how to calculate antioxidant activity by dpph

Other radical probes, such as TRAP (total peroxyl radical trapping antioxidant parameter) monitoring the reaction between peroxyl radicals and the sample quenched the chemiluminescence, CUPRAC (cupric reducing antioxidant power) by which the reduction of Cu2+ is observed, Fremys salt (galvinoxyl radical), aroxyl radical (2,6-di-tert-butyl-4(4-methoxyphenyl)phenoxyl radical) [42,60,63,64]. Phull A., Abbas Q., Ali A., Raza H., Ja S., Zia M., Haq I. Antioxidant, cytotoxic and antimicrobial activities of green synthesized silver nanoparticles from crude extract of Bergenia ciliata. Beside the application of AgNPs in diverse areas, the large number of articles dealing with antioxidant properties of silver nanoparticles have been published spreading in the recent decade. The DPPH scavenging assay was done for . Environmentally friendly procedure for in-situ coating of regenerated cellulose fibres with silver nanoparticles. Other theory was presented by Vijayan et al. The authors explained the results based on the hypothesis that in the presence of H2O2 the dispersed nanoparticles may induce the formation of reactive oxygen species and hydrogen peroxide inside a cell. Bal M., Daneu N., Balov ., Dutkov E., Tkikov ., Brianin J., Vargov M., Balov M., Zorkovsk A., Bal P. Bio-mechanochemical synthesis of silver nanoparticles with antibacterial activity. Alves T.F., Chaud M.V., Grotto D., Jozala A.F., Pandit R., Rai M., Alves C. Association of silver nanoparticles and curcumin solid dispersion: Antimicrobial and antioxidant properties. Tan D., Garca F. Main group mechanochemistry: From curiosity to established protocols. The antioxidant activity is measured by determining the loss of fluorescence and fluorescein is usually used as a probe [59,60]. The analyte flows through column in a liquid mobile phase to a detector (UV/Vis, DAD, MS, etc.,). The results of DPPH and superoxide anion radical scavenging assays lead to reduced antioxidant capacity of prepared nanoparticles in comparison to plant extract, but the total reducing power of AgNPs indicated more reducing activity than plant extract. Hydrogen peroxide and DPPH scavenging activity of AgNPs was also compared with ascorbic acid. AZ1 and its antimicrobial activity against several human pathogenetic bacteria. Bookshelf The present phytochemicals (flavonoids) and silver ions could serve as antioxidants through single electron and hydrogen atom transfer [127]. In addition to plant extract, bacterial strains are also used for AgNPs biosynthesis. The free radical scavenging activity of Teucrium polium extract, and of AgNPs produced chemically and by green synthesis was studied by DPPH and FRAP assays in [163]. Wei L., Lu J., Xu H., Patel A., Chen Z.-S., Guofang C., Chen Z. Performed by Mohammad Shah Hafez Kabir Founder and CEO, GUSTO A Research GroupB. Paste experimental data into the box on the right. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (, silver nanoparticles, antioxidant activity, silver nanoparticles synthesis. Khoshnamvand M., Huo C., Liu J. As the name implies, antioxidants are substances that are capable of counteracting the damaging, but normal, effects of the physiological process of oxidation in animal tissue. Promising antioxidant properties were reported by Sivasankar et al. Frstermann U. Nitric oxide and oxidative stress in vascular disease. . DPPH (2,2-diphenyl-1-picrylhydrazyl) is a purple stable free radical reacting with hydrogen donor (Scheme 1). FRAP values are calculated from increasing absorbances measured at 593 nm [39,42,43]. Chen S.-F., Zhang H. Aggregation kinetics of nanosilver in different water conditions. It is necessary to note that oxidative stress has not only negative effects on human body, as free radicals have an irreplaceable function in living organisms. There is no universal antioxidant, as different antioxidants react with different reactive species by various mechanisms, at various locations and protect specific molecular targets [35,36]. want to know how to. The antioxidant capacities of structurally different phenolic compounds were evaluated. Epub 2018 Sep 6. For the basic study of antioxidant activity, the use of various in vitro methods is a suitable choice, either with a mechanism of action such as hydrogen transfer or electron transfer. Sivasankar P., Seedevi