Antioxidant activity of Camellia sinensis leaves and tea from a lowland plantation in Malaysia

Methanol extracts of fresh tea leaves from a lowland plantation in Malaysia were screened for total phenolic content (TPC) and antioxidant activity (AOA). AOA evaluation included 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical-scavenging ability, ferric-reducing antioxidant power (FRAP), and ferrous-ion chelating (FIC) ability. Ranking, based on TPC and AOA, was as follows: shoots > young leaves > mature leaves. TPC and AOA of lowland leaves were comparable to those of highland plants. A green tea produced by drying young leaves in a household microwave oven for 4 min showed significantly higher TPC and AOA than did four commercial brands of green and black tea.     

Antioxidant activity of various extracts of old tea leaves and black tea wastes (Camellia sinensis L.)

The antioxidant activity of various extracts of old tea leaves (OTL) and black tea wastes (BTW) in comparison with that of green tea leaves (GTL) was evaluated. The highest extraction yield and antioxidant activity were found in hot water extracts of GTL. Hot water extracts of OTL and BTW showed the same statistically significant antioxidant activities (P < 0.05). Also, there was no statistically significant difference between the antioxidant activities of OTL and BTW extracts obtained by the other two extraction methods (ethyl acetate and methanolic methods) and, in some cases, BTW extracts acted even better than OTL extracts. The antioxidant activity of tea extracts was not concomitant with the development of their reducing power. This suggested that the antioxidant activity of tea extracts likely involves other mechanisms in addition to those of reductones. Totally, BTW extracts had antioxidant activities comparable with or even better than those of OTL extracts. Hence, OTL and BTW, which are often considered as agricultural wastes, can be used as potent natural antioxidative sources.     

Antioxidant activity of some phenolic constituents from green pepper (Piper nigrum L.) and fresh nutmeg mace (Myristica fragrans)

Antioxidant potential of phenolic compounds from green pepper (Piper nigrum L.) and lignans from fresh mace (Myristica fragrans) were evaluated for their ability to scavenge 1,1′-diphenyl-2-picrylhydrazyl (DPPH) radical, inhibit lipid peroxidation and protect plasmid DNA damage upon exposure to gamma radiation. EC₅₀ values of the major phenolic compounds of green pepper namely, 3,4-dihydroxyphenyl ethanol glucoside, 3,4-dihydroxy-6-(N-ethylamino) benzamide and phenolic acid glycosides were found to be 0.076, 0.27 and 0.12 mg/ml, respectively, suggesting a high radical scavenging activity of these phenolics. These results were further confirmed with cyclic voltammetry. Acetone extract of nutmeg mace and its subsequent TLC isolated fractions constituted mainly of lignans as revealed by GC–MS analysis. The major compounds were tentatively identified from their mass spectral fragmentation pattern. DPPH radical scavenging capacity of the acetone extract as well as its fractions was comparatively lower than that of green pepper phenolics. In contrast, these fractions had a greater ability to inhibit lipid oxidation than phenolics from pepper as revealed by β-carotene–linoleic acid assay. A DNA protecting role of these compounds even at doses as high as 5 kGy further suggested the potential use of green pepper and fresh nutmeg mace and their extracts as a nutraceutical in preventing oxidative damage to cells.     

Effects of iron,ascorbate, meat and casein on the antioxidant capacity of green tea under conditions of in vitro digestion

The hypothesis that interactions of dietary polyphenols with dietary iron occur during digestion and result in a decrease of the post-absorptive antioxidant properties of polyphenols was investigated. The hypothesis was tested in vitro, under conditions that simulate gastrointestinal digestion. Mixtures of green tea, iron, and three dietary factors that modify the form of iron in the lumen, namely ascorbic acid, meat or casein, were subjected to an in vitro gastrointestinal digestion. Antioxidant capacity (FRAP assay), iron concentration (ferrozine assay) and polyphenol concentration (Folin–Ciocalteau assay) were measured in the in vitro digests. The presence of iron decreased the antioxidant capacity and the polyphenol concentration of green tea digests. The presence of ascorbic acid increased, while meat and casein decreased the antioxidant capacity of green tea. The factorial analysis of the data suggests that protein and iron interact with green tea polyphenols during the in vitro digestion and decrease their antioxidant capacity. These results support the aforementioned hypothesis.     

