Search results for: DPPH assays

Authors: Fatemeh Cheraghali, Saeedeh Shojaee-Aliabadi, Seyede Marzieh Hosseini, Leila Mirmoghtadaie

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In recent years, the field of natural antimicrobial and antioxidant compounds is one of the main research topics in the food industry. Application of agricultural residues is mainly cheap, and available resources are receiving increased attention. Walnut green husk is one of the agricultural residues that is considered as natural compounds with biological properties because of phenolic compounds. In this study, maltodextrin 10% was used for microencapsulation of walnut green husk extract. At first, the extract was examined to consider extraction yield, total phenolic compounds, and antioxidant activation. The results showed the extraction yield of 81.43%, total phenolic compounds of 3997 [mg GAE/100 g], antioxidant activity [DPPH] of 84.85% for walnut green husk extract. Antioxidant activity is about 75%-81% and by DPPH. At the next stage, microencapsulation was done by spry-drying method. The microencapsulation efficiency was 72%-79%. The results of SEM tests confirmed this microencapsulation process. In addition, microencapsulated and free extract was more effective on gram-positive bacteria’s rather than the gram-negative ones. According to the study, walnut green husk can be used as a cheap antioxidant and antimicrobial compounds due to sufficient value of phenolic compounds.

Keywords: biopolymer, microencapsulation, spray-drying, walnut green husk

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Authors: Ririn Lestari Sri Rahayu, Mustofa Ahda

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The use of synthetic antioxidants often causes a negative effect on health and increases the incidence of carcinogenesis. Development of the natural antioxidants should be investigated. However, natural antioxidants have a low toxicity and are safe for human consumption. Ethanol extract of mangosteen rind (Garcinia mangostana) contains natural antioxidant compounds that have various pharmacological activities. Antioxidants from the ethanol extract of mangosteen rind have free radicals scavenging activities. The scavenging activity of ethanol extract of mangosteen rind was determined by DPPH method. The phenolic compound from the ethanol extract of mangosteen rind is determined with Folin-Ciocalteu method. The results showed that the absolute ethanol extract of mangosteen rind has IC₅₀ of 40.072 ug/mL. The correlation of total phenols content with free radical scavenging activity has an equation y: 5.207x + 205.51 and determination value (R²) of 0.9329. Total phenols content from the ethanol extract of mangosteen rind has a good correlation with free radicals scavenging activity of DPPH.

Keywords: Antioxidant, Garcinia mangostana, Inhibition concentration 50%, Phenolic.

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Authors: Asma Temagoult, Bariza Zitouni, Yassin Noui

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Cactus fruit contains different nutritional and functional components, which are used because of their benefits to human health, such as flavonoids, phenolic compounds, carotenoids and vitamins C. It has hypoglycemic and hypolipidemic action, and antioxidant properties related to anticarcinogenic, antiulcerogenic and immunomodulatory effects. The antioxidant and nutritional properties have been characterized in cactus prickly pear (Opuntia ficus-indica Mill.), cultivar yellow, grown in Arris area; Eastern of Algeria. The antioxidant properties of this cactus cultivar were higher than the others cactus cultivar in the world. The amount of fruit phenolic compounds revealed contents between 20.65 and 45.70 mg / 100 g of FW for total polyphenols and 0.519 - 0.591 mg / 100 g of FW for the flavonoids. The antioxidant activity was evaluated by DPPH radical scavenging and FRAP (ferric reducing antioxidant power) methods. The average recorded to the potassium content is about 1070 mg / 100 g of the fresh weight; sodium is 60.7 mg / 100 g of the fresh weight and 80 mg / 100g for the calcium. According to the high value of this cactus, it was considered as a good nutrient and important pharmaceutical resource. It could be used as a natural additive or substituted food supplement in many foodstuffs production, to benefit from these benefits.

Keywords: antioxidant properties, DPPH, FRAP, nutritional properties, Opuntia ficus indica

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Authors: M. Pourshirazi, M. Esmaelifar, A. Aliahmadi, F. Yazdian, A. S. Hatamian Zarami, S. J. Ashrafi

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Medicinal fungi are the new potential source of drugs to improve the treatment of diseases with association to oxidative agents such as cancers. Monascus purpureus contains functional components potentially effective in improving human health. In the present work, ethanolic extract of Monascus purpureus (EEM) was evaluated for health improving potential mainly focusing on antioxidant and anticancer activities. Ferric ion reducing power (FRAP), scavenging of DPPH radicals and determining viability of breast carcinoma MCF-7 and cervical carcinoma HeLa cells with MTT assay were evaluated. Our data showed a significant antioxidant activity of EEM with 142.45 µg/ml inhibition concentration of 50% DPPH radicals and 2112.33 µg eq.Fe2+/mg extract of FRAP assay. These results might be caused by antioxidant components such as pigments and phenolic compounds. Further, the results demonstrated that EEM caused significant reduction in the viability of MCF-7 with IC50 of 7 µg/ml but not have good effect against viability of HeLa cells. Accordingly, Monascus purpureus is presented as a strong potential of breast cancer treatment. In further study, the mechanistic studies are needed to determine the mechanisms of anticancer activity of EEM.

Keywords: Monascus purpureus, antioxidant, cancer, ethanolic extract

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Authors: Ghavamaldin Asadian, Aptin Rahnavard, Mariamalsadat Taghavi

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Achillea millefolium L. is one of the most important medicinal plants with antioxidant compounds. The use of compounds derived from plants reduces the incidence of many chronic diseases. The purpose of this investigation is study of total phenolic content and antioxidant activity some of ecotypes yarrow grown in natural habitats in Iran. This experimental study was conducted in 2013 at the Islamic Azad University, Tonekabon Branch. After identifying the natural sites, we have attempted to harvest of aerial part and after drying in lab temperature, essential oil was extracted by steam distillation. In this research for evaluate the antioxidant properties was used of three method, DPPH, Antioxidant capacity ferro revival and phosphomolybdenum, that all mechanism is based on the electron donating. All ecotypes had antioxidant activity and ecotypes grown in Kandovan region were measured with the most total phenolic (89.5 mg GA/g dew) and flavonoid (20.4 µg/g dew) and the lowest in Saveh (71.3 mg GA/g dew, 17.4 µg/g dew). Variation of the antioxidant properties were significant (P≤0.01) in areas and were accounted Kandovan with highest value and the lowest in Save. As a result, yarrow essential oil grown in Kandovan in terms of amount of total phenolic, flavonoid and antioxidant property, it was determined the best natural habitat.

