Chemical composition and in vitro antioxidant evaluation of a water-soluble Moldavian balm (Dracocephalum moldavica L.) extract

The chemical composition and antioxidant properties of a water-soluble extract of Moldavian balm (Dracocephalum moldavica L., syn. Moldavian dragonhead) prepared by hydrodistillation are presented in this study. The total phenol content was estimated as gallic acid equivalents by the Folin-Ciocalteu reagent method, while the qualitative-quantitative composition of the extract was determined by high performance liquid chromatography coupled with photodiode array detection. The antioxidant properties assessed included iron(III) reduction and iron(II) chelation and 1,1-diphenyl-2-picrylhydrazyl, 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonate) and superoxide anion free radical scavenging. In addition, the ability of the extract to protect 2-deoxy-d-ribose and bovine brain-derived phospholipids against hydroxyl radical-mediated degradation was assessed. The extract principally contained polar compounds including hydroxycinnamic acids and flavonoids, with caffeic and ferulic acids, luteolin-7-O-glucoside, rosmarinic acid, luteolin and apigenin being identified from their chromatographic behavior and spectral characteristics. The Moldavian balm extract demonstrated activity in all the antioxidant assays; however, it was not as potent as the positive control except in the phospholipid-based assay where its hydroxyl radical scavenging activity was statistically indistinguishable from that demonstrated by Pycnogenol.     

Chemical composition and in vitro antioxidative activity of a lemon balm (Melissa officinalis L.) extract

The leaf material of lemon balm (Melissa officinalis L.) was extracted with 450 ml/l aqueous ethanol by medium pressure liquid-solid extraction. The total phenolic content of the extract was estimated as gallic acid equivalents by Folin-Ciocalteu reagent method and a qualitative-quantitative compositional analysis was carried out using high performance liquid chromatography coupled with photodiode array detection. The lemon balm extract contained hydroxycinnamic acid derivatives and flavonoids with caffeic acid, m-coumaric acid, eriodictyol-7-O-glucoside, naringin, hesperidin, rosmarinic acid, naringenin, hesperetin being identified based on their chromatographic behaviour and spectral characteristics. The extract was also investigated for potential in vitro antioxidant properties in iron(III) reduction, iron(II) chelation, 1,1-diphenyl-2-picrylhydrazyl, 2,2′-azinobis(3-ethylbenzothiazoline-6-sulphonate), superoxide anion and nitric oxide free-radical scavenging, and inhibition of β-carotene-linoleic acid bleaching assays. The extract demonstrated antioxidant activity in all the assays. However, it was not as potent as the positive controls except in the β-carotene-linoleic acid bleaching assay, where its activity was superior to that of gallic and caffeic acids and statistically indistinguishable from quercetin and BHA. The exceptionally high antioxidant activity and the fact that this assay is of biological relevance warrants further investigation of lemon balm extract in ex vivo and in vivo models of oxidative stress.     

Bioprospecting traditional Pakistani medicinal plants for potent antioxidants

Antioxidant potential of four methanol extracts from three selected plant species, namely Salvia nubicola (Lamiaceae), Acer oblongifolium (Aceraceae) and Hedera nepalensis (Araliaceae) was measured using assays in aqueous and lipid systems. Antioxidant activities were investigated in aqueous systems by using DPPH radical-scavenging assay, ABTS radical-scavenging assay and DNA protection assay, while antioxidant activity in a lipid system was determined by using the thiobarbituric acid-reactive substances (TBARS) assay. Additionally, the Folin-Ciocalteu method was used to measure total phenolic content. Methanol extracts of leaves and flowers of S. nubicola showed the highest Trolox equivalent (TE) values in the case of the DPPH assay, 2484 ± 4.9 mmol TE/g extract, as well as total phenolic content, 139 ± 0.2 mg gallic acid equivalents/g extract. Three fractions (A-C) of the methanol extract of S. nubicola leaves and flowers were produced by semi-preparative HPLC. Fraction B was found to be the most active in the DPPH radical-scavenging assay and had the highest total phenol content. HPLC-DAD and LC-MS revealed rosmarinic acid in S. nubicola extracts and chlorogenic acid and rutin in H. nepalensis extracts as the main phenolic antioxidants.     

