Antioxidant activity of enzymatic extracts from a brown seaweed Ecklonia cava by electron spin resonance spectrometry and comet assay

The potential antioxidant activity of enzymatic extracts from Ecklonia cava was evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl and alkyl radical scavenging using a electron spin resonance (ESR) spectrophotometer compared with ESR signal intensity and inhibitory effect of DNA damage on comet assay. E. cava was enzymatically hydrolyzed with five food industrial carbohydrases (AMG, Celluclast, Termamyl, Ultraflo and Viscozyme) from Novozyme Nordisk. All the tested extracts exhibited strong radical scavenging activities and the values were dose-dependent. The DPPH and hydroxyl radical scavenging activities of the enzymatic extracts were compatible with the vitamin C, which the alkyl radical scavenging activity was even higher than that of vitamin C. Also in the present study, the enzymatic extracts showed strong inhibitory effect against DNA damage in a dose dependent manner. These results indicated that the E. cava might be valuable natural antioxidative source.     

Determination of physicochemical properties of sulphated fucans from sporophyll of Undaria pinnatifida using light scattering technique

The weight average molecular weight (M_w) of the intact sulphated fucans extracted with water from the sporophyll of Undaria pinnatifida without acid treatment was determined using the high performance size exclusion chromatography coupled to multiangle laser light scattering and refractive index detection (HPSEC-MALLS-RI) system. The effects of different heating conditions on the determination of M_w were investigated. The extracted intact fucoidans mostly consisted of carbohydrates (54.9%) and sulphate (41.5%) with monosaccharide composition of 78.8% fucose and 21.2% galactose. The M_w of the intact fucoidans was reduced from 23,600 to 5200 kDa when heated in boiling water for 1–15 min. Microwave heating for 30 s decreased the M_w of fucoidans to 2400 kDa despite no significant polymer degradation. The results indicate that the 30 s-microwave heating yielded a more accurate M_w value of the intact fucoidans than any other heat treatments used in this study.     

Free radical scavenging activity of porcine plasma protein hydrolysates determined by electron spin resonance spectrometer

A study was conducted to investigate the antioxidant capacity of porcine plasma protein before and after enzymatic hydrolysis. Porcine plasma protein was hydrolyzed by using Alcalase with degree of hydrolysis (DH) ranged from 0 to 17.8%. The free radical scavenging effects of porcine plasma protein hydrolysates (PPH) were evaluated by electron spin resonance (ESR) spectrometer. The reducing power of PPH increased with increasing of DH (P < 0.05). The 5-h PPH exhibited the strongest inhibition of lipid oxidation, as indicated by lowest thiobarbituric acid-reactive substance values in a liposome-oxidizing system, and the strongest free radical scavenging ability on 1,1-diphenyl-2-picrylhydrazyl radicals (DPPH), hydroxyl (OH) and superoxide (O₂-) radicals. The increase of protein concentration enhanced (P < 0.05) free radical scavenging effect of PPH. Although non-hydrolyzed plasma protein displayed an antioxidative effect, it was far less potent than PPH. The results indicated that the antioxidant capacity of porcine plasma protein could be enhanced by enzymatic hydrolysis of Alcalase.     

Fucoxanthin content and antioxidant properties of Undaria pinnatifida

This study investigated the fucoxanthin content of New Zealand (NZ) Undaria pinnatifida harvested from two locations in the Marlborough Sounds, New Zealand across its growing season. Fucoxanthin content and antioxidant properties of processed New Zealand U. pinnatifida and commercial wakame from Japan and Korea were further compared. Results showed that U. pinnatifida harvested from Port Underwood had higher fucoxanthin content in the blade compared to Pelorus Sound. The sporophyll also contained a significant amount of fucoxanthin throughout the harvest season, although lower than in the blade. Two antioxidant measurement methods, DPPH and CUPRAC, were utilised to measure antioxidant activities. Processed NZ U. pinnatifida had a lower fucoxanthin content and antioxidant activity than freeze-dried Undaria. Fucoxanthin content and antioxidant activities of NZ processed U. pinnatifida were not significantly different from other commercial samples from Japan and Korea. In conclusion, U. pinnatifida in New Zealand has a great potential to be a food and nutraceutical resource.     

