Neuronal cell protective and antioxidant effects of phenolics obtained from Zanthoxylum piperitum leaf using in vitro model system

In order to obtain basic data for the utilisation of Zanthoxylum piperitum leaf as a functional substance in the food industry, the antioxidative and neuronal cell protective effects of silica-gel open column chromatographic fractions, obtained from Z. piperitum leaf, were examined. ABTS and FRAP assays indicated that the ZP4 fraction (eluted with methanol/chloroform, 1:4, v/v) of Z. piperitum leaf was a more potent radical-scavenger and reducing agent than the other five fractions. The ZP4 fraction also presented protective effects against H₂O₂-induced neurotoxicity in a dose-dependent manner. Therefore, our study verified that the ZP4 fraction has strong antioxidative and neuronal protective effects which are correlated with its high level of phenolics, particularly quercitrin, afzelin, and hyperoside. These phenolics of Z. piperitum leaf can be utilised as effective and safe functional food substances, i.e., natural antioxidants, and may reduce the risk of neurodegenerative disorders.     

The antioxidant activity and the bioactive compound content of Stevia rebaudiana water extracts

Stevia rebaudiana (SR), a chrysanthemum herb, has been used as a vegetable-based sweetening additive in health drinks and in other foods. This study was conducted to investigate the antioxidant activity and the bioactive compounds found in water extracts taken from SR leaves and calli. Analysis of vitamins in the leaves showed that folic acid (52.18 mg/100 g) was a major component, followed by vitamin C. The total phenolic and flavonoid contents were found to be 130.76 μg catechin and 15.64 μg quercetin for leaves and 43.99 μg catechin and 1.57 μg quercetin for cellus at mg of water extracts, respectively. Pyrogallol was the major material among the phenolic compounds in both leaf and callus extracts. Furthermore, our results showed that the leaf extracts contained higher amounts of free radicals, hydroxyl radicals and superoxide anion radical scavenging activities than those of the callus extracts.     

Antioxidant and anti-proliferative activity of Rhizoma Smilacis Chinae extracts and main constituents

Rhizoma Smilacis Chinae (RSC) is a widely used herbal material in functional food and folk medicine. In this study, methanol extract (ME), water extract (WE), polysaccharide fraction and ethyl acetate fraction (EF) of RSC were prepared and the constituents were analysed by HPLC. Different antioxidant tests were employed to evaluate the antioxidant activities of RSC extracts and its main constituents, astilbin and chlorogenic acid. The results showed that RSC extracts possessed comparable antioxidant activity to butylated hydroxyanisole in a dose-dependent manner. The radical-scavenging capacity of ME and EF was even stronger than astilbin and chlorogenic acid. The EF and ME of RSC also showed stronger anti-proliferative activity on HepG2 cells than astilbin and chlorogenic acid, with IC₅₀ values of 47 and 32 μg/mL for 24 h treatment, respectively. Flow cytometric analysis revealed that RSC extracts induced cell cycle arrest at G2/M phase and a late apoptosis of the cells.     

In vitro antioxidant activities of methanol extracts of five Phyllanthus species from India

In this study, the antioxidant activity of methanol extracts of five plants from the genus Phyllanthus was evaluated by various antioxidant assays, including total antioxidant, free radical scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging, nitric oxide scavenging, reducing power and metal ion chelating activities. The various antioxidant activities were compared to standard antioxidants such as butylated hydroxytoluene and ascorbic acid. All the extracts showed strong antioxidant activity in all the tested methods. Among the five plants Phyllanthus debilis has been found to possess the highest activity in all tested models, the activity decreased in the order Phyllanthus debilis>Phyllanthus urinaria>Phyllanthus virgatus>Phyllanthus maderaspatensis>Phyllanthus amarus. In addition to the antioxidant activity of these plants, the total phenolic compounds, flavonoids and flavonols were measured in the extracts. A correlation between the antioxidant activity and total phenolic content was observed.     

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.     

Screening of the antioxidant potentials of six Salvia species from Turkey

This study was designed to examine the in vitro antioxidant activities of the methanol extracts of six Salvia species [Salvia caespitosa Montbret & Aucher ex Bentham (ENDEMIC), Salvia hypargeia Fisch. & Mey. (ENDEMIC), Salvia euphratica subsp. euphratica Montbret & Aucher ex Bentham (ENDEMIC), Salvia sclarea L., Salvia candidissima subsp. candidissima Montbret & Aucher ex Bentham and Salvia aethiopis L.] from Turkey. The extracts were screened for their possible antioxidant activities by two complementary test systems, namely DPPH free radical-scavenging and β-carotene/linoleic acid systems. Non-polar subfractions of the methanol extracts of Salvia species studied did not show any antioxidant activity in both test systems. In the first case, the most active plant was S. euphratica subsp. euphratica, an endemic species, with an IC₅₀ value of 20.7 ± 1.22 μg/ml, followed by S. sclarea (IC₅₀ = 23.4 ± 0.97 μg/ml) among the polar subfractions. In the β-carotene/linoleic acid test system, polar extract of S. hypargeia was superior to the polar extracts of other Salvia species studied (69.2% ± 1.90%). This activity was followed by S. sclarea with 63.5% ± 4.24% inhibition rate. The inhibition rate of the synthetic antioxidant, buthylated hydroxytoluene (BHT), was also determined to be 96%. Since the polar extracts of Salvia species dealt with here exhibited excellent antioxidant activities when compared to BHT, it seems possible to keep perishable fat-containing food longer by direct addition of an extract of sage.     

