Antioxidant and radical scavenging activities of the pyroligneous acid from a mangrove plant, Rhizophora apiculata

Total phenolics content, free radical scavenging activity, reducing power, and antioxidant activity of the pyroligenous acid from a mangrove plant, Rhizophora apiculata were evaluated. Dichloromethane extraction of the raw pyroligneous acid successfully yield 2 extracts, i.e. concentrated pyroligneous acid (CPA) and concentrated pyroligneous acid extract (CPAE). Phenolic contents in CPAE and CPA, expressed as (±)-catechin equivalents/g of the sample were 5465 ± 367 mg and 2502 ± 152 mg, and expressed as gallic acid equivalents/g of the sample were 2919 ± 209 mg and 1348 ± 90 mg, respectively. CPAE exhibited superior free radical scavenging activity with EC₅₀ value = 0.1235 mg/ml, or 80.96% of free radical scavenging capability. The ferric reducing power of CPAE was approximately 3.7, 5.1, 6.1, and 21.3 times higher than that of ascorbic acid, BHA, BHT and alpha-tocopherol. In phosphomolybdenum assay, CPAE showed the greatest antioxidant efficacy (A₆₉₅ = 1.278) compared to those of CPA and different standards. In addition, the free radical scavenging activity, ferric reducing power and total antioxidative activity of CPAE and CPA showed positive correlation with their total phenolic content with R² values ranging from 0.9624 to 0.9979.     

Antioxidant activity of compounds isolated from the pyroligneous acid, Rhizophora apiculata

The polyphenols (CPAE II) was isolated from the dichloromethane extract of the pyroligneous acid, Rhizophora apiculata by simultaneous acid base and solvent extraction method. Its qualitative and quantitative composition was studied by gas chromatography mass spectroscopy (GC/MS) and out of 57 peaks, 52 compounds were identified, representing 95.47% of the total polyphenols. The CPAE II was then fractionated to four fractions (F1–F4) by means of thin layer chromatography and silica gel column chromatography with dichloromethane, dichloromethane/chloroform/ethyl acetate mixture (8:1:1; 4:3:3, v/v/v), and ethyl acetate, respectively. The antioxidant properties of the CPAE II and the fractions were evaluated. Among the four fractions, fraction 1 (F1) was the most potent in DPPH radical scavenging activity and molybdenum (VI) reducing power. It was subjected to further purification by means of silica gel column chromatography with hexane, hexane/diethyl ether mixture (9:1, 6:1, 3:1, v/v), and diethyl ether, respectively. 2,6-Dimethoxyphenol (syringol) and dihydroxybenzenes (catechol and 3-methoxycatechol) were isolated and identified by GC/MS, ¹H NMR, ¹³C NMR spectral analyses, and confirmed by GC co-injection with authentic standards. Syringol, catechol and 3-methoxycatechol constitute 39.08, 4.21 and 1.10% of F1, respectively. Their antioxidant activities were evaluated by DPPH radical scavenging activity, ABTS radical cation scavenging activity, phosphomolybdenum and ferric reducing antioxidant power (FRAP). The trend in antioxidant capacity was similar in all the four assays, with dihydroxybenzenes > 2,6-dimethoxyphenols, although discrepancies in the ranking within the dihydroxybenzenes were present. These three compounds which showed significant antioxidant activities were isolated for the first time from the pyroligneous acid, R. apiculata.     

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 and antibacterial activities of Nephelium lappaceum L. extracts

Ether, methanolic and aqueous extracts of lyophilized rambutan (Nephelium lappaceum L.) peels and seeds were evaluated for phenolic contents, antioxidant and antibacterial activities. High amounts of phenolic compounds were found in the peel extracts and the highest content was in the methanolic fraction (542.2 mg/g dry extract). Several potential antioxidant activities, including reducing power, β-carotene bleaching, linoleic peroxidation and free radical scavenging activity, were evaluated. The peel extracts exhibited higher antioxidant activity than the seed extracts in all methods determined (P < 0.05). The methanolic fraction was found to be the most active antioxidant as shown by their 50% DPPH inhibition concentration, 4.94 μg/mL. The results indicated this fraction exhibited greater DPPH radical scavenging activity than BHT and ascorbic acid (0.32 g dry extract/g BHT or ascorbic acid). Antibacterial activity against eight bacterial strains was assessed by disc diffusion and broth macrodilution methods. All peel extracts exhibited antibacterial activity against five pathogenic bacteria. The most sensitive strain, Staphylococcus epidermidis, was inhibited by the methanolic extract (MIC 2.0 mg/mL).     

