Inhibitory effect of some natural and semisynthetic phenolic lipids upon acetylcholinesterase activity

The effect of phenolic lipids isolated from rye grains and cashew nut shell liquid (CNSL) from Anacardium occidentale and their semisynthetic derivatives on erythrocyte ghost’s acetylcholinesterase activity was studied. It has been shown that all tested compounds decreased the enzymatic activity of acetylcholinesterase. This effect depends on the type of studied compounds. Three of them completely inhibit acetylcholinesterase activity at the micromolar concentration.     

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.     

Effect of steam treatment on soluble phenolic content and antioxidant activity of the Chaga mushroom (Inonotus obliquus)

The effect of steam treatment on free phenolic acids in Chaga mushrooms (Inonotus obliquus) was investigated. Untreated and steam-treated (120 °C, 3 h) samples of I. obliquus were extracted with organic solvents and free phenolic acid-containing fractions were isolated. Free phenolic acids were determined by LC/PDA (liquid chromatography/photodiode array), ESI LC/MS (electrospray ionisation liquid chromatography/mass spectrometry), and GC/MS (gas chromatography/mass spectrometry). After the steam treatment, the soluble phenolic content determined by modified Folin–Ciocalteu method was increased and antioxidant activity was enhanced, as confirmed by a DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity assay. The amounts of vanillic acid, protocatechuic acid, syringic acid, and 2,5-dihydroxyterephthalic acid were increased significantly as the result of the steam treatment, suggesting that the liberation of low molecular weight free phenolics was enhanced by the steaming process. Consequently, the radical scavenging activity was also significantly enhanced by free phenolics produced using this method.     

Free radical scavenging activity and phenolic content in achenes and thalamus from Fragaria chiloensis ssp. chiloensis, F. vesca and F. x ananassa cv. Chandler

The total phenolic, flavonoid and anthocyanin content of achenes (true fruit) and thalamus (receptacle) from the native South American Fragaria chiloensis ssp. chiloensis (f. patagonica and f. chiloensis), Fragaria vesca and Fragaria x ananassa cv. Chandler was determined by spectrophotometric means. Highest phenolic content was found in F. vesca while lowest content was measured for white strawberry (F. chiloensis ssp. chiloensis, f. chiloensis). The total anthocyanin and total flavonoid contents in the samples investigated was lower for the white strawberry and higher in F. x ananassa cv. Chandler. Total flavonoid content showed a better correlation than total anthocyanins with the free radical scavenging effect of the extracts measured by means of the DPPH discoloration assay. In the superoxide anion assay all the acetone extracts of strawberries showed similar activity. The data presented in this study demonstrate that the amount of phenolic compounds differ significantly between species and subspecies and determine the free radical scavenging activity of fruits. On a w/w basis, higher total phenolics including flavonoids was found in achenes. The highest total anthocyanin content was found in the achenes of F. chiloensis and F. vesca, while F. ananassa presented higher antocyanin content in thalamus. The main anthocyanin in thalamus of F. ananassa (95%) were pelargonidin derivatives which were also present in F. chiloensis ssp. chiloensis f. patagonica (62.6%) but were not detected in F. vesca and F. chiloensis ssp. chiloensis f. chiloensis. In F. vesca and F. chiloensis ssp. chiloensis f. chiloensis only cyanidin derivatives were found both in thalamus and achenes.     

Wine extracts from Sardinian grape varieties attenuate membrane oxidative damage in Caco-2 cell monolayers

One of the most important sites of polyphenol action seems to be in the gastrointestinal system before absorption. We investigated the ability of three wine phenolic extracts, obtained from grape varieties grown in Sardinia, Cannonau (red), Vermentino and Malvasia (white), to exert an antioxidant action against tert-butyl hydroperoxide (TBH)-induced oxidative damage to Caco-2 cell monolayers as a model system of the human intestine. TBH treatment caused the disruption of epithelial integrity, measured as transepithelial electrical resistance, and markers of the peroxidation process of membrane lipids, MDA, fatty acid hydroperoxides and 7-ketocholesterol. All wine extracts were able to counteract the oxidising action of TBH and, in spite of the differences in phenolic composition, exerted a comparable activity. Our findings point out a direct antioxidant action of the wine extracts on enterocytes exposed to oxidising species and further support the opinion that total phenolic content is not essential for antioxidant activity.     