Terpenoids and hexenes from the leaves of Crataegus pinnatifida

Crataegus pinnatifida have long been used in traditional Chinese medicine and European herbal medicine, and are widely consumed as food, in the form of juice, drink, jam and canned fruit. Four new compounds, a sesquiterpene and its glycoside (1–2), two monoterpene glycosides (3–4), together with eight known compounds (5–12), were isolated from the leaves of C. pinnatifida. Their structures were elucidated as (5Z)-6-[5-(2-hydroxypropan-2-yl)-2-methyltetrahydrofuran-2-yl]-3-methylhexa-1,5-dien-3-ol (1), (5Z)-6-[5-(2-O-β-d-glucopyranosyl-propan-2-yl)-2-methyl tetrahydrofuran-2-yl]-3-methylhexa-1,5-dien-3-ol (2), 5-ethenyl-2-[2-O-β-d-glucopyranosyl-(1″ → 6′)-β-d-glucopyranosyl-propan-2-yl]-5-methyltetrahydrofuran-2-ol (3), 4-[4β-O-β-d-xylopyranosyl-(1″ → 6′)-β-d-glucopyranosyl-2,6,6-trimethyl-1-cyclohexen-1-yl]-butan-2-one (4), (Z)-3-hexenyl O-β-d-glucopyranosyl-(1″ → 6′)-β-d-glucopyranoside (5), (Z)-3-hexenyl O-β-d-xylopyranosyl-(1″ → 6′)-β-d-glucopyranoside (6), (Z)-3-hexenyl O-β-d-rhamnopyranosyl-(1″ → 6′)-β-d-glucopyranoside (7), (3R,5S,6S,7E,9S)-megastiman-7-ene-3,5,6,9-tetrol (8), (3R,5S,6S,7E,9S)-megastigman-7-ene-3,5,6,9-tetrol9-O-β-d-glucopyranoside (9), (6S,7E,9R)-6,9-dihydroxy-4,7-megastigmadien-3-one 9-O-[β-d-xylopyranosyl-(1″ → 6′)-β-d-glucopyranoside] (10), Linarionoside C (11), and (3S,9R)-3,9-dihydroxy-megastigman-5-ene 3-O-primeveroside (12), using a combination of mass spectroscopy, 1D and 2D NMR spectroscopy and chemical analysis. Cytotoxicity of the new compounds was assayed against selected human glioma (U87) cell lines.     

Antioxidant activity of Rhizoma Smilacis Glabrae extracts and its key constituent-astilbin

Rhizoma Smilacis Glabrae is widely consumed by Chinese as functional food and in folk medicine for its medicinal properties. In this study, methanol and water extracts of Rhizoma Smilacis Glabrae were prepared. The water extract was further divided into polysaccharide and supernatant fractions. Constituents in different extracts were analysed by capillary electrophoresis, and levels of total phenolics were also determined using the Folin-Ciocalteu method. Astilbin, the main constituent in the herb, was isolated and purified. Different antioxidant tests were employed to evaluate the antioxidant activities of the extracts and the isolated astilbin, and the results were compared with two commonly used synthetic antioxidants-butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT). Methanol, water extract and supernatant fraction showed concentration dependent antioxidant activity while polysaccharide didn’t show any antioxidant activity. Purified astilbin showed the strongest antioxidant activity in comparison to any other extracts.     

Molecular characteristics of partially hydrolyzed fucoidans from sporophyll of Undaria Pinnatifida and their in vitro anticancer activity

The effects of molecular characteristics on the anticancer activity of fucoidans were investigated after hydrolysis by copper acetate and then fractionation with 30 and 5 kDa membranes, which produced three fucoidan fractions: F_>30 K, F_5–30 K and F_<5 K. The F_>30 K and F_5–30 K consisted of mostly carbohydrate (58.2–61.3%) and sulphate (31.7–35.5%) with small amounts of proteins (1.2–6.4%). However, the major constituents of F_<5 K were sulphate (31.8%) and ash (37.5%) with smaller amounts of carbohydrate (15.5%) and protein (1.2%). The molecular weights (M_w) of F_>30 K, F_5–30 K and F_<5 K, obtained by a light scattering technique, were 262, 5.6 and 1.6 kDa, respectively. The observed anticancer activities were 18.0–28.5% for F_>30 K, 19.2–57.5% for F_5–30 K and 26.5–36.5% for F_<5 K, respectively, in the concentration range of 0.2–0.8 mg/mL. The results suggest that the anticancer activity of fucoidans could be considerably improved by lowering their M_w and by improving the binding properties of sulphate groups possibly through changing the molecular conformation.     

Antioxidant properties of peptide fractions from silver carp (Hypophthalmichthys molitrix) processing by-product protein hydrolysates evaluated by electron spin resonance spectrometry

Silver carp processing by-product protein is usually discarded as an industrial solid waste. In this study the protein was recovered using a pH-shift method, after which seven commercial proteases were separately employed to prepare antioxidative hydrolysates. Among the hydrolysates, pepsin hydrolysates, which had the highest free radical-scavenging activity, were further separated into five peptide fractions, SCPH-I (>10 kDa), SCPH-II (5–10 kDa), SCPH-III (3–5 kDa), SCPH-IV (1–3 kDa), and SCPH-V (<1 kDa), by using ultrafiltration. The antioxidative properties of the peptide fractions were investigated, using a free radical-scavenging assay, by electron spin resonance. The results show that SCPH-V had the highest scavenging effects on DPPH (1,1-diphenyl-2-picrylhydrazyl), hydroxyl and superoxide anion radicals. SCPH-V had potent antioxidant activity in the prevention of the peroxidation of linoleic acid and alleviation of H₂O₂-induced oxidative stress in human intestinal epithelial caco-2 cells. The results indicated that the antioxidant capacity of silver carp by-product hydrolysates could be enhanced by ultrafiltration.     