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 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 polymeric proanthocyanidins from fruit stones and pericarps of Litchi chinensis Sonn

Fruit stones and pericarps of Litchi chinensis, waste products of the food, were studied as a source of polymeric proanthocyanidins. The structures of the polymeric proanthocyanidins isolated from Litchi chinensis were characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) coupled with high performance liquid chromatography electrospray ionization mass spectrometry (HPLC-ESI-MS) analysis. The spectra obtained through MALDI-TOF MS analysis revealed that the dominant A-type procyanidin polymers occurred in each polymer length with one or more A-type linkages. The polymeric proanthocyanidins of litchi fruit stones exhibited longer polymer length than those of fruit pericarps, with polymerization degrees up to 20 and 11 for fruit stones and pericarps, respectively. After depolymerization with toluene-??-thiol, HPLC-ESI-MS analysis showed that epicatechin and A-type dimer were the major constituent units, and the mean polymerization degrees were 15.4 and 5.8 for polymeric proanthocyanidins of fruit stones and pericarps, respectively. The antioxidant properties investigated using DPPH, ABTS and FRAP methods showed that the higher polymerization degree of polymeric proanthocyanidins from litchi fruit stones exhibited higher antioxidant activities than those from litchi pericarps.     

Antioxidant activities of Salvia miltiorrhiza and Panax notoginseng

Traditional Chinese medicinal herb Salvia miltiorrhiza (SM) and Panax notoginseng (PN) have been widely used for the prevention and treatment of vascular diseases in the clinics. To better understand their mechanisms of pharmacological actions, the in vitro antioxidant activities of extract of Salvia miltiorrhiza (ESM) and extract of Panax notoginseng (EPN) were evaluated with different antioxidant testing systems. Their activities of scavenging superoxide anion radicals, DPPH radicals, hydroxyl radicals, and hydrogen peroxide, chelating Ferrous ion, and ferric ion reducing power were assessed. The results showed that the mechanisms of their antioxidant effects were distinct and diverse. ESM possessed strong reducing power and high scavenging activities against free radicals including superoxide anion, hydroxyl and DPPH radicals, but a weaker scavenging activity for hydrogen peroxide. ferrous ion chelating activity of ESM was undetectable at the tested concentrations. In contrary, EPN exhibited strong ferrous ion chelating activity and high scavenging activities against hydrogen peroxide, hydroxyl radicals, and a weak activity against superoxide anion and DPPH free radicals. EPN did not show any ferric ion reducing power. Since their antioxidant mechanisms are complementary, the combined use of ESM and EPN might be even more beneficial. These antioxidant properties of SM and PN are likely part of the reasons that they are effective in the prevention and treatment of vascular diseases.     

Evaluation of antioxidant activities of various solvent extracts of Carissa opaca fruits

The chloroform and aqueous fractions of Carissa opaca fruit, a traditional medicinal fruit in Pakistan possessed a high amount of total phenolic and flavonoid contents as compare to other solvent fractions with potent antioxidant activities in scavenging DPPH, superoxides, hydroxyl, hydrogen peroxide, ABTS radicals, and had strong iron chelating activity. On the other hand, the ethyl acetate fraction showed the highest inhibition of β-carotene linoleic acid peroxidation and phosphomolybdate assay. A high correlation coefficient existed between EC₅₀ values of DPPH, superoxides, hydroxyl, hydrogen peroxide, ABTS radicals, total phenolics and flavonoids, but a non significant correlation was found in the case of iron chelaters, β-carotene and phosphomolybdate assay. This study verified that the chloroform and aqueous fractions have strong antioxidant activities which were correlated with its high level of phenolics and flavonoids. These fractions can be used as a source of potential antioxidant or functional food material.     

