Phenolics from monofloral honeys protect human erythrocyte membranes against oxidative damage

The aim of the present work was to analyze the phenolic extracts from two monofloral Cuban honeys for their in vitro total antioxidant capacity, phenolic compounds content and free radical scavenging activity. The phenolic extracts, rich in lipophilic compounds, were tested further for their ability to inhibit AAPH-induced oxidative damage (hemolysis, lipid peroxidation and cytosolic depletion of reduced glutathione and decrease of superoxide dismutase activity) in erythrocytes. Results indicate an important total antioxidant capacity measured by TEAC and ORAC assays, as well as a relevant radical scavenging activity performed by EPR. Moreover, 13 phenolic compounds were identified using HPLC–LC/MS with quercetin as the most abundant flavonoid. The results also show that both extracts were able to inhibit erythrocytes oxidative damage, and that this may likely be due to their incorporation into cell membranes and their ability to cross it and reach the cytosol. In fact, flavonoid uptake by erythrocytes was further confirmed by testing quercetin, which efficiently incorporated into erythrocytes. Overall, this study indicates that honey contains relevant antioxidant compounds responsible, at least in part, for its biological activity and that uptake of its flavonoids may provide defense and promote cell functions in erythrocytes.     

Enhanced antioxidant and antityrosinase activities of longan fruit pericarp by ultra-high-pressure-assisted extraction

The health benefits of fruits acting against chronic diseases are ascribed to their antioxidant activities which are mainly responsible due to the presence of phenolic compounds. The use of ultra-high-pressure-assisted extraction (UHPE) has shown great advantages for the extraction of these phenolic compounds from longan fruit pericarp (LFP). Studies were carried out to investigate the effects of UHPE at pressures of 200, 300, 400 and 500 MPa on total phenolic contents, extraction yield, antioxidant and antityrosinase activities from LFP. The antioxidant activities of these extracts were analyzed, using various antioxidant models like 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, total antioxidant capacity and superoxide anion radical scavenging activity. Extract from ultra-high-pressure-assisted extraction at 500 MPa (UHPE-500) showed the highest antioxidant activities of all the tested models. In addition, it also showed moderate tyrosinase inhibitory activity. Three phenolic acids, namely gallic acid, ellagic acid, and corilagin were identified and quantified by HPLC. Corilagin content was the highest compared to other phenolic acids identified. UHPE-500 obtained the higher phenolic acid contents compared to other high pressure processing and conventional extractions (CE). Compared with CE, UHPE-500 exhibited good extraction effectiveness in terms of higher extraction yields with high phenolic contents and also with higher antioxidant and antityrosinase activities.     

Effect of poplar-type propolis on oxidative stability and rheological properties of O/W emulsions

Propolis is known to possess antioxidant activity. However, there is no information on this activity in emulsions O/W. The protective effect of propolis on the oxidation and rheological properties of emulsions O/W containing wheat germ and almond oils was evaluated. Emulsions O/W were prepared with different concentration of propolis extract, almond oil and wheat germ oil. All emulsions physically stable without phase separation were stored at 37?°C for 9?weeks. Chemical composition of propolis was established by Gas chromatography coupled to mass spectrometry. Rheological characterization of different emulsions was performed evaluating consistency index and flow behavior index. The oxidation was monitored by measuring the lipid hydroperoxides and thiobarbituric acid-reactive substances (TBARS) methods. Flavonoids, phenolic acid esters, and aromatic acids were the main groups of compounds found in propolis. The results showed that popolis was good antioxidant in the concentration of 0.02 and 0.04% when lipid phase was constituted by almond oil. The rheological behavior is typical of a non-Newtonian fluid, being almond oil more adequate for having a higher stable O/W emulsion.     

