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.     

Estimation of neuroprotective effects of Laurocerasus officinalis Roem. (cherry laurel) by in vitro methods

Dichloromethane, ethyl acetate, acetone, methanol, and water extracts prepared from the fruits and leaves of Laurocerasus officinalis Roem. (LO) (Rosaceae) were screened for their cholinesterase inhibitory activity against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), the key enzymes in pathogenesis of Alzheimer's disease (AD), using ELISA microplate reader at 50, 100, and 200 ??g mL⁻¹. As AD is associated with oxidative stress, the antioxidant activity of the extracts was also tested by radical-forming methods against 2,2-diphenyl-1-picrylhydrazyl (DPPH), N,N-dimethyl-p-phenylendiamine (DMPD), and superoxide radicals as well as iron-related methods; iron-chelating capacity and ferric-reducing antioxidant power (FRAP) assays. Total phenol and flavonoid quantification was achieved using Folin-Ciocalteau and AlCl₃ reagents, respectively. The highest AChE (44.01 ± 1.75%) and BChE (19.91 ± 0.37%) inhibition was caused by the LO-leaf-methanol extract 200 ??g mL⁻¹, while it showed the best radical-scavenging activity against DPPH at 2000 ??g mL⁻¹. Only, the dichloromethane and water extracts of the fruits and the leaf water extract had an iron-chelating capacity, while the leaf methanol extract displayed the highest FRAP. The leaf methanol extract (113.45 ± 0.71 mg g⁻¹ extract) was found to be the richest in total phenols, while the leaf acetone extract (139.90 ± 4.64 mg g⁻¹ extract) had the most abundant amount of total flavonoids.     

Neuroprotective potential of some terebinth coffee brands and the unprocessed fruits of Pistacia terebinthus L. and their fatty and essential oil analyses

In the current study, neuroprotective effects of the ethyl acetate and methanol extracts of four terebinth coffee brands and the fruits of Pistacia terebinthus L. were investigated through enzyme inhibition tests against acetylcholinesterase, butyrylcholinesterase, and tyrosinase as well as antioxidant test systems. Antioxidant activity was measured using radical scavenging activity tests and metal-related tests including metal-chelation capacity, ferric-reducing antioxidant power (FRAP), and phosphomolybdenum reducing power (PRAP). The fatty oils of the coffee brands and the fruits and the fruit essential oil were examined by GC–MS. Total phenol and flavonoid contents were calculated spectrophotometrically. The extracts had moderate inhibition against butyrylcholinesterase (9.78–45.74% at 200 μg mL⁻¹) and potent scavenging activity against DPPH. They exerted strong activity in FRAP and metal-chelation tests and modest activity in PRAP test. Oleic acid was identified as the major fatty acid in the fatty oils, while α-pinene (26.31%) was dominant in the essential oil.     

Antioxidant activity and rosmarinic acid changes in salicylic acid-treated Thymus membranaceus shoots

In order to characterise the antioxidant activity of Thymus membranaceus Boiss. subsp. membranaceus, an endemic species in Southeast Spain, five different analytical methods were used. Water, methanol and hexane extracts obtained from 60-day-old in vitro-grown shoots were assayed for their antioxidative properties using DPPH (1,1-diphenyl-2-picrylhydrazyl radical), ABTS (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonate), FRAP (ferric reducing antioxidant power), reducing power and conjugated diene assays. Total soluble phenol content, as well as rosmarinic acid, in the different extracts were also determined. Methanolic extracts exhibited the best antioxidant activity and the highest amounts of total soluble phenolics. A single application of salicylic acid (10 μM) on culture media resulted in an increase in rosmarinic acid and phenolic levels, which in turn improved the extracts’ antioxidant properties. These current findings open new opportunities for obtaining valuable natural antioxidants for commercial exploitation by using tissue culture systems.     

Polyphenol contents and in vitro antioxidant activities of lyophilised aqueous extract of kiwifruit (Actinidia deliciosa)

The aim of this study was to determine the antioxidant potency and total phenolic and flavonoid contents of kiwifruit (Actinidia deliciosa) in vitro by analysing the radical scavenging activity of lyophilised water extract from kiwifruit (LEK) for 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), 1,1-diphenyl-2-picryl-hydrazyl (DPPH), N,N-dimethyl-p-phenylenediamine (DMPD), and superoxide anion radical (O₂⁻) as well as the total reducing power by FRAP and CUPRAC assays and the metal chelating activities. LEK showed efficient radical scavenging activity with DPPH, ABTS, DMPD, and O₂⁻ radicals; ferric (Fe³⁺) and cupric (Cu²⁺) ion reducing power and metal chelating activities. Moreover, the amounts of phenolic compounds, such as caffeic acid, ferulic acid, syringic acid, ellagic acid, catechol, pyrogallol, p-hydroxy benzoic acid, vanillin, p-coumaric acid, gallic acid, quercetin, ??-tocopherol and ascorbic acid, in LEK were quantified by LC-MS-MS. The results show that pyrogallol (2070.0 mg/kg LEK) is the main phenolic compound responsible for the antioxidant and radical scavenging activities of LEK. Finally, total phenolic and flavonoid contents were determined as gallic acid (GAE) and quercetin equivalents (QE). The GAE and QE values in LEK were 16.67 ± 2.83 ??g GAE/mg and 12.95 ± 0.52 ??g QE/mg, respectively. The results suggest that consumption of kiwifruit (A. deliciosa) can be beneficial effects due to its antioxidant properties.     

