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).     

Evaluation of the antioxidant activity of five endemic Ligustrum species leaves from Taiwan flora in vitro

Leaves from the plant species belonging to the genus Ligustrum are widely used as tea or herbal medicine in Europe, China, and Japan. The antioxidant properties of five Ligustrum species from Taiwan were compared using in vitro antioxidant methods such as DPPH radical scavenging, TEAC, and FRAP assays. Cell-based antioxidant methods were used, including Fe²⁺/ascorbate-induced lipid peroxidation on brain homogenate and AAPH-induced erythrocyte haemolysis. The amounts of major phenolic compounds from the Ligustrum species, including phenylpropanoids, flavonoids, and iridoids, were determined by spectrophotometric methods. The results showed that all Ligustrum species exhibited antioxidant, radical-scavenging, anti-haemolytic, and lipid peroxidation-inhibiting activities at different magnitudes of potency. A significant correlation was found between antioxidant activity and the amount of antioxidant components, in particular, total phenolics and phenylpropanoids. Among all Ligustrum species from Taiwan, Ligustrum morrisonense is presented as potential source of natural antioxidants.     

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).     

In vitro antioxidant activities of the methanol extracts of four Helichrysum species from Turkey

This study was designed to examine the in vitro antioxidant activities of the methanol extracts of four Helichrysum species (Helichrysum noeanum Boiss., H. chionophilum Boiss. & Bal., H. plicatum DC. subsp. plicatum, H. arenarium (L.) Moench. subsp. aucheri (Boiss.) Davis & Kuphicha). The extracts were screened for their possible antioxidant activitiy by two complementary test systems, namely DPPH free radical-scavenging and β-carotene/linoleic acid systems. In the first case, non-polar subfractions of the methanol extracts of Helichrysum species studied did not show any antioxidant activity, while the most active one was H. chionophilum (IC₅₀ =40.5 μg/ml) among the polar subfractions. In the β-carotene/linoleic acid test system, inhibition rates of the oxidation of linoleic acid of H. noeanum and H. arenarium were very close to each other. The inhibition rate of the synthetic antioxidant BHT was determined to be 96%. Since the polar extracts of Helichrysum species dealt with here exhibited interesting antioxidant activities when compared to BHT, we believe that it would be useful to take the results into consideration as an alternative for food processing industries.     

Evaluation of antioxidant,antibacterial and anti-tyrosinase activities of four Macaranga species

The methanolic fresh leaf extracts of Macaranga gigantea, Macaranga pruinosa, Macaranga tanarius and Macaranga triloba were screened for their antioxidant properties (AOP), tyrosinase inhibition and antibacterial activities. Total phenolic content (TPC), 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging, ferric-ion reducing power (FRAP), ferrous-ion chelating (FIC) and lipid peroxidation inhibition (LPI) activities were used to evaluate the AOP. Modified 3,4-dihydroxy-L-phenylalanine (L-DOPA) method was used to determine tyrosinase inhibition activity, whereas antibacterial activity was determined using the disc-diffusion technique. TPC screening of the same species from different collection sites showed no significant difference between sites. M.triloba showed the highest ascorbic acid equivalent antioxidant activity (AEAC), FRAP and LPI values. M. tanarius, which showed the lowest TPC, AEAC, FRAP and LPI activities, exhibited the best FIC activity. M. pruinosa showed the best tyrosinase inhibition activity, whereas M. triloba showed the best antibacterial activity against Gram-positive bacteria species, with minimal inhibition dosage (MID) values as low as 10 μg/disc.     

Phenolic content, antioxidant, anti-inflammatory and anticancer activities of the edible halophyte Suaeda fruticosa Forssk

Suaeda fruticosa is an edible and medicinal halophyte known for its hypoglycaemic and hypolipidaemic activities. In this study, novel biological activities of the shoot extracts related to their phenolics were investigated. Results showed an appreciable total phenolic (31.8 mg GAE/g DW) in shoot extracts. The estimation of antioxidant capacities using oxygen radical absorbance capacity (ORAC method) and a cell based-assay (WS1) of four extracts (hexane, dichloromethane, methanol and water) showed that shoot methanol extract exhibit the highest antioxidant activities. The same extract displayed the utmost anti-inflammatory activity, inhibiting nitric oxide (NO) release, by 66.4% at 160 μg/ml in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Besides, the dichloromethane extract showed the highest anticancer activity against human lung carcinoma (A-549) and colon adenocarcinoma cell lines (DLD-1, Caco-2 and HT-29) with specificity against DLD-1 (IC₅₀ = 10 ± 1 μg/ml). These findings demonstrate the remarkable potentiality of this edible halophyte as valuable source of antioxidants which exhibit original and interesting anti-inflammatory and anticancer capacities.     