Propolis suppresses CdCl₂-induced cytotoxicity of COS7 cells through the prevention of intracellular reactive oxygen species accumulation

Propolis is a natural product made by honeybees and contains various compounds, including flavonoids, amino acids and fatty acids. These compounds are considered to have antiviral, antibacterial and antioxidative properties. On the other hand, cadmium (Cd), an industrial and environmental pollutant, preferentially accumulates in the kidney and induces kidney injury. We previously reported that exposure to CdCl? induced cell death though intracellular reactive oxygen species (ROS) generation in kidney tubule epithelial COS7 cells. Here, we investigated whether propolis extracts suppress CdCl?-induced cytotoxicity. Predictably, pretreatment with propolis extracts significantly suppressed CdCl?-induced cytotoxicity and intracellular ROS generation. Propolis extracts not only showed superoxide dismutase and antioxidative activities, but also increased the expression of heme oxygenase-1 (HO-1), an antioxidative enzyme. Moreover, we determined the involvement of hypoxia inducible factor-1α in propolis extract-derived HO-1 induction. We demonstrate for the first time the utility of propolis for Cd-related COS7 cytotoxicity, and these novel findings are considered to contribute to the control of ROS-derived disorders.     

Estimation of phytochemical content and antioxidant activity of some selected traditional Indian medicinal plants

The powder samples and methanol extract of 11 medicinal plants were subjected to analysis of proximate composition and measurement of antioxidant activity. Different parameters studied include phenolic contents, moisture, ash, crude fiber, fats and waxes. The assays employed were ferric reducing antioxidant power, trolox equivalent antioxidant capacity and scavenging effect on the 1,1-diphenyl-2-picrylhydrazyl free radical. Results obtained indicate that the antioxidant potential varied significantly from plant to plant. The total phenolic contents were determined spectrophotometrically using Folin-Ciocalteu reagent. Significant correlation is observed between ferric reducing antioxidant power and phenolic contents (R(2) = 0.96). These findings show that the polyphenolic constituents in the extracts are responsible for free radical scavenging capacity.     

Assessment of the antioxidant activities of Brazilian extracts of propolis alone and in topical pharmaceutical formulations

The antioxidant activity of extracts of propolis and of formulations added with these extracts were measured by scavenging different radicals in different systems. For the ethanolic extract of propolis (EEP) and the glycolic extract of propolis (GEP) the IC50 observed were respectively of 0.024 and 0.035 microL/mL in scavenging hydroxyl radical, 0.016 and 0.012 microL/mL in inhibiting lipid peroxidation, 0.22 and 0.24 microL/mL in inhibiting chemiluminescence produced in the H2O2/luminol/horseradish peroxide (HRP) system and about 0.005 microL/mL for both extracts in inhibiting chemiluminescence produced in the xanthine/luminol/xanthine oxidase (XOD) system. The antioxidant activity of extracts of propolis in the formulations was not able to be assessed neither using the deoxyribose assay, since the formulation components interfered in the assay measurements, nor using chemiluminescence in the H2O2/luminol/HRP system, since this method did not show to be sensitive for the extract of propolis evaluation. However, the antioxidant activity of extracts of propolis could be successfully evaluated in the formulations using both lipid peroxidation and chemiluminescence generated in the xanthine/luminol/XOD system inhibitions.     

Antioxidant activities and free radical scavenging potential of Bauhinia microstachya (RADDI) MACBR. (Caesalpinaceae) extracts linked to their polyphenol content

Aqueous and hydro-ethanolic extracts of Bauhinia microstachya leaves (AEBM and HEBM) were investigated for their phenolic content and phytochemical profile (by spectrophotometry and HPLC), and for their antioxidant activities and free radical scavenging potential in different in vitro systems (TRAP, TEAC, TBARS, nitric oxide, superoxide and hydroxyl radical). HEBM presented a 27.4% higher content of phenolics when compared to AEBM and a distinct phytochemical profile was observed. Our work suggests that both extracts have potent antioxidant activities and that their antioxidant capacity and efficiency vary according to the radical-generating system. In general, HEBM was more effective than AEBM in avoiding ROS-generating damage and in scavenging the various radicals formed. Nevertheless, when results were normalized to total phenolic content, a different profile of antioxidant activities and free radical scavenging potential was observed, particularly against oxidative lipid damage and superoxide radical. B. microstachya extracts may be considered an interesting source of natural antioxidants as well as other phenolic-rich plants.     