Antioxidant activity of cichoric acid and alkamides from Echinacea purpurea, alone and in combination

The antioxidant activity of extracts of the stems, leaves, and roots of Echinacea purpurea was compared with the antioxidant activity of purified cichoric acid and alkamides, both constituents of Echinacea purpurea. The antioxidant activity was determined using different methods: effect on oxygen consumption rate of a peroxidating lipid emulsion, and scavenging of radicals, i.e. 2,2-diphenyl-1-picrylhydrazyl (DPPH), measured by two different techniques. The efficacy of the extracts in the reaction with DPPH correlated well with the amount of cichoric acid present in the various extracts. The alkamides alone showed no antioxidant activity in any of the tests. Alkamides present in the extract increased, however, the antioxidative effect of cichoric acid in the peroxidating lipid emulsion. The activity was further compared with that of rosmarinic acid, a well-characterised antioxidant, and the extracts as well as cichoric acid were found to be efficient scavengers of radicals with an activity comparable to that of rosmarinic acid. Cichoric acid was found to have a stoichiometric factor of 4.0 in scavenging DPPH and to react in a second-order reaction with DPPH with a rate constant of 40 l/mol/s at 25 °C in methanol.     

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.     

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.     

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 activities of mangrove Rhizophora apiculata bark extracts

Depolymerisation of mangrove Rhizophora apiculata bark extracts in the presence of phloroglucinol nucleophiles in ethanol was carried out. The flavan-3-ols and their phloroglucinol adducts were separated using reversed phase liquid chromatography (HPLC). The HPLC analysis of mangrove R. apiculata showed that catechin was the most common component of the flavanoid monomers. The antioxidant activities of these mangrove tannins were evaluated and compared with several commercial tannins by using reducing power, DPPH and ABTS assays with butylated hydroxytoluene, BHT and l-(+)-ascorbic acid as standards. All tannins had reducing power and percentage scavenging activities similar to the (+)-catechin and l-(+)-ascorbic acid standards. In the DPPH assay, >90% of the maximum scavenging activity was attained at 30 μg ml⁻¹. Mangrove tannins had stronger antioxidant activity than the BHT standard in the DPPH assay. The results of the ABTS assay were correlated with the DPPH assay. Scavenging activity in the ABTS assay increased as the tannin concentration increased, up to a plateau at 50 μg ml⁻¹.     

Evaluation of reducing power and radical scavenging activities of water and ethanol extracts from sumac (Rhus coriaria L.)

The purpose of this study was to determine the reducing power, metal chelating, and radical scavenging capabilities of water and ethanol extracts of sumac (Rhus coriaria L.), comparatively. The water and ethanol extracts of sumac were evaluated for their radical scavenging activities by means of the DPPH and DMPD assays. Water extract of sumac (R. coriaria L.) scavenged radicals effectively with EC₅₀ values of 36.4 ??g/ml for DPPH free radical and 44.7 ??g/ml for DMPD cation radical. Similarly, the total reducing power of water extract was found higher than ethanol extract in both potassium ferricyanide reduction (FRAP) and cupric ions reduction capacity methods (CUPRAC). 2,2′-Bipyridine was used to determine the metal chelating activity and the result of water extract was found higher than ethanol extract. Total phenolic and total flavonoid contents of both extracts were studied as well. The values of water extract were found to be higher than that of ethanol extract. The present study found that water extracts of sumac (R. coriaria L.) have effective antioxidant and radical scavenging activities as compared to ethanol extracts.     

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.     

