Search results for: DPPH assays

Authors: Saeid Jafari, Khursheed Ahmad Sheikh, Randy W. Worobo, Kitipong Assatarakul

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In this study, the extraction of cocoa shell powder (CSP) was optimized, and the optimized extracts were spray-dried for encapsulation purposes. Temperature (45-65 ◦C), extraction time (30–60 min), and ethanol concentration (60–100%) were the extraction parameters. The response surface methodology analysis revealed that the model was significant (p ≤ 0.05) in interactions between all variables (total phenolic compound, total flavonoid content, and antioxidant activity as measured by 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP assays), with a lack of fit test for the model being insignificant (p > 0.05). Temperature (55 ◦C), time (45 min), and ethanol concentration (60%) were found to be the optimal extraction conditions. For spray-drying encapsulation, some quality metrics (e.g., water solubility, water activity) were insignificant (p > 0.05). The microcapsules were found to be spherical in shape using a scanning electron microscope. Thermogravimetric and differential thermogravimetric measurements of the microcapsules revealed nearly identical results. The gum arabic + maltodextrin microcapsule (GMM) showed potential antibacterial (zone of inhibition: 11.50 mm; lower minimum inhibitory concentration: 1.50 mg/mL) and antioxidant (DPPH: 1063 mM trolox/100g dry wt.) activities (p ≤ 0.05). In conclusion, the microcapsules in this study, particularly GMM, are promising antioxidant and antibacterial agents to be fortified as functional food ingredients for the production of nutraceutical foods with health-promoting properties.

Keywords: functional foods, coco shell powder, antioxidant activity, encapsulation, extraction

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Authors: Tatjana Kadifkova Panovska, Svetlana Kulevanova, Blagica Jovanova

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Biological systems possess the ability to neutralize the excess of reactive oxygen species (ROS) and to protect cells from destructive alterations. However, many pathological conditions (cardiovascular diseases, autoimmune disorders, cancer) are associated with inflammatory processes that generate an excessive amount of reactive oxygen species (ROS) that shift the balance between endogenous antioxidant systems and free oxygen radicals in favor of the latter, leading to oxidative stress. Therefore, an additional source of natural compounds with antioxidant properties that will reduce the amount of ROS in cells is much needed despite their broad utilization; many plant species remain largely unexplored. Therefore, the purpose of the present study is to investigate the antioxidant activity of twenty-five selected medicinal and aromatic plant species. The antioxidant activity of the ethanol extracts was evaluated with in vitro assays: 2,2’-diphenyl-1-pycryl-hydrazyl (DPPH), ferric reducing antioxidant power (FRAP), non-site-specific- (NSSOH) and site-specific hydroxyl radical-2-deoxy-D-ribose degradation (SSOH) assays. The Folin-Ciocalteu method and AlCl3 method were performed to determine total phenolic content (TPC) and total flavonoid content (TFC). All examined plant extracts manifested antioxidant activity to a different extent. Cinnamomum verum J.Presl bark and Ocimum basilicum L. Herba demonstrated strong radical scavenging activity and reducing power with the DPPH and FRAP assay, respectively. Additionally, significant hydroxyl scavenging potential and metal chelating properties were observed using the NSSOH and SSOH assays. Furthermore, significant variations were determined in the total polyphenolic content (TPC) and total flavonoid content (TFC), with Cinnamomum verum and Ocimum basilicum showing the highest amount of total polyphenols. The considerably strong radical scavenging activity, hydroxyl scavenging potential and reducing power for the species mentioned above suggest of a presence of highly bioactive phytochemical compounds, predominantly polyphenols. Since flavonoids are the most abundant group of polyphenols that possess a large number of available reactive OH groups in their structure, it is considered that they are the main contributors to the radical scavenging properties of the examined plant extracts. This observation is supported by the positive correlation between the radical scavenging activity and the total polyphenolic and flavonoid content obtained in the current research. The observations from the current research nominate Cinnamomum verum bark and Ocimum basilicum herba as potential sources of bioactive compounds that could be utilized as antioxidative additives in the food and pharmaceutical industries. Moreover, the present study will help the researchers as basic data for future research in exploiting the hidden potential of these important plants that have not been explored so far.

Keywords: ethanol extracts, radical scavenging activity, reducing power, total polyphenols.

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Authors: Leila Najafian

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Sarcoplasmic protein hydrolysates (SPHs) and myofibrillar protein hydrolysates (MPHs) from patin (Pangasius sutchi) were produced using two types of proteases: Papain and Alcalase. 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS) radical scavenging activities and metal chelating activity assays for antioxidant activities were carried out on the SPHs and MPHs. The hydrolysates were isolated and purified by ultrafiltration, gel filtration and reverse phase high-performance liquid chromatography (RP-HPLC) and liquid chromatography with tandem mass spectrometry detection (LC-MS/MS) was used in identifying peptide sequences. The results showed that when the DH of MPHs increased, the protein solubility increased, while the highest amount of the protein solubility of SPHs was after 60 min incubation. The effect of DH on antioxidant activities of SPHs and MPHs was investigated. Among the hydrolysates, papain-MPH and Alcalase-SPH, which had the highest antioxidant activities, were purified. The potent fractions obtained from RP-HPLC of sarcoplasmic (SI 3 fraction) and myofibrillar (MI 4 fraction) hydrolysates showed the highest DPPH radical scavenging activity. The FVNQPYLLYSVHMK peptide for MPH and the LVVDIPAALQHA peptide for SPH exhibited the highest antioxidant activity. The presence of hydrophobic and hydrophilic amino acids, namely leucine (L), valine (V), phenylalanine (F), histidine (H) and proline (P), in the peptide sequences of SPH and MPH are believed to contribute to high antioxidant activity. Hence, SPH and MPH from patin have the potential as a natural functional ingredient in food and pharmaceutical industry.