Keywords: achillea millefolium L., antioxidant compounds, DPPH, total phenolic, flavonoid natural habitats

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Authors: Imad Abdelhamid El-Haci, Wissame Mazari, Fayçal Hassani, Fawzia Atik Bekkara

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Much attention has been paid to the antioxidants, which are expected to prevent food and living systems from peroxidative damage. Incorporation of synthetic antioxidants in food products is under strict regulation due to the potential health hazards caused by such compounds. The use of plants as traditional health remedies is very popular and important for 80% of the world’s population in African, Asian, Latin America and Middle Eastern Countries. Their use is reported to have minimal side effects. In recent years, pharmaceutical companies have spent considerable time and money in developing therapeutics based upon natural products extracted from plants. In other part, due to the continuous emergence of antibiotic-resistant strains there is continual demand for new antibiotics. Chemical compounds from medicinal plant especially are targeted by many researches. In this light, genus Polygonum (Polygonaceae), comprising about 45 genera (300 species), is distributed worldwide, mostly in north temperate regions. They have been reported to have uses in traditional medicine, such as anti-inflammation, promoting blood circulation, dysentery, diuretic, haemorrhage and many other uses. In our study, Polygonum maritimum (from Algerian coast) was extracted with 80% methanol to obtain a crude extract. P. maritimum extract (PME) had a very high content of total phenol, which was 352.49 ± 18.03 mg/g dry weight, expressed as gallic acid equivalent. PME exhibited excellent antioxidant activity, as measured using DPPH and H2O2 scavenging assays. It also showed a high antibacterial activity against gram positive bacterial strains: Bacillus cereus, Bacillus subtilis and Staphylococcus aureus with an MIC 0,12 mg/mL.

Keywords: Polygonum maritimum, crude extract, antioxidant activity, antibacterial activity

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Authors: E. Chiam, E. Koh, W. Teh, M. Prabhakaran

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Fruits and vegetables (F&V) are widely eaten for their nutritional value and associated health benefits being an immense source of bioactive compounds. However, F&V peels are often discarded, and it accounts for a higher proportion of food waste. Incorporation of F&V peels as functional ingredients can add more value to food due to the higher amounts of phytochemicals present in them. In this research, methanolic extracts of different F&V peels, namely apple, orange, kiwi, grapefruit, dragon fruit, pomelo, and pumpkin are investigated for their total phenolic content (TPC) by Folin-Ciocalteau (FC) assay and the antioxidant capacity was evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and phosphomolybdenum assay using UV-Vis spectroscopy. Evaluation of the α-glucosidase inhibitory assay was carried out during this study to determine the antidiabetic potential of F&V peels. Results of our study showed that grapefruit peels contained the highest total phenolic content of 477.81 ± 0.01 mg gallic acid equivalent per gram dry weight of the sample, and kiwi peel had the highest antioxidant capacity (90.51 ± 0.10 % inhibition of DPPH radical) among the different F&V peels studied. Fruit peels exhibited high α-glucosidase inhibitory activity. Comparing fruit peels with vegetable peels, it was found that fruit peels had high total phenolic content, antioxidant capacity and anti-diabetic potential compared to vegetable peels.

Keywords: polyphenolics, fruit peels, antioxidant, antidiabetic

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Authors: Sakinah Abdullah, Rosna Mat Taha

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Antioxidants are compounds that protect cells against the damaging effects of reactive oxygen species such as singlet oxygen, superoxide, peroxyl radicals, and peroxynitrite which result in oxidative stress leading to cellular damage. Natural antioxidant are in high demand because of their potential in health promotion and disease prevention and their improved safety and consumer acceptability. Plants are rich sources of natural antioxidant. Dioscorea alata L. known as 'ubi badak' in Malaysia were well known for their antioxidant content, but this plant was seasonal. Thus, tissue culture technique was used to mass propagate this plant. In the present work, a comparative study between in vitro (from tissue culture) and in vivo (from intact plant) samples of Dioscorea alata L. for their antioxidant potential by 2,2-diphenil -1- picrylhydrazyl (DPPH) radical scavenging activity method and their total phenolic and flavonoid contents were carried out. All samples had better radical scavenging activity but in vivo samples had the strongest radical scavenging activity compared to in vitro samples. Furthermore, tubers from in vivo samples showed the greatest free radical scavenging effect and comparatively greater phenolic content than in vitro samples.

Keywords: Dioscorea alata, tissue culture, antioxidant, in vivo, in vitro, DPPH

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Authors: Bilal Ahmad Ashwar, Inam u nisa, Asima Shah, Adil Gani, Farooq Ahmad Masoodi

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The effects of 5% BHT and green tea extracts (GTE) on the physical, barrier, mechanical, thermal and antioxidant properties of potato starch films were investigated. Results showed both BHT and GTE significantly lowered solubility of films. Addition of BHT significantly decreased water vapour transmission rate. Both BHT and GTE promoted significant increase in the elastic modulus but a decrease in % EAB, however BHT was more effective in increasing elastic modulus. Increase in glass transition temperature (Tg) and enthalpy of transition (ΔH) of films was observed with the incorporation of GTE and BHT. Scanning electron microscopy (SEM) revealed smooth surface of the films. The DPPH radical scavenging ability of both BHT and GTE films were stronger in fatty food stimulant (95% ethanol. The GTE and BHT films were individually applied to fresh beef samples and were stored at 4 0C and room temperature for 10 days. Metmyoglobin formation and lipid oxidation (TBARS) were monitored periodically. The addition of GTE extracts and BHT resulted in decreases in metmyoglobin and TBARS values. We conclude that extracts of GTE and BHT have potential as preservatives for fresh beef.