In vitro antioxidant and antimicrobial activities of the extract of Pericarpium Citri Reticulatae of a new Citrus cultivar and its main flavonoids

To provide the base for the application of a new cultivar of Citrus reticulata Blanco, whose family is Rutaceae, a flavonoid extract of Pericarpium Citri Reticulatae (FEPCR), the dried rind of the ripe citrus, was obtained with 80% aq. ethanol. Total flavonoid content of FEPCR was determined by a colorimetric method. Total phenol content was estimated as gallic acid equivalents. The major constituents of FEPCR, including Hesperidin, Nobiletin and Tangeretin, were determined by HPLC analysis. The antioxidant activities of FEPCR, Hesperidin, Nobiletin and Tangeretin were evaluated by various antioxidant assays, including DPPH scavenging, hydroxyl radical scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging and reducing power. All samples showed antioxidant activities to some degree in all the tested methods. In addition to the antioxidant activity, the antimicrobial assay was measured as well. Six strains of microorganisms including Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Salmonella typhi and Enterobacter cloacae were used in the tests. FEPCR and Hesperidin displayed a broad antimicrobial spectrum and exerted antimicrobial effects in antimicrobial tests. But Tangeretin and Nobiletin exhibited low antimicrobial activities.     

Flavonoid contents and antioxidant activities from Cinnamomum species

Cinnamomum has long been regarded as a food or medicinal plant. Leaves of five species of Cinnamomum, namely C. burmanni, C. cassia, C. pauciflorum, C. tamala and C. zeylanica, were chosen to investigate their antioxidant activities in this study. C. zeylanica exhibited the highest total phenolic content while C. burmanni had the highest flavonoid content among the five species. These five species were then screened for their antioxidant potentials using various in-vitro models such as total antioxidant capability, 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging activity, reducing power and superoxide anion scavenging activity at various concentrations. C. zeylanica showed the highest DPPH radical scavenging activity, total antioxidant activity and reducing power, while C. tamala exhibited the highest superoxide anion scavenging activity. By the analysis of the high performance liquid chromatography coupled to diode array detector (HPLC-DAD), three flavonoid compounds namely quercetin, kaempferol and quercetrin were identified and quantified. This study suggested that Cinnamomum leaf can be used potentially as a readily accessible source of natural antioxidants.

Industrial relevance

This study was focused to evaluate the antioxidant activities of five species of Cinnamomum leaf which is normally used in medicine and also used in food preparation. This study provided an alternative of utilizing Cinnamomum leaf as a readily accessible source of natural antioxidants in food and pharmaceutical industry.     

In vitro evaluation of antioxidant activities of aqueous extracts from natural and cultured mycelia of Cordyceps sinensis

Cordyceps sinensis, one of the best known traditional Chinese medicines and health foods, has been highly valued for the treatment of a wide range of diseases and reported to have antioxidant properties. In the present study, the antioxidant activities of hot-water extracts from natural and cultured mycelia of C. sinensis were investigated and evaluated using six in vitro assays, including inhibition of linoleic acid peroxidation; scavenging abilities on DPPH, hydroxyl and superoxide anion radicals; the reducing power and the chelating ability on ferrous ions. Among these assays, the extracts showed the best effect on the inhibition of linoleic peroxidation with the lowest IC50 values and with an inhibition rate over 90% at concentration of 0.8-1.6 mg/ml, more stable than that of α-tocopherol, a recognised natural antioxidant. The scavenging activities on superoxide anion and hydroxyl radicals of the two extracts were slightly lower than that of butylated hydroxytoluene. DPPH scavenging activities of both extracts reached over 80% inhibition at 4-8 mg/ml. Both extracts showed moderate reducing power and ferrous ion chelating activity. The IC50 value of the extract from cultured mycelia in all the tests, except for linoleic acid peroxidation, was significantly lower than that of natural mycelia. There was no evident correlation between the antioxidant activity and the content of protein, polysaccharides and mannitol of extracts from C. sinensis; the antioxidant activity may be due to a combined effect of these or some other compounds. These results suggested that both the extracts from cultured and natural mycelia have direct and potent antioxidant activities and that the cultured mycelia of the fungus could be used for the antioxidant activity to reduce the human demands on the natural resources of the fungus, an endangered species.     