Antioxidant properties of various solvent extracts of mulberry (Morus indica L.) leaves

The antioxidant properties and total phenolic contents of methanol, acetone and water extracts of mulberry (Morus indica L.) leaves were examined. Various experimental models including iron (III) reducing capacity, total antioxidant capacity, DPPH radical scavenging activity and in vitro inhibition of ferrous sulphate-induced oxidation of lipid system were used for characterization of antioxidant activity of extracts. The three extracts showed varying degrees of efficacy in each assay in a dose-dependent manner. Methanolic extract with the highest amount of total phenolics, was the most potent antioxidant in all the assays used. In addition, the effect of temperature (50 °C and 100 °C), pH (3, 5, 7, 9 and 11) and storage (5 °C) on the antioxidant activity of methanolic extract was investigated. The antioxidant activity of the extract remained unchanged at 50 °C and was maximum at neutral pH. The extract stored at 5 °C in the dark was stable for 30 days after which the antioxidant activity decreased (p ? 0.05) gradually. On the basis of the results obtained, mulberry leaves were found to serve as a potential source of natural antioxidants due to their marked antioxidant activity.     

Antioxidant assessment of an anthocyanin-enriched blackberry extract

Gel filtration of black berry (Rubus fruticosus sp) ethanolic extracts was employed to obtain an anthocyanin-enriched extract. The anthocyanin profile identified cyanidin-3-glucoside as the primary (e.g., 90% of total) anthocyanin present in blackberry. Gel filtration of crude extracts resulted in a 20-fold increase in total anthocyanin content, with no change in the proportion of cyanidin-3-glucoside. Antioxidant activities of both the crude and anthocyanin-enriched blackberry extracts were determined using cell-free (ORAC) and cell-based (INT-407 intracellular) antioxidant assays. Antioxidant activity, assessed by the ORAC assay, indicated a 7-fold increase in activity for the anthocyanin-enriched fraction. Similar results were obtained for the anthocyanin-enriched extract using the intracellular antioxidant assay with INT-407 cells. Our results indicate that the anthocyanin content, and more specifically the presence of cyanidin-3-glucoside, in blackberry, contributes a major part of the antioxidant ability to suppress both peroxyl radical-induced chemical and intracellular oxidation.     

Antioxidant activities of Sideritis congesta Davis et Huber-Morath and Sideritis arguta Boiss et Heldr: Identification of free flavonoids and cinnamic acid derivatives

Different solvent extracts of endemic Sideritis (Labiatae) species, Sideritis congesta Davis et Huber-Morath and Sideritis arguta Boiss et Heldr, were analyzed for free flavonoids (quercetin, apigenin, myricetin and kaempferol) and cinnamic acid derivatives (rosmarinic acid, ferulic acid, caffeic acid, p-coumaric acid and chlorogenic acid) using HPLC-DAD. All the phenolics were quantified in acid-hydrolyzed extracts, except rosmarinic acid, chlorogenic acid and myricetin which were quantified in raw samples. Antioxidant activities of extracts of these two plants and many of their components in pure form were evaluated based on DPPH^. and ABTS^.+ assays. In general, S. arguta extracts displayed higher antioxidant activity than S. congesta extracts possibly due to their richness in antioxidant components of strong activity. Acetone extract of S. arguta, with its strikingly high TEAC value of 3.2 mM trolox and low IC₅₀ value of 38.3 ??g/mL showed the highest antioxidant potency among all extracts. ??-tocopherol, the positive control, displayed IC₅₀ and TEAC values of 33.8 ??g/mL and 2.9 mM trolox, respectively. No direct correlation was found between antioxidant activities and total phenolic contents of the plant extracts studied.     

Non-toxic Salvia sclareoides Brot. extracts as a source of functional food ingredients: Phenolic profile,antioxidant activity and prion binding properties

Salvia sclareoides is an aromatic herb native to Portugal, of which phenolic content (Folin–Ciocalteau method), chemical profile (HPLC/DAD), antioxidant activity (DPPH, β-carotene/linoleic acid assays), acute toxicity (MTT method, adapted for non-adherent cells), genotoxicity (short-term chromosomal aberration assay) and prion binding properties were evaluated in the acetone, water, ethanol, methanol and n-butanol extracts. The latter presented the highest phenolic content and antioxidant activity (DPPH assay), and was the single one with the flavonoids (+)-catechin, kaempferol O-glucoside and quercetin. Vanillic acid was the major component of all extracts but gallic, gentisic, caffeic, syringic, coumaric and ferulic acids were also found in some extracts. Only the n-butanol extract had components binding to the cellular form of human prion protein detected by NMR which showed specificity for two regions of the folded domain and for the unstructured N-terminal region. Extracts were not cytotoxic nor genotoxic, reinforcing the potential of S. sclareoides for nutraceutical purposes.     

