Antioxidant Power Test Research Articles

Chemical composition, antioxidant and antimicrobial activities of Lantana camara Linn leaves essential oil from Burkina Faso

The objective of this study was to determine chemical composition and biological activity of essential oil of the leaves of Lantana camara from Burkina Faso. The essential oil was obtained by hydrodistillation and analyzed by GC and GC/MS. Antioxidant activity was determined by 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay and Ferric reduction antioxidant power (FRAP) test. Antimicrobial activity was assessed by agar disc diffusion method and microdilution method. The main components of essential oil of L. camara were caryophyllene oxide (23.015%), spathulenol (13.421%), humulen-1, 2-epoxide (8.046%), β-caryophyllene (7.93%), E-nerolidol (6.933%) and α-humulene (4.925%). The oil showed good radical scavenging power and moderate reducing power compared with quercetin, ascorbic acid and butylhydroxytoluene (BHT). Essential oil of L. camara showed antibacterial activity with inhibition diameters between 08±00 mm and 23.5±2.12 mm. Minimum inhibitory concentration (MIC) values were ranging from 4% to 8% and minimum bactericidal concentration (MBC) were determined for the strains of Escherichia coli (8%) only. L. camara essential oil had inhibitory action on all fungal strains with MICs of 2% to 4% and minimum fungicidal concentration (MFC) of 4% for Saccharomyces cerevisiae. Hence L. camara essential oil could be used as antifungal agent, as antioxidant and could be a potential antibacterial agent especially against Escherichia coli.

Effects of deproteinization methods on primary structure and antioxidant activity of Ganoderma lucidum polysaccharides

Ganoderma lucidum polysaccharides (GLP) as one of water-soluble polysaccharides has attracted much attention because of its bioactivities, especially antioxidant activity. Deproteinization is an essential step in the purification process of polysaccharides. In this study, three classic deproteinization methods, including neutral protease method, TCA precipitation and CaCl2 salting out, were evaluated for crude GLP processing. The methods had ability to remove proteins (71.50–87.36%), and meanwhile polysaccharide loss (8.35–11.39%) was observed. Structure analysis indicated that these deproteinization methods had no significant effect (p > 0.05) on molecular weight of polysaccharides, but led to varying degrees of glycoside bond losses (1.14–64.05%). Moreover, the antioxidant activities of deproteinized polysaccharides were measured in vitro by hydroxyl radical, reducing power, 2, 2-diphenyl-1-picryl-hydrazyl (DPPH) free radical and ferric-reducing antioxidant power tests. Purified GLP by enzymolysis maintained the strongest antioxidants activities with the retention rate of over 47.40%, and its deproteinization efficiency and polysaccharide loss ratio were 74.03% and 11.39% respectively. In view of relatively high purity and antioxidant activity, enzymolysis was a suitable deproteinization method for GLP production.

Assessment of anticholinesterase and antioxidant properties of the extracts and (+)-catechin obtained from Arceuthobium oxycedri (D.C.) M. Bieb (dwarf mistletoe)

The hexane, chloroform, ethyl acetate, water, and n-butanol extracts of Arcethobium oxycedri (DC.) M.Bieb. (Santalaceae) were evaluated for their inhibitory effects against acetyl- (AChE) and butyrylcholinesterase (BChE), related to Alzheimer's disease at 200 μg/mL using microplate assays. Since oxidative stress contributes much to neurodegeneration, antioxidative effect of the extracts was determined in metal-chelating and ferric-reducing antioxidant power (FRAP) test systems as well as antiradical effect against 2,2-diphenyl-1-picrylhydrazyl (DPPH). Total phenol and flavonoid contents of the extracts were calculated spectrophotometrically. Our findings indicated that the EtOAc extract, the richest one in total phenol and flavonoid amount (455.22 ± 4.35 and 194.45 mg/g extract, respectively) displayed the highest cholinesterase inhibition (45.37 ± 1.23% for AChE and 94.74 ± 2.43% for BChE at 200 μg/mL). Fractionation of the EtOAc led to isolation of (+)-catechin as the major compound. Catechin exerted remarkable antioxidant effect, whereas it did not cause cholinesterase inhibition. Therefore, the EtOAc extract deserves further investigation for its high BChE inhibition.