In vitro antioxidant activities of Rosmarinus officinalis extracts treated with supercritical carbon dioxide

The leaves of Rosmarinus officinalis (rosemary) were subjected to supercritical CO₂ extraction (SFE). Different sources of variability, including location (Izmir, Canakkale and Mersin) and harvesting time (December, March, June and September), were considered. Among active constituents of rosemary, carnosic acid, carnosol and rosmarinic acid were analyzed by HPLC. Variability of the amounts of active constituents appears to be due to different geographical locations of growth and seasonal variations. The levels of the constituents were higher in the months of December 2003 and September 2004. In addition to this, 12 SFE extracts were screened for their radical-scavenging capacities and antioxidant activities by various in vitro assays, namely total phenol assay, DPPH radical-scavenging activity and trolox equivalent antioxidant capacity (TEAC).     

Antioxidant properties of Phyllanthus amarus extracts as affected by different drying methods

The total phenolic content (TPC) and antioxidant activity of fresh and dried Phyllanthus amarus plant materials were evaluated using the Folin-Ciocalteau method, 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity and ferric reducing antioxidant power (FRAP) assays. Different drying treatments led to significant reduction (P<0.05) in antioxidant properties of P. amarus methanolic extracts, with microwave drying causing the highest decrease in TPC and antioxidant activity exhibited by the reduction in both radical scavenging activity and FRAP. On the other hand, boiling water extracts appeared to exhibit significantly stronger antioxidant potentials (P<0.05) even in dried plant materials due to greater solubility of compounds, breakdown of cellular constituents as well as hydrolysis of tannins. Its strong free radical scavenging activity suggests that it has great potential in the food industry as functional food ingredient.     

Antioxidant activities and phenolic compounds of pigmented rice bran extracts

This study was carried out to investigate the antioxidant activities and phenolic compounds of pigmented rice (black, red, and green rice) and brown rice brans. Antioxidant activity was determined by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical assay, 2,2-azino-bis-(3-ethylenebenzothiozoline-6-sulfonic acid) (ABTS) radical cation assay, reducing power, and chelating ability. Phenolic compounds were measured by using HPLC. Pigmented rice brans were extracted by using aqueous mixtures of acetone, ethanol, and methanol to determine the most effective extraction solvent. Of all solvents examined, extract from 40:60 acetone-water mixtures (v/v) provided the highest DPPH radical assay as well as the highest total phenolic and flavonoid content. We finally selected 40% acetone as an extraction solvent for antioxidant study of pigmented rice bran. Antioxidant activities of 40% acetone extracts of pigmented rice bran, measured in the range of 0 to 1500 μg/mL. At 500 μg/mL concentration, red rice bran, which had the highest total phenolic (259.5 μg/mg) and total flavonoid (187.4 μg/mg) contents, showed the highest antioxidant activity: 83.6%, 71.5%, 1.2%, and 16.4% for DPPH radical assay, ABTS radical cation assay, reducing power, and chelating ability, respectively. Red rice bran showed a lower EC(50) value (112.6 μg/mL) than that of butylated hydroxytoluene (144.5 μg/mL) from the DPPH radical assay. The major phenolic acids of red rice bran were ferulic, vanillic and p-coumaric acids. The results indicated pigmented rice bran might be used as a natural antioxidant. PRACTICAL APPLICATION: The present study revealed black and red rice bran shows high antioxidant activities and they contain high amount of phenolic compounds. Indeed, black and red rice bran could be better raw materials for manufacturing the food with high antioxidant activity.     

Radical scavenging and antibacterial properties of the extracts from different Thymus pulegioides L. chemotypes

Radical scavenging and antibacterial properties of large thyme extracts isolated from five chemotypes of Thymus pulegioides L. growing wild in Lithuania were studied. The chemotypes were defined according to the main essential oil components: linalool (L), geranial/geraniol/neral (G/G/N), thymol (T), carvacrol/γ-terpinene/p-cymene (C/γT/pC) and thymol/carvacrol/γ-terpinene/p-cymene (T/C/γT/pC). The contents of phenolic compounds, flavonoids and flavonols were determined. It was found that the extracts of phenolic chemotypes containing remarkable concentrations of thymol and/or carvacrol were stronger DPPH and ABTS free radical scavengers in the model systems. The antibacterial activity of the extracts depended on the plant chemotype, extract preparation, solvent used and finally the sensitivity of bacteria. Bacillus cereus, Micrococcus luteus, Staphylococcus epidermidis and Staphylococcus aureus were the most sensitive to the all extracts applied, whereas Escherichia coli, Salmonella typhimurium and Enterobacter aerogenes remained resistant.     

