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.     

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.     

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.     

Antioxidant and free radical scavenging activities of wild bitter melon (Momordica charantia Linn. var. abbreviata Ser.) in Taiwan

Momordica charantia Linn. var. abbreviata Ser. (Cucurbitaceae), also known as “Shan Ku Gua”, is a wild variety of bitter melon (BM) in Taiwan. The size of its fruits is only about one-fifth of the commonly seen BM. It is commonly consumed as vegetable and also used as a popular folk medicine. In this study, the antioxidant and free radical scavenging activities of BM aqueous (BM-H₂O) and ethanol (BM-EtOH) extracts were evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH), metal chelation, cytochrome c and xanthine oxidase inhibition (XOI) assays, as well as FeCl₂-ascorbic acid induced lipid peroxidation (thiobarbituric acid reactive substances, TBARS) assay in rat liver homogenates in vitro. Total flavonoid and phenol contents of BM extracts were also analyzed. Results showed that both BM-H₂O (IC₅₀=129.94 μg/ml) and BM-EtOH (IC₅₀=156.78 μg/ml) possess potent DPPH radical scavenging activity, which was better than vitamin E (IC₅₀=172.21 μg/ml). These extracts also showed better iron chelating activity than vitamin E. However, they were weaker than vitamin E in free radical scavenging, xanthine oxidase inhibitory and anti-lipid peroxidation activities. With the exception of XOI activity [IC₅₀=7.90 μg/ml (BM-H₂O) vs. 7.69 μg/ml (BM-EtOH)], BM-H₂O showed a lower IC₅₀ value in free radical scavenging [IC₅₀=6.15 μg/ml (BM-H₂O) vs. 7.08 μg/ml (BM-EtOH)] and anti-lipid peroxidation [IC₅₀=53.72 μg/ml (BM-H₂O) vs. 88.51 μg/ml (BM-EtOH) for liver; 82.53 μg/ml (BM-H₂O) vs. 91.83 μg/ml (BM-EtOH) for brain] activities than BM-EtOH. Both BM extracts showed a weak anti-lipid peroxidation activity in plasma. BM-H₂O (62.0 mg/g) possessed a significant higher concentration of total flavonoids than BM-EtOH (44.0 mg/g), but was lower in the total phenol content (BM-H₂O: 51.6 mg/g vs. BM-EtOH: 68.8 mg/g). In conclusion, BM extracts possess potent antioxidant and free radical scavenging activities. These antioxidant activities could have contributed, at least partly, to the therapeutic benefits of the certain traditional claims of wild BM.     

Polyphenolic compounds characterization and reactive nitrogen species scavenging capacity of Oenothera paradoxa defatted seed extracts

Biological investigations have revealed high scavenging capacity of Oenothera paradoxa defatted seed extract on reactive nitrogen species such as NO and ONOO⁻. The characteristics of the polyphenols present in the extracts were checked using high performance liquid chromatography coupled with electrospray negative ionization ion trap mass spectrometry. Extracts contained five groups of compound: phenolic acids (gallic acid, ethyl gallate, ellagic acid and ferulic acid pentoside), flavanols (catechin, catechin gallate) and oligomeric procyanidins, flavonols (quercetin-3-O-glucoside, quercetin-3-O-glucuronide, quercetin-3-O-pentoside and quercetin), and gallotannins (tetragalloyl glucose, pentagalloyl glucose and hexagalloyl glucose). Penta-O-galloyl-β-d-glucose were present in the extracts in concentrations from 9.44 to 16.75 mg/g, which demonstrated a significant NO and ONOO⁻ scavenging activity with IC₅₀ 0.20 and 0.06 μM, respectively, may be considered as an O. paradoxa extract quality marker.     

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.     