Keywords: patin (Pangasius sutchi), protein hydrolysates, antioxidative peptides, mass spectrometry

Procedia PDF Downloads 238

Authors: Djemai Zoughlache Soumia, Yahia Mouloud, Lekbir Adel, Meslem Meriem, Maouchi Madiha, Bahi Ahlem, Benbia Souhila

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Natural extracts is known for their contents of biologically active molecules. In this context, we attempted to evaluate the antioxidant activity of the methanolic extract prepared from the bark of the roots of Zizyphus lotus. The quantitative analysis based on the dosage, phenolic compounds, flavonoids and tannins provided following values: 0.39 ± 0.007 ug EAG/mg of extract for phenolic compounds, 0.05 ± 0.02ug EQ/mg extract for flavonoids and 0.0025 ± 7.071 E-4 ECT ug/mg extract for tannins. The study of the antioxidant activity by the DPPH test in vitro showed a powerful antiradical power with an IC50 = 8,8 ug/ml. For the DPPH test in vivo we used two rats lots, one lot with a dose of 200 mg/kg of the methanol extract and a control lot. We found a significant difference in antiradical activity with p < 0.05.

Keywords: Zizyphus lotus, antioxidant activity, DPPH, phenolic compounds, flavonoids, tannins

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Authors: T. Ould Bellahcen, A. Amiri, I. Touam, F. Hmimid, A. El Amrani, M. Cherki

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The antioxidant activity of three extracts (water, lipidic and ethanolic) prepared from the microalgae Spirulina platensis isolated from Moroccan lake, using 2, 2 diphenyl-1-picrylhydrazyl (DPPH) and 2,2’-azino-bis ethylbenzthiazoline-6-sulfonic acid (ABTS) radical assay, was studied and compared. The obtained results revealed that the IC₅₀ found using DPPH were lower than that of ABTS for all extracts from these planktonic blue-green algae. The high levels of phenolic and flavonoid content were found in the ethanolic extract 0,33 ± 0,01 mg GAE/g dw and 0,21 ± 0,01 mg quercetin/g dw respectively. In addition, using DPPH, the highest activity with IC₅₀ = 0,449 ± 0,083 mg/ml, was found for the ethanolic extract, followed by that of lipidic extract (IC₅₀ = 0,491 ± 0,059 mg/ml). The lowest activity was for the aqueous extract (IC₅₀ = 4,148 ± 0,132 mg/ml). For ABTS, the highest activity was observed for the lipidic extract with IC₅₀ = 0,740 ± 0,012 mg/ml, while, the aqueous extract recorded the lowest activity (IC₅₀ = 6,914 ± 0, 0067 mg/ml). A moderate activity was showed for the ethanolic extract (IC₅₀ = 5,852 ± 0, 0171 mg/ml). It can be concluded from this first study that Spirulina platensis extracts show an interesting antioxidant and antiradicals properties suggesting that this alga could be used as a potential source of antioxidants. A qualitative and quantitative analysis of polyphenol and flavonoids in the extracts using HPLC is in progress so as to study the correlation between the antioxidant activity and chemical composition.

Keywords: Spirulina platensis, antioxidant, DPPH, ABTS

Procedia PDF Downloads 143

Authors: Sharon N. Nuñal, May Flor S. Muegue, Nizzy Hope N. Cartago, Raymund B. Parcon, Sheina B. Logronio

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The antioxidant and anti-inflammatory potentials of Perna Viridis, Modiolus philippinarum, and Mytella charruana found in the Philippines were assessed. Mussel protein samples were hydrolyzed using trypsin, maturase, alcalase and pepsin at 1% and 2% concentrations and then fractionated through membrane filtration (<10 kDa and <30 kDa). Antioxidant assays showed that pepsin hydrolysate at 2% enzyme concentration exhibited the maximum activities for both 2,2-Diphenyl-1-picrylhydrazyl (DPPH) Radical Scavenging Activity (155-176 µM TE/mg protein) and 2,2-azinobis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging (67-68 µM TE/mg protein) assays while trypsin hydrolysate dominated the Ferric Reducing Antioxidant Power (FRAP) for the three mussel species. Lower molecular weight peptide fractions at <10 kDa exhibited better antioxidant activities than the higher molecular weight fractions. The anti-inflammatory activities of M. philippinarum and M. charruana showed comparable protein denaturation inhibition potentials with the highest in P. Viridis samples (98.93%). The 5-Lipoxygenase (5-LOX) inhibitory activities of mussel samples showed no significant difference with inhibition exceeding 70%. P. Viridis demonstrated the highest inhibition against Cyclooxygenase-2 (COX-2) at 56.19%, while the rest showed comparable activities. This study showed that the three mussel species are potential sources of bioactive peptides and lipids with antioxidant and anti-inflammatory properties.

Keywords: anti-inflammatory, antioxidant, bioactive properties, mussel

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Authors: Luís R. Silva, Ana C. Gonçalves, Catarina Bento, Fábio Jesus, Branca M. Silva

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Prunus avium Linnaeus, more known as sweet cherry, is one of the most appreciated fruit worldwide. Most of these quantities are produced in Fundão region, being Saco the cultivar most produced. Saco is very rich in bioactive compounds, especially phenolics, and presents great antioxidant capacity. The purpose of the present study was to investigate the chemical profile and biological potential, concerning antioxidant, anti-diabetic activity and protective effects towards erythrocytes by Saco sweet cherry collected from Fundão region (Portugal). The hydroethanolic extracts were prepared and passed through a C18 solid-phase extraction column. The phenolic profile analyzed by LC-DAD method allowed to the identification of 22 phenolic compounds, being 16 non-phenolics and 6 anthocyanins. In respect to non-coloured phenolics, 3-O-caffeoylquinic and ρ-coumaroylquinic acids were the main ones. Concerning to anthocyanins, cyanidin-3-O-rutinoside was found in higher amounts. Relatively to biological potential, Saco showed great antioxidant potential, through DPPH and NO radical assays, with IC50 =16.24 ± 0.46 µg/mL and IC50 = 176.69 ± 3.35 µg/mL for DPPH and NO, respectively. These results were similar to those obtained for ascorbic acid control (IC50 = 16.92 ± 0.69 and IC50 = 162.66 ± 1.31 μg/mL for DPPH and NO, respectively). In respect to antidiabetic potential, Saco revealed capacity to inhibit α-glucosidase in a dose-dependent manner (IC50 = 10.79 ± 0.40 µg/mL), being much active than positive control acarbose (IC50 = 306.66 ± 0.84 μg/mL). Additionally, Saco extracts revealed protective effects against ROO•-mediated toxicity generated by AAPH in human blood erythrocytes, inhibiting hemoglobin oxidation (IC50 = 38.57 ± 0.96 μg/mL) and hemolysis (IC50 = 73.03 ± 1.48 μg/mL), in a concentration-dependent manner. However, Saco extracts were less effective than quercetin control (IC50 = 3.10 μg/mL and IC50 = 0.7 μg/mL for inhibition of hemoglobin oxidation and hemolysis, respectively). The results obtained showed that Saco is an excellent source of phenolic compounds. These ones are natural antioxidant substances, which easily capture reactive species. This work presents new insights regarding sweet cherry antioxidant properties which may be useful for the future development of new therapeutic strategies for preventing or attenuating oxidative-related disorders.