Partial characterization of white cabbages (Brassica oleracea var. capitata f. alba) from different regions by glucosinolates, bioactive compounds, total antioxidant activities and proteins

Glucosinolates (GLS), antioxidative compounds, total radical scavenging activities (TRSAs) and proteins of white cabbage samples derived from different regions of Europe, collected in the spring and autumn, were studied. Glucobrassicin and sinigrin were the dominating GLS in all analyzed cabbage samples. Depending on origin, these two GLS accounted for ∼30% to ∼70% of the total. The total GLS content ranged from 3.3 to 7.7 μmol/g dw in lyophilized vegetables. Assays based on electron transfer [total phenols by Folin-Ciocalteu reagent (FCR), 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)] radical cation (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH)] were used to compare the TRSAs and the main bioactive compounds in cabbage. Total polyphenols varied from 2.4 to 4.9 GAE/g dw. The TRSAs ranged from 2.7 to 8.2 μmol TE/g dw in ABTS test and from 2.4 to 5.4 μmol TE/g dw in DPPH assay. The maximum amount of polyphenol compounds, antioxidant activity, as well as total GLS content, were recorded in Belgian cabbage harvested in the autumn and the lowest ones were found for Poland 2 cabbage harvested in the spring. In extracted and separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) proteins from cabbage leaves only in samples from England and Belgium some differences in patterns were found in the regions of 60 and 97 kDa. The calculated correlations between antioxidative potency and the abundance of bioactive compounds were highly statistically significant. This suggests that TRSA could serve as a means of standardization of natural mixtures, at least in the case of cabbage, necessary to compare results of biological studies carried out for vegetable derived samples.     

Antioxidant and hepatoprotective activity of ethanolic extract of leaves of Solidago microglossa containing polyphenolic compounds

The antioxidative and hepatoprotective potential of Solidago microglossa D.C, a widely used medicinal plant from Brazil was investigated. The leaf extract showed inhibition against thiobarbituric acid reactive species (TBARS) induced by different prooxidants (10 μM FeSO₄ and 5 μM sodium nitroprusside SNP) in rat liver, brain and phospholipid homogenates from egg yolk. Moreover, the free radical scavenging activities of the extract was evaluated by the scavenging of 2,2-diphenyl-1-picrylhydrazyl (DPPH) (IC_50, 3.8 ± 0.5 μg/ml) and hydroxyl radical on benzoic acid hydroxylation (IC_50, 32.3 ± 1.3 μg/ml) and deoxyribose (IC_50, 39.1 ± 2.4 μg/ml) assays. The ethanolic extract showed significant hepatoprotective activity against paracetamol (250 mg/kg) induced liver damage in mice in a dose dependent manner. The phenolic composition and their quantification by high performance liquid chromatography (HPLC) resulted in the identification of gallic acid and flavonoids: quercetrin (quercetin-3-O-rhamnoside), rutin (quercetin-3-O-rutinoside) and quercetin.     

The thermostability, bioactive compounds and antioxidant activity of some vegetables subjected to different durations of boiling: Investigation in vitro