Keywords: starch film, WVTR, tensile properties, SEM, thermal analysis, DPPH scavenging activity, TBARS, metmyoglobin

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Authors: Harleen Kaur, Richa

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Plants produce a large variety of secondary metabolites. Phenolics are the compounds that contain hydroxyl functional group on an aromatic ring. These are chemically heterogeneous compounds. Some are soluble only in organic solvents, some are water soluble and others are large insoluble polymers. Flavonoids are one of the largest classes of plant phenolics. The carbon skeleton of a flavonoid contains 15 carbons arranged in two aromatic rings connected by a three carbon ridge. Both phenolics and flavonoids are good natural antioxidants. Four Indian aromatic plants were selected for the study i.e, Achillea species, Jasminum primulinum, Leucas cephalotes and Leonotis nepetaefolia. All the plant species were collected from Chail region of Himachal Pradesh, India. The identifying features and anatomical studies were done of the part containing the essential oils. Phenolic cotent was estimated by Folin Ciocalteu’s method and flavonoids content by aluminium chloride method. Antioxidant property was checked by using DPPH method. Maximum antioxidant potential was found in Achillea species, followed by Leonotis nepetaefolia, Jaminum primulinum and Leucas cephalotes. Phenolics and flavonoids are important compounds that serve as defences against herbivores and pathogens. Others function in attracting pollinators and absorbing harmful radiations.

Keywords: antioxidants, DPPH, flavonoids, phenolics

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Authors: Naiduwadura Ivon Upekala De Silva, Nathan Lehman, Talia Fargason, Trenton Paul, Zihan Zhang, Jun Zhang

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SRSF1 governs splicing of over 1,500 mRNA transcripts. SRSF1 contains two RNA-recognition motifs (RRMs) and a C-terminal Arg/Ser-rich region (RS). It has been thought that SRSF1 RRMs exclusively recognize single-stranded exonic splicing enhancers, while RS lacks RNA-binding specificity. With our success in solving the insolubility problem of SRSF1, we can explore the unknown RNA-binding landscape of SRSF1. We find that SRSF1 RS prefers purine over pyrimidine. Moreover, SRSF1 binds to the G-quadruplex (GQ) from the ARPC2 mRNA, with both RRMs and RS being crucial. Our binding assays show that the traditional RNA-binding sites on the RRM tandem and the Arg in RS are responsible for GQ binding. Interestingly, our FRET and circular dichroism data reveal that SRSF1 unfolds the ARPC2 GQ, with RS leading unfolding and RRMs aiding. Our saturation transfer difference NMR results discover that Arg residues in SRSF1 RS interact with the guanine base but with other nucleobases, underscoring the uniqueness of the Arg/guanine interaction. Our luciferase assays confirm that SRSF1 can alleviate the inhibitory effect of GQ on gene expression in the cell. Given the prevalence of RNA GQ and SR proteins, our findings unveil unexplored SR protein functions with broad implications in RNA splicing and translation.

Keywords: SR, SRSF!, RNA G-quadruplex, unfolding, RNA binding

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Authors: Neera Kapoor

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Background: Knockdown Resistance (KDR) is one of the mechanisms of insecticide resistance in insects caused by the reduced target site sensitivity i.e. voltage gated sodium channel (VGSC) rendering it less sensitive to the toxic effects of DDT and pyrethroids. In this study, we evaluated insecticide susceptibility and its underlying KDR mechanism in eight Ae. aegypti and five Ae. albopictus field populations. Methodology: Field population was collected from four different geographical regions of India covering 18 districts of ten states. For genotyping of twelve KDR alleles in Ae. aegypti field populations, three PCR based assays were used; with DNA sequencing; ASPCR; PCR-RFLP. Genomic DNA was isolated, and three partial domains (II, III, and IV) of VGSC were amplified and sequenced. Results: Molecular screening for common KDR mutations, revealed the presence of five mutations viz. S989P, V1016G, T1520I, F1534C/L. Two novel mutations were observed, first at T1520 (ACC) residue where a C > T substitution at the second position of codon results in amino acid change to Isoleucine (ATC). Second mutation was an alternative point mutation at F1534 (TTC) residue where a substitution of T > C at the first position of codon results in an amino acid change to Leucine (CTC). ASPCRs were not accurate, so three PCR-RFLP assays were developed for genotyping of five KDR alleles in Ae. aegypti; viz. T1520I, F1534C/L. Representative samples of all genotypes (n=200) were sequenced to validate the newly developed PCR based assays for Ae. aegypti. Genotyping results showed that 989P is linked to 1016G and novel mutation 1520I was always found with 1534C allele. Conclusion: Present study confirmed the presence of DDT and pyrethroid resistance among Ae. aegypti populations in India and for the first time reported KDR mutations in this species from India including two novel mutations. Results of present study lead us to infer that, at least five KDR mutations (S989P, V1016G, T1530I, F1534C, and F1534L) can be seen as a potential marker for DDT/pyrethroid resistance.

Keywords: F1534C, F1534L, S989P, T1530I, V1016G

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Authors: Joe Modise, Bamidel Joseph Okoli, Nas Molefe, Imelda Ledwaba

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The present study describes the chemical profile and antioxidant potential of the stem bark of Acacia decurrens. The methanol fraction of A. decurrens stem bark gave the highest yield (20 %), while the hexane fraction had the lowest yield (0.2 %). The GC-MS spectra of the hexane, chloroform and ethyl acetate fractions confirm the presence of fifty two major compounds and the ICP-OES analysis of the stem bark was found to contain Co(0.41), Zn(1.75), Mn(3.69), Ca(8.67), Ni(10.54), Mg(12.98), Cr(24.38), K(47.88), Fe(154.62) ppm; which is an indication of hyper-accumulation capacity. The UV-Visible spectra of showed four absorption maxima for hexane fraction at 665 (0.028), 410 (0.116), 335 (0.278) and 250 (0.007) nm, three for chloroform fraction at 665 (0.028), 335 (0.278) and 250 (0.007) nm , three for ethyl acetate fraction at 665 (0.070), 390 (0.648) and 345 (0.663) nm and three for methanol fraction at 385 (0.508), 310 (0.886) and 295 (0.899) nm respectively. Quantitative phytochemical screening indicated that the alkaloid (0.6-3.3) % and saponins (5.1-8.6) % contents of the various fractions were significantly lower than the tannin (30.9-55.8) mg TAE/g, steroid(13.92-41.2) %, phenol (40.6-65.5) mgGAE/g and flavonoids (210.2 -284.9) mg RUE/g contents. The antioxidant activity of the fractions was analysed by different methods and revealed good to moderate antioxidant potential with different IC50 values viz. (42.2-49.6) mg/mL for ABTS and (37.8-75.0) μg/ml for DPPH respectively, compared to standard antioxidants. Based on obtained results, the A.decurrens stem bark fractions can be a source of safe, sustainable natural antioxidant drug and can be exploited as a source of controlled green-heavy metal cleaner.

