaluated by measuring the inhibition of nitric oxide (NO) production by murine macrophage-like RAW264.7 cells. SPRE exhibited a potent NO inhibitory effect, with an IC₅₀ of 10.7 μg/ml. Evaluation of the anti-allergic activity showed that SPRE inhibited the release of β-hexosaminidase from antigen-stimulated rat basophilic leukemia (RBL-2H3) cells with an IC₅₀ of 20.9 μg/ml. In addition, SPRE exhibited only moderate cytotoxicity on human keratinocyte cells, with CC₅₀ of 33.6 μg/ml. These findings support the potential use of SPRE as a nutraceutical for antibacterial, anti-inflammatory and anti-allergic proposes.