This research was performed in order to compare the water and acetone extracts of raw and boiled for 10, 20, 40 and 60 min Korean lotus roots (KLR) and Polish white onion (PWO) in the contents of their bioactive compounds, antioxidant activity and thermostability.It was found that polyphenols (mg GAE/g), flavanols (μg GAE/g), flavonoids (mg CE/g), anthocyanins (mg CGE/kg) and tannins (mg CE/g) in water extract of raw lotus roots were 14.18 ± 0.7, 8.41 ± 0.5, 1.09 ± 0.06, 21.3 ± 1.2 and 7.29 ± 0.4, and of white onion - 11.11 ± 0.6, 6.78 ± 0.3, 0.71 ± 0.03, 17.00 ± 0.9 and 1.64 ± 0.08, respectively, and significantly higher in KLR (P < 0.05). The antioxidant activity of raw KLR water extract (139.4 ± 6.1, 53.1 ± 3.6 and 89.3 ± 4.6 μmol TE/g for DPPH, CUPRAC and ABTS, respectively) was significantly higher than in white onion (23.84 ± 1.8, 31.9 ± 2.1 and 38.14 ± 2.6 for DPPH, CUPRAC and ABTS, respectively, P < 0.05).The thermostability of the water KLR extract’s of polyphenols, flavanols, flavonoids, anthocyanins and tannins was high and even after 60 min of boiling remains as 40.0, 42.3, 50.5, 41.4 and 41.0%, respectively. After 60 min of boiling the most thermostable compounds were flavonoids - remaining at 50.5% in water extract of KLR. Also after 60 min of boiling the thermostability of the antioxidant activity of water extracts of KLR remained significantly high: 40.6, 42.3, 46.3 and 43.6%, according to DPPH, FRAP, ABTS and CUPRAC assays, respectively.Similar relationship was obtained with acetone extracts, but the value was lower than with the water ones. In conclusion, the contents of some bioactive compounds, the antioxidant activity and the thermostability in water and acetone extracts of KLR are significantly higher than the same indices in PWO. FTIR and fluorimetry can be used as additional markers for the characterization of bioactive compounds in vegetables.     

Isolation and identification of compounds from the ethanolic extract of flowers of the tea (Camellia sinensis) plant and their contribution to the antioxidant capacity

While beneficial health properties of tea leaves have been extensively studied, less attention has been given to that of flowers of the tea (Camellia sinensis) plant. In this work, the ethanolic extract and its ethyl acetate-soluble fraction (EEA) from the tea flowers were found to possess the potent antioxidant activity using 2, 2-diphenyl-1-picrylhydrazyl (DPPH) free radical-scavenging assay. The compounds present in EEA had comparatively strong DPPH scavenging activity and strongly contributed to the antioxidant activity of the tea flowers. From EEA, besides eight catechins, five flavonol glycosides were isolated and their structures were elucidated on the basis of mass spectrometry and nuclear magnetic resonance spectroscopy as myricetin 3-O-β-d-galactopyranoside, quercetin 3-O-β-d-galactopyranoside, kaempferol 3-O-β-d-galactopyranoside, kaempferol 3-O-β-d-glucopyranoside, and kaempferol 3-O-[α-l-rhamnopyranosyl-(1-6)-β-d-glucopyranoside]. In addition, epigallocatechin gallate and epicatechin gallate were found as the major active components responsible for the antioxidant activity of tea flowers through the use of a combination of preparative liquid chromatography separation and DPPH assay.     

Bolus ingestion of white and green tea increases the concentration of several flavan‐3‐ols in plasma,but does not affect markers of oxidative stress in healthy non‐smokers

White tea (WT) is rich in flavan‐3‐ols as green tea (GT) and might provide health protective effects due to the strong antioxidant properties of flavan‐3‐ols. Since intervention studies with WT are lacking, we evaluated the effects of WT consumption on antioxidant status, antioxidant capacity and biomarkers of oxidative stress compared to water and GT. After an overnight fast, 70 healthy non‐smokers were randomized to consume 600 mL of WT, GT or water (control). Plasma (epi‐)catechin and epi(gallo)catechingallate, antioxidant capacity (Folin assay, trolox equivalent antioxidant capacity test), 8‐iso‐prostaglandin F_2α, ascorbic acid and uric acid were determined before and several times within 8 h after consumption. DNA strand breaks were measured in vivo and ex vivo (H₂O₂ stimulation) in leukocytes. Plasma flavan‐3‐ols significantly increased after WT and GT ingestion. Trolox equivalent antioxidant capacity was lower after 5 h in controls versus WT (p=0.031) and GT (p=0.005). Folin‐Ciocalteu reducing capacity, ascorbic and uric acid as well as markers of oxidative stress (8‐iso‐prostaglandin‐F_2α, DNA strand breaks) were not affected by the beverages. A short‐term increase of catechins does not change plasma antioxidant capacity in healthy subjects. Conclusions with respect to health protective effects of WT and GT on the basis of these biomarkers can, thus, not be drawn.     