Evernia prunastri and Pseudoevernia furfuraceae lichens and their major metabolites as antioxidant,antimicrobial and anticancer agents

The aim of this study is to investigate chemical composition of acetone extracts of the lichens Evernia prunastri and Pseudoevernia furfuraceae and in vitro antioxidant, antimicrobial, and anticancer activities of these extracts and some their major metabolites. HPLC–UV method was used for identification of secondary metabolites. Antioxidant activity was evaluated by free radical scavenging, superoxide anion radical scavenging, reducing power and determination of total phenolic compounds. As a result of the study physodic acid had largest antioxidant activities. Total content of phenol in extracts was determined as pyrocatechol equivalent. The antimicrobial activity was estimated by determination of the minimal inhibitory concentration by the broth microdilution method. The most active was also physodic acid. Anticancer activity was tested against FemX (human melanoma) and LS174 (human colon carcinoma) cell lines using MTT method.     

Wheat Seedlings as Food Supplement to Combat Free Radicals: An In Vitro Approach

The present study was designed to evaluate the antioxidant activity of 5 organic solvent extracts (petroleum ether, n-hexane, chloroform, ethyl acetate and methanol) of wheat grains, 3, 5 and 7 days old wheat seedlings. To determine the antioxidant activity of five extracts of four different samples, 1,1-diphenyl-2-picrylhydrazyl and 2,2’-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity, total phenolic content and ferrous reducing power ability were carried out. 1,1-Diphenyl-2-picrylhydrazyl radical scavenging effect of chloroform and ethyl acetate extracts of 3 days old wheat seedlings was higher than wheat grains. Chloroform, ethyl acetate and methanol extracts of 3 days old wheat seedlings exhibited higher 2,2’-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging effcet than extracts of other samples. The phenolic content was high in chloroform, ethyl acetate and methanol extract of 5 days old wheat seedlings. When compared with wheat grain, reducing power ability was high in chloroform, ethyl acetate and methanol extract of wheat seedlings, especially in 3 and 5 days old wheat seedlings. From the above results, it was concluded that chloroform, ethyl acetate and methanol extract of 3, 5 and 7 days old wheat seedlings showed better antioxidant activity than the wheat grain extracts. Hence, the results of the present study suggest the intake of wheat seedlings as a food supplement to combat the diseases caused by free radicals.     

Polyphenol profile by UHPLC-MS/MS,anti-glycation,antioxidant and cytotoxic activities of several samples of propolis from the northeastern semi-arid region of Brazil

Context: Propolis has promising biological activities. Propolis samples from the Northeast of Bahia, Brazil – sample A from Ribeira do Pombal and B, from Tucano – were investigated, with new information regarding their biological activities.

Objective: This paper describes the chemical profile, antioxidant, anti-glycation and cytotoxic activities of these propolis samples.

Material and methods: Ethanol extracts of these propolis samples (EEP) and their fractions were analyzed to determine total phenolic content (TPC); antioxidant capacity through DPPH^?, FRAP and lipid peroxidation; anti-glycation activity, by an in vitro glucose (10?mg/mL) bovine serum albumine (1?mg/mL) assay, during 7?d; cytotoxic activity on cancer (SF295, HCT-116, OVCAR-8, MDA-MB435, MX-1, MCF7, HL60, JURKAT, MOLT-4, K562, PC3, DU145) and normal cell lines (V79) at 0.04–25?μg/mL concentrations, for 72?h. The determination of primary phenols by ultra high-pressure liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) and volatile organic compounds content by gas chromatography-mass spectrometry (GC-MS) were also performed.