Antioxidant capacity of cinnamon extract for palm oil stability

BackgroundSpices and their bioactive components are more promising attractions for their inclusion in diet-based regimes to improve human health. These are sources of natural antioxidants and play an important role in the chemoprevention of diseases and aging. The aim of the current study was to explore the antioxidant potential of cinnamon; a widely used spice throughout the world.MethodsThe current research was aimed to investigate the antioxidant potential of cinnamon extract. For the purpose, cinnamon sticks were procured from local super market, while palm oil was obtained from local oil industry. The resultant extract was analyzed for its antioxidant activity through total phenolic content (TPC), free radical scavenging activity (DPPH assay), and total antioxidant activity was measured by ferric reducing antioxidant power (FRAP) test. The shelf life of palm oil was checked by adding cinnamon extract in oil at different levels i.e., 0.05, 0.10, 0.15, 0.20 and 0.25%, to compare the antioxidant potential of the extract whereas, To acted as control and TBHA @ 0.1% was used as synthetic antioxidant in the oil samples. The oil samples were analyzed for rancidity check during storage (after every seven days for a storage period of four weeks).ResultsThe results indicated that total phenolic contents (TPC); 1,1-diphenyl-2-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) values of cinnamon extract were as 355.01 ± 8.34 gallic acid equivalent per gram (mg GAE/g), 90.18 ± 2.12 (%) and 132.82 ± 3.12 (μmol/g), respectively. The oxidative parameters for treatments i.e., To, TBHA, T1, T2, T3, T4, T5 were recorded as peroxide value (2.61 ± 0.07, 2.42 ± 0.08, 2.57 ± 0.05, 2.56 ± 0.03, 2.54 ± 0.02, 2.54 ± 0.01, 2.46 ± 0.06 meq/kg, respectively), free fatty acids (0.601 ± 0.05, 0.522 ± 0.02, 0.580 ± 0.07, 0.572 ± 0.03, 0.56 ± 00.07, 0.552 ± 0.03, 0.536 ± 0.05%, respectively), TBA value (22.4 ± 1.45, 20.1 ± 0.73, 21.8 ± 0.42, 21.2 ± 1.56, 20.7 ± 0.48, 20.5 ± 0.59, 20.2 ± 0.91 μg/kg, respectively) and iodine value (52.82 ± 2.12, 52.71 ± 2.38, 52.68 ± 2.96, 52.97 ± 2.14, 52.93 ± 2.12, 53.15 ± 2.38, 52.71 ± 2.96, respectively). Overall, the statistical analysis indicated that all parameters regarding oil stability i.e., peroxide value (PV), free fatty acid content, (FFA) thiobarbituric acid (TBA) value and iodine value (IV) were significant with respect to treatments and storage.ConclusionFrom the present study, it can be concluded that the cinnamon extract proved effective in reducing the lipid oxidation of palm oil and it can be successfully used in place of synthetic antioxidants in food preparations.

Variability of the chemical composition and the antioxidant activity of the essential oils of two subspecies of Artemisia campestris L. growing in Algeria

The aim of the present work was to investigate the chemical composition variability of the essential oils (EOs) of two subspecies of Artemisia campestris L. The studied subspecies were ssp. eu-campestris (Briq. & Cav.) and ssp. glutinosa (J. Gay ex Besser) Batt. Several samples coming from different regions of collection were adopted. The plant material was harvested at full flowering phenological status. Essential oils obtained by hydrodistillation of the aerial parts of the studied plant were analyzed by GC and GC–MS. Subsequently, the antioxidant activity of these EOs were also determined in vitro using three different assays: DPPH· (2,2-diphenyl-1-picrylhydrazyl) free radicals scavenging activity, ferric reducing antioxidant power and β-carotene bleaching test. The results of the EOs analysis showed the presence of monoterpenes hydrocarbons in high amounts in the majority of studied subspecies samples. The EOs samples presented large variability of the chemical compositions for both studied subspecies; besides, a new chemotype (capillene/capillin) was identified for the ssp. eu-campestris of this plant which grows in the location of Aflou (Laghouat).