Antioxidant activity of polyphenolic compounds extracted from defatted raw and dry heated Tamarindus indica seed coat

The antioxidative properties and total phenolic contents of raw and dry heated seed coat of Tamarindus indica were examined. The raw and dry heated samples were extracted with methanol followed by 70% acetone, after treating with petroleum ether, solvents were removed using rotary vacuum evaporator and the extracts which contain residual moisture were freeze-dried, respectively. Methanol extracts for both raw and dry heated seed coat samples contained higher level of total phenolics and tannins than the aqueous acetone extracts. The extracts were screened for their potential antioxidant activities using such as O₂⁻, OH, α,α-diphenyl-β-picrylhydrazyl (DPPH), ABTS⁺, FRAP and linoleic acid emulsion model systems. At different concentration of respective solvent extracts, the maximum level of superoxide anion radical scavenging activity (79-85%) was observed at 200 μg of both the raw and dry heated seed coat extracts in the reaction mixture. The DPPH radical and ABTS cation radical scavenging activities were well proved with the ferric reducing antioxidant capacity of the extracts. Interestingly, among the extracts, methanol and aqueous acetone extracts of dry heated sample showed the highest hydroxyl radical scavenging activity of 56.6 and 45.7%, respectively. All extracts, exhibited good antioxidant activity (64.5-71.7%) against the linoleic acid emulsion system and the values were lower and higher than the synthetic antioxdiant, BHA and ascorbic acid, respectively.     

Synthesized oversulphated,acetylated and benzoylated derivatives of fucoidan extracted from Laminaria japonica and their potential antioxidant activity in vitro

Three sulphated polysaccharide derivatives (oversulphated, acetylated and benzoylated fucoidan) were successfully synthesized, and their antioxidant activities were investigated employing various established in vitro systems. A novel catalyst N-bromosuccinimide (NBS) was used in acetylation and benzoylation reaction, and the degree of acetylation was evaluated using IR and NMR spectra. Both fucoidan derivatives possessed considerable antioxidant activity, and had stronger antioxidant ability than fucoidan in certain tests. The benzoylated fucoidan showed strongest superoxide and hydroxyl radical scavenging activity, however, the acetylated fucoidan exhibited strongest activity on scavenging DPPH radical and reducing power. Available data obtained with in vitro models suggested that substituted groups of fucoidan played an important role on antioxidant activity. The mechanism on the antioxidant activity of sulphonyl, acetyl and benzoyl group is different.     

Investigations of anti-inflammatory,antinociceptive, antioxidant and aldose reductase inhibitory activities of phenolic compounds from Sideritis brevibracteata

The Sideritis species are widely used in the treatment of gastrointestinal disorders, rheumatism, cough, the common cold and as a diuretic, and they are commonly used as an herbal tea in folk medicine in Turkey. The teas of Sideritis have unique tastes, pleasant aromas and a transparent yellow colour, and are preferred as a hot beverage. In order to evaluate this information, the crude methanolic extract and the water soluble part of the methanolic extract after partition with chloroform and n-butanol were investigated for their anti-inflammatory and antinociceptive activities. Phytochemical studies performed on the n-butanol extract of the overground parts of Sideritis brevibracteata, which is rich in phenolic compounds with promising activities, resulted in the isolation of six acetylated allose containing 8-hydroxyflavone glycosides and a phenylethanoid glycoside. The structures of the compounds were established by spectroscopic evidence (UV, IR, NMR, MS). Furthermore, these phenolic compounds of S. brevibracteata were studied for anti-inflammatory, antinociceptive, antioxidant and aldose reductase inhibitory activities. The experimental data demonstrated that S. brevibracteata displayed remarkable anti-inflammatory, antinociceptive and antioxidant activities.     