Keywords: antioxidant capacity, health benefits, phenolic compounds, saco

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Authors: Thi Huong Vu, Vijay Jayasena, Zhongxiang Fang, Gary Dykes

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Carob kibble has recently received attention due to the presence of high level of polyphenol antioxidants. The capacity of microorganisms to improve antioxidant activities and total phenolics in carob kibble was investigated in the study. Two types of microorganisms including lactic acid bacteria Lactobacillus plantarum (L. plantarum) and yeast Saccharomyces cerevisiae (S. cerevisiae) were used in single and in their combination as starters. The total phenolic content was determined by the Folin–Ciocalteu method. Antioxidant activities were assessed scavenging capacity using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS). The study found that S. cerevisiae alone considerably improved 55% total phenolics content at 15 h, while L. plantarum caused in a loss of 20% through the process. Antioxidant capacity of the yeast-fermented samples significantly increased by 43 % and 10 % in ABTS and DPPH assays, respectively. However, reduction of 13 % and 32 % inhibition were recorded in the carob treated with L. plantarum. In the combination of S. cerevisiae and L. plantarum (1:1), both total phenolic content and antioxidant activity of carob kibble were a similar trend as these of S. cerevisiae single, but a lower improvement. The antioxidant power of the extracts was linearly correlated to their total phenolic contents (R=0.75). The results suggested that S. cerevisiae alone was the better for enhancement of both total phenolic content and antioxidant activity in carob kibble using submerged fermentation. The efficiency of fermentation reached the highest at 15h. Thus submerged fermentation with S. cerevisiae offers a tool with simple and cost effective to further increase the bioactive potential of carob kibble, which is in use for food, cosmetic and pharmaceutical industries.

Keywords: antioxidant activity, carob kibble, lactobacillus plantarum, saccharomyces cerevisiae, total phenolics

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Authors: Hawraa Zbeeb, Hala Khalifeh, Mohamad Khalil, Francesca Storace, Francesca Baldini, Giulio Lupidi, Laura Vergani

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Sarcopoterium spinosum, a widely distributed spiny shrub belonging to the Rosaceae family, is rich in essential and beneficial constituents. In fact, S. spinosum fruits and roots are traditionally used as herbal medicine in the eastern Mediterranean landscape, and this shrub is mentioned as a medicinal plant in a large number of ethnobotanical surveys. Aqueous root extracts from S. spinosum are used by traditional medicinal practitioners for weight loss treatment of diabetes and pain. Moreover, the anti-diabetic activity of S. spinosum root extract has been reported in different studies, but the beneficial effects of aerial parts, especially fruits, have not been elucidated yet. The aim of the present study was to investigate the in vitro antioxidant and lipid-lowering properties of an ethanolic extract from S. spinosum fruits using both hepatic (FaO) and endothelial (HECV) cells in an attempt to evaluate its possible employment as a nutraceutical supplement. First of all, in vitro spectrophotometric assays were employed to characterize the extract. The total phenol content (TPC) was evaluated by Folin–Ciocalteu spectrophotometric method and the radical scavenging activity was tested by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2, 2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assays. After that, the beneficial effects of the extract were tested on cells. FaO cells treated for 3 hours with 0.75 mM oleate/palmitate mix (1:2 molar ratio) mimic in vitro a moderate hepato-steatosis. HECV cells exposed for 1 hour to 100 µM H₂O₂ mimic an oxidative insult leading to oxidative stress conditions. After the metabolic and oxidative insult, both cell lines were treated with increasing concentrations of the S. spinosum extract (1, 10, 25 µg/mL) for 24 hours. The results showed the S. spinosum ethanolic extract is rather rich in phenols (TPC of 18.6 mgGAE/g dry extracts). Moreover, the extract showed a good scavenging ability in vitro (IC₅₀ 15.9 µg/ml and 10.9 µg/ml measured by DPPH and ABTS assays, respectively). When the extract was tested on cells, the results showed that it could ameliorate some markers of cell dysfunction. The three concentrations of the extract led to a significant decrease in the intracellular triglyceride (TG) content in steatotic FaO cells measured by spectrophotometric assay. On the other hand, HECV cells treated with increasing concentrations of the extract did not result in a significant decrease in both lipid peroxidation measured by the Thiobarbituric Acid Reactive Substances (TBARS) assay, and in reactive oxygen species (ROS) production measured by fluorometric analysis after DCF staining. Interestingly, the ethanolic extract was able to accelerate the wound repair of confluent HECV cells with respect to H₂O₂-insulted cells as measured by T-scratch assay. Taken together, these results seem to indicate that the ethanol extract from S. spinosum fruits is rich in phenol compounds and plays considerable lipid-lowering activity in vitro on steatotic hepatocytes and accelerates wound healing repair on endothelial cells. In light of that, the ethanolic extract from S. spinosum fruits could be a potential candidate for nutraceutical applications.