Activity guided fractionation of pomegranate extract and its antioxidant, antidiabetic and antineurodegenerative properties

Pomegranate (Punica granatum L.) peel is a rich source of polyphenols, especially tannins, and concentrations of these compounds may be influenced by extraction processes. This study was aimed to examine phenolic composition and biological activities of pomegranate peel 70% ethanolic extract and its fractions (petroleum ether, ethyl acetate, butanol and water) obtained by liquid/liquid extractions. Fractions were analyzed for polyphenolic content and the highest total phenolic content was obtained in ethyl acetate fraction (200.9 mg gallic acid equivalent/g). This fraction had the highest amounts of gallic acid and ellagic acid, while water fraction contained the highest contents of punicalin and punicalagin. Antioxidant activity was evaluated by four different methods, and butanol and ethyl acetate fractions were the most active in 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis (3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS) assays, whereas water fraction showed the strongest activity in ferric reducing antioxidant power (FRAP) and β-carotene tests. All fractions showed better inhibitory effect on α-glucosidase, with inhibition concentration IC50 = 0.26–4.57 μg/mL, than α-amylase (IC50 = 23.6–284.3 μg/mL), and the most active was ethyl acetate fraction. Antineurodegenerative activity was tested using acetylcholinesterase (AChE) and tyrosinase (TYR) inhibitory activity assays, and the most effective inhibitor of both enzymes was 70% ethanolic extract. Pomegranate peel could be considered as a valuable agro-industrial waste product which could be used as low-cost natural source of biologically active compounds.

Interactive effects of aluminum and cadmium on phenolic compounds, antioxidant enzyme activity and oxidative stress in blueberry (Vaccinium corymbosum L.) plantlets cultivated in vitro

To evaluate the potential role of phenolic compounds in Al and Cd stress tolerance mechanisms, Vaccinium corymbosum cv. Legacy plantlets were exposed to different metal concentrations. The present study used an in vitro plant model to test the effects of the following treatments: 100μM Al; 100μMAl + 50μMCd; and 100μMAl + 100μMCd during periods of 7, 14, 21 and 30 days. The oxidative damage was determined by the accumulation of malondialdehyde (MDA) and hydrogen peroxide (H2O2). The antioxidant activity values were determined using 1,1-diphenyl-2-picrylhydrazine (DPPH) and the ferric reducing antioxidant power test (FRAP). Additionally, the phenolic compound concentrations were determined using HPLC-DAD. The exposure to Al and Cd increased the MDA and H2O2 contents differentially, while the antioxidant capacity values showed differences between DPPH and FRAP with the largest changes in FRAP relative to Cd. SOD had the highest activity in the first 7 days, leading to a significant increase in phenolic compounds observed after 14 days, and chlorogenic acid was the major compound identified. Our results revealed that phenolic compounds seem to play an important role in the response to ROS. Therefore, the mechanisms of tolerance to Al and Cd in V. corymbosum will be determined by the type of metal and time of exposure.

Argan Oil-Mediated Attenuation of Organelle Dysfunction, Oxidative Stress and Cell Death Induced by 7-Ketocholesterol in Murine Oligodendrocytes 158N.

Argan oil is widely used in Morocco in traditional medicine. Its ability to treat cardiovascular diseases is well-established. However, nothing is known about its effects on neurodegenerative diseases, which are often associated with increased oxidative stress leading to lipid peroxidation and the formation of 7-ketocholesterol (7KC) resulting from cholesterol auto-oxidation. As 7KC induces oxidative stress, inflammation and cell death, it is important to identify compounds able to impair its harmful effects. These compounds may be either natural or synthetic molecules or mixtures of molecules such as oils. In this context: (i) the lipid profiles of dietary argan oils from Berkane and Agadir (Morocco) in fatty acids, phytosterols, tocopherols and polyphenols were determined by different chromatographic techniques; and (ii) their anti-oxidant and cytoprotective effects in 158N murine oligodendrocytes cultured with 7KC (25–50 µM; 24 h) without and with argan oil (0.1% v/v) or α-tocopherol (400 µM, positive control) were evaluated with complementary techniques of cellular and molecular biology. Among the unsaturated fatty acids present in argan oils, oleate (C18:1 n-9) and linoleate (C18:1 n-6) were the most abundant; the highest quantities of saturated fatty acids were palmitate (C16:0) and stearate (C18:0). Several phytosterols were found, mainly schottenol and spinasterol (specific to argan oil), cycloartenol, β-amyrin and citrostadienol. α- and γ-tocopherols were also present. Tyrosol and protocatechic acid were the only polyphenols detected. Argan and extra virgin olive oils have many compounds in common, principally oleate and linoleate, and tocopherols. Kit Radicaux Libres (KRL) and ferric reducing antioxidant power (FRAP) tests showed that argan and extra virgin olive oils have anti-oxidant properties. Argan oils were able to attenuate the cytotoxic effects of 7KC on 158N cells: loss of cell adhesion, cell growth inhibition, increased plasma membrane permeability, mitochondrial, peroxisomal and lysosomal dysfunction, and the induction of oxiapoptophagy (OXIdation + APOPTOsis + autoPHAGY). Altogether, our data obtained in 158N oligodendrocytes provide evidence that argan oil is able to counteract the toxic effects of 7KC on nerve cells, thus suggesting that some of its compounds could prevent or mitigate neurodegenerative diseases to the extent that they are able to cross the blood-brain barrier.