Antioxidant activity of Spirulina platensis extracts by supercritical carbon dioxide extraction

Supercritical CO₂ extraction of antioxidants from Spirulina platensis was optimized using response surface methodology. About 10.26 g/kg of extracts from S. platensis could be obtained under the optimum conditions of 48 °C at 20 MPa over a 4 h period. The antioxidant activity of the extracts prepared under the optimized condition, determined by linoleic acid peroxidation inhibition method, was lower compared with BHT and Trolox, but significantly higher than α-tocopherol in 300 min and became similar to α-tocopherol after that. The components of the extracts were further analyzed, and the results showed that the extracts contained 85.1 g/kg of flavonoids, 77.8 g/kg of β-carotene, 113.2 g/kg of vitamin A and 3.4 g/kg of α-tocopherol, which may contribute greatly to their high antioxidant activity. The main fatty acids in the extracts were palmitic acid (35.32%), linolenic acid (21.66%) and linoleic acid (20.58%).     

Antioxidant activity and proline content of leaf extracts from Dorystoechas hastata

Dorystoechas hastata (D. hastata) is a monotypic plant endemic to Antalya province of Turkey. D. hastata leaves are used to make a tea locally called “çalba tea”. Diethyl ether (E), ethanol (A), and water (W) were used for the sequential preparation of extracts from dried D. hastata leaves. A hot water extract (S) was also prepared by directly boiling the powdered plant in water. The antioxidant activities of the extracts were tested by ferric thiocyanate (FTC) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging methods. E extract exhibited the greatest antioxidant activity with FTC method, whereas S extract exhibited the lowest IC₅₀ value (6.17 ± 0.53 μg/ml) for DPPH radical scavenging activity. Total phenolic contents of the extracts were estimated by Folin–Ciocalteu method and S extract was found to contain the highest amount (554.17 ± 20.83 mg GAE/g extract) of phenolics. Extract A contained highest flavonoid content and there was a inverse linear correlation (R² = 0.926) between IC₅₀ values for DPPH radical scavenging activity and flavonoid contents of all extracts. Reducing power of extracts increased in a concentration-dependent manner. S extract was found to possess higher reducing power than equivalent amount of ascorbic acid at 20 and 25 μg/ml concentrations. Linear correlation between reducing power and concentration of E, A, and W extracts (R² > 0.95) was observed. A, W, and S extracts contained relatively high levels of proline. The results presented suggest that D. hastata may provide a natural source of antioxidants and proline.     

Antioxidant properties of extract and fractions from Enteromorpha prolifera, a type of green seaweed

The ethanol extract and its solvent subfractions, partitioned by n-hexane (HX), chloroform (CF) and ethylacetate (EA), from Enteromorpha prolifera were measured for antioxidant activities, and a structural identification of the active compound was performed using spectroscopic techniques. The CF fraction showed the most potent 2,2-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl (OH) radical scavenging activities with strong reducing ability. The DPPH and hydroxyl radical scavenging capacities of the CF fraction were comparable to the capacities of the positive controls, BHA and α-tocopherol, at concentrations ranging from 0.25 to 1.0 mg/mL. However, little correlation (r² = 0.03–0.48) was observed between antioxidant activities and total phenolic contents of the extracts. Further fractionation and spectroscopic analysis of the CF fraction suggested that the strong antioxidant activity of the extracts from E. prolifera was because of a chlorophyll compound, pheophorbide a, rather than phenolic compounds.     

Antioxidant properties of three extracts from Pleurotus citrinopileatus

Pleurotus citrinopileatus Sing. (Lentinaceae) was successfully cultivated and commercially available in Taiwan. The ethanolic, cold and hot water extracts were prepared from P. citrinopileatus fruit bodies, mycelia and fermentation filtrate and their antioxidant properties studied. For three samples, ethanolic extracts exhibited higher antioxidant activities than water extracts. Reducing powers of three extracts from fruit bodies were effective and 1.03-1.10 at 5 mg/ml. With regard to EC₅₀ values of scavenging abilities on 1,1-diphenyl-2-picrylhydrazyl radicals, the effectiveness was in a descending order: ethanolic>hot water>cold water extracts. Scavenging abilities of water extracts from three samples on hydroxyl radicals were 53.4-80.1% at 20 mg/ml. Chelating abilities of cold and hot water extracts on ferrous ions were higher than those of ethanolic extracts. Contents of total phenols were in the descending order: fruit bodies (8.62-12.38 mg/g)>mycelia (5.84-7.85 mg/g)>filtrate (4.80-5.57 mg/g). Overall, three extracts from fruit bodies were more effective in antioxidant properties assayed than those from mycelia and filtrate. Ethanolic extracts were more effective in antioxidant properties assayed, except for scavenging abilities on hydroxyl radicals.     