Keywords: antioxidant activity, ethanolic extract, lipid-lowering activity, phenolic compounds, Sarcopoterium spinosum fruits

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Authors: Renee P. Baang, Romeo M. del Rosario, Nenita D. Palmes

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The crude methanol extracts of five indigenous vegetables namely, Amarathus tricolor, Basella rubra L, Chochurus olitorius L., Ipomea batatas, and Momordica chuchinensis L., were examined for their phytochemical profile and antioxidant activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical. The values for DPPH radical scavenging activity ranged from 7.6-89.53% with B. rubra and I. batatas having the lowest and highest values, respectively. The total flavonoid content of all five indigenous vegetables ranged from 74.65-277.3 mg quercetin equivalent per gram of dried vegetable material while the total phenolic content ranged from 1.93-6.15 mg gallic acid equivalent per gram dried material. Phytochemical screening revealed the presence of steroids, flavonoids, saponins, tannins, carbohydrates and reducing sugars, which may also be associated with the antioxidant activity shown by these indigenous vegetables.

Keywords: antioxidant, DPPH radical scavenging activity, Philippine İndigenous vegetables, phytochemical screening

Procedia PDF Downloads 308

Authors: Hajir Abdllha, Alaa Mohamed, Khansa Almoniem, Naga Adam, Wdeea Alhaadi, Ahmed Elshikh, Ahmed Ali, Ismail Makuar, Anas Elnazeer, Nagat Elrofaei, Samir Abdoelftah, Monier Hemidan

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The present study was designed to investigate antimicrobial, and antioxidant activities of five species from Acacia (Acacia albidia, Acacia mellifera, Acacia nubica, Acacia seyal var. seyal and Acacia tortilis). Phytochemical study was piloted to detect the bioactive compounds, which have been responsible from the biological activities. The ethanol, chloroform and acetone plant extracts were seasoned against standard bacteria strains of gram +ve bacteria Staphylococcus aureus (ATCC 25923), Gram -ve bacteria Pseudomonas aeruginosa (ATCC 27853) and standard fungi Candida albicans (ATCC 90028), using cup-plate method. The antioxidant activities were conducted via DPPH radical scavenging and metal chelating assays. Prospective activity against the five species was observed in acetone extract. Ethanol extract showed highest activities against Staphylococcus aureus, and Candida albicans. Potential antioxidant activity was presented by ethanol. Cholorophorm and acetone extracts via DPPH, the radical scavenging activities were found to be 91±0.03, 88±0.01 and 85±0.04 respectively. The results of phytochemical screening showed that all extracts of studied plant contain flavonoids, saponins, terpenoids, steroids, alkaloids, phenols and tannins. This study gives rise to antioxidant, antimicrobial properties of studied plant, and showed interesting correlation with the phytochemical constituents and biological activities.

Keywords: antimicrobial, antioxidant, Acacia albidia, Acacia mellifera, Acacia nubica, Acacia seyal var. seyal, Acacia tortilis

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Authors: K. Fellah, H. Benmehdi, A. Amrouche, H. Malainine, F. Memmou, H. Dalile, W. Siata

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This work aims to evaluate the antioxidant of flavonoids extracted from the leaves of Prunus persica L. A phytochemical screening allowed us to highlight the different phytochemicals present in the leaves of the studied plant. The selective extraction of flavonoids gave yields of 0.71, 1.5, and 4.8% for the fractions ethyl ether, ethyl acetate and n- butanol, respectively. The reading of the antioxidant activity of different extracts of flavonoids by HPLTC method revealed positive reaction (yellow spots) on the TLC plates sprayed with DPPH. Using the DPPH method, the fractions of flavonoids (bunanol, ethyl acetate and Diethyl ether) showed a potent scavenging activity with IC50 = 0.22; 0.27 and 0.76 mg / ml, respectively. Furthermore, our findings revealed the extracts under study exhibited higher reducing potential which depends upon extract concentration. These results obtained from this investigation confirm that the Prunus persica remains a major resource of bioactive molecules.

Keywords: Prunus persica L., phytochemical study, flavonoids, antioxidant activity, TLC bioautographic, FRAP, DPPH

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Authors: Hajir, B. Abdllha, , Alaa, I. Mohamed, Khansa, A. Almoniem, Naga, I. Adam, Wdeea, Alhaadi, Ahmed, A. Elshikh, Ahmed, J. Ali, Ismail, G. Makuar, Anas, M. Elnazeer, Nagat, A. Elrofaei, Samir, F. Abdoelftah, Monier, N. Hemidan

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The present study was designed to investigate antimicrobial, and antioxidant activities of five species from Acacia (Acacia albidia, Acacia mellifera, Acacia nubica, Acacia seyal var.seyal and Acacia tortilis). Phytochemical study was piloted to detect the bioactive compounds, which have been responsible from the biological activities. The ethanol, chloroform and acetone plant extracts were seasoned against standard bacteria strains of gram +ve bacteria Staphylococcus aureus (ATCC 25923) ,Gram -ve bacteria Pseudomonas aeruginosa (ATCC 27853) and standard fungi Candida albicans (ATCC 90028), using cup-plate method. The antioxidant activities were conducted via DPPH radical scavenging and metal chelating assays. Prospective activity against the five species was observed in acetone extract. Ethanol extract showed highest activities against Staphylococcus aureus, and Candida albicans. Potential antioxidant activity was presented by ethanol. Cholorophorm and acetone extracts via DPPH, the radical scavenging activities were found to be 91±0.03, 88±0.01 and 85±0.04 respectively. The results of phytochemical screening showed that all extracts of studied plant contain flavonoids, saponins, terpenoids, steroids, alkaloids, phenols and tannins. This study give rise to antioxidant, antimicrobial properties of studied plant, and showed interesting correlation with the phytochemical constituents and biological activities.