Keywords: Acacia decurrens, antioxidant, DPPH, ABTS, hyperaccumulation, Menstruum, ICP-OES, GC-MS, UV/visible

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Authors: Cristian Moisa, Andreea Lupitu, Adriana Csakvari, Dana G. Radu, Leonard Marian Olariu, Georgeta Pop, Dorina Chambre, Lucian Copolovici, Dana Copolovici

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Green synthesis of nanoparticles (NPs) represents a promising, accessible, eco-friendly, and safe process with significant applications in biotechnology, pharmaceutical sciences, and farming. The aim of our study was to obtain silver nanoparticles, using plant wastes extracts resulted in the essential oils extraction process: Thymus vulgaris L., Origanum vulgare L., Lavandula angustifolia L., and in hemp processing for seed and fibre, Cannabis sativa. Firstly, we obtained aqueous extracts of thyme, oregano, lavender, and hemp (two monoicous and one dioicous varieties), all harvested in western part of Romania. Then, we determined the chemical composition of the extracts by liquid-chromatography coupled with diode array and mass spectrometer detectors. The compounds identified in the extracts were in agreement with earlier published data, and the determination of the antioxidant activity of the obtained extracts by DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) assays confirmed their antioxidant activity due to their total polyphenolic content evaluated by Folin-Ciocalteu assay. Then, the silver nanoparticles (AgNPs) were successfully biosynthesised, as was demonstrated by UV-VIS, FT-IR spectroscopies, and SEM, by reacting AgNO₃ solution and plant extracts. AgNPs were spherical in shape, with less than 30 nm in diameter, and had a good bactericidal activity against Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli, Klebsiella pneumoniae, Pseudomonas fluorescens).

Keywords: plant wastes extracts, chemical composition, high performance liquid chromatography mass spectrometer, HPLC-MS, scanning electron microscopy, SEM, silver nanoparticles

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Authors: Cristian Soitu, Alexander Feuerborn, Cyril Deroy, Alfonso Castrejon-Pita, Peter R. Cook, Edmond J. Walsh

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Microfluidics now stands as an academically mature technology after a quarter of a century research activities have delivered a vast array of proof of concepts for many biological workflows. However, translation to industry remains poor, with only a handful of notable exceptions – e.g. digital PCR, DNA sequencing – mainly because of biocompatibility issues, limited range of readouts supported or complex operation required. This technology exploits the domination of interfacial forces over gravitational ones at the microscale, replacing solid walls with fluid ones as building blocks for cell micro-environments. By employing only materials used by biologists for decades, the system is shown to be biocompatible, and easy to manufacture and operate. The method consists in displacing a continuous fluid layer into a pattern of isolated chambers overlaid with an immiscible liquid to prevent evaporation. The resulting fluid arrangements can be arrays of micro-chambers with rectangular footprint, which use the maximum surface area available, or structures with irregular patterns. Pliant, self-healing fluid walls confine volumes as small as 1 nl. Such fluidic structures can be reconfigured during the assays, giving the platform an unprecedented level of flexibility. Common workflows in cell biology are demonstrated – e.g. cell growth and retrieval, cloning, cryopreservation, fixation and immunolabeling, CRISPR-Cas9 gene editing, and proof-of-concept drug tests. This fluid-shaping technology is shown to have potential for high-throughput cell- and organism-based assays. The ability to make and reconfigure on-demand microfluidic circuits on standard Petri dishes should find many applications in biology, and yield more relevant phenotypic and genotypic responses when compared to standard microfluidic assays.

Keywords: fluid walls, micro-chambers, reconfigurable, freestyle

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Authors: Prasanna Ramachandran, Kareem Graham, Helena Kiefel, Sunit Jain, Todd DeSantis

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Microbes that reside inside the mammalian GI tract, commonly referred to as the gut microbiome, have been shown to have therapeutic effects in animal models of disease. We hypothesize that specific proteins produced by these microbes are responsible for this activity and may be used directly as therapeutics. To speed up the discovery of these key proteins from the big-data metagenomics, we have applied machine learning techniques. Using amino acid sequences of known epitopes and their corresponding binding partners, protein interaction descriptors (PID) were calculated, making a positive interaction set. A negative interaction dataset was calculated using sequences of proteins known not to interact with these same binding partners. Using Random Forest and positive and negative PID, a machine learning model was trained and used to predict interacting versus non-interacting proteins. Furthermore, the continuous variable, cosine similarity in the interaction descriptors was used to rank bacterial therapeutic candidates. Laboratory binding assays were conducted to test the candidates for their potential as therapeutics. Results from binding assays reveal the accuracy of the machine learning prediction and are subsequently used to further improve the model.

Keywords: protein-interactions, machine-learning, metagenomics, microbiome

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Authors: Amessis-Ouchemoukh Nadia, Salhi Rim, Ouchemoukh Salim, Ayad Rabha, Sadou Dyhia, Guenaoui Nawel, Hamouche Sara, Madani Khodir

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The cooking method is a key factor influencing the quality of vegetables. In this study, the effect of the most common cooking methods on the nutritional composition, phenolic content, pigment content and antioxidant activities (evaluated by DPPH, ABTS, CUPRAC, FRAP, reducing power and phosphomolybdene method) of fresh, steamed, and boiled red beet was investigated. The fresh samples showed the highest nutritional and bioactive composition compared to the cooked ones. The boiling method didn’t lead to a significant reduction (p< 0.05) in the content of phenolics, flavonoids, flavanols and DPPH, ABTS, FRAP, CUPRAC, phosphomolybdeneum and reducing power capacities. This effect was less pronounced when steam cooking was used, and the losses of bioactive compounds were lower. As a result, steam cooking resulted in greater retention of bioactive compounds and antioxidant activity compared to boiling. Overall, this study suggests that steam cooking is a better method in terms of retention of pigments and bioactive compounds and antioxidant activity of beetroot.