Antioxidant activity of Mammea longifolia bud extracts

Mammea longifolia buds (nagkesar) are extensively used in India as a minor spice. The antioxidant activity of its methanol (NM) and aqueous-ethanol (NW) extracts were evaluated by several in vitro experiments, e.g., DPPH, hydroxyl, superoxide radicals and H₂O₂-scavenging assays as well as inhibition of Fe(II)-induced lipid peroxidation of rat liver mitochondria. The extracts were found to possess impressive antioxidant activity in all the tests, the activity of NW being higher than that of NM in most of the assays. The differential activities of NW and NM could be correlated with their respective total phenolic, flavonoid and proanthocyanidin contents.     

A comparison between bark extracts from Pinus pinaster and Pinus radiata: Antioxidant activity and procyanidin composition

The composition and antiradical activity of procyanidins from the bark of two kinds of pine, Pinus pinaster and Pinus radiata, were compared. Both the total bark extract and the fraction soluble in both water and ethyl acetate (OW) were evaluated, because of their promising results in previous experiments.     

Antioxidant activities of Sechium edule (Jacq.) Swartz extracts

The antioxidant activities (AA) of Sechium edule extracts were tested by three established in vitro methods, namely reducing power, β-carotene linoleate model and 1,1- diphenyl-2 picrylhydrazyl (DPPH) radical-scavenging. Leaf ethanolic extracts and leaf and seed water extracts showed strong inhibitory activity by β-carotene bleaching (AA values of 90%). Furthermore, these extracts exerted hydrogen-donating ability in the presence of DDPH stable radical (IC₅₀ 2 μg/ml). These extracts also showed strong reducing power by the potassium ferricyanide reduction method. Leaf and seed extracts may be exploited as biopreservatives in food applications as well as for health supplements or functional food, to aleviate oxidative stress.     

Antioxidant properties of hot water extracts from Agrocybe cylindracea

Agrocybe cylindracea (DC: Fr.) Mre. was available in the form of fruit bodies, mycelia and fermentation filtrate. From these three forms, hot-water extracts were prepared and their antioxidant properties were studied. Antioxidant activities of hot-water extracts from fruit bodies, mycelia and filtrate were 63.6%, 81.6% and 56.8% at 20 mg ml⁻¹, respectively. EC₅₀ values in reducing power were 2.72, 3.97 and 3.09 mg ml⁻¹ whereas those in scavenging abilities of 1,1-diphenyl-2-picrylhydrazyl radicals were 0.62, 1.66 and 0.82 mg ml⁻¹ for fruit bodies, mycelia and filtrate, respectively. At 20 mg ml⁻¹, the scavenging abilities of hydroxyl radicals were 80.1%, 57.0% and 54.3% for fruit bodies, mycelia and filtrate, respectively. With regard to EC₅₀ values in chelating abilities on ferrous ions, the hot-water extract from filtrate was better than that from mycelia. Total phenols were the major naturally occurring antioxidant components found in hot-water extracts and in the range of 23.74–30.16 mg g⁻¹. From EC₅₀ values obtained, it can be concluded that hot-water extracts from three forms of A. cylindracea were good in antioxidant properties.     

Antioxidant properties of water extracts from Monascus fermented soybeans

Solid-state bioprocessing of soybean by Monascus is a biotechnological strategy to produce Monascus-fermented soybeans (MFS) with more beneficial components. The objective of this study was to evaluate the antioxidant properties of cold and hot water extracts from MFS as compared to uninoculated soybeans. With regard to the EC₅₀ values of antioxidant activities, the effectiveness of cold water extracts was in a descending order: soybeans > MFS-31527 > MFS-31499 whereas that of the hot water extracts: soybeans ∼ MFS-31499 > MFS31527. Cold water extracts showed higher reducing power and lower scavenging ability on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals than hot water extracts. For both extracts, MFS and soybeans showed comparable effective chelating abilities on ferrous ions. Total phenols were the major naturally occurring antioxidant components found. For both extracts, soybeans were more effective in antioxidant activity and scavenging ability on DPPH radicals whereas MFS-31499 and MFS-31527 were more effective in reducing power and scavenging ability on hydroxyl radicals. Based on the results obtained, MFS-31499, MFS-31527 and soybeans were relatively effective in the antioxidant properties assayed and might be potential antioxidants for application in food products.