Cytoprotective and antioxidant activity of free,conjugated and insoluble-bound phenolic acids from swallow root (Decalepis hamiltonii)

Swallow root (Decalepis hamiltonii) was extracted for free (SRFP), conjugated (SRCP) and insoluble-bound phenolic acids (SRIBP), and evaluated for cytoprotectivity, 1,1,diphenyl-2-picrylhydrazyl (DPPH) scavenging ability, reducing power and protection to DNA damage. In addition, the constituent phenolic acids in the extracts were also analysed. Results indicated a total phenol content of 20.72, 7.97 and 11.52 mg gallic acid equivalents (GAE)/g for SRFP, SRCP and SRIBP extracts, respectively. At 0.12 μg/mL concentration SRCP showed 87% cytoprotection (on NIH 3T3 cells) compared to SRFP (47%) and SRIBP (65%). DPPH radical scavenging activity indicated an IC₅₀ of 0.046, 0.06 and 0.128 μg/mL for SRCP, SRIBP and SRFP, respectively. Also, SRCP showed higher reducing power and DNA protectivity (80%). HPLC analysis of phenolic acid extracts showed the presence of hydroxybenzoate and cinnamate derivatives. Among the phenolics identified gallic, gentisic, protocatechuic and p-coumaric acids were the major contributors to antioxidant activity.     

Direct characterisation of phenolic antioxidants in infusions from four Mapuche medicinal plants by liquid chromatography with diode array detection (HPLC-DAD) and electrospray ionisation tandem mass spectrometry (HPLC-ESI-MS)

A simple, fast and direct method was developed for the qualitative analysis of phenolic constituents from infusions of Mapuche medicinal plants. Teas made of Linum chamissonis Schiede, Quinchamalium chilensis Mol., Adesmia emarginata Clos. and Escallonia illinita K. Presl. were analysed by high-performance liquid chromatography with diode array detector (HPLC-DAD) and electrospray mass spectrometry (ESI-MS). This technique allowed for the first time the tentative identification of 16 phenolic compounds in E. illinita, 27 in Q. chilensis, 10 in L. chamissonis and 19 in A. emarginata. The compounds were mainly phenolic acids, flavonoid glycosides, anthocyanins and tannins. The total phenolic and flavonoid content of the infusions was assessed as well as the free radical scavenging capacity measured by the bleaching of a solution of the 2,2-diphenyl-1-picrylhydrazyl (DPPH⁻) radical. From the four species, Q. chilensis exhibited the strongest antioxidant activity with highest total phenolic and flavonoid content.     

The distribution of phenolic acids in pumpkin’s hull-less seed,skin, oil cake meal,dehulled kernel and hull

The phenolic acids in whole hull-less seed, its skin and oil cake meal, dehulled kernel and hull of pumpkin (Cucurbita pepo) were separated into free, esterified and insoluble-bound forms, which were then identified and quantified by high-performance liquid chromatography with a photodiode array detector. In all samples, protocatechuic, p-hydroxybenzoic, vanillic, trans-p-coumaric, ferulic, trans-sinapic acids and p-hydroxybenzaldehyde were quantified. Caffeic acid was present in all samples except in hulls, while syringic acid was not detectable only in skin and oil cake meal. p-Hydroxybenzoic acid was the dominant phenolic compound in all investigated samples, with 34.7%, 52.0%, 51.4%, 67.4% and 51.8% found in hull-less seed, oil cake meal, skin, dehulled kernels and hulls, respectively, based on total phenolic acid content. Most phenolic acids were present in bound (esterified and insoluble) form, from 50.6% in skin to 84.1% in hull-less seed.     

Effect of phenolic acids on glucose and organic acid metabolism by lactic acid bacteria from wine

The influence of phenolic (p-coumaric, caffeic, ferulic, gallic and protocatechuic) acids on glucose and organic acid metabolism by two strains of wine lactic acid bacteria (Oenococcus oeni VF and Lactobacillus hilgardii 5) was investigated. Cultures were grown in modified MRS medium supplemented with different phenolic acids. Cellular growth was monitored and metabolite concentrations were determined by HPLC-RI. Despite the strong inhibitory effect of most tested phenolic acids on the growth of O. oeni VF, the malolactic activity of this strain was not considerably affected by these compounds. While less affected in its growth, the capacity of L. hilgardii 5 to degrade malic acid was clearly diminished. Except for gallic acid, the addition of phenolic acids delayed the metabolism of glucose and citric acid in both strains tested. It was also found that the presence of hydroxycinnamic acids (p-coumaric, caffeic and ferulic) increased the yield of lactic and acetic acid production from glucose by O. oeni VF and not by L. hilgardii 5. The results show that important oenological characteristics of wine lactic acid bacteria, such as the malolactic activity and the production of volatile organic acids, may be differently affected by the presence of phenolic acids, depending on the bacterial species or strain.     

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.     