Determination of free phenolic acids and antioxidant activity of methanolic extracts obtained from fruits and leaves of Chenopodium album

In this study, determination of phenolic acids as well as investigation of antioxidant activity of methanolic extracts from the fruits and leaves of Chenopodium album is described. Extracts were subjected to acidic hydrolysis in order to obtain total free phenolic acids. However, some of phenolic acids were identified and quantified by HPLC-DAD. The results were confirmed by LC-MS equipped with MS-ESI. In addition, Folin–Ciocalteu method was applied to determine the total phenolic contents. The antioxidant activity of C. album extracts was examined by using DPPH and hydroxyl radical-scavenging activity assays. Results revealed that the leaves extract exhibits better performance in antioxidant assays and in the higher total phenolic contents (3066 mg of GAE/100 g) when compared to fruits extract (1385 mg of GAE/100 g). From these results it has been revealed that the methanolic extracts of C. album from fruits and leaves have great potential as a source for natural health products.     

Antioxidant activity and phenolic compounds of Holotrichia parallela Motschulsky extracts

Insects have been relatively unexplored as potential sources of natural antioxidants. We report the antioxidant activity of extracts of the adult large black chafer beetle Holotrichia parallela Motschulsky, a common crop pest in China. The antioxidant activity of the ethanolic extract (EE) and the water extract (WE) of adult H. parallela were evaluated by four different in vitro assays. EE showed potent metal-chelating activity and inhibition of lipid peroxidation. WE proved to be an excellent antioxidant in the scavenging of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals and metal-chelating activity. Catechin was identified in the ethanolic extract and proteins were the main components in the water extracts. Both compounds could contribute to the antioxidant activity of the species. These results suggest that adult H. parallela might be used as a nutraceutical to alleviate oxidate-induced diseases and as a natural antioxidant additive in the food industry.     

A detailed study on the antioxidant activity of the stem bark of Dalbergia sissoo Roxb., an Indian medicinal plant

A detailed study was performed on the antioxidant activity of the aqueous and methanol extracts (AED and MED respectively) of the stem bark of the plant, Dalbergia sissoo Roxb. (Fam. Fabaceae), also known as Indian Rosewood. The antioxidant activity of the extracts was measured by in vitro chemical analyses involving the assays of (1) 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity (2) ferric ion reducing power (3) ferrous ion chelating activity and (4) Au nanoparticle formation potential. A simplified method was developed based on Au nanoparticles formation to assess the antioxidant activity of any plant extract, and was used for the first time to assay the antioxidant activity of AED and MED. In all the assays, AED showed significantly greater activity over MED. This work provides a scientific support for the high antioxidant activity of this plant and thus it may find potential applications in the treatment of the diseases caused by ROS.     

The hydroxycinnamic acid content of barley and brewers’ spent grain (BSG) and the potential to incorporate phenolic extracts of BSG as antioxidants into fruit beverages

The hydroxycinnamic acid (HA) content of starting barley for brewers’ spent grains (BSG), whole BSG and phenolic extracts from BSG was measured using high performance liquid chromatography (HPLC) and correlated with antioxidant potential. The effect of BSG phenolic extracts on antioxidant activity of fruit beverages was also assessed (using the total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays). The concentration of HA present in barley extract and BSG was in the order of ferulic acid (FA), p-coumaric acid (p-CA) derivatives, FA derivatives, p-CA, caffeic acid (CA) and CA derivatives. Results suggested that brewing and roasting decreased the HA content. Antioxidant activity was significantly (P < 0.05) correlated with caffeic acid (R² = 0.8309) and total HA (R² = 0.3942) concentrations. Addition of extracts to fruit beverages resulted in a significant (P < 0.05) increase in antioxidant activity of cranberry juice, measured by the FRAP assay. In vitro digestion significantly (P < 0.05) reduced TPC, DPPH and FRAP activity of the fruit beverages.     