Chromatography Analysis of Fatty Acids, Volatile Compounds and Alkaloids of Ephedra alata Growing Wild in Southern Tunisia and Evaluation of their Antioxidant Activity

In order to provide more information about phytochemical composition of Tunisian Ephedra alata known for their medicinal uses, fatty acids, volatile compounds and alkaloids have been analyzed in fresh or dry plant materials whether powdered or not. Extraction was performed with hexane and ethanol in soxhlet extractor and all extracts were analyzed by GC-MS. Phosphomolybdenum, DPPH and FRAP methods were used to evaluate their antioxidant activities. The dry matter present a more diversified fatty acids composition dominated by palmitic acid, oleic acid and linolelaidic acid. Hydrocarbons represent the largest group of volatile compounds, followed by alcohols. The most known detected ephedrine-type alkaloid characteristic of the Ephedra genus are dominated by pseudoephedrine, norephedrine, ethylephedrine. Some other compounds are mentioned for the first time in E. alata alkaloids such as azetidine, 1,2- dimethyl-3-phenylaziridine, N-ethyl benzamide and N-methyl-mandelamide. Lower total antioxidant and anti-radicular effects were shown with hexane extracts and alkaloid fractions instead the ferric reducing antioxidant power test showed the highest results with obtained alkaloid from powdered fresh and dry plant materials. Lipids, volatile compounds and alkaloids in E. alata that have been assessed could provide possible roles in case of human breast cancer and health benefits and therefore a research on this plant might be of value in drug industry.

A Multitarget Approach toward the Development of 8-Substituted Purines for Photoprotection and Prevention of UV-Related Damage.

Ultraviolet (UV) light is the most abundant and significant modifiable risk factor for skin cancer and many other skin diseases such as early photo-aging. Across the solar radiation spectrum, UV light is the main cause behind skin problems. In the search for novel photoprotective compounds, a new series of 8-substituted purines were synthesized from commercially available 6-hydroxy-4,5-diaminopyrimidine hemisulfate or 4,5-diaminopyrimidine. All title compounds were investigated for their UV filtering, antioxidant, antifungal, and antiproliferative activities. For the photoprotection assays we used a diffuse transmittance technique to determine the sun protection factor (SPF) in vitro, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric ion reducing antioxidant power (FRAP) tests for evaluating the antioxidant activity of the more potent compounds. Among them, 8-(2,5-dihydroxyphenyl)-7H-purin-6-ol (compound 26) proved to be a good radical scavenger and is also endowed with broad-spectrum UVA filtering capabilities, suitable for further development as a protective molecule.

Inhibitors of ceramide de novo biosynthesis rescue damages induced by cigarette smoke in airways epithelia.