Keywords: antimicrobial, Antioxidant, Acacia albidia, Acacia mellifera, acacia nubica, acacia seyal var.seyal, Acacia tortilis

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Authors: Jitpisute Chunthorng-Orn, Thana Juckmeta, Onmanee Prajuabjinda, Arunporn Itharat

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Inflammation and oxidative stress work together to produce symptoms in cancer patients. The whole part of it is used as a preparation to treat cancer patients in Khampramong temple which has been a place of treatment and palliative care for cancer patients since 2005. Thus, the objective of this study was to investigate anti-inflammatory and antioxidant activities of Heliotropium indicum extracts. Dried plant materials were extracted in a similar manner to those practiced by the Khampramong Temple i.e. maceration in 95% ethanol and boiling in water. For anti-inflammation activity, both extracts were tested for suppression of nitric oxide (NO) production in LPS-induced RAW 264.7 cells. They were also tested for antioxidant activity by DPPH radical scavenging assay. This study found that the ethanolic extract of Heliotropium indicum exhibited higher inhibitory activity of NO release than Indomethacin as a positive control (IC50 value of 24.17±2.12 and 34.67±6.23 μg/mL, respectively). For DPPH radical scavenging assay, the ethanolic extract also exhibited antioxidant activity but less than BHT as a antioxidant compound (EC50 values = 28.91±4.26 and 13.08±0.29 μg/mL, respectively). In contrast, its water extract had no inhibitory activity on NO release (IC50 > 100 μg/mL) and no inhibitory activity on DPPH radicals (EC50 values > 100 μg/mL). The results showed correlation between anti-inflammation and antioxidant activity and these results also support using this plant to treat cancer patients.

Keywords: Heliotropium indicum, RAW 264.7, DPPH, Khampramong Temple

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Authors: Asima Shah, F. A. Masoodi, Adil Gani, Bilal Ahmad Ashwar

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The present study was designed to evaluate the effect of γ-rays on the antioxidant and antiproliferative potential of β-glucan isolated from oats. The β-glucan was irradiated with 0, 2, 6, and 10 kGy by gamma ray. The samples were characterized by FT-IR, GPC, and quantitative estimation by Megazyme β-glucan assay kit. The average molecular weight of non-irradiated β-glucan was 199 kDa that decreased to 70 kDa at 10 kGy. Both FT-IR spectrum and chemical analysis revealed that the extracted β-glucan was pure having minor impurities. Antioxidant activity was evaluated by DPPH, lipid peroxidation, reducing power, metal chelating ability and oxidative DNA damage assays. Results revealed that the antioxidant activity of β-glucan increased with the increase in irradiation dose. Irradiated β-glucan also exhibited dose dependent cancer cell growth inhibition with irradiation doses. The study revealed that low molecular weight β-glucan with enhanced antioxidant and antiproliferative activities can be produced by a simple irradiation method.

Keywords: γ-irradiation, antioxidant activity, antiproliferative activity, β-glucan, oats

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Authors: Muhammad Mukhtar

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The aim of this study is to evaluate the antioxidant activity and acute toxicity of the extracts of Ficus iteophylla by reactions with 1, 1-diphenyl-2-picryhydrazyl radical (DPPH) and method developed by Lork 1983, respectively. Stem bark of Ficus iteophylla was collected, air dried, pulverized to fine powdered and sequentially extracted using acetone, methanol and water in order of increasing polarity. The result shows strong radical scavenging activity against DPPH for all the extracts when compared with ascorbic acid. The LD50 of 316 mg/kg was calculated for all the three extras, and the values were found to be within the practically toxic range, and therefore, care should be taken when using the plants in traditional medicine.

Keywords: antioxidant, acute toxicity, Ficus iteophylla

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Authors: Houria Djebar, Amina Saib, Malika Berredjem, Khaoula Bechlem, Mohammed-Reda Djebar

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Reactive oxygen species (ROS) have a high potential to damage almost all types of cellular components of the body, which explains their involvement in the induction and/or amplification of several pathologies. Supplementation of the body by exogenous antioxidants is very useful against these harmful species. In this context, we attempted to evaluate the in vitro and in vivo antioxidant activities of three newly synthesized amidophosphonates (AP1, AP2, and AP3). The results relating to the in vitro tests for DPPH radical scavenging activity shows that these amidophosphonates have a modest antiradical power (ARP) less effectively pronounced compared with an analogue marketed in Algeria: (Dursban) Clorpiryphos ethyl. However, in vivo effects were evaluated on some antioxidant systems (LP intensity, CAT activity and GSH content), or in combination with 2, 2-diphenyl-picrylhydrazyle (DPPH) radical in paramecium tetraurelia used as a complementary system to rapidly elucidate the cytotoxicity. On the basis of the results obtained it can be concluded that amidophosphonates studied exhibited a mild protective effect. The mechanism for how they influenced the antioxidant activities was discussed.

Keywords: Paramecium tetraurelia, amidophosphonates, antioxidant activity, DPPH free radical, in vitro experiments, biochemical parameters

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Authors: S.Kauroo, J. Govinden Soulange, D. Marie