Profiling of in vitro neurobiological effects and phenolic acids of selected endemic Salvia species

The ethyl acetate and methanol extracts from 16 Salvia L. species were screened for their inhibitory activity against acetylcholinesterase, butyrylcholinesterase, lipoxygenase, and tyrosinase; the enzymes linked to neurodegeneration. Their antioxidant activity was also tested using DPPH radical scavenging, metal-chelation, and ferric-reducing antioxidant power (FRAP) assays. Total flavonoid content of the extracts was determined by AlCl₃ reagent, while HPLC technique was applied for analysis of various phenolic acids in the extracts. The extracts exerted weak cholinesterase and tyrosinase inhibition, and remarkable inhibition against lipoxygenase (13.07 ± 2.73-74.21 ± 5.61%) at 100 μg ml⁻¹. The methanol extracts showed higher antioxidant activity in DPPH radical scavenging and FRAP assays. The extracts were analyzed for their gallic, protocateuchic, p-hydroxy-benzoic, vanillic, caffeic, chlorogenic, syringic, o- and p-coumaric, ferulic, rosmarinic, and tr-cinnamic acid contents and the methanol extract of Salvia ekimiana (153.50 mg 100 g⁻¹) was revealed to be the richest in terms of rosmarinic acid.     

Phenolic composition,antioxidant and acetylcholinesterase inhibitory activities of Sclerocarya birrea and Harpephyllum caffrum (Anacardiaceae) extracts

Total phenolic content, proanthocyanidins, gallotannins, flavonoids, and antioxidant activities of Sclerocarya birrea and Harpephyllum caffrum methanolic extracts were evaluated using in vitro assays. S. birrea young stem extract contained the highest levels of total phenolic content (14.15 ± 0.03 mg GAE/g), flavonoids (1.21 ± 0.01 mg CE/g) and gallotannins (0.24 ± 0.00 mg GAE/g). H. caffrum stem bark extract had the highest content of proanthocyanidins (1.47%). The EC₅₀ values of the extracts in the DPPH free radical scavenging assay ranged from 4.26 to 6.92 μg/ml, compared to 6.86 μg/ml for ascorbic acid. A dose-dependent linear curve was obtained for all extracts in the ferric-reducing power assay. Dichloromethane and methanol extracts exhibited dose-dependent acetylcholinesterase inhibitory activity. Similarly, all extracts exhibited high antioxidant activity comparable to butylated hydroxytoluene based on the rate of β-carotene bleaching (84.1–93.9%). The two Anacardiaceae species provide a source of natural antioxidants and acetylcholinesterase inhibitors, and may be beneficial to the health of consumers.     

Antioxidant potential of crocins and ethanol extracts of Gardenia jasminoides ELLIS and Crocus sativus L.: A relationship investigation between antioxidant activity and crocin contents

Crocins are water-soluble carotenoids responsible for the colour of saffron and gardenia. In this study, we isolated and identified three major crocins from gardenia, and then evaluated their antioxidant potential using four in vitro antioxidant tests in comparison with saffron ethanol extract (SE), gardenia ethanol extract (GE) and gardenia resin fraction (GRF). The relationship between total crocin contents and antioxidant activity of ethanol extracts of two herbs was investigated and the antioxidant potentials of three different polar crocins were compared. The crocins appeared to possess antioxidant activity when tested by four in vitro antioxidant models. However, in anti-hemolysis, DPPH radical-scavenging and lipid peroxidation assays, GRF exhibited significantly stronger antioxidant activity than crocins and no correlation between total crocin contents and antioxidative function was revealed, which implied that ingredients other than crocins in gardenia gave markedly strong antioxidant activity. In the phosphomolybdenum assay, antioxidant capacities of fractions and extracts correlated with total crocin contents (R = 0.93). Moreover, comparison of results indicated that sugars attached to the crocetin moiety seemed to be beneficial for the antioxidant activity of these water-soluble pigments.     