Enhancement of antioxidant activity,total phenolic and flavonoid content of black soybeans by solid state fermentation with Bacillus subtilis BCRC 14715

In the present study, a solid state fermentation of black soybeans with Bacillus subtilis BCRC 14715 was performed. The effect of fermentation on the changes of total phenolic and flavonoid content and antioxidant activities including 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging effect, and Fe²⁺-chelating ability exerted by various solvent (water, 80% methanol, 80% ethanol, 80% acetone) extracts of black soybeans was examined. It was found that fermentation enhanced the total phenolic and flavonoid content as well as antioxidant activity of the black soybean extract. Among the various extracts examined, the acetone extract of fermented black soybeans showed the highest total phenolic and flavonoid content. The acetone extract and the methanol extract of fermented black soybeans showed the highest DPPH free radical-scavenging effect and Fe²⁺-chelating ability, respectively. Analysis of extraction yields showed that the active principle associated with the DPPH radical-scavenging effect was most efficiently extracted from black soybeans using water, regardless of fermentation. Water and methanol effectively extract the Fe²⁺-chelating principles from non-fermented and fermented black soybeans, respectively.     

Flavonoids and phenolic acids from Labisia pumila (Kacip Fatimah)

Both total phenolic content (TPC) and total flavonoid content (TFC) of Labisia pumila extracts were determined spectrophotometrically. L. pumila leaves extracted in 60% methanol (MeOH) were fractionated on C18 cartridge and the antioxidant property of each fraction was determined by measuring free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity. The 40% MeOH fraction exhibited the highest scavenging activity. Nine flavonols (quercetin, myricetin and kaempferol), two flavanols (catechin and epigallocatechin) and nine phenolic acids were identified from this active fraction by UPLC–ESI-MS/MS, and confirmed by comparison with the mass spectra of standard aglycones, theoretical fragments generated from MS Fragmenter software, and literature values.     

Assessment of the antioxidant potential and fatty acid composition of four Centaurea L. taxa from Turkey

This paper focused on the assessment of antioxidant property and fatty acid composition of four Centaurea species. The antioxidant activity of its methanol extract was evaluated by several in vitro experiments including phosphomolybedum assay, DPPH assay, β-carotene/linoleic acid, ferric and cupric reducing power. Total phenolic and flavonoid contents were also evaluated. The methanol extract of Centaurea pulcherrima var. pulcherrima showed the superior free radical scavenging activity, linoleic acid inhibition capacity, reducing power and also had the highest total phenolic content. A significant relationship between antioxidant capacity and total phenolic components was found. The oils of Centaurea taxa were also analysed for fatty acid concentration by gas chromatography. The principal fatty acids in the species were palmitic acid (23.38–30.49%) and linoleic acid (20.19–29.93%). These findings suggest that the Centaurea species could be used as a potential source of new natural antioxidants and unsaturated fatty acids in food industry, cosmetics and pharmaceutical preparations.     

Development and validation of an HPLC-method for determination of free and bound phenolic acids in cereals after solid-phase extraction

Whole cereal grains are a good source of phenolic acids associated with reduced risk of chronic diseases. This paper reports the development and validation of a high-performance liquid chromatography–diode array detection (HPLC–DAD) method for the determination of phenolic acids in cereals in either free or bound form. Extraction of free phenolic acids and clean-up was performed by an optimised solid-phase extraction (SPE) protocol on Oasis HLB cartridges using aqueous methanol as eluant. The mean recovery of analytes ranged between 84% and 106%. Bound phenolic acids were extracted using alkaline hydrolysis with mean recoveries of 80–95%, except for gallic acid, caffeic acid and protocatechuic acid. Both free and bound phenolic extracts were separated on a Nucleosil 100 C₁₈ column, 5 μm (250 mm × 4.6 mm) thermostated at 30 °C, using a linear gradient elution system consisting of 1% (v/v) acetic acid in methanol. Method validation was performed by means of linearity, accuracy, intra-day and inter-day precision and sensitivity. Detection limits ranged between 0.13 and 0.18 μg/g. The method was applied to the analysis of free and bound phenolic acids contents in durum wheat, bread wheat, barley, oat, rice, rye, corn and triticale.     

Antioxidant, antiacetylcholinesterase and antimicrobial activities of Cymbopogon schoenanthus L. Spreng (lemon grass) from Tunisia

Aqueous extract, proanthocyanidin rich extract, and organic extracts of Cymbopogon schoenanthus L. Spreng (lemon grass) shoots from three different locations in South Tunisia were screened for their antioxidant, acetylcholinesterase and antimicrobial activities. In addition to the evaluation of these activities, the contents of flavonoids and total phenolic compounds were determined.Antioxidant activity measured by DPPH assay showed that the proanthocyanidin extract exhibited higher antioxidant activity than the aqueous extract. Extract concentration providing 50% inhibition (IC₅₀) ranged from 16.4 ± 6.8 μg/mL to 26.4 ± 6.8 μg/mL. The antioxidant activity was also determined using the β-carotene/linoleic acid bleaching test. The best results (IC₅₀ = 0.11 ± 0.10 mg/mL) were obtained with the proanthocyanidin extract of the plants collected from the desert region (Dhibat).The greatest acetylcholinesterase inhibitory activity (IC₅₀ = 0.23 ± 0.04 mg/mL) was exhibited by the ethyl acetate and methanol extracts of the plants collected from the mountainous region. It seems that extracts obtained with more polar solvents gave better results.The proanthocyanidin extracts showed a good antimicrobial activity against Streptococcus sobrinus at low concentration (MIC = 4 mg/mL). Therefore, these extracts could be used to prevent carious lesions by inhibiting S. sobrinus growth.     