Keywords: beta vulgaris, cooking methods, bioactive compounds, antioxidant activities

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Authors: Rachmat Mauludin, Irda Fidrianny, Dita Sasri Primaviri, Okti Alifiana

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Natural products such as propolis, green tea and corncob are containing several compounds called antioxidant. Antioxidant can be used in topical application to protect skin against free radical, prevent skin cancer and skin aging. Previous study showed that the extract of propolis that has the highest antioxidant activity was ethanolic extract of propolis (EEP). It is important to make a dosage form that could keep the stability and could protect the effectiveness of antioxidant activity of the extracts. In this research, nanoemulsion (NE) was chosen to formulate those natural products. NE is a dispersion system between oil phase and water phase that formed by mechanical force with a lot amount of surfactants and has globule size below 100 nm. In pharmaceutical industries, NE was preferable for its stability, biodegradability, biocompatibility, its ease to be absorbed and eliminated, and for its use as carrier for lipophilic drugs. First, all of the natural products were extracted using reflux methods. Green tea and corncob were extracted using 96% ethanol while propolis using 70% ethanol. Then, the extracts were concentrated using rotavapor to obtain viscous extracts. The yield of EEP was 11.12%; green tea extract (GTE) was 23.37%; and corncob extract (CCE) was 17.23%. EEP contained steroid/triterpenoid, flavonoid and saponin. GTE contained flavonoid, tannin, and quinone while CCE contained flavonoid, phenol and tannin. The antioxidant activities of the extracts were then measured using DPPH scavenging capacity methods. The values of DPPH scavenging capacity were 61.14% for EEP; 97.16% for GTE; and 78.28% for CCE. The value of IC50 for EEP was 0.41629 ppm. After the extracts were evaluated, NE was prepared. Several surfactants and co-surfactants were used in many combinations and ratios in order to form a NE. Tween 80 and Kolliphor RH40 were used as surfactants while glycerin and propylene glycol were used as co-surfactants. The best NE consists of 26.25% of Kolliphor RH40; 8.75% of glycerin; 5% of rice bran oil; 3% of extracts; and 57% of water. EEP NE had globule size around 23.72 nm; polydispersity index below 0.5; and did not cause any irritation on rabbits. EEP NE was proven to be stable after passing stability test within 63 days at room temperature and 6 cycles of Freeze and Thaw test without separated. Based on TEM (Transmission Electron Microscopy) test, EEP NE had spherical structure with most of its size below 50 nm. The antioxidant activity of EEP NE was monitored for 6 weeks and showed no significant difference. The value of DPPH scavenging capacity for EEP NE was around 58%; for GTE NE was 96.75%; and for CCE NE was 55.69%.

Keywords: propolis, green tea, corncob, antioxidant, nanoemulsion

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Authors: Sifi Ibrahim, Benaddou Fatima Zohra, Yousfi Mohamed

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The Marrubium deserti and Ammodaucus leucotrichus L. an Algerian endemic species, has several applications in traditional medicine for example as a remedy for asthma and diabetes, and was found to have antibacterial properties. In this work, an antioxidant and antimicrobial activities was performed on phenolic extracts of Marrubium deserti, Ammodaucus leucotrichus plants. The yield of methanol maceration of these plants is 12.4% and 20.4% respectively. The content of total polyphenols, flavonoids and anthocyanin in methanolic extracts, are varied between 19.52±1.88 and 59.24±3.45 mg/g gallic acid equivalent, and 2.08±0.29 to 1.46±0.39 mg/g quercetin equivalent, and 0.395 to 1.934µmol/g respectively. The total chlorophylls and carotenoids were be ranged from 0.149±0.20 to 1.537±0.20 g/ml and 1.537±0.20 to 0.149 ± 0.20 g/ml, respectively. According to DPPH and FRAP test, the values of EC50 was shows a higher activity of Marrubium deserti than Ammodaucus leucotrichus with EC50 values (DPPH) were 34.53±0.71 μg/mL and 258.60±15.67 mg/ml respectively. The TEAC values of FRAP test was a highly superior for Marrubium deserti 209.66±0.26 mg Equivalent Trolox/g dry residue than Ammodaucus leucotrichus 45.88±2.93 mg Trolox Equivalent/g dry residue. The antimicrobial activity against nine strains of bacteria (Staphylococcus aureus(+), Staphylococcus aureus (-), Bacillus cereus, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae and Salmonella typhi), was showed that the tested extracts are a significant antibacterial activity with inhibition zones ranging from 10 to 50 mm. the value of CMI were ranging from 0.89 to 14.29 mg/ml.

Keywords: phenolic extract, antioxidant activity, antimicrobial activity, Marrubium deserti, Ammodaucus leucotrichus

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Authors: Stela Papa, Klementina Puto, Migena Pllaha

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HCV is a hepatotropic RNA virus, transmitted primarily via the blood route, which causes progressive disease such as chronic hepatitis, liver cirrhosis, or hepatocellular carcinoma. HCV nowadays is a global healthcare problem. A variety of immunoassays including old and new technologies are being applied to detect HCV in our country. These methods include Immunochromatography assays (ICA), Fluorescence immunoassay (FIA), Enzyme linked fluorescent assay (ELFA), and Enzyme linked immunosorbent assay (ELISA) to detect HCV antibodies in blood serum, which lately is being slowly replaced by more sensitive methods such as rapid automated analyzer chemiluminescence immunoassay (CLIA). The aim of this study is to estimate HCV infection in carriers and chronic acute patients and to evaluate the use of new diagnostic methods. This study was realized from September 2016 to May 2018. During this study period, 2913 patients were analyzed for the presence of HCV by taking samples from their blood serum. The immunoassays performed were ICA, FIA, ELFA, ELISA, and CLIA assays. Concluding, 82% of patients taken in this study, resulted infected with HCV. Diagnostic methods in clinical laboratories are crucial in the early stages of infection, in the management of chronic hepatitis and in the treatment of patients during their disease.

Keywords: CLIA, ELISA, Hepatitis C virus, immunoassay

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Authors: Najafi S., Heidai R., Jamei R., Tofigh F.