Juniperus sibirica Burgsdorf. as a novel source of antioxidant and anti-inflammatory agents

In order to examine antioxidant properties of methanol extracts of cones and needles of the unexplored Juniperus sibirica Burgsdorf. (Cupressaceae) species, various assays which measure free radical scavenging ability were carried out: DPPH, hydroxyl, superoxide anion and nitric oxide radical scavenger capacity test and reducing power (FRAP) assay. In all of the tests the extracts showed a potent antioxidant effect compared with BHT, a well-known synthetic antioxidant. In addition, the anti-inflammatory activity considering inhibitory potency toward COX-1 and 12-LOX was observed. Both extracts showed markedly anti-inflammatory activity, with needles having somewhat higher potency concerning both assays, reaching IC₅₀ at 1.29 mg/mL towards COX-1 and IC₅₀ at 1.34 mg/mL towards 12-LOX. Besides, in the extracts examined the total phenolic and flavonoid amounts were also determined, together with presence and content of the selected flavonoids: luteolin-7-O-glucoside, apigenin-7-O-glucoside, luteolin, apigenin, rutin and quercetin, which were studied using LC–MS/MS technique. LC–MS/MS analysis showed a noticeable content of natural products according to which the examined J. sibirica Burgsdorf. species could well be regarded as a promising new source of bioactive natural compounds, which can be used both as a food supplement and a remedy.     

Antioxidant versus anti-diabetic properties of leaves from Vernonia amygdalina Del. growing in Malaysia

The antioxidant and anti-diabetic properties of the sequential extracts of Vernonia amygdalina based on the chemical composition of the most effective anti-diabetic extract were studied. Using DPPH and ABTS radical scavenging as well as FRAP assays, the extracts showed a consistent dose-dependent trend of potent antioxidant activity in the following solvents: water extract > methanol extract > chloroform extract > and petroleum ether extracts. In the oral glucose tolerance test, the chloroform extract exerted the highest response (33.3%), similar to metformin (27.2%), after 2 h compared to the control (50.8%, P < 0.05). After a 14-day administration in diabetic rats, the chloroform extract recorded the highest blood (23.5%) and serum (21.4%) glucose-lowering effects (P < 0.05). GC–MS analysis of the chloroform extract revealed high levels of linoleic acid (4.72%), α-linolenic acid (10.8%) and phytols (12.0%), as well as other compounds.     

Free radical scavenging properties and phenolic characterization of some edible plants

Antioxidant activity of the ethanolic extracts of the fruits of Fritillaria pontica Wahlenb. (Liliaceae), Euonymus latifolius (L.) Mill. ssp. latifolius (Celastraceae), and Vicia sativa L. ssp. nigra (L.) Ehrh. var. nigra L. (Fabaceae), the aerial parts of Turritis laxa (Sibth & Sm.) Hayek (Brassicaceae), Vicia cracca L. (Fabaceae), and Oxyria digyna (L.) Hill. (Polygonaceae) was screened by 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and ferric-reducing antioxidant power (FRAP) assays at 0.5, 1.0, and 2.0 mg ml⁻¹ concentrations. Total phenolic contents of the extracts were determined using Folin-Ciocalteau reagent. T. laxa was also tested for its anti-acetylcholinesterase activity. The extracts were analyzed by LC–DAD–MS for their flavonoid content and the ethanolic extract of T. laxa has been found to contain rutin in appreciable amounts (7.63 ± 0.2%). Rutin and hyperoside were detected qualitatively in F. pontica, where vitexin was identified in O. digyna. It was also the most active in the antioxidant tests.     

Determination of isoflavone content and antioxidant activity in Psoralea corylifolia L. callus cultures

Relation between isoflavones production and antioxidant activity in Psoralea corylifolia cell cultures was studied. High performance liquid chromatography analysis revealed that root-derived callus cultures produced maximum amount of daidzein whereas genistein by leaf-derived callus. Cell cultures grown under continuous illumination (40-μmol m⁻² s⁻¹) produced several-fold more isoflavones daidzein (2.28% dry wt) and genistein (0.21% dry wt) than that of field grown plants. The antioxidant activity of extracts was determined using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, phosphomolybdenum assay and correlated with the content of total phenolics in the extracts. Calli grown under continuous illumination exhibited strong antioxidant activities compared to dark grown callus cultures and explants materials.     

Major flavonoids with antioxidant activity from Teucrium polium L.

Teucrium polium L. (Lamiaceae) aerial parts are used widely in the daily diet and for medicinal purposes. This plant is used also as a spice and refreshing beverage. Phytochemical and bioactivity studies of this plant have been carried out. Aerial parts of the plant were extracted with petroleum ether, chloroform, methanol and water successively. Fractionation of the methanol extract yielded four major flavonoids. The crude extracts and isolated compounds were screened for their antioxidant and free radical scavenging activities using DPPH radical-scavenging, beta-carotene/linoleic acid and ammonium thiocyanate methods. Methanol extract, rutin and apigenin were found to be the most active fractions as radical-scavengers with IC₅₀ values of 20.1 ± 1.7, 23.7 ± 1.9 and 30.3 ± 2.1 μg/ml, respectively. The samples with the highest inhibition of oxidation of beta-carotene and lipid peroxidation in ammonium thiocyanate methods were also found to be methanol extract, rutin and apigenin. Methoxylated flavonoids exhibited a lesser antioxidant activity.