Variation of antioxidant activity and the levels of bioactive compounds in lipophilic and hydrophilic extracts from hot pepper (Capsicum spp.) cultivars

Peppers (Capsicum spp.) are a rich source of diverse bioactive compounds with potential health-promoting properties. This study investigated the extraction efficiency of five solvents on antioxidant activities from cayenne (CA408 and Mesilla), jalapeño (Ixtapa) and serrano (Tuxtlas) pepper cultivars. Freeze-dried peppers were extracted using a Soxhlet extractor with five solvents: hexane, ethyl acetate, acetone, methanol, and methanol:water (80:20). The levels of specific bioactive compounds (phenolics, capsaicinoids, carotenoids and flavonoids) were determined by HPLC and antioxidant activities were assayed by three methods. For all pepper cultivars tested, hexane extracts had the highest levels of capsaicinoids and carotenoids, but methanol extracts had the maximum levels of flavonoids. Hexane extracts showed higher 2,2-diphenyl-1-pricrylhydrozyl (DPPH) radical-scavenging activity and higher reducing power, and acetone extracts (from Mesilla pepper) had a high reducing power. All pepper extracts, except hexane, were effective in preventing deoxyribose degradation, and the inhibition was increased by high concentrations of extracts. The results of the present study indicated that, among the different measures of antioxidant activity, DPPH radical-scavenging activity was strongly correlated with total bioactive compounds (capsaicinoids, carotenoids, flavonoids and total phenolics) in pepper cultivars.     

Antioxidant activity of cichoric acid and alkamides from Echinacea purpurea, alone and in combination

The antioxidant activity of extracts of the stems, leaves, and roots of Echinacea purpurea was compared with the antioxidant activity of purified cichoric acid and alkamides, both constituents of Echinacea purpurea. The antioxidant activity was determined using different methods: effect on oxygen consumption rate of a peroxidating lipid emulsion, and scavenging of radicals, i.e. 2,2-diphenyl-1-picrylhydrazyl (DPPH), measured by two different techniques. The efficacy of the extracts in the reaction with DPPH correlated well with the amount of cichoric acid present in the various extracts. The alkamides alone showed no antioxidant activity in any of the tests. Alkamides present in the extract increased, however, the antioxidative effect of cichoric acid in the peroxidating lipid emulsion. The activity was further compared with that of rosmarinic acid, a well-characterised antioxidant, and the extracts as well as cichoric acid were found to be efficient scavengers of radicals with an activity comparable to that of rosmarinic acid. Cichoric acid was found to have a stoichiometric factor of 4.0 in scavenging DPPH and to react in a second-order reaction with DPPH with a rate constant of 40 l/mol/s at 25 °C in methanol.     

Identification,quantification and antioxidant activity of acylated flavonol glycosides from sea buckthorn (Hippophae rhamnoides ssp. sinensis)

A novel acylated flavonol glycoside: isorhamnetin (3-O-[(6-O-E-sinapoyl)-β-d-glucopyranosyl-(1 → 2)]-β-d-glucopyranosyl-7-O-α-l-rhamnopyranoside) (1), together with two known acylated flavonol glycosides: quercetin (3-O-[(6-O-E-sinapoyl)-β-d-glucopyranosyl-(1 → 2)]-β-d-glucopyranosyl-7-O-α-l-rhamnopyranoside) (2) and kaempferol (3-O-[(6-O-E-sinapoyl)-β-d-glucopyranosyl-(1 → 2)]-β-d-glucopyranosyl-7-O-α-l-rhamnopyranoside) (3) were isolated from the n-butanol fraction of sea buckthorn (Hippophae rhamnoides ssp. sinensis) berries for the first time by chromatographic methods, and their structures were elucidated using UV, MS, ¹H and ¹³C NMR, and 2D NMR. Compounds 1–3 showed good scavenging activities, with respective IC₅₀ values of 8.91, 4.26 and 30.90 μM toward the 2,2′-diphenyl-1-picrylhydrazyl (DPPH) radical; respective Trolox equivalent antioxidant capacities of 2.89, 4.04 and 2.44 μM μM⁻¹ toward 2,2′-azino-bis-3-ethyl-benzothiazoline-6-sulphonate (ABTS) radical. The quantitative analysis of the isolated acylated flavonol glycosides was performed by HPLC–DAD method. The contents of compounds 1–3 were in the range of 12.2–31.4, 4.0–25.3, 7.5–59.7 mg/100 g dried berries and 9.1–34.5, 75.1–182.1, 29.2–113.4 mg/100 g dried leaves, respectively.     