Main nutrients,phenolics, antioxidant activity,DNA damage protective effect and microstructure of Sphallerocarpus gracilis root at different harvest time

Sphallerocarpus gracilis roots harvested at two different times (T1: 12-10-2009 and T2: 15-04-2010) were analysed for their main nutrients, phenolics, antioxidant activity, DNA damage protective effect and microstructure. Results revealed that higher amounts of starch were detected in T1 S. gracilis root, whereas higher levels of reducing sugar, protein, amino acids and phenolic compounds were determined in T2 S. gracilis root. The antioxidant activity was assessed by DPPH, FRAP and TEAC assay and T2 S. gracilis root was found to possess higher antioxidant activity. Also, results of the correlation analysis indicated that caffeic, p-coumaric and gallic acids contributed substantially to the antioxidant property and there was significant correlation between DPPH radicals scavenging capacity and FRAP and TEAC. The highest and lowest DNA damage protective effects were found of free phenolic extracts from T2 and T1 S. gracilis roots, respectively. The microstructure of S. gracilis root varied greatly with the harvest time.     

Content of polyphenolic constituents and antioxidant activity of some Stachys taxa

Seven Croatian Stachys taxa (S. alpina, S. officinalis, S. palustris, S. recta subsp. recta, S. recta subsp. subcrenata, S. salviifolia and S. sylvatica) have been investigated in order to determine their content of biologically active compounds (polyphenols, tannins, phenolic acids and flavonoids) as well as their antioxidant activity. All taxa tested had a high content of total polyphenols, medium content of total phenolic acids, and a rather low content of tannins and flavonoids. The total phenolic acids content correlated significantly with total polyphenols content and the content of polyphenols unadsorbed on hide powder. A low correlation between plant phenolic acids and tannins was observed. Methanolic, ethanolic and dichloromethane extracts were investigated using DPPH, lipid peroxidase and xanthine oxidase assays. The extracts showed no inhibitory effects against lipid peroxidation and xanthine oxidase, but they had the ability to scavenge DPPH. The most effective in the DPPH assay were methanolic extracts of S. recta sub. recta and S. palustris whose radical scavenging activity was higher then that of the reference rutin. A low correlation between radical scavenging capacities of extracts with total flavonoids content was observed. The results indicate that investigated Stachys taxa exhibit potent antiradical activity and therefore could be a potential material for extracting free radical scavengers.     

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.     

Comparison of constituents, antioxidant potency, and acetylcholinesterase inhibition in Lentinus edodes, Sparassis crispa, and Mycoleptodonoides aitchisonii

This study evaluates three edible mushrooms: Lentinus edodes, Sparassis crispa, and Mycoleptodonoides aitchisonii, in terms of acetylcholinesterase (AChE) inhibition, antioxidant potency and constituents free amino acids and mineral. The concentration of essential amino acids was found to be 34.10 mg/g in M. aitchisonii, 26.25 mg/g in S. crispa, 25.99 mg/g in L. edodes. S. crispa displayed the highest DPPH scavenging activity and phenolic contents. The best results for AChE inhibition were obtained from M. aitchisonii. These results suggest that M. aitchisonii has high potential for cognitive improvement by AChE inhibition and antioxidant potency.     

Synergistic and suppressive effects of dietary phenolic acids and other phytochemicals from cereal extracts on nuclear factor kappa B activity

In this study the combinatorial action of the main phenolic acids (ferulic, caffeic, p-coumaric and sinapic acids) found in extracts of free (ME) and bound phenolic acids (HE) from oat, barley and wheat flour on the modulation of NF-κB activity was investigated. The results show that combination of phenolic acids in low concentrations (<0.1 μg/ml) has a significant synergistic, or enhanced effect, on NF-κB activity, while their combination in high concentrations (0.3–70 μg/ml) helps to suppress the strong effect obtained by individual solutions of ferulic and p-coumaric acids. The modulation of NF-κB activity observed by HE extracts is the effect of combinatorial action of the phenolic acids present therein, while the modulation of NF-κB activity observed with ME extracts is the result of combinatorial action of phenolic acids together with other phenolic compounds present in the extracts. These results increase the knowledge about the health promoting effect from cereal consumption and dietary phenolic acids.