Results: The EEP polar fractions exhibited up to 90% protection against lipid peroxidation. The IC₅₀ value for anti-glycation activity of EEP was between 16.5 and 19.2?μg/mL, close to aminoguanidine (IC₅₀?=?7.7?μg/mL). The use of UHPLC-MS/MS and GC-MS allowed the identification of 12 bioactive phenols in the EEP and 24 volatile compounds, all already reported.

Conclusions: The samples present good antioxidant/anti-glycation/cytotoxic activities and a plethora of biologically active compounds. These results suggest a potential role of propolis in targeting ageing and diseases associated with oxidative and carbonylic stress, aggregating value to them.     

Antioxidant and radical scavenging properties of <Emphasis Type="Italic">Iris germanica</Emphasis>

The antioxidant activity of aqueous and ethanol extracts of iris (Iris germanica L., family Iridaceae) has been evaluated in vitro using various antioxidant assays, including reducing power, free radical scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging, and metal chelating activities. Both aqueous and ethanol extracts exhibit strong total antioxidant activity, showing 95.9, 88.4, 79.9% and 90.5, 78.0, 65.3% inhibition on peroxidation of linoleic acid emulsion in concentrations of 10, 30, and 50 μg-ml, respectively. Both extracts also possess effective reducing power and exhibit free radical scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging, and metal chelating activities in concentrations of 20, 40, and 60 μg-ml. The antioxidant properties were compared to those of reference antioxidants (BHA, BHT, and α-tocopherol). In addition, the total content of phenolic compounds in both aqueous and ethanol iris extracts has been determined as gallic acid equivalent. The results indicate that iris has in vitro antioxidant properties, which can be the major factor responsible for the inhibition of lipid peroxidation. Published in Khimiko-Farmatsevticheskii Zhurnal, Vol. 41, No. 8, pp. 13–18, August, 2007.     

Phenols and antioxidant activity of hydro-alcoholic extracts of propolis from Algarve,South of Portugal

Propolis is a natural honeybee product known to be beneficial for human health, with a complex chemical composition, highly dependent on the collection site. The objective of the present research was to evaluate phenols and antioxidant activity of propolis samples collected in three main areas of Algarve, South of Portugal. Water revealed to be less effective for extracting phenolic compounds from propolis than the methanol and water/ethanol. The last two were good extraction solvents of phenols. Nevertheless water/ethanol was the solvent chosen because it was able to extract phenols in considerable amounts being less toxic than methanol.     

Antioxidant and free radical scavenging activities of Terminalia chebula

Free radicals react with biological molecules and destroy the structure of cells, which eventually causes free-radical induced disease such as cancer, renal failure, aging, etc. In this study, 6 extracts and 4 pure compounds of Terminalia chebula RETZ. were investigated for anti-lipid peroxidation, anti-superoxide radical formation and free radical scavenging activities. The superoxide radical scavenging of the 4 pure compounds was further evaluated using electron spin resonance (ESR) spectrometry. The results showed that all tested extracts and pure compounds of T. chebula exhibited antioxidant activity at different magnitudes of potency. The antioxidant activity of each pure compound was derived from different pathways and was suggested to be specific.     

Antimutagenic, antioxidant and free radical scavenging activity of ethyl acetate extracts from white, yellow and red onions.

The beneficial effects of red, yellow and white onion extracts have been assessed by antioxidant activity and antimutagenic activity. And the effects compared to BHT and ascorbic acid. Total phenolic compounds and flavonoids in onion extracts were determined. Yellow onion extract had more organic acid and free sugar than those detected in the white and red onion extract. The scavenging activity of DPPH radical and H(2)O(2) were increased depending on the concentration. The antioxidant activities using beta-carotene-linoleate system and reducing power were increased but the effect was small to that of BHT and ascorbic acid. After digested, extracts showed antimutagenic activities, and it seems that they inhibit the mutagenicity for digesting. This study demonstrated that the antimutagenicities and antioxidant properties of ethyl acetate extract against mutagens were related to their phenols and flavonoids, which are heat stable and losses digestive juices are relatively low.     