Loss of rutin and antioxidant activity of asparagus juice caused by a pectolytic enzyme preparation from Aspergillus niger

We found that a commercial pectolytic enzyme preparation from Aspergillus niger (pectinase AN) contained laccase activity that decreased rutin content and antioxidant activity of asparagus juice. This research investigated the effects of pH, temperature, and concentration of pectinase AN on pectinase AN’s laccase activity to decrease rutin content and antioxidant activity of asparagus juice. Asparagus juice was incubated with pectinase AN at different pHs (3.2, 4.5 and 5.8), temperatures (25, 37, and 50 °C) and enzyme concentrations (0.1%, 0.5% and 1%). Rutin content and antioxidant activity of samples was determined by HPLC and 2,2′-diphenyl-1-picrylhydrazyl (DPPH) free radical method, respectively. The rate of loss of rutin and antioxidant activity of asparagus juice was smaller at pH 3.2 than at pH 4.5 and pH 5.8, smaller for 0.1% pectinase AN than 0.5% and 1% pectinase AN. The rate of loss of rutin of asparagus juice was greater at 25 °C than at the other two temperatures. Pectinase AN can decrease rutin content and antioxidant activity of asparagus juice at the selected conditions. But rutin content and antioxidant activity of asparagus juice produced using pectinase AN could be less decreased at pH 3.2 and 0.1% of enzyme with less than 2 h of incubation time. This information was helpful for juice industry to produce juices with high antioxidant activity using pectinase AN.     

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.     

In vitro studies to assess the antidiabetic,anti-cholinesterase and antioxidant potential of Spergularia rubra

Spergularia rubra is distributed all over the world, being its infusion used as diuretic. In spite of its large use, the antidiabetic, anti-cholinesterase and antioxidant activities of this species have not been assessed and its chemical composition is scarcely known. In the work herein a hydromethanolic extract was studied.     

Antioxidant activity of the essential oil and various extracts of Nepeta flavida Hub.-Mor. from Turkey

This study was designed to examine the chemical composition and in vitro antioxidant activity of the essential oil and various extracts (hexane, dichloromethane and methanol sub-fractions) of Nepeta flavida. GC and GC–MS analyses of the essential oil resulted in the identification of 68 compounds, representing 96.4% of the oil; 1,8-cineole (38.9%) and linalool (25.1%) were the main components, comprising 64.0% of the total oil. The samples were subjected to a screening for their possible antioxidant activities by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and β-carotene-linoleic acid assays. In the first case, the IC₅₀ value of the N. flavida essential oil was determined to be 42.8 ± 2.19 μg/ml. Among the extracts, the strongest activity was exhibited by the polar sub-fraction of the methanol extract with an IC₅₀ value of 63.2 ± 1.75 μg/ml. In the β-carotene-linoleic acid system, N. flavida essential oil exhibited 86.3% ± 1.69 inhibition against linoleic acid oxidation. Among the extracts prepared with various solvents, a correlation was observed between the polarity and antioxidant activity. The extracts exhibited the same activity pattern in this system the most active one is the polar sub-fraction, 79.7% ± 0.89. On the other hand, 1,8-cineole, a major compound of the essential oil, exhibited marked antioxidant activity in both systems, whereas the other compound, linalool, did not show any activity. The amount of total phenolics was highest in the polar and non-polar sub-fractions. Particularly, a positive correlation was observed between the total phenolic content and the antioxidant activity of the extracts. As estimated from the results, amounts of phenolic compounds were less in hexane and dichloromethane extracts than in the others. In conclusion, antioxidant potentials of polar and non-polar methanol sub-fractions could be attributed to their high phenolic contents. In both systems, antioxidant capacities of BHT, ascorbic acid, curcumin and α-tocopherol were also determined in parallel experiments.     