Exposure to cigarette smoke represents the most important risk factor for the development of chronic obstructive pulmonary disease (COPD). COPD is characterized by chronic inflammation of the airways, imbalance of proteolytic activity resulting in the destruction of lung parenchyma, alveolar hypoxia, oxidative stress, and apoptosis. Sphingolipids are structural membrane components whose metabolism is altered during stress. Known as apoptosis and inflammation inducer, the sphingolipid ceramide was found to accumulate in COPD airways and its plasma concentration increased as well. The present study investigates the role of sphingolipids in the cigarette smoke-induced damage of human airway epithelial cells. Lung epithelial cells were pre-treated with sphingolipid synthesis inhibitors (myriocin or XM462) and then exposed to a mixture of nicotine, acrolein, formaldehyde, and acetaldehyde, the major toxic cigarette smoke components. The inflammatory and proteolytic responses were investigated by analysis of the mRNA expression (RT-PCR) of cytokines IL-1β and IL-8, and matrix metalloproteinase-9 and of the protein expression (ELISA) of IL-8. Ceramide intracellular amounts were measured by LC-MS technique. Ferric-reducing antioxidant power test and superoxide anion radical scavenging activity assay were used to assess the antioxidant power of the inhibitors of ceramide synthesis. We here show that ceramide synthesis is enhanced under treatment with a cigarette smoke mixture correlating with increased expression of inflammatory cytokines and matrix metalloproteinase 9. The use of inhibitors of ceramide synthesis protected from smoke induced damages such as inflammation, oxidative stress, and proteolytic imbalance in airways epithelia.

The estimation of the traditionally used yarrow (Achillea millefolium L. Asteraceae) oil extracts with anti-inflamatory potential in topical application

Ethnopharmacological relevanceOlive or sunflower oil yarrow extracts have been extensively used against inflammatory disorders and skin wound healing in traditional medicine. Aim of the studyTo evaluate oil yarrow extracts traditional use in treatment of topical/dermatological skin impairments, sodium lauryl sulfate test was applied, and in vivo measuring of the biophysical parameters (erythema index, skin capacitance and the pH of the skin) in the artificially irritated skin was performed. As traditionally olive and sunflower oil have been used equally for extracts production, the experiment was carried out to investigate whether any of the oil extractants has the advantage over the other, and if the method of extraction might influence the desired activity. The observed activity has been connected to the chemical profile of the investigated extracts and their antioxidative properties. Materials and methodsIn vivo measurements were performed using the appropriate probes for measuring skin capacitance, pH of the skin and erythema index (EI). The designed experiment enabled the evaluation of the anti-inflammatory effects of a seven-day application of oil yarrow extracts known in traditional medicine, on artificially irritated skin of volunteers. The chemical profile for the investigated samples was achieved applying the HPLC and UHPLC-MS methods. Also, ferric reducing antioxidant power (FRAP) test was performed to assess the antioxidant properties of the investigated samples. ResultsThe application of tested oil extracts on artificially irritated skin in vivo demonstrated the ability to re-establish their optimal pH and hydration of skin to the values measured prior to the irritation. Considering the EI transition, the investigated samples succeeded in re-establishing the baseline values, with no significant difference after three- and seven-day application. ConclusionsThe data obtained in the study showed that the oil yarrow extracts had an evident anti-inflammatory property. Namely, the investigated extracts demonstrated significant anti-inflammatory effect in an in vivo double blind randomized study, using a sodium lauryl sulfate test. The skin parameters assessed in the study (skin capacitance, pH and EI) were restored to the basal values after three- and seven-day treatment with the tested extracts. The shown effects were attributed to yarrow oil extracts composition. The yarrow oil extracts might be used as promising base in the phytopreparations designed for dermatological application as anti-inflammatory agents with a positive impact on the skin pH and its moisture content.

Niosomes Consisting of Tween-60 and Cholesterol Improve the Chemical Stability and Antioxidant Activity of (-)-Epigallocatechin Gallate under Intestinal Tract Conditions.

In order to improve the chemical stability and antioxidant activity of (-)-epigallocatechin gallate (EGCG) in the gastrointestinal tract, niosomes composed of Tween-60 and cholesterol were developed to encapsulate EGCG in this investigation. EGCG loaded niosomes with encapsulation efficiency around 76% exhibited a small Z-average diameter about 60 nm. Compared to free EGCG, the EGCG remaining in dialysis tubes was significantly improved for niosomes at pH 2 and 7.4. Meanwhile, the residual EGCG for niosomes increased from 3% to 49% after 2 h incubation in simulated intestinal fluid (SIF). Pancreatin was found to impact the stability of niosomes in SIF mainly. Furthermore, the results from ferric reducing antioxidant power and cellular antioxidant activity tests indicated that EGCG loaded niosomes exhibited stronger antioxidant ability than free EGCG during intestinal digestion. Thus, we can infer that niosomal encapsulation might be a promising approach to improve the oral bioavailability of EGCG in the body.