Determination of antioxidant activities of various extracts and essential oil compositions of Thymus praecox subsp. skorpilii var. skorpilii

The aim of this work is to examine the chemical composition and antioxidant activity of separated essential oils and different solvent extracts of Thymus praecox subsp. skorpilii var. skorpilii (TPS). The ethanol, acetone, methanol, hexane, aqueous extracts and separated essential oils of TPS were assessed for their antioxidant activities. Antioxidant activities were evaluated by reduction of Mo(VI) to Mo(V), reducing power, superoxide scavenging activity, free radical-scavenging activity, metal chelating activity, linoleic acid peroxidation, hydrogen peroxide scavenging activity and peroxide scavenging activity. Essential oils were characterized in total to be 41 components, whereas 9 components were isolated by column chromatography for antioxidant activity. TPS essential oil was found to contain thymol (40.31%) and o-cymene (13.66%) as the major components. The ethanol, methanol and water extracts exerted significant free radical-scavenging activity. The methanol and water extracts displayed highest superoxide scavenging activity. The water extract has the highest total phenolics (6.211 mg gallic acid (GAE)/g DW) and flavonoids (0.809 mg quercetin/g DW).     

Polyphenols and triterpenes from Chaenomeles fruits: Chemical analysis and antioxidant activities assessment

Mugua, fruit of the genus Chaenomeles, is a valuable source of health food and Chinese medicine. To elucidate the bioactive compounds of five wild Chaenomeles species, extracts from fresh fruits were investigated by HPLC-DAD/ESI-MS/MS. Among the 24 polyphenol compounds obtained, 20 were flavan-3-ols (including catechin, epicatechin and procyanidin oligomers). The mean polymerisation degree (mDP) of procyanidins was examined by two acid-catalysed cleavage reactions; the mDP value was the highest in Chaenomeles sinensis and the lowest in Chaenomeles japonica. Total polyphenol content (TPC) reached 46.92 and 46.28 mg/g gallic acid equivalents (GAE) in Chaenomeles speciosa and Chaenomeles thibetica, respectively, although their main bioactive compounds were different (being epicatechin and procyanidin B2 in the former, and catechin and procyanidin B1 in the latter). These two species also exhibited equally strong free radical scavenging activities. Our results further showed that the antioxidant ability of Chaenomeles fruits was significantly correlated to their total polyphenol contents. Two triterpenes (oleanolic acid and ursolic acid) were the highest quantity in Chaenomeles cathayensis and C. thibetica, respectively.     

Structural characterisation of polysaccharides from Tricholoma matsutake and their antioxidant and antitumour activities

In this study, polysaccharides from Tricholoma matsutake (TM-P) were purified using a DEAE-Sepharose fast flow column and three polysaccharide fractions (TM-P1, TM-P2 and TM-P3) were obtained. The chemical composition and structural characteristics of TM-P2 were quite different from those of TM-P1 and TM-P3. TM-P2 consisted of glucose, galactose and mannose with a molar ratio of 5.9:1.1:1.0. The glycosidic linkages were mainly composed of 1,6- and 1-linked glucose. Furthermore, TM-P2 showed the strongest in vitro antioxidant and antitumour activities. The oxygen radical absorbance capacity (ORAC) of TM-P2 was 2100.44 μmol Trolox/g. The antiproliferative activities of TM-P2 (4.0 mg/ml) on the growth of HepG2 and A549 cells were 67.98% and 59.04%, respectively.     

In vitro determination of antioxidant activity of proteins from jellyfish Rhopilema esculentum

In this study, the antioxidant activity of proteins isolated from jellyfish, Rhopilema esculentum Kishinouye (R. esculentum), was determined by various antioxidant assays, including superoxide anion radical-scavenging, hydroxyl radical-scavenging, total antioxidant activity, reducing power and metal chelating activity. Butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), α-tocopherol, vitamin C and mannitol were used as standards in those various antioxidant activities. The crude protein (CP) and the protein fractions isolated by Sephadex chromatography, first peak (FP) and second peak (SP), had very low reductive power and metal chelating abilities compared to EDTA, but they showed strong scavenging effects on the superoxide anion radical, hydroxyl radical and varying total antioxidant activity. FP and SP exhibited stronger scavenging effects on the superoxide anion radical than BHA, BHT or α-tocopherol. The EC₅₀ values of FP and SP were 6.12 and 0.88 μg/ml, respectively, while values EC₅₀ of BHA, BHT and α-tocopherol were 31, 61 and 88 μg/ml, respectively. CP, FP and SP showed far higher hydroxyl radical-scavenging activities than did vitamin C or mannitol. The EC₅₀ values of CP, FP and SP were 48.76, 45.42 and 1.52 μg/ml, but EC₅₀ values of vitamin C and mannitol were 1907 and 4536 μg/ml, respectively. In a β-carotene–linoleate system, SP and CP showed antioxidant activity, but lower than BHA. Of the three samples, SP had the strongest antioxidant activity. So, SP may have a use as a possible supplement in the food and pharmaceutical industries.     