Antiacetylcholinesterase and antioxidant activities of Plectranthus barbatus tea,after in vitro gastrointestinal metabolism

This study describes the in vitro metabolism, by the gastrointestinal tract, together with the biological activity of the herbal tea obtained from Plectranthus barbatus leaves. The results showed that there was no appreciable degradation and that the activity was kept constant after gastric juice digestion. In the presence of pancreatic juice, degradations varied from negligible, in the case of flavonoid glucuronides, to almost complete, in the case of the abietane diterpenoid. The digested decoction contained only 50% of its initial biological activity, after pancreatic digestion. The action of Caco-2 cells on the extract revealed that neither rosmarinic acid, the main compound of the extract, nor the other components present in minute quantities were metabolised by the intact cells. Rosmarinic acid could be found inside Caco-2 cells although in trace amount. Glucoronidase from Escherichia coli, a gut bacterium, was able to hydrolyse the flavonoid derivatives, thus the aglycones were formed and permeate the Caco-2 cells.     

Enhanced antioxidant activity of Monascuspilosus fermented products by addition of ginger to the medium

The fermented products of Monascus sp. are known for their antihypercholesterolaemic effects, however, their antioxidant activities are different from those of many plant-derived foods. To evaluate the effect of ginger addition into the medium on the antioxidant activity of Monascuspilosus fermented products, we cultured uninoculated PDB medium (PDB), inoculated PDB medium (MP), uninoculated ginger-containing medium (PDBG), and inoculated ginger-containing medium (MPG). The broth and mycelia were collected, freeze-dried, and extracted to evaluate their free radical scavenging activities, inhibition of peroxidation, phenolic content, inhibition of DNA damage, cellular antioxidant activity, and expression of the antioxidant enzymes. The results showed that MPG had significantly higher antioxidant activity than PDB, MP, and PDBG at all fermentation time points. Moreover, the fermentation process significantly increased the antioxidant activities of MPG. After the inherent level of antioxidant capacity was increased, the modified M. pilosus fermented product demonstrated a higher anti-atherosclerotic value than the unmodified product.     

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.     

Chemical composition, mineral content and antioxidant activity of Verbena officinalis L.

Aqueous and hydroalcoholic extracts from Verbena officinalis L. were obtained and characterised. The analysis by HPLC-DAD and LC-MS allowed the detection and identification of three iridoids, fifteen flavonoids and four phenolic acid derivatives. Four flavonoids, scutellarein 7-diglucuronide (9), scutellarein 7-glucuronide (13), pedalitin 6-galactoside (15) and scutellarein 7-glucoside (19) are reported for the first time from this plant. In addition, three new flavonoids have been isolated: scutellarein 7-O-(2-O-feruloyl)-diglucuronide (5), pedalitin 6-O-diglucuronide (6) and pedalitin 6-O-(2-O-feruloyl)-diglucuronide (13). To our knowledge, these flavonoids have not been reported as natural products. Both extracts showed significant antioxidant activity using three in vitro model systems and the results have been correlated with total phenolic and total flavonoid contents. The results have allowed establishing an important relation structure-activity and significant correlations have also been found between the mineral content and the flavonoids present in both extracts.     

Evaluation of the antioxidant activity of Ruellia tuberosa

The antioxidant activity of Ruellia tuberosa L. (Acanthaceae) was investigated by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical-scavenging assay and the hydrogen peroxide-induced luminol chemiluminescence assay. The methanolic extract (ME) and its four fractions of water (WtF), ethyl acetate (EaF), chloroform (CfF), and n-hexane (HxF) were prepared and then subjected to antioxidant evaluation. The results of both methods revealed that R. tuberosa possesses potent antioxidant activity. The antioxidant activities of the different fractions tested decreased in the order of EaF > CfF > ME > WtF > HxF according to the hydrogen peroxide-induced luminol chemiluminescence assay, and results were the same with the exception of the rank order of HxF and WtF according to the DPPH free radical-scavenging assay. The results provide useful information on the pharmacological activities associated with free radicals of this traditional folk remedy.     

Flavonols, betacyanins content and antioxidant activity of cactus Opuntia macrorhiza fruits

Flavonols, betacyanins content and antioxidant activity of red-purple cactus Opuntia macrorhiza fruits, a promising cactus pear species, were evaluated in comparison to the well known cactus pear fruits of Opuntia ficus-indica. Flavonol profile was evaluated by HPLC-DAD prior to and after enzymatic hydrolysis (glycosides vs. aglycons). In addition betacyanins, responsible for the purple to red color of cactus pear fruits were also estimated and compared to Beta vulgaris ssp. (roots) and red O. ficus-indica fruits. In all cases, cactus fruit pulps showed no flavonols at all. While different derivatives of isorhamnetin glycosides were found in O. ficus-indica fruit's peel, isorhamnetin-3-O-rutinoside was the only compound in O. macrorhiza fruit. Correspondingly, antioxidant activity assays showed a high antioxidant activity of both, O. macrorhiza fruit's peel and pulp. With regard to betacyanins, O. macrorhiza fruit's peel and pulp provide a deep red-purple color, whose average impact is higher compared to red beet (B. vulgaris spp.) and about 8-fold higher than red fruits from O. ficus-indica. Supporting these results, estimation of color attributes (L*, a*, b*, C and H°) showed also a high color impact of both O. macrorhiza fruit's peel and pulp.