Purification,preliminary structural characterization and in vitro antioxidant activity of polysaccharides from Acanthus ilicifolius

Crude Acanthus ilicifolius (A. ilicifolius) polysaccharides (CAIP) were obtained by hot water extraction and deproteinated. Two major polysaccharide fractions, the neutral A. ilicifolius polysaccharide (NAIP) and the acid A. ilicifolius polysaccharide B (AAIP-B), were isolated from CAIP by chromatography using DEAE-Sepharose Fast Flow and Sepharose CL-6B. The molecular weights (M_w) of NAIP and AAIP-B, determined by high performance gel-filtration chromatography (HPGFC), were 11,775 and 23,161 Da, respectively. AAIP-B contained 51.23% uronic acid, characteristic of a pectin-type hetero-polysaccharide. Analysis of the neutral monosaccharide composition indicated that NAIP contained high proportions of arabinose, galactose and glucose. However, AAIP-B was mainly composed of rhamnose, arabinose and galactose. Their structure properties were confirmed by FT-IR.     

Main nutrients,phenolics, antioxidant activity,DNA damage protective effect and microstructure of Sphallerocarpus gracilis root at different harvest time

Sphallerocarpus gracilis roots harvested at two different times (T1: 12-10-2009 and T2: 15-04-2010) were analysed for their main nutrients, phenolics, antioxidant activity, DNA damage protective effect and microstructure. Results revealed that higher amounts of starch were detected in T1 S. gracilis root, whereas higher levels of reducing sugar, protein, amino acids and phenolic compounds were determined in T2 S. gracilis root. The antioxidant activity was assessed by DPPH, FRAP and TEAC assay and T2 S. gracilis root was found to possess higher antioxidant activity. Also, results of the correlation analysis indicated that caffeic, p-coumaric and gallic acids contributed substantially to the antioxidant property and there was significant correlation between DPPH radicals scavenging capacity and FRAP and TEAC. The highest and lowest DNA damage protective effects were found of free phenolic extracts from T2 and T1 S. gracilis roots, respectively. The microstructure of S. gracilis root varied greatly with the harvest time.     

Purification,composition analysis and antioxidant activity of a polysaccharide from the fruiting bodies of Ganoderma atrum

A water-soluble protein-bound polysaccharide was extracted from the fruiting bodies of Ganoderma atrum and isolated by gel-filtration chromatography. Its primary structural features and molecular weight were characterized by infrared spectrometry, gas chromatography, size exclusion chromatography, amino acid analyzer and high-performance liquid chromatography (HPLC). The data obtained indicated that the glycoprotein contains 10.1% of protein and 17 general amino acids and it is rich in glutamic acid, asparagic acid, alanine, glycine, threonine, and serine. It was mainly composed of mannose, galactose and glucose in a molar ratio of 1:1.28:4.91, with an average molecular weight of about 1013 kDa. The existence of an O-glycosidic linkage in PSG-1 (polysaccharide1) was demonstrated by a β-elimination reaction. The antioxidant activity of the purified polysaccharides was evaluated in vitro by 1,1-diphenyl-2-picryl-hydrazyl (DPPH) free radical scavenging assay, self-oxidation of 1,2,3-phentriol assay. Those various antioxidant activities were compared to standard antioxidants vitamin C and BHT. It was found that the scavenging effects of the purified polysaccharides increased with measuring concentration. The results indicated that the purified polysaccharides showed strong DPPH free radical and superoxide anion radical scavenging activities. This study suggested that the purified polysaccharides could potentially be used as natural antioxidants.