Evaluation of polyphenolic acid esters as potential antioxidants

Six polyphenolic acid esters were synthesized and their antioxidative properties were evaluated in three model systems [2,2'-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay, 2,2'-azobis(2-amidinopropane)dihydrochloride (AAPH)-induced lipid peroxidation system, and the dye-bleaching assay of peroxynitrite radical]. Among these compounds, we found that compounds 4 [3,4-dihydroxy-benzoic acid-(2-phenoxyethyl ester)], and 5 [3,4-dihydroxy-cinnamic acid-(2-phenoxyethyl ester)] provided comparable activity to caffeic acid phenethyl ester (CAPE) in the DPPH model. Compound 3 [2,5-dihydroxy-benzoic acid-(2-phenoxyethyl ester)], was found to be more active than CAPE in the AAPH system, it also displayed about 2-fold greater activity than CAPE in the peroxynitrite radical model. These results suggest that these phenolic acid ester derivatives, with their potent anti-oxidant activities, may have useful applications as antioxidants.     

In vitro antioxidant activity and total phenolic content of the inflorescences, leaves and fruits of Sorbus torminalis (L.) Crantz

The antioxidant potential of 70% methanolic extracts from the inflorescences, leaves and fruits of Sorbus torminalis (L.) Crantz was evaluated using three in vitro test systems: the DPPH (2,2-diphenyl-1-picryl-hydrazyl) and the ABTS [2,2-azino-bis(3-ethylbenzo-thiazoline-6-sulfonic acid)] free radical scavenging assays, and the AAPH [2,2'-azobis-(2-amidinopropane) dihydrochloride]-induced [corrected] linoleic acid (LA) peroxidation test. The results were compared with the activity of the extracts obtained from the model antioxidant Sorbus species (Sorbus aucuparia L.), and also with the activity of phenolic standards such as quercetin, Trolox [(+/-)-6-hydroxy-2,2,7,8-tetramethylchroman-2-carboxylic acid], BHA (butylated hydroxyanisole), BHT (butylated hydroxytoluene) and TBHQ (tert-butylhydroquinone). The radical scavenging capacities of the S. torminalis extracts towards the DPPH radical were in the range of 62.0-244.1 micromolar Trolox equivalents/g d.w. of plant material. They were significantly (p < 0.05) correlated with the results of the ABTS test (r = 0.8535), and with the chain-breaking activities determined in the LA-peroxidation test (r = 0.9831). In comparison with the synthetic standards, the free radical scavenging capacity of the Sorbus extracts was remarkably higher than their chain-breaking activity. Both kinds of antioxidant effects of the extracts were significantly (R2 > 0.8097, p < 0.05) influenced by the total phenolic content (TPC) as determined by the Folin-Ciocalteu method. The plant tissues derived from S. torminalis exhibited lower antioxidant potentials than those of S. aucuparia by a factor of 1.5-3.2, partially due to the lower TPC levels (multiplicity factors of 1.2-1.9). After the original antioxidant capacities of the extracts were recalculated according to the TPC levels, the resulting antioxidant capacities of the phenolic fractions in the S. torminalis extracts were lower than those from S. aucuparia by a factor of 1.1-1.6, suggesting that the distinctive chemistry of the phenolic constituents also influences the antioxidant power of the two species.     