Antioxidant capacities of Artemisia annua L. leaves and enzymatically treated Artemisia annua L. in vitro and in broilers

The antioxidant capacities of Artemisia annua L. leaves (AL) and enzymatically treated A. annua L. (EA) were investigated both in vitro and in broilers in the current study. Total phenolic and flavonoid contents were determined by Folin-Ciocalteu and aluminium chloride methods, respectively. Antioxidant capacities were evaluated by scavenging activities against 2,2-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP) test and reducing power (RP) test. The total phenolic and flavonoid contents, ABTS and DPPH radical scavenging activities, FRAP and RP of AL were higher (P<0.05) than EA. The effects of AL and EA on growth performance and antioxidant capacity of broilers were investigated by using 192 one-d-old broilers. Birds were randomly allotted into three groups with eight replicates of eight birds each, and received a basal diet and basal diet supplemented with either 5g/kg AL or 1g/kg EA until 42 d of age, respectively. Compared to control, broilers in AL group had lower (P<0.05) average daily gain (ADG) in 1–21 d. EA increased (P<0.05) ADG in 1–21 d and 1–42 d compared to control and AL groups. Both AL and EA increased (P<0.05) activities of total superoxide dismutase (T-SOD) in serum at 42 d and in liver at 21 d, glutathione peroxidase (GSH-Px) in liver at 42 d, and catalase (CAT) in serum and liver at 42 d, and decreased (P<0.05) malondialdehyde (MDA) concentration in liver at 21 d. But only EA increased (P<0.05) activities of T-SOD in serum at 21 d and in liver at 42 d, GSH-Px in serum at 21 d and 42 d and in liver at 21 d, and reduced (P<0.05) MDA concentration in serum at 21 d and 42 d and in liver at 42 d. In conclusion, EA would be more effective than AL in improving growth performance and antioxidant capacity in broilers.

Effect of cadmium on phenolic compounds, antioxidant enzyme activity and oxidative stress in blueberry (Vaccinium corymbosum L.) plantlets grown in vitro

Cadmium (Cd2+) can affect plant growth due to its mobility and toxicity. We evaluated the effects of Cd2+ on the production of phenolic compounds and antioxidant response of Vaccinium corymbosum L. Plantlets were exposed to Cd2+ at 50 and 100µM for 7, 14 and 21 days. Accumulation of malondialdehyde (MDA), hydrogen peroxide (H2O2) and the antioxidant enzyme SOD was determined. The profile of phenolic compounds was evaluated using LC-MS. The antioxidant activity was measured using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and the ferric reducing antioxidant power test (FRAP). Cd2+ increased the content of MDA, with the highest increase at 14 days. The presence of Cd2+ resulted in changes in phenolic compounds. The main phenolic compound found in blueberry plantlets was chlorogenic acid, whose abundance increased with the addition of Cd2+ to the medium. The changes in the composition of phenolic compounds showed a positive correlation with the antioxidant activity measured using FRAP. Our results suggest that blueberry plantlets produced phenolic compounds with reducing capacity as a selective mechanism triggered by the highest activity of Cd2+.

Effects of extracts and isolated compounds from safflower on some index of promoting blood circulation and regulating menstruation