Antioxidant potentials and rosmarinic acid levels of the methanolic extracts of Salvia verticillata (L.) subsp. verticillata and S. verticillata (L.) subsp. amasiaca (Freyn & Bornm.) Bornm

This study was designed to examine the in vitro antioxidant activities and rosmarinic acid levels of the methanol extracts of Salvia verticillata subsp. verticillata and S. verticillata subsp. amasiaca. The extracts were screened for their possible antioxidant activity by two complementary test systems, namely DPPH free radical-scavenging and β-carotene/linoleic acid systems. In the first case, S. verticillata subsp. verticillata was superior to the subsp. amasiaca with an IC₅₀ value of 14.5 ± 1.21 μg mg⁻¹. In the β-carotene/linoleic acid test system, inhibition capacity of S. verticillata subsp. verticillata was 74.4 ± 1.29%. Antioxidant activities of BHT, ascorbic acid, curcumin and α-tocopherol were determined in parallel experiments. Activity of rosmarinic acid was also screened for better establishing the relationship between rosmarinic acid level and antioxidant activity for the plant extracts. S. verticillata subsp. verticillata had the highest rosmarinic acid level with a value of 28.7 ± 0.89 μg mg⁻¹. There is a strong correlation between the rosmarinic acid level and antioxidant activity potential. Our results showed that rosmarinic acid and its derivatives are more likely to be responsible for most of the observed antioxidant activities of Salvia species.     

Antioxidant properties of aqueous extracts from Terminalia catappa leaves

Aqueous extracts were prepared from green, yellow fallen and red fallen leaves of Terminalia catappa L. (Combretaceae) and their antioxidant activity, scavenging and chelating abilities were evaluated. Aqueous extracts from three different leaves showed high antioxidant activities and moderate scavenging abilities on hydroxyl radicals at 1 mg/ml. EC₅₀ values in antioxidant activity were 0.549-0.557 mg/ml whereas those in scavenging ability on hydroxyl radicals were 0.631-0.686 mg/ml for aqueous extracts prepared from three leaves with boiling water for 3 min. EC50 values in reducing power were 0.15-0.23 mg/ml. EC50 values in scavenging abilities on superoxide anion and 1,1-diphenyl-2-picrylhydryl radicals were 0.36-0.44 and 10.4-35.3 mg/ml, respectively. EC50 values in chelating abilities on ferrous and cupric ions were 0.41-2.50 and 8.96-9.89 mg/ml, respectively. Based on the results obtained, the aqueous extracts displayed higher antioxidant properties. Six phenolic compounds identified in the aqueous were p-hydroxybenzoic acid, 4-hydroxyphenylpropionic acid, m-coumaric acid, 3,4-dihydroxybenzoic acid, p-coumaric acid and gallic acid. Therefore, the aqueous extracts from three different leaves of T. catappa might be a potential antioxidant supplement for application in food products or as a drink.     

Antioxidant properties of methanolic extracts from Agrocybe cylindracea

Agrocybe cylindracea (DC: Fr.) Mre. (Bolbitiaceae) was available in the form of fruit bodies, mycelia and fermentation filtrate. From these three forms, methanolic extracts were prepared and their antioxidant properties studied. The methanolic extract from fruit bodies showed high inhibitory ability on lipid oxidation (90.0-97.3%) at 5-20 mg/ml and had an EC₅₀ value of 0.94 mg extract/ml. EC₅₀ values in reducing power were 2.36, 6.89 and 5.00 mg extract/ml for fruit bodies, mycelia and filtrate, respectively. The scavenging ability of the methanolic extract from fruit bodies on 1,1-diphenyl-2-picrylhydrazyl radicals was 89.0% at 1 mg/ml whereas those from mycelia and filtrate was 91.4% and 94.9% at 10 mg/ml, respectively. The chelating abilities of methanolic extracts from fruit bodies, mycelia and filtrate on ferrous ions were 90.6%, 84.6% and 96.3% at 5 mg/ml, respectively. Total phenols were the major detected naturally occurring antioxidant components found in methanolic extracts from A. cylindracea and in the range of 15.55-23.47 mg/g. Based on EC₅₀ values, A. cylindracea was good in antioxidant properties, except for the scavenging ability on hydroxyl radicals.