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Psiadia species from the Asteraceae are traditionally used in the folk medicine of Mauritius to treat cutaneous and bronchial infections. The present study aimed at validating the phytomedicinal properties of the selected species from the Asteraceae family, namely Psiadia arguta, Psiadia viscosa, Psiadia lithospermifolia, and Distephanus populifolius. Dried hexane, ethyl acetate, and methanol leaf extracts were studied for their antioxidant properties using the DPPH (1, 1-diphenyl-2-picryl-hydrazyl), FRAP (Ferric Reducing Ability of Plasma), and Deoxyribose assays. Antibacterial activity against human pathogenic bacteria namely Escherichia coli (ATCC 27853), Staphylococcus aureus (ATCC 29213), Enterococcus faecalis (ATCC 29212), Klebsiella pneumonia (ATCC27853), Pseudomonas aeruginosa (ATCC 27853), and Bacillus cereus (ATCC 11778) was measured using the broth microdilution assay. Qualitative phytochemical screening using standard methods revealed the presence of coumarins, tannins, leucoanthocyanins, and steroids in all the tested extracts. The measured phenolics level of the selected plant extracts varied from 24.0 to 231.6 mg GAE/g with the maximum level in methanol extracts in all four species. The highest flavonoids and proanthocyanidins content was noted in Psiadia arguta methanolic extracts with 65.7±1.8 mg QE/g and 5.1±0.0 mg CAT/g dry weight (DW) extract, respectively. The maximum free radical scavenging activity was measured in Psiadia arguta methanol and ethyl acetate extracts with IC50 11.3±0.2 and 11.6± 0.2 µg/mL, respectively and followed by Distephanus populifolius methanol extracts with an IC50 of 11.3± 0.8 µg/mL. The maximum ferric reducing antioxidant potential was noted in Psiadia lithospermifolia methanol extracts with a FRAP value of 18.8 ± 0.4 µmol Fe2+/L/g DW. The antioxidant capacity based on DPPH and Deoxyribose values were negatively related to total phenolics, flavonoid and proanthocyanidins content while the ferric reducing antioxidant potential were strongly correlated to total phenolics, flavonoid and proanthocyanidins content. All four species exhibited antimicrobial activity against the tested bacteria (both Gram-negative and Gram-positive). Interestingly, the hexane and ethyl acetate extracts of Psiadia viscosa and Psiadia lithospermifolia were more active than the control antibiotic Chloramphenicol. The Minimum inhibitory concentration (MIC) values for hexane and ethyl acetate extracts of Psiadia viscosa and Psiadia lithospermifolia against the tested bacteria ranged from (62.5 to 500 µg/ml). These findings validate the use of these tested Asteraceae in the traditional medicine of Mauritius and also highlight their pharmaceutical potential as prospective phytomedicines.

Keywords: antibacterial, antioxidant, DPPH, flavonoids, FRAP, Psiadia spp

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Authors: Chebbah Kaoutar, Marchioni Eric, Menad Ahmed, Mekkiou Ratiba, Sarri Djamel, Ameddah Souad, Boumaza Ouahiba, Seghiri Ramdane, Benayache Samir, Benayache Fadila

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This study was designed to estabilish a preliminary phytochemical screening, evaluate the phenolic and flavonoid content according to the Folin-Ciocalteu procedure, and aluminum chloride method respectively and to determine qualitatively, using HPLC-UV method, the most important products present in ethyl acetate (EtOAc) and n-butanol (n-BuOH) extracts of the aerial parts of Genista cephalantha Spach. from East Algeria. The antioxidant activity of these extracts was spectrophotometrically tested by measuring their ability to scavenge a stable DPPH free radical and by β-Carotene/linoleic acid bleaching assay. Evaluated extracts showed a good activity in both antioxidant system assays.

Keywords: phenolic compounds, flavonoids, HPLC-DAD-UV, antioxidant activity, genista cephalantha, fabaceae

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Authors: Karolis Leonavičius, Dalius Kučiauskas, Dangiras Lukošius, Arnoldas Jasiūnas, Kostas Zdanys, Rokas Stanislovas, Emilis Gegevičius, Žana Kapustina, Juozas Nainys

Abstract:

Screening throughput is a common bottleneck in many research areas, including functional genomics, drug discovery, and directed evolution. High-throughput screening techniques can be classified into two main categories: (i) affinity-based screening and (ii) functional screening. The first one relies on binding assays that provide information about the affinity of a test molecule for a target binding site. Binding assays are relatively easy to establish; however, they reveal no functional activity. In contrast, functional assays show an effect triggered by the interaction of a ligand at a target binding site. Functional assays might be based on a broad range of readouts, such as cell proliferation, reporter gene expression, downstream signaling, and other effects that are a consequence of ligand binding. Screening of large cell or gene libraries based on direct activity rather than binding affinity is now a preferred strategy in many areas of research as functional assays more closely resemble the context where entities of interest are anticipated to act. Droplet sorting is the basis of high-throughput functional biological screening, yet its applicability is limited due to the technical complexity of integrating high-performance droplet analysis and manipulation systems. As a solution, the Droplet Genomics Styx platform enables custom droplet sorting workflows, which are necessary for the development of early-stage or complex biological therapeutics or industrially important biocatalysts. The poster will focus on the technical design considerations of Styx in the context of its application spectra.

Keywords: functional screening, droplet microfluidics, droplet sorting, dielectrophoresis

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Authors: Hyun Young Kim, Min Jung Kim, Ji Hyun Kim, Sanghyun Lee, Eun Ju Cho

Abstract:

Oxidative stress that results from overproduction of free radicals can lead to pathogenesis of human diseases including cancer, neurodegenerative diseases, and cardiovascular disease. Aster yomena (Kitam.) Honda (A. yomena) belonging to Compositae family is a perennial plant, and it has anti-inflammatory, anti-asthmatic and anti-obesity effects. In this study, we investigated the antioxidative effect of A. yomena by measuring 2, 2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl radical (˙OH) and superoxide radical (O₂⁻) scavenging activities in vitro. A. yomena was extracted with ethanol and then partitioned with n-hexane, methylene chloride (CH₂Cl₂), ethyl acetate (EtOAc) and n-butanol (n-BuOH). In DPPH radical scavenging assay, the concentration of A. yomena from 10 to 100μg/mL dose-dependently raised the inhibition of DPPH oxidation. Especially, EtOAc fraction of A. yomena showed the highest DPPH radical scavenging activity among other fractions. The ˙OH radical scavenging activities of the extract and four fractions of A. yomena were increased by over 80% at a concentration of 50μg/mL. Especially, the IC50 value of EtOAc fraction was 0.03 μg/mL that is the lowest value compared with the values of other fractions. In addition, we found that the EtOAc fraction of A. yomena was showed to be better at O₂⁻ radical scavenging than other fractions. Taken together these results, we suggested that A. yomena, especially EtOAc fraction, can be used as a natural antioxidant against free radicals. Acknowledgements: This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (NRF-2016R1D1A1B03931593).