Ethnopharmacological relevanceCarthamus tinctorius is used as one of the Traditional Chinese Medicine (TCM) materials in prescriptions and composite to promote blood circulation to remove blood stasis, regulate menstruation and alleviate pain for over 2500 years. Modern pharmacological experiments have demonstrated that safflower has wide-reaching biological activities, including dilating coronary artery, modulating immune system, improving myocardial ischemia, anticoagulation and thromboprophylaxis, antioxidation, antihypoxic, antiaging, antifatigue, antiinflammation, anti-hepatic fibrosis, antitumor, analgesia, etc. Materials and methodsPlatelet aggregation of safflower extract and main constituents in safflower were determined by PAF-induced or ADP-induced platelet aggregation in vitro. Anticoagulation activity was measured by clotting assay of thrombin time (TT), prothrombin time (PT) and activated partial thromboplastin time (APTT) according to the methods provided by the biological reagents provider (Sun Biochemical). Antioxidant effects of safflower were assessed using DPPH radical-scavenging activity test, ABTS radical-scavenging activity test and ferric reducing antioxidant power test. In addition, rats ovary granulosa cell proliferation activity was used for the bio-activity index on regulate menstruation of safflower. ResultsSafflower extract at the concentration of 0.7g/mL (P<0.001) and 0.5g/mL (P<0.01) had significantly antagonistic effect on PAF-induced platelet aggregation, compared with negative control. And the anti-platelet aggregation of 0.7g/mL safflower extract was significantly stronger than that of positive control (P<0.001). 0.7g/mL of hydroxysafflor yellow A (P<0.01), anhydrosafflor yellow B (P<0.05), 6-hydroxykaempferol-3-O-rutinoside (P<0.05), keampferol-3-O-β-rutinoside (P<0.01) had significant effect on platelet aggregation compared with negative control. Safflower extract at the concentration of 0.5g/mL (P<0.001) and 0.125g/mL (P<0.01) could significantly inhibit ADP-induced platelet aggregation, compared with negative control. And antagonistic effect of safflower extract was significantly stronger than the effect of positive control (P<0.001). Adenosine (P<0.001), anhydrosafflor yellow B (P<0.01) and 6-hydroxykaempferol-3-O-rutinoside (P<0.01) at the concentration of 0.5g/mL had significant effect on ADP-induced platelet aggregation compared with negative control. 0.125g/mL of adenosine (P<0.05) had significant effect on ADP-induced platelet aggregation compared with negative control. The effect of 0.5g/mL adenosine (P<0.01) and 6-hydroxykaempferol-3-O-rutinoside (P<0.05) was significantly stronger than that of positive control. Safflower extract at the concentration of 0.7mg/mL (P<0.001) and 0.5mg/mL (P<0.001) had significantly anticoagulation activity in PT, TT and APTT, compared with negative control. However, the respective compound didn’t have significant effect on PT and TT at experiment concentration. At the concentration of 0.7mg/mL, hydroxysafflor yellow A (P<0.01), 6-hydroxykaempferol-3,6,7-tri-O-β-d-glucoside (P<0.05), 6-hydroxyapigenin-6-O-glucoside-7-O-glucuronide (P<0.01), anhydrosafflor yellow B (P<0.001), 6-hydroxykaempferol-3-O-rutinoside (P<0.05) and keampferol-3-O-β-rutinoside (P<0.05) significantly prolonged APTT, compared with negative control. At the concentration of 0.5mg/mL, hydroxysafflor yellow A (P<0.05), 6-hydroxyapigenin-6-O-glucoside-7-O-glucuronide (P<0.05), anhydrosafflor yellow B (P<0.001), 6-hydroxykaempferol-3-O-rutinoside (P<0.05) and keampferol-3-O-β-rutinoside (P<0.05) could significantly prolong APTT, compared with negative control. From the results of DPPH, ABTS radical scavenging activity test and Fe3+ reduction power test, 5mg/mL, 2.5mg/mL and 1.25mg/mL safflower extract had antioxidant effects. Every compound with each concentration (5mg/mL, 2.5mg/mL and 1.25mg/mL) had significant effect on Fe3+ reduction power (P<0.001 vs. negative control). Safflower extract, cytidine, 6-hydroxy-kaempferol-3,6-di-O-β-d-glucoside-7-O-β-d-glucuronide, 6-hydroxykaemp-ferol-3,6,7-tri-O-β-D-glucoside and keampferol-3-O-β-rutinoside significantly promoted ovarian granulosa cell proliferation. ConclusionBased on previous researches, the activities of safflower extract and pure compounds isolated from safflower were studied in this paper. This study found some compounds with the effects of anti-platelet aggregation, anticoagulation, antioxidation and ovarian granulosa cell proliferation, and further revealed the possible pharmacological mechanism of safflower.

Antioxidant activity of Echinophora platyloba DC essential oil: a comparative study on four different methods

The present study is conducted in order to investigate the antioxidant capacity of Echinophora platyloba DC (Duncan) essential oil (EO), growing wild in west Azerbaijan, Iran, with four different assays. The aerial parts of Echinophora platyloba DC were provided from Maraghe city district, northwest of Iran and its phytochemicals were determined using GC-MS analysis. In order to evaluate and compare the antioxidant activity of the EO, various concentrations (1, 2.5, 5 and 10 mg.ml-1) of the oil and reference antioxidants (ascorbic acid and BHT) were analyzed by four different assays such as 2,2’-azinobis 3-ethylbenzothiazoline-6-sulphonic acid (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), Ferric reducing antioxidant power (FRAP) and β-carotene bleaching test (BCBT). The oil exhibited moderate antioxidant activity with a dose-response in DPPH and ABTS assays and it exhibited strong antioxidant activity in FRAP and BCBT assays. It is concluded that FRAP and BCBT assays are more suitable spectrometric assays for antioxidant capacity evaluation of Echinophora platyloba.