Chemical characterization,antioxidant and cytotoxic activities of Brazilian red propolis

Propolis is known for a long time for its health benefits and biological activities. Here, the red variety from the northeast of Brazil was chemically analyzed and extracts were investigated regarding their antioxidant and antitumor activity. Hydroalcoholic extracts, obtained from the red propolis, revealed polyphenol content, 2,2-diphenyl-1-picrylhydrazyl scavenging potential and enzymatic activities for catalase-like and superoxide dismutase-like. Cytotoxic activity was evaluated for human laryngeal epidermoid carcinoma cell (Hep-2), human cervical adenocarcinoma (HeLa) and human normal epithelial embryonic kidney (Hek-293). Survival analysis for non-tumor cell line showed greater IC50 compared to tumor cell lines, suggesting an increased sensitivity that may correlate with the higher proliferative index of the tumor vs. normal cells. Our results indicate that the Brazilian red propolis is capable of inhibiting cancer cell growth and constitutes an excellent source of antioxidant and antitumor natural agent.     

Propolis influence on erythrocyte membrane disorder (hereditary spherocytosis): A first approach

Propolis is a resinous substance collected from plants by bees. Its composition depends on the vegetation, the season, and the source area. It usually contains many chemical compounds such as polyphenols, steroids and amino acids.The hereditary spherocytosis (HS) is a type of anaemia characterized by microcytic and hyperchromic red cells, spherical in shape and without central pallor. Clinically, subjects present from asymptomatic conditions to severe haemolytic anaemia.In this study it was evaluated the effect of two propolis extracts in the osmotic fragility of HS patient red blood cell (RBC) membrane. It was found that propolis decreases the erythrocytes membrane fragility, being the effect of Bornes propolis more pronounced than Fundão propolis’. This effect was related with the higher phenolic content of the former propolis. The results obtained in vitro suggest that the membrane fragility increases under oxidative stress conditions for the patient RBC’s and the protection effect of propolis is due to its antioxidant properties. These results open doors for future investigations in order to elucidate the mechanisms, identify the main compounds involved in this fragility protection of the erythrocyte membrane. This is the first work reporting an evaluation of the propolis effect in a blood disease.     

别克参提取物抗氧化活性研究

目的：研究别克参提取物的体外抗氧化活性。方法：采用清除二苯代苦味酰基（DPPH）自由基、2,2’-氨基-（3-乙基-苯并噻唑啉-6-磺酸）二氨盐（ABTS）自由基与超氧阴离子（O2^-）自由基法,对别克参不同提取物的体外抗氧化活性进行评价。结果：别克参不同提取物清除DPPH·自由基、ABTS·自由基的能力强弱顺序为：乙酸乙酯提取物〉石油醚提取物〉正丁醇提取物〉水提取物。别克参不同提取物对O2^-自由基有一定的清除能力,但不明显。结论：别克参不同提取物抗氧化活性差异可能与各部位中所含抗氧化成分的种类和结构有关。     

Antioxidant activity of Hypericum androsaemum infusion: scavenging activity against superoxide radical,hydroxyl radical and hypochlorous acid

Hypericum androsaemum is a medicinal plant species containing many polyphenolic compounds, namely flavonoids and phenolic acids. Since polyphenolic compounds have high antioxidant potential, the ability of H. androsaemum infusion to act as a scavenger of reactive oxygen species (superoxide radical, hydroxyl radical and hypochlorous acid) was investigated. Superoxide radical was generated by the xanthine/xanthine oxidase and phenazine methosulphate/NADH systems. The infusion-mediated prevention of nitroblue tetrazolium reduction by the superoxide radical was used as the measured endpoint. Hydroxyl radical was generated by the Fe3+-EDTA/ascorbate Fenton system, and assayed by evaluating deoxyribose degradation using the thiobarbituric acid method. Hypochlorous acid scavenging activity was tested by measuring the inhibition of hypochlorous acid-induced 5-thio-2-nitrobenzoic acid oxidation to 5,5'-dithiobis(2-nitrobenzoic acid). The tested infusion mainly exhibited a potent scavenging effect on superoxide radicals (although a noncompetitive inhibitory effect on xanthine oxidase was also observed). The infusion also acted as a moderate scavenger of hydroxyl radicals and hypochlorous acid. A phytochemical study of the infusion was also undertaken, and nine phenolic compounds were identified.