Keywords: Aster yomena (Kitam.) Honda (A. yomena), free radicals, antioxidant, EtOAc fraction

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Authors: Fatiha Bedjou, Meriem Meddas, Tadjajikt Chekkal

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The present work aims to evaluate the antioxidant activity of ethanolic extracts and essential oils of aromatic plants of the Lamiaceae family: Thymus algeriensis and Salvia rosmarinus, and Anacardiaceae: Pistacia lentiscus, Myrtaceae: Eucalyptus polybracetea. The polyphenols were measured using the Folin-Ciocalteu method; the results showed that the essential oils studied as well as the ethanolic extracts are relatively rich in polyphenols. Their antioxidant properties were tested by the synthetic DPPH radical trapping method. The IC50 values were determined according to the graph representing the percentage of inhibition of the DPPH radical by essential oils and by ethanolic extracts, according to our results there is a correlation between the level of polyphenols present in the different essential oils and different ethanolic extracts and their ability to neutralize free radicals. Several combinations were carried out between the essential oils and also between the ethanolic extracts in order to determine the type of interactions existing between the combined substances, the results were represented in the form of isobolograms. Additive and super-additive effects were observed in combinations of essential oils, and super-additive and sub-additive effects were observed for combinations of ethanolic extracts.

Keywords: essential oils, ethanolic extracts, DPPH, combination

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Authors: Ntokozo Dambuza, Maryna Van De Venter, Trevor Koekemoer

Abstract:

Oxidative stress contributes to the pathogenesis of many diseases and consequently antioxidant therapy has attracted much attention as a potential therapeutic strategy. Regardless of the quantities ingested, antioxidants need to reach the diseased tissues at concentrations sufficient to combat oxidative stress. Bioavailability is thus a defining criterion for the therapeutic efficacy of antioxidants. In addition, therapeutic antioxidants must possess biologically relevant characteristics which can target the specific molecular mechanisms responsible for disease related oxidative stress. While many chemical antioxidant assays are available to quantify antioxidant capacity, they relate poorly to the biological environment and provide no information as to the bioavailability. The present comparative study thus aims to characterise green and fermented rooibos extracts, well recognized for their exceptional antioxidant capacity, in terms of antioxidant bioavailability and efficacy in a disease relevant cellular setting. Chinese green tea antioxidant activity was also evaluated. Chemical antioxidant assays (FRAP, DPPH and ORAC) confirmed the potent antioxidant capacity of both green and fermented rooibos, with green rooibos possessing antioxidant activity superior to that of fermented rooibos and Chinese green tea. Bioavailability was assessed using the PAMPA assay and the results indicate that green and fermented rooibos have a permeation coefficient of 5.7 x 10-6 and 6.9 x 10-6 cm/s, respectively. Chinese green tea permeability coefficient was 8.5 x 10-6 cm/s. These values were comparable to those of rifampicin, which is known to have a high permeability across intestinal epithelium with a permeability coefficient of 5 x 10 -6 cm/s. To assess the antioxidant efficacy in a cellular context, U937 and red blood cells were pre-treated with rooibos and Chinese green tea extracts in the presence of a dye DCFH-DA and then exposed to oxidative stress. Green rooibos exhibited highest activity with an IC50 value of 29 μg/ml and 70 μg/ml, when U937 and red blood cells were exposed oxidative stress, respectively. Fermented rooibos and Chinese green tea had IC50 values of 61 μg/ml and 57 μg/ml for U937, respectively, and 221 μg/ml and 405 μg/ml for red blood cells, respectively. These results indicate that fermented and green rooibos extracts were able to permeate the U937 cells and red blood cell membrane and inhibited oxidation of DCFH-DA to a fluorescent DCF within the cells.

Keywords: rooibos, antioxidants, permeability, bioavailability

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Authors: Shivankar Agrawal, Sunil Kumar Deshmukh, Colin Barrow, Alok Adholeya

Abstract:

A variety of genetic and environmental factors cause various cosmetics and dermatological problems. There are already claimed drugs available in market for treating these problems. However, the challenge remains in finding more potent, environmental friendly, causing minimal side effects and economical cosmeceuticals. This leads to an increased demand for natural cosmeceutical products in the last few decades. Plant derived ingredients are limited because plants either contain toxic metabolites, grow too slow or seasonal harvesting is a problem. To identify new bioactive cosmetics ingredients of marine microbial bioresource, we screened 35 marine fungi isolated from marine samples collected from Andaman Island and west coast of India. Fungal crude extracts were investigated for their antityrosinase, antioxidant and antibacterial activities for the purpose of identifying anti-aging, skin-whitening and anti-acne biomolecule with the potential in cosmetics. In the tyrosinase inhibition and 2, 2-Diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assays, two fungal extracts, including “P2”, Talaromyces stipitatus and “D4”, Aspergillus terreus showed high inhibitory activity at 1mg/mL for tyrosinase inhibition and 0.5mg/mL for DPPH scavenging. The in vitro antimicrobial activity was investigated by the agar well diffusion method. In the tyrosinase inhibition assay, 8 extracts showed significant antibacterial activity against bacteria causing skin and wound infection in humans. In the course of systematic screening program for bioactive marine fungi, strain “D5” was found to be most potent strain with MIC value of 1mg/mL, which was morphologically identified as Simplicillium lamellicola. The effects of the most active crude extracts against their susceptible test microorganisms were also investigated by SEM analysis. Further investigations will focus on purification and characterization major active components responsible for these activities.