Abstract:

In order to study the effects of magnetic field on the root system and growth of Phaseolus vulgaris, an experiment was conducted in 2012. The possible involvement of magnetic field (MF) pretreatment in physiological factors of Phaseolus vulgaris was investigated. Seeds were subjected to 10 days with 1.8 mT of magnetic field for 1h per day. MF pretreatment decreased the plant height, fresh and dry weight, length of root and length of shoot, Chlorophyll a, Chlorophyll b and carotenoid in 10 days old seedling. In addition, activity of enzymes such as Catalase and Guaiacol peroxidase was decreased due to MF exposure. Also, the total Protein and DPPH content of the treated by magnetic field was not significantly changed in compare to control groups, while the flavonoid, Phenol and prolin content of the treated of the treated by magnetic field was significantly changed in compare to control groups. Lateral branches of roots and secondary roots increased with MF. The results suggest that pretreatment of this MF plays important roles in changes in crop productivity. In all cases there was observed a slight stimulating effect of the factors examined. The growth dynamics were weakened. The plants were shorter. Moreover, the effect of a magnetic field on the crop of Phaseolus vulgaris and its structure was small.

Keywords: carotenoid, Chlorophyll a, Chlorophyll b, DPPH, enzymes, flavonoid, germination, growth, phenol, proline, protein, magnetic field, phaseolus vulgaris

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Authors: Borokini Funmilayo Boede

Abstract:

B. alba L., commonly called ‘Amunututu’ and Solanecio biafrae called ‘Worowo’ among the Yoruba tribe in the southwest part of Nigeria are reported to be of great ethnomedicinal importance but are among many underutilized green leafy vegetables in the country. Many studies have established the nutritional values of these vegetables, utilization are very poor and indepth information on their chemical profiles is scarce. The aqueous, methanolic and ethanolic extracts of these vegetables were subjected to phytochemical screening and phenolic profiles of the alcoholic extracts were characterized by using high-performance liquid chromatography coupled with diode array detector (HPLC-DAD). Total phenol and flavonoid contents were determined, antioxidant activities were evaluated using five in vitro assays to assess DPPH, nitric oxide and hydroxyl radical-scavenging abilities, as well as reducing power with ferric reducing antioxidant assay and phosphomolybdate method. The antibacterial activities of the extracts against Staphylococcus aureus, Pseudomonas aeruginosa, and Salmonella typhi were evaluated by using agar well diffusion method and the antifungal activity evaluated against food-associated filamentous fungi by using poisoned food technique with the aim of assessing their nutraceutical potentials to encourage their production and utilization. The results revealed the presence of saponnin, steroids, tannin, terpenoid and flavonoid as well as phenolic compounds: gallic acid, chlorogenic acid, caffeic acid, coumarin, rutin, quercitrin, quercetin and kaemferol. The vegetables showed varying concentration dependent reducing and radical scavenging abilities from weak to strong compared with gallic acid, rutin, trolox and ascorbic acid used as positive controls; the aqueous extracts which gave higher concentrations of total phenol displayed higher ability to reduce Fe (lll) to Fe (ll) and stronger inhibiting power against hydroxyl radical than the alcoholic extracts and in most cases exhibited more potency than the ascorbic acids used as positive controls, at the same concentrations, whereas, methanol and / or ethanol extracts were found to be more effective in scavenging 2, 2-diphenyl-1-picryl hydrazyl radical and showed higher ability to reduce Mo (VI) to Mo (V) in total antioxidant assay than the aqueous extracts. However, the inhibition abilities of all the extracts against nitric oxide were comparable with the ascorbic acid control at the same concentrations. There were strong positive correlations with total phenol (mg GAE/g) and total flavonoid (mg RE/g) contents in the range TFC (r=0.857- 0999 and r= 0.904-1.000) and TPC (r= 0.844- 0.992 and r= 0.900 -0.999) for Basella alba and Senecio biafrae respectively. Inhibition concentration at 50 % (IC50) for each extract to scavenge DPPH, OH and NO radicals ranged from 32.73 to 1.52 compared with control (0.846 - -6.42) mg/ml. At 0.05g/ml, the vegetables were found to exhibit mild antibacterial activities against Staphylococcus aureus, Pseudomonas aeruginosa and Salmonella typhi compared with streptomycin sulphate used as control but appreciable antifungi activities against (Trichoderma rubrum and Aspergillus fumigates) compared with bonlate antibiotic positive control. The vegetables possess appreciable antioxidant and antimicrobial properties for promoting good health, their cultivation and utilization should be encouraged especially in the face of increasing health and economic challenges and food insecurity in many parts of the world.

Keywords: antimicrobial, antioxidants, extracts, phytochemicals

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Authors: Eman Abdullah Morsi, Hend Okasha, Heba Abdel Hady, Mortada El-Sayed, Mohamed Abbas Shemis

Abstract:

Context: Linum usitatissimum (Linn), known as Flaxseed, is one of the most important medicinal plants traditionally used for various health as nutritional purposes. Objective: Estimation of total phenolic and flavonoid contents as well as evaluate the antioxidant using α, α-diphenyl-β-picrylhydrazyl (DPPH), 2-2'azinobis (3-ethylbenzthiazoline-6-sulphonic acid (ABTS) and total antioxidant capacity (TAC) assay and investigation of anti-inflammatory by Bovine serum albumin (BSA) and anticancer activities of hepatocellular carcinoma cell line (HepG2) and breast cancer cell line (MCF7) have been applied on hexane, ethyl acetate, n-butanol and methanol extracts and also, fractions of methonal extract (hexane, ethyl acetate and n-butanol). Materials and Methods: Phenolic and flavonoid contents were detected using spectrophotometric and colorimetric assays. Antioxidant and anti-inflammatory activities were estimated in-vitro. Anticancer activity of extracts and fractions of methanolic extract were tested on (HepG2) and (MCF7). Results: Methanolic extract and its ethyl acetate fraction contain higher contents of total phenols and flavonoids. In addition, methanolic extract had higher antioxidant activity. Butanolic and ethyl acetate fractions yielded higher percent of inhibition of protein denaturation. Meanwhile, ethyl acetate fraction and methanolic extract had anticancer activity against HepG2 and MCF7 (IC50=60 ± 0.24 and 29.4 ± 0.12µg.mL⁻¹) and (IC50=94.7 ± 0.21 and 227 ± 0.48µg.mL⁻¹), respectively. In Gas chromatography-mass spectrometry (GC-MS) analysis, methanolic extract has 32 compounds, whereas; ethyl acetate and butanol fractions contain 40 and 36 compounds, respectively. Conclusion: Flaxseed contains totally different biologically active compounds that have been found to possess good variable activities, which can protect human body against several diseases.