The antioxidant and free-radical scavenging activities of extract and fractions from corn silk (Zea mays L.) and related flavone glycosides

This study was designed to evaluate both the antioxidant and free-radical scavenging activities of extract and fractions from corn silk. N-butanol fraction (BF) demonstrated the highest total phenolic content (164.1 ± 9.7 μg GAE/g DCS) and total flavonoids content (69.4 ± 5.1 μg RE/g DCS), accompanied with the highest antioxidant activity compared to other fractions through all antioxidant assays. Two flavone glycosides showing potent antioxidant activity were isolated from BF and identified, by comparing spectral data (UV, FAB-MS and NMR) with literature values, to be isoorientin-2″-O-α-l-rhamnoside and 3′-methoxymaysin. The two isolated flavone glycosides, particularly isoorientin-2″-O-α-l-rhamnoside, demonstrated significant total antioxidant activity, DPPH radical scavenging activity, reducing power and iron-chelating capacity, with EC₅₀ values of 14.24 ± 1.49, 22.69 ± 2.33, 6.58 ± 1.07 and 30.25 ± 3.05 μg/ml, respectively. Results obtained indicated that corn silk extracts can be used potentially as a ready accessible and valuable bioactive source of natural antioxidants.     

The efficacy of cashew nut (Anacardium occidentale L.) skin extract as a free radical scavenger

The free radical scavenging activity of ethanolic extracts of cashew nut (Anacardium occidentale, L.) skin powder (CSP) was evaluated by employing various in vitro antioxidant assay systems. The yield of the extract as well as the total phenolic content was also determined. The yield of ethanolic extract of the skin powder was quite high (0.45 g/g powder) with a total phenolic content of 243 mg/g extract. The cashew nut skin extract (CSE) demonstrated promising antioxidant activity with EC₅₀ of 1.30 ± 0.02 μg/ml in 2,2′-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radical scavenging assay, 10.69 ± 1.13 μg/ml in superoxide scavenging assay, 17.70 ± 0.05 μg/ml in deoxyribose oxidation assay, 24.66 ± 0.32 μg/ml in lipid peroxidation (LPO) assay and 6.00 mg/ml in iron chelation assay. To identify the compounds in the CSE responsible for the antioxidant activity, thin layer chromatography (TLC) was performed with the extract. The spot showing protection towards β-carotene bleaching was extracted and analyzed by high performance liquid chromatography (HPLC); epicatechin was found to be the major polyphenol present. The results of the present study suggest that cashew nut skin, a byproduct of cashew processing industry, can be used as an economical source of natural antioxidants.     

Antioxidant,anticholinesterase and antimicrobial constituents from the essential oil and ethanol extract of Salvia potentillifolia

The essential oil of Salvia potentillifolia was analysed by GC and GC–MS. Totally, 123 components were detected in both hydrodistilled and steam-distilled oils, α- and β-pinenes being major compounds. The antioxidant activities were determined by using complementary tests, namely, DPPH radical-scavenging, β-carotene-linoleic acid and reducing power assays. The ethanol extract also showed better activity (IC₅₀ = 69.4 ± 0.99 μg/ml) than that of BHT in the DPPH system, and showed great lipid peroxidation inhibition in the β-carotene-linoleic acid system (IC₅₀ = 30.4 ± 0.50 μg/ml). The essential oil showed meaningful butyrylcholinesterase activity (65.7 ± 0.21%), and α-pinene showed high acetylcholinesterase inhibitory activity (IC₅₀ = 86.2 ± 0.96 μM) while β-pinene was inactive. Antimicrobial activity was also investigated on several microorganisms, and the essential oil showed high activity against Bacillus subtilis and B. cereus. It also exhibited remarkable anticandidal activity against Candida albicans and C. tropicalis with MIC values of 18.5 and 15.5 μg/ml, respectively, while α- and β-pinenes showed moderate activity.     