P., Poongodi S. Characterization, antimicrobial and antioxidant property of exopolysaccharide mediated silver nanoparticles synthesized by Streptomyces violaceus MM72. Popular answers (1) Ali Mojaddar Langroodi Urmia University 10th Aug, 2017 DPPH scavenging effect %= AD-AS/ AD *100 Where AD is absorbance value at 517 nm of the methanolic solution of DPPH and. Docea A.O., Calina D., Buga A.M., Zlatian O., Popescu E.L., Stoica A.E., Catalina A. 2023 Apr 11;9(4):e15444. Al-Bahrani R., Raman J., Lakshmanan H., Hassan A.A., Sabaratnam V. Green synthesis of silver nanoparticles using tree oyster mushroom Pleurotus ostreatus and its inhibitory activity against pathogenic bacteria. The site is secure. Rajput S., Kumar D., Agrawal V. Green synthesis of silver nanoparticles using Indian Belladonna extract and their potential antioxidant, anti-inflammatory, anticancer and larvicidal activities. Generating of ABTS+ and its reaction with an antioxidant [52]. In order for the substance to be considered as an antioxidant, it must be active at low concentration (phenolic antioxidants often loose activity at high concentration and act as prooxidant), its amount needs to be satisfactory high to deactivate the target molecule, it must react with oxygen or nitrogen free radicals, and the final product of the reaction should be less toxic than removed radical. In view of the mentioned fact, the AgNPs are studied as antibacterial agents to inhibit resistant strains by multiple mechanisms of action, involving induction of oxidative stress, inhibition of DNA replication, or interaction with enzymes and proteins [5,6]. The authors observed excellent nitric oxide scavenging activity of AgNPs in a concentration-dependent manner, but the activity was in general lower than that of ascorbic acid standard. Unauthorized use of these marks is strictly prohibited. This method is based on the reduction of DPPH radical at a glassy carbon electrode [65,66,67]. Because of this, there is an enormous need to develop a uniform protocol for determining the antioxidant capacity of nanoparticles to precisely assess their potential. 2. measurement of extinction, scattering, and absorption cross sections. 2013 Feb;65(2):271-9. doi: 10.1111/j.2042-7158.2012.01591.x. They speculated that improved antioxidant properties of AgNPs are due to the simultaneous activity of polyphenols as antioxidant agents and AgNPs as a catalyst [124]. [Detection of antioxidative capacity of bamboo leaf extract by scavenging organic free radical DPPH]. Before The level of disappearance of purple color depends on the concentration of the antioxidant. THG-LS1. Hayyan M., Hashim M.A., Alnashef I.M. 2014;14(6):494-504. doi: 10.2174/1389557514666140622204037. Gahlawat G., Shikha S., Chaddha B.S., Chaudhuri S.R., Mayilraj S. Microbial glycolipoprotein-capped silver nanoparticles as emerging antibacterial agents against cholera. According to the authors, the antioxidant ability of AgNPs is caused by the presence of phenolic compounds, terpenoids, and flavonoids in plants which allow nanoparticles to act as singlet oxygen quenchers, hydrogen donors, and reducing agents [129]. Saravanakumar K., Wang M. Microbial Pathogenesis Trichoderma based synthesis of anti-pathogenic silver nanoparticles and their characterization, antioxidant and cytotoxicity properties. When comparing the activity with the extract, the choice of concentration seems to be a key factor influencing the result, as the antioxidant activity is concentration-dependent. Mittal [157] and his co-workers published a study dealing with the synthesis of AgNPs using Syzygium cumini extract. The antioxidant capacity was determined at different concentrations (100, 200, 300, 400 and 500 mM). An official website of the United States government. antioxidant Any agente.g., vitamin A, vitamin C, seleniumwhich is capable of reducing highly histotoxic oxygen-reduction products and reactive oxygen species (e.g., hydroxyl radical), which derive from superoxide anion (O2-) and H2O2, the univalent and bivalent reduction products of oxygen, and are generated during the normal intermediary metabolism of the respiratory chain. Saifuddin N., Wong C.W., Yasumira a.a.N., Nur Yasumira A.A. The chosen composition of the mobile phase (eluent) depends on the used stationary phase and interactions between analytes. This is the standard form in which the