Antioxidant activity of ethanolic extract of Cortex fraxini and use in peanut oil

Cortex fraxini was extracted with 95% ethanol to obtain a crude antioxidant extract. The antioxidant activity was evaluated using the linoleic acid peroxidation method and the free radical scavenging assays, namely 2,2′-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl radicals. Cortex fraxini extract (CFE) showed high inhibition of peroxidation of linoleic acid when compared with butylated hydroxytoluene (BHT). CFE also exhibited excellent scavenging activity on DPPH and hydroxyl radicals. Total antioxidant activity was measured by the reduction of Mo(VI) to Mo(V) by the extract, and subsequent formation of a green phosphate/Mo(V) complex at acid pH. CFE had significant total antioxidant activity and the effects were increased with increasing reaction time. The total phenolic content of the sample, analyzed by using Folin–Ciocalteu’s reagent, was 91.33 mg/g dry weight expressed as pyrocatechol equivalents. Then the suitability of CFE as an antioxidant was determined in peanut oil, and the decrease of lipid oxidation was monitored using thiobarbituric acid-reactive substances (TBARS) assay. CFE treatment significantly (P < 0.05) reduced lipid oxidation in peanut oil compared to the control. No significant differences (P = 0.05) in lipid oxidation were detected between CFE antioxidant and BHT antioxidant samples.     

Evaluation of antioxidant ability of ethanolic extract from dill (Anethum graveolens L.) flower

Antioxidant activities of ethanolic extract from dill flower and its various fractions were evaluated with 2,2-diphenyl-1-picrylhydrazyl radical scavenging, Trolox equivalent antioxidant capacity, reducing power, chelating power, and β-carotene bleaching assays. The flower extract was successively separated into n-hexane, ethyl acetate and ethanol soluble fractions by liquid–liquid partition. Dill leaf and seed extracts were used for comparison. In all assays, the flower extract showed higher antioxidant activity than the leaf and seed extracts. With regard to various fractions of the flower extract, the sequence for antioxidant activity was ethyl acetate fraction > ethanol fraction > original flower extract > n-hexane fraction. Phenols including flavonoids and proanthocyanidins should be responsible for antioxidant abilities of the flower extract. Chlorogenic acid, myricetin, and 3,3’,4′,5,7-pentahydoxyflavan (4 → 8)-3,3′,4′,5,7-pentahydoxyflavan were the major phenolic acid, flavonoid, and proanthocyanidin, respectively, in the dill flower extract.     

Concentration of biologically active compounds extracted from Ilex paraguariensis St. Hil. by nanofiltration

The aim of this study was to characterise the bioactive compounds in mate (Ilex paraguariensis St. Hil) extract and in concentrated mate extract obtained by nanofiltration (NF). Also, the impact of NF on the antioxidant activity of both mate extracts was evaluated in vitro and using eukaryotic cells of Saccharomyces cerevisiae (yeast assay). The results showed a significant increase in the contents of total phenolics (338%), chlorogenic acid (483%), theobromine (323%), caffeine (251%), chlorophyll (321%), condensed tannins (278%) and saponins (211%) in the concentrated mate extract. The concentrated mate extract showed higher in vitro antioxidant activity than the mate extract. According to the results obtained, it can be stated that the use of nanofiltration membrane is a valid approach for the concentration of biologically active compounds in aqueous extract of mate.     

Antioxidant and antimicrobial activity of Cynara cardunculus extracts

The whole, fresh involucral bracts of cardoon, Cynara cardunculus L. (Compositae), were extracted with EtOH and an aqueous suspension of the obtained EtOH extract was partitioned successively with CHCl₃, EtOAc and n-BuOH, leaving a residual water extract. All obtained extracts were evaluated for their antioxidant and antimicrobial properties. The antioxidant potential was evaluated using following in vitro methods: FRAP (ferric reducing antioxidant power) assay, and scavenging of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical. Antimicrobial activity was estimated using a microdilution technique against food-borne, mycotoxin producers and human pathogenic bacteria and micromycetes. The following bacteria were tested: Salmonella typhimurium, Escherichia coli, Bacillus subtilis, Staphylococcus epidermidis, Staphylococcus aureus, as well as micromycetes: Aspergillus niger, Aspergillus ochraceus, Aspergillus flavus, Penicillium ochrochloron, Penicillium funiculosum, Trichoderma viride, Fusarium tricinctum and Alternaria alternata. Results showed that all extracts possessed concentration-dependent antioxidant activity. In biological assays, C. cardunculus extracts showed antimicrobial activity comparable with standard antibiotics.