Antioxidant activity of microwave-assisted extract of Buddleia officinalis and its major active component

Buddleia officinalis Maxim, commonly used as rice dye for festivals, was extracted with ethanol using microwave-assisted extraction and Soxhlet extraction. The antioxidant activities of microwave-assisted extract of B. officialis (MEB) and Soxhlet extract of B. officianils (SEB) at the optimum extraction conditions were evaluated and compared with synthetic antioxidant butylated hydroxytoluene (BHT) employing DPPH free radical assay, ABTS assay, total antioxidant activity and reducing power. MEB and SEB had stronger antioxidant activities than BHT in all assays except reducing power, and the effects decreased as follows: MEB > SEB > BHT. The total phenolic contents of MEB and SEB reached 113.56 mg/g and 100.94 mg/g dry weight of extract, respectively, expressed as pyrocatechol equivalents, while the total flavonoids contents were 75.33 mg/g and 62.56 mg/g dry weight of extract, respectively, expressed as catechin equivalents (P < 0.05). Higher phenolic and flavonoids compounds may be major contributors to their higher antioxidant activities. Following activity-oriented separation, luteolin was isolated as an active principle, which exhibited excellent free radical scavenging activities with DPPH IC₅₀ 3.09 μg/ml and ABTS IC₅₀ 2.20 μg/ml.     

Standardised Mangifera indica extract is an ideal antioxidant

The standardised ethanolic and aqueous extracts of Mangifera indica leaf were prepared and analysed for their free radicals scavenging activity. The IC₅₀ values using the DPPH assay were 0.17 ± 0.02 and 0.49 ± 0.4 mg/ml, respectively. Standardised ethanolic extracts of the M. indica leaf had a solid content of 9.1 ± 0.7%, mangiferin concentration of 73 ± 0.17 mg/g of dry weight of the extract, free radical scavenging activity (IC₅₀) of 0.17 ± 0.02 mg/ml and total phenolic content of 590 ± 48 mg/g of extract. The protection exhibited by these extracts against lipid peroxidation was superior to butylated hydroxytoluene (BHT) and commercial grape seed extract. These extracts at higher concentration did not exhibit pro-oxidant activities when compared to vitamin C. Our findings also show that the aqueous and ethanolic extracts of M. indica leaf protect NIH/3T3 cells from oxidant-induced cell death.     

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).     

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.     

Free radical scavenging activities and bioactive substances of Jerusalem artichoke (Helianthus tuberosus L.) leaves

The total phenolic content and radical scavenging activities of Jerusalem artichoke (Helianthus tuberosus L.) leaves were investigated. Results indicated that the ethyl acetate fraction contained the highest total phenolic content (266.69 ± 2.51 mg GAE/g dry extract) accompanied with strongest free radical scavenging abilities. Following an in vitro radical scavenging activity-guide fractionation procedure, six phenolic compounds which strongly quenched free radicals were separated from ethyl acetate fraction. Among them, 3-O-caffeoylquinic acid and 1,5-dicaffeoylquinic acid played a dominant role due to their strong free radical scavenging abilities and their high contents. The content of 3-O-caffeoylquinic acid in n-butanol fraction was 74.58 ± 1.05 mg/g, while 1,5-dicaffeoylquinic acid in ethyl acetate fraction was 104.51 ± 2.86 mg/g. The results imply that the leaves of Jerusalem artichoke might be a potential source of natural antioxidants.     

Antimicrobial and antioxidant properties of the essential oils and methanol extract from Mentha longifolia L. ssp. longifolia

This study was designed to evaluate antimicrobial and antioxidant activities of the essential oil and methanol extract from Mentha longifolia ssp. longifolia. The essential oil showed strong antimicrobial activity against all 30 microorganisms tested whereas the methanol extract almost remained inactive. In contrast, the extract showed much better activity than the essential oil in antioxidant activity assays employed, e.g. in the inhibition of free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) and β-carotene/linoleic acid systems. In the former, the extract was able to reduce the stable free radical DPPH with an IC₅₀ of 57.4 μg/ml while that of the oils was 10 700 μg/ml. When compared to BHT, a synthetic antioxidant, both showed weaker antioxidative potential. Similarly, in β-carotene/linoleic acid assay, these samples were not effectively able to inhibit the linoleic acid oxidation; exhibiting only 24% and 36% inhibitions at 2 mg/ml, respectively; both were far below than that of BHT. Total phenolic constituent of the extract was 4.5 g/100 g as gallic acid equivalent. GC–MS analysis of the oil resulted in the identification of 45 constituents, cis-piperitone epoxide, pulegone and piperitenone oxide being the main components.