In vitro antioxidant activities of the methanol extracts of five Allium species from Turkey

This study was designed to examine the in vitro antioxidant activities of the methanol extracts of five Allium species, namely Allium nevsehirense, A. sivasicum, A. dictyoprosum, A. scrodoprosum subsp. rotundum and A. atroviolaceum; the former two are endemic for the Turkish flora. The extracts were screened for their possible antioxidant activities by two complementary tests; DPPH free radical-scavenging and β-carotene/linoleic acid assays. In the first case, non-polar subfractions of the extracts did not show any antioxidant potential, while the polar subfractions exhibited marked activity. Among the polar ones, the most active one was A. atroviolaceum with an IC₅₀ of 79.0 ± 2.75 μg/ml. In the β-carotene/linoleic acid assay, the inhibition ratios of the oxidation of linoleic acid by A. atroviolaceum and A. dictyoprosum were too close to each other (71.2 ± 2.20% and 72.3 ± 1.20%, respectively), while that of the synthetic antioxidant, BHT, was 96%.     

GC/MS analysis and in vitro antioxidant activity of essential oil and methanol extracts of Thymus caramanicus Jalas and its main constituent carvacrol

Chemical composition of the essential oil, antioxidant activity (DPPH and β-carotene/linoleic acid assays), and total phenolic content (Folin–Ciocalteu assay) of aerial parts of Thymus caramanicus were determined. The highest radical-scavenging activity (DPPH test) was shown by the polar subfraction of the methanol extract (IC₅₀ = 43.0 μg/ml) which was also higher than that of butylated hydroxytoluene (BHT, IC₅₀ = 19.7 μg/ml). However, it was the nonpolar subfraction of the methanol extract that showed the highest inhibition (84.4%), as assessed by the β-carotene/linoleic acid assay, which was only slightly lower than that shown by BHT (93.3%). The antioxidant activities of the essential oil main component (carvacrol) were also evaluated for comparison. Total phenolic content of the polar subfraction, as gallic acid equivalents, was 124.3 μg/mg. Essential oil extracted from the aerial parts by hydrodistillation was analysed by GC and GC/MS. Fifteen constituents, representing 99.3% of the oil, were identified, of which the major ones, carvacrol (85.9%), thymol (3.3%), p-cymene (3.2%), γ-terpinene (1.8%) and borneol (1.3%), accounted for 95.6% of the oil.     

Concordance between antioxidant activities and flavonol contents in different extracts and fractions of Cuscuta chinensis

Chinese herbs employed in traditional Chinese medicine (TCM) have been used for centuries in the practice of medicated diet and dietetic therapy. The seed of Cuscuta chinensis Lam. (Convolvulaceae), a commonly used traditional Chinese herb, is frequently added in Chinese cooking and preparation of refreshments, including porridge and alcoholic beverages, to nourish the human body. In the present study, we compared the antioxidant activities of water and ethanol extracts from the seeds C. chinensis and also of its different organic fractions, including n-hexane, ethyl acetate, n-butanol and organic water, by assessing their DPPH (1,1-diphenyl-2-picryl hydrazine) free radical-scavenging, superoxide anion scavenging, anti-superoxide anion formation and anti-lipid peroxidation abilities. The flavonol contents of all test samples were analyzed by high-performance liquid chromatography with an ultraviolet (HPLC–UV) detector. The results showed that there is a direct correlation of the flavonol content with the antioxidant activities from the extracts and fractions of C. chinensis. Moreover, the ethyl acetate fraction demonstrated significantly better and higher antioxidant effects, and also had a higher flavonol content than had the remaining samples (P < 0.05). The water fractions, however, exhibited the weakest antioxidant activity, and had low concentrations of flavonols. Thus, we suggest that the ethanol extract of C. chinensis, but not its water extract, could be used as a dietary nutritional supplement to promote human health and prevent oxidation-related diseases, due to its antioxidant properties.