Keywords: phenolic content, flavonoid content, HepG2, MCF7, hemolysis-assay, flaxseed

Procedia PDF Downloads 127

Authors: M. Shemis, O. Maher, G. Casterou, F. Gauffre

Abstract:

Hepatitis C virus (HCV) is a major cause of chronic liver disease with a global 170 million chronic carriers at risk of developing liver cirrhosis and/or liver cancer. Egypt reports the highest prevalence of HCV worldwide. Currently, two classes of assays are used in the diagnosis and management of HCV infection. Despite the high sensitivity and specificity of the available diagnostic assays, they are time-consuming, labor-intensive, expensive, and require specialized equipment and highly qualified personal. It is therefore important for clinical and economic terms to develop a low-tech assay for the direct detection of HCV RNA with acceptable sensitivity and specificity, short turnaround time, and cost-effectiveness. Such an assay would be critical to control HCV in developing countries with limited resources and high infection rates, such as Egypt. The unique optical and physical properties of gold nanoparticles (AuNPs) have allowed the use of these nanoparticles in developing simple and rapid colorimetric assays for clinical diagnosis offering higher sensitivity and specificity than current detection techniques. The current research aims to develop a detection assay for HCV RNA using gold nanoparticles (AuNPs). Methods: 200 anti-HCV positive samples and 50 anti-HCV negative plasma samples were collected from Egyptian patients. HCV viral load was quantified using m2000rt (Abbott Molecular Inc., Des Plaines, IL). HCV genotypes were determined using multiplex nested RT- PCR. The assay is based on the aggregation of AuNPs in presence of the target RNA. Aggregation of AuNPs causes a color shift from red to blue. AuNPs were synthesized using citrate reduction method. Different sets of probes within the 5’ UTR conserved region of the HCV genome were designed, grafted on AuNPs and optimized for the efficient detection of HCV RNA. Results: The nano-gold assay could colorimetrically detect HCV RNA down to 125 IU/ml with sensitivity and specificity of 91.1% and 93.8% respectively. The turnaround time of the assay is < 30 min. Conclusions: The assay allows sensitive and rapid detection of HCV RNA and represents an inexpensive and simple point-of-care assay for resource-limited settings.

Keywords: HCV, gold nanoparticles, point of care, viral load

Procedia PDF Downloads 206

Authors: Farzad Tofigh, Saeideh Najafi, Reza Heidari, Rashid Jamei

Abstract:

In order to study the effects of magnetic field on the root system and growth of Phaseolus vulgaris, an experiment was conducted in 2012. The possible involvement of magnetic field (MF) pretreatment in physiological factors of Phaseolus vulgaris was investigated. Seeds were subjected to 10 days with 1.8 mT of magnetic field for 1h per day. MF pretreatment decreased the plant height, fresh and dry weight, length of root and length of shoot, Chlorophyll a, Chlorophyll b and carotenoid in 10 days old seedling. In addition, activity of enzymes such as Catalase and Guaiacol peroxidase was decreased due to MF exposure. Also, the total Protein and DPPH content of the treated by magnetic field was not significantly changed in compare to control groups, while the flavonoid, Phenol and prolin content of the treated of the treated by magnetic field was significantly changed in compare to control groups. Lateral branches of roots and secondary roots increased with MF. The results suggest that pretreatment of this MF plays important roles in changes in crop productivity. In all cases there was observed a slight stimulating effect of the factors examined. The growth dynamics were weakened. The plants were shorter. Moreover, the effect of a magnetic field on the crop of Phaseolus vulgaris and its structure was small.

Keywords: carotenoid, chlorophyll a, chlorophyll b, DPPH, enzymes, flavonoid, germination, growth, phenol, proline, protein, magnetic field

Procedia PDF Downloads 504

Authors: Chungkham Sarojnalini, Wahengbam Sarjubala Devi

Abstract:

Antioxidant properties and nutritive values of raw and cooked Pool barb, Puntius sophore (Hamilton-Buchanan) of Eastern Himalayas, India were determined. Antioxidant activity of the methanol extract of the raw, steamed, fried and curried Pool barb was evaluated by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging assay. In DPPH scavenging assay the IC50 value of the raw, steamed, fried and curried Pool barb was 1.66 microgram/ml, 16.09 microgram/ml, 8.99 microgram/ml, 0.59 microgram/ml whereas the IC50 of the reference ascorbic acid was 46.66 microgram/ml. This results shows that the fish have high antioxidant activity. Protein content was found highest in raw (20.50±0.08%) and lowest in curried (18.66±0.13%). Moisture content in raw, fried and curried was 76.35±0.09, 46.27±0.14 and 57.46±0.24 respectively. Lipid content was recorded 2.46±0.14% in raw and 21.76±0.10% in curried. Ash content varies from 12.57±0.11 to 22.53±0.07%. The total aminoacids were varied from 36.79±0.02 and 288.43±0.12 mg/100 g. Eleven essential mineral elements were found abundant in all the samples. The samples had a considerable amount of Fe ranging from 152.17 to 320.39 milligram/100 gram, Ca 902.06 to 1356.02 milligram/100 gram, Zn 91.07 to 138.14 milligram/100 gram, K 193.25 to 261.56 milligram/100 gram, Mg 225.06 to 229.10 milligram/100 gram. Ni was not detected in the curried fish. The Mg and K contents were significantly decreased in frying method; however the Fe, Cu, Ca, Co and Mn content were increased significantly in all the cooked samples. The Mg and Na contents were significantly increased in curried sample and the Cr content was decreased significantly (p<0.05) in all the cooked samples.