Keywords: antioxidant, antimicrobial activity, tyrosinase, cosmeceuticals, marine fungi

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Authors: Amel Bouaziz, Seddik Khennouf, Mussa Abu Zarga, Shtayway Abdalla, Saliha Djidel, Assia Bentahar, Saliha Dahamna, Smain Amira

Abstract:

The present study aimed to evaluate the hypotensive and the in vitro antioxidant activities of Crataegus azarolus L. (Rosaceae), a plant widely used as natural remedy for hypertension in folk medicine. The antioxidant potential of methanolic extract (ME)and its three fractions of Chloroform (CHE), ethyl acetate (EAE)and water (AqE) have been investigated using several assays, including the DPPH scavenging, ABTS scavenging, hydroxyl radical scavenging. Inhibition of lipid peroxidation was performed by the β-carotene bleaching assay, ferric thiocyanate method and thiobarburic acid method. Total phenolic and total flavonoid contents of the extracts were estimated using Folin-Chiocalteu reagent and AlCl3, respectively. EAE extract showed the highest polyphenolic and flavonoids contents (396,04±1.20 mg GAE/g of dry extract and 32,73 ± 0.03mg QE/g of dry extract) respectively. Similarly, this extract possessed the highest scavenging activity for DPPH radical (IC 50 = 0,006±0,0001mg /ml), ABTS radical (IC50=0.0035±0,0007 mg/ml) and hydroxyl radical(IC 50=0,283± 0.01 mg/ml). In addition, the EAE exhibited the highest antioxidant activity in the inhibition of linoleic acid/ß-carotene coupled oxidation (89,21%), lipid peroxidation in the ferric thiocyanate(FTC) method (90.13%), and thio-barbituric acid (TBA) method (74.23%). Intravenous administration of Me and EAE decreased mean arterial blood pressure, systolic and diastolic blood pressure in anesthetized rats dose-dependently, at the dose range of 0.4 to 12 mg/kg. The mean arterial blood pressure dropped by 27.58 and 39.37% for ME and EAE, respectively. In conclusion, The present study supported the significant potential to use C. azarolus by-products as a source of natural antioxidants and provides scientific justification for its traditional uses as cardio-protective and anti-hypertensive remedy.

Keywords: Crataegus azarolus, polyphenols, flavonoids, hypertension, antioxidant activity, free radicals, peroxidation

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Authors: Abdelaaty A. Shahat, Mansour S. Alsaid

Abstract:

Four Acetylated flavonol glycosides were isolated from Centaurium spicatum (L.) Fritsch (Gentianaceae). Structure elucidation, especially the localization of the acetyl groups, and complete 1H and 13C NMR assignments of these biologically active compounds were carried out using one- and two-dimensional NMR methods, including CNMR, DEPT-135 and DEPT-90 and gradient-assisted experiments such as DQF-COSY, TOCSY, HSQC and HMBC experiments. The antioxidant activities of the new acetylated flavonoid glycosides using DPPH• assay were determined. The compounds tested showed a good DPPH• activity compared with control, but their activity was lower than that of their corresponding aglycone, quercetin.

Keywords: Centaurium spicatum, flavonoids, biological activity, isolation, glycosides

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Authors: Mohammed Falalu Hamza

Abstract:

A phytochemical investigation conducted on the leaves extract of Cryptocarya latifolia (Lauraceae) revealed the presence of two major essential oils; Nerolidol (1) and Copaene (2) with the aid of gas chromatography-mass spectrometry (GC-MS). The compounds exhibited good anti-oxidant capacity using 2,2-diphenyl-1-picryl-hydrazyl (DPPH) radical scavenging assay. The result shows that the anti-oxidant capacity of the compounds is dependent on concentration similar to the standard (ascorbic acid). This study shows that the leaves extract of C. latifolia is a good source of important natural antioxidants.

Keywords: broad-leaved quince, phytochemical, anti-oxidant, essential oils

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Authors: Supriyono, Sumardiyono, Puti Pertiwi

Abstract:

Crude palm oil (CPO) is a vegetable oil that came from a palm tree bunch. Palm oil tree was known as highest vegetable oil yield. It was grown across Equatorial County, especially in Malaysia and Indonesia. The greenish red color on CPO was came from carotenoid antioxidant, which could be extracted and use separately as functional food and other purposes as antioxidant source. Another antioxidant that also found in CPO is tocopherol. The aim of the research work is to find antioxidant activity on CPO comparing to the synthetic antioxidant that available in a market. On this research work, antioxidant was extracted by using a mixture of acetone and n. hexane, while activity of the antioxidant extract was determine by DPPH method. The extracted matter was shown that their antioxidant activity was about 45% compare to pure tocopherol and beta carotene.

Keywords: antioxidant, , beta carotene, , crude palm oil, , DPPH, , tocopherol

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Authors: F. T. Senberber, N. Tugrul, E. Moroydor Derun

Abstract:

As a nutraceutical, sumac (Rhus Coriaria) was extracted by using different solvents of methanol, ethanol, and water. The DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) method of free radical scavenging capacity was used to determine the effects of solvent on antioxidant activities of the plant. The total phenolic content was studied by The Folin Ciocalteu Reagent method. The antioxidant activities of extracts exhibit minor changes in different solvents and varied in the range of 84.3–86.4 %. The total phenolic contents are affected by the selected solvent. The highest total phenolic content was determined at the liquid phase of water and it was estimated as 26.3 mg/g in gallic acid.

Keywords: DPPH, solvent, sumac, total phenolic content

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Authors: Hanaa H. Abd El Baky, Gamal S. El Baroty, Eman A. Ibrahem

Abstract:

Microalgae Scenedesmus obliquus, the carotenoides (astaxanine and β-caroteine) were identified as the major bioactive constituents. In this work we prepared functional pre-biotic cakes to increase general mental health. Functional cakes were formulated by adding algal caroteinods at 2 and 4 mg/100g to flower and the cakes were storage for 20 days. Oxidative stability of both function cakes products were examined during storage periods by DPPH and TBA assays, and the results revealed that both values in function food products were significantly much low than that in untreated food products. Data of sensory evaluation revealed that treated biscuit and cakes with algae or algae extracts were significantly acceptable as control for main sensory characteristics (colour, odour/aroma, flavour, texture, the global appreciation, and overall acceptability). Thus, it could be concluded that functional biscuits and cakes (very popular and well balanced nutritional food) had good sensory and nutritional profiles and can be developed as new niche food market.

Keywords: Scenedesmus obliquus, carotenoids, functional cakes antioxidant, nutritional profiles

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