Phenolic Compounds in Chilean Mistletoe (Quintral, Tristerix tetrandus) Analyzed by UHPLC-Q/Orbitrap/MS/MS and Its Antioxidant Properties.

Mass spectrometry has become a method of choice to characterize bioactive compounds in biological samples because of its sensitivity and selectivity. Hybrid ultra-HPLC hyphenated with Orbitrap mass analyzer is an innovative state of the art technology that allows fast and accurate metabolomic analyses. In this work the metabolites of a Chilean mistletoe endemic to the VIII region of Chile were investigated for the first time using UHPLC mass analysis (UHPLC-PDA-HESI-Orbitrap MSn). The anthocyanins, together with the non-pigmented phenolics were fingerprinted and correlated with the antioxidant capacities measured by the bleaching of the DPPH radical, the ferric reducing antioxidant power (FRAP), the superoxide anion scavenging activity assay (SA), and total content of phenolics, flavonoids and anthocyanins measured by spectroscopic methods. Six anthocyanins were identified, and among them, the 3-O-glycosides of delphinidin and cyanidin were the major ones. In addition, several phenolic acids (including feruloylquinic acid, feruloyl glucose, chlorogenic acid) and several flavonols (luteolin, quercetin, apigenin, isorhamnetin and glycoside derivatives) were also identified. The mistletoe leaves showed the highest antioxidant activity as measured by the DPPH radical bleaching, ferric reducing antioxidant power and superoxide anion scavenging activity tests (13.38 ± 0.47 µg/mL, 125.32 ± 5.96 µmolTE/g DW and 84.06 ± 4.59 at 100 µg/mL, respectively).

Antioxidant activity of various extracts of selected gourd vegetables.

Study was conducted to evaluate the antioxidative activity of methanolic (ME), ethanolic (EE) and butanolic extracts (BE) of selected gourd vegetables. The antioxidant activity was investigated using different assays namely ferric thiocyanate test (FTC), thiobarbituric acid test (TBA), ferric reducing antioxidant power (FRAP) and DPPH free radicals scavenging test. A densitometric HPTLC analysis was performed for the analysis of phenolic acids and flavonoids. Different extracts of the selected gourd vegetables revealed different antioxidant activity. Different extracts of Lagenaria siceraria, Momordica charantia and Luffa cylindrica revealed significantly higher (p<0.05) concentrations of total phenols, flavonids, tannins and carotenoids content and also the antioxidant activity in comparison to remaining vegetable extracts. Correlation studies indicated that FRAP test best described the antioxidant activity of phenols, flavonoids and carotenoids (r=0.854, 0.692 and 0.915 respectively). HPTLC profiles revealed the presence of maximum number of phenolic acids and flavonoids in L. siceraria and M. charantia.

Neuroprotective and Antioxidant Activities of 4-Methylcoumarins: Development of Structure-Activity Relationships.

Coumarins are a major class of polyphenols that are abundantly present in many dietary plants and possess different biological activities. Neuroprotective effect of 28 variously substituted 4-methylcoumarins was evaluated in a cell model of oxidative stress-induced neurodegeneration, which measures viability in PC12 cells challenged with hydrogen peroxide by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The inhibitory activity of these compounds against intracellular reactive oxygen species (ROS) formation was also determined by 2',7'-dichlorofluorescein diacetate method in the same cells. Chemical redox-based assays including 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) tests were employed to explore structure-antioxidant activity relationships in a cell-free environment. The results demonstrated that 4-methylcoumarins containing ortho-dihydroxy or ortho-diacetoxy substituents on the benzenoid ring possess considerable neuroprotective effects. ortho-Dihydroxy compounds inhibited cytotoxicity (44.7-62.9%) and ROS formation (41.6-71.1%) at 50 µM and showed considerable antioxidant effects. We conclude that 4-methylcoumarins are promising neuroprotective and antioxidant scaffolds potentially usefull for management of neurodegenerative diseases.