Keywords: antioxidant property, pool barb, minerals, aminoacids, proximate composition, cooking methods

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Authors: Anupalli Roja Rani, Saraswathi Jaggali

Abstract:

Rose-scented geranium (Pelargonium graveolens L’Hér.) is an erect, much-branched shrub. It is indigenous to various parts of southern Africa, and it is often called Geranium. Pelargonium species are widely used by traditional healers in the areas of Southern Africa by Sotho, Xhosa, Khoi-San and Zulus for its curative and palliative effects in the treatment of diarrhea, dysentery, fever, respiratory tract infections, liver complaints, wounds, gastroenteritis, haemorrhage, kidney and bladder disorders. We have used Plant materials for extracting active compounds from analytical grades of solvents methanol, ethyl acetate, chloroform and water by a soxhlet apparatus. The phytochemical screening reveals that extracts of Pelargonium graveolens contains alkaloids, glycosides, steroids, tannins, saponins and phenols in ethyl acetate solvent. The antioxidant activity was determined using 1, 1-diphenyl-2-picrylhydrazyl (DPPH) bleaching method and the total phenolic content in the extracts was determined by the Folin–Ciocalteu method. Due to the presence of different phytochemical compounds in Pelargonium the anti-microbial activity against different micro-organisms like E.coli, Streptococcus, Klebsiella and Bacillus. Fractionation of plant extract was performed by column chromatography and was confirmed with HPLC analysis, NMR and FTIR spectroscopy for the compound identification in different organic solvent extracts.

Keywords: Pelargonium graveolens L’Hér, DPPH, micro-organisms, HPLC analysis, NMR, FTIR spectroscopy

Procedia PDF Downloads 503

Authors: Harshada Vaidya-Kannur, Dattatraya Naik

Abstract:

Propolis, one of the beehive products also referred as bee-glue is sticky dark coloured complex mixture of compounds. The volatile oil can be isolated from the propolis by hydrodistillation. The mark that is left behind after the removal of volatile oil is referred as the de-oiled propolis. Antioxidant as well as anti-inflammatory properties of total ethanolic extract of de-oiled propolis (TEEDP) was investigated. Another lot of deoiled propolis was successively exacted with hexane, ethyl acetate and ethanol. Activities of these fractions were also determined. Antioxidant activity was determined by studying ABTS, DPPH and NO radical scavenging. Determination of anti-inflammatory activity was carried out by topical TPA induced mouse ear oedema model. It is noteworthy that ethyl acetate fraction of deoiled propolis (EAFDP) exhibited 49.45 % TEAC activity at the concentration 0.2 mg/ml which is equivalent to the activity of trolox at the concentration 0.2 mg/ml. Its DPPH scavenging activity (72.56%) was closely comparable to that of trolox (75%). However its NO scavenging activity was comparatively low. From IC50 values it could be concluded that the efficiency of scavenging ABTS radicals by the de-oiled propolis was more pronounced as compared to scavenging of other radicals. Studies by TPA induced mouse ear inflammation model indicated that the de-oiled propolis of Indian origin had significant topical anti-inflammatory activity. The EAFDP was found to be the most active fraction for this activity also. The purification of EAFP yielded six pure crystalline compounds. These compounds were identified by their physical data and spectral data.

Keywords: anti-inflammatory activity, anti-oxidant activity, column chromatography, de-oiled propolis

Procedia PDF Downloads 289

Authors: Sonja Djilas, Gordana Ćetković, Jasna Čanadanović-Brunet, Vesna Tumbas Šaponjac, Slađana Stajčić, Jelena Vulić, Milica Vinčić

Abstract:

In recent years, interest in food rich in bioactive compounds, such as polyphenols, increased the advantages of the functional food products. Bioactive components help to maintain health and prevention of diseases such as cancer, cardiovascular and many other degenerative diseases. Recent research has shown that the fruit pomace, a byproduct generated from the production of juice, can be a potential source of valuable bioactive compounds. The use of fruit industrial waste in the processing of functional foods represents an important new step for the food industry. Sour cherries have considerable nutritional, medicinal, dietetic and technological value. According to the production volume of cherries, Serbia ranks seventh in the world, with a share of 7% of the total production. The use of sour cherry pomace has so far been limited to animal feed, even though it can be potentially a good source of polyphenols. For this study, local variety of sour cherry cv. ‘Feketićka’ was chosen for its more intensive taste and deeper red color, indicating high anthocyanin content. The contents of total polyphenols, flavonoids and anthocyanins, as well as radical scavenging activity on DPPH radicals and reducing power of sour cherry juice and pomace were compared using spectrophotometrical assays. According to the results obtained, 66.91% of total polyphenols, 46.77% of flavonoids, 46.77% of total anthocyanins and 47.88% of anthocyanin monomers from sour cherry fruits have been transferred to juice. On the other hand, 29.85% of total polyphenols, 33.09% of flavonoids, 53.23% of total anthocyanins and 52.12% of anthocyanin monomers remained in pomace. Regarding radical scavenging activity, 65.51% of Trolox equivalents from sour cherries were exported to juice, while 34.49% was left in pomace. However, reducing power of sour cherry juice was much stronger than pomace (91.28% and 8.72% of Trolox equivalents from sour cherry fruits, respectively). Based on our results it can be concluded that sour cherry pomace is still a rich source of natural antioxidants, especially anthocyanins with coloring capacity, therefore it can be used for dietary supplements development and food fortification.

Keywords: antioxidants, polyphenols, pomace, sour cherry

Procedia PDF Downloads 325

Authors: Dana A. Thal, Heike Kahlert, Fritz Scholz

Abstract:

Thiol molecules are known to easily form self-assembled monolayers (SAM) on Au surfaces. Depending on the thiol’s structure, surface modifications via SAM can be used for electrode sensor development. In the presented work, 1-decanethiol coated polycrystalline Au electrodes were applied to indirectly assess the radical scavenging potential of plant compounds and extracts. Different plant compounds with reported antioxidant properties as well as an extract from the plant Gynostemma pentaphyllum were tested for their effectiveness to prevent SAM degradation on the sensor electrodes via photolytically generated radicals in aqueous media. The SAM degradation was monitored over time by differential pulse voltammetry (DPV) measurements. The results were compared to established antioxidant assays. The obtained data showed an exposure time and concentration dependent degradation process of the SAM at the electrode’s surfaces. The tested substances differed in their capacity to prevent SAM degradation. Calculated radical scavenging activities of the tested plant compounds were different for different assays. The presented method poses a simple system for radical scavenging evaluation and, considering the importance of the test system in antioxidant activity evaluation, might be taken as a bridging tool between in-vivo and in-vitro antioxidant assay in order to obtain more biologically relevant results in antioxidant research.

Keywords: alkanethiol SAM, plant antioxidant, polycrystalline Au, radical scavenger

Procedia PDF Downloads 299