potential compound or extract is mixed with a mixture of DPPH and absorption spectrum after a given time is reported. Careers, Unable to load your collection due to an error. Mittal A.K., Kaler A., Banerjee U.C. In addition, an excessive amount of reactive nitrogen species is formed during the oxidative stress. Biogenic synthesis of silver nanoparticles from white rot fungi: Their characterization and antibacterial studies. The preferred wavelength is 734 nm because the interference with other absorbing components is minimized. There are many studies dealing with mentioned problem and herein we summarize the knowledge about methods evaluating the antioxidant activity of silver nanoparticles reported so far. HPLC is a valuable method for antioxidant capacity determination. How do you calculate the antioxidant activity of food using DPPH method? Physical methods are useful because of speed and not using toxic chemicals, but on the other hand disadvantages are also there which are previously mentioned [80]. Lower values of silver nanoparticles (prepared by Aesculus hippocastanum) antioxidant capacity was also obtained by Kp [126] who used three different method of scavenging activity determinationDPPH, total reducing power, and superoxide anion radical scavenging assay. ) radical scavenging activity is generally quantified in terms of inhibition percentage of the pre-formed free radical by antioxidants, and the EC (50) (concentration required to obtain a 50% antioxidant effect) is a typically employed parameter to express the antioxidant capacity and to compare the activity of different compounds. [154] who determined antioxidant properties of silver nanoparticles prepared by exopolysaccharides from Lactobacillus brevis. Crude antioxidant samples are first subjected to HPLC (normally reversed-phase) separation, and the eluate corresponding to each peak is reacted with DPPH radical at a certain flow rate for a constant time period (e.g., 60 s), resulting in loss of DPPH radical and the antioxidant itself, which can be recorded by a UV detector (Kedare & Singh . Epub 2015 Oct 1. Oxidative stress, caused by endogenous factors such as reactive oxygen species (ROS) including the hydroxyl radical, superoxide anion radical, hydrogen peroxide, singlet oxygen, nitric oxide radical, hypochlorite radical, etc., and exogenous factors such as smoking, ionizing radiation, pollution, organic solvents, pesticides, etc., are able to a. These products may initiate new radical reactions leading to damage of biomolecules (nitrosylation of DNA or proteins) [19]. This methodology, also called mechanochemical synthesis has recently attracted a significant attention of the research world [85,86,87,88,89,90,91]. Genesis and development of DPPH method of antioxidant assay. Halliwell B. Antioxidants and human disease: A general introduction. Mello L.D., Kubota L.T. 4 and their antimicrobial application. Heliyon. The presence of delocalized spare electrons on the whole molecule prevents dimerization and also gives the color to the molecule of DPPH with absorption maximum at around the value of 520 nm in UV/Vis spectra. Silver nanoparticles synthesized and coated with pectin: An ideal compromise for anti-bacterial and anti-biofilm action combined with wound-healing properties. Bhat R., Deshpande R., Ganachari S.V., Huh D.S., Venkataraman A. Photo-irradiated biosynthesis of silver nanoparticles using edible mushroom Pleurotus florida and their antibacterial activity studies. 2. Silver nanoparticles: Synthesis methods, bio-applications and properties. The scavenging activity against DPPH radical is higher than that against ABTS and NOx. A comparative study of stability, antioxidant, DNA cleavage and antibacterial activities of green and chemically synthesized silver nanoparticles. Compounds (even without antioxidant properties) with redox potential lower than the redox potential of Fe3+/Fe2+ pair may reduce Fe3+ to Fe2+ and can increase the FRAP value to obtain false high results. Ecotoxicology and environmental safety antioxidant activity of chemically synthesized AgNPs and biosynthesized pongamia pinnata leaf extract mediated AgNPs A comparative study. In general, contradictory results can be found in literature. First, the separation of sample is promoted and then is mixed with radical solution (ABTS, DPPH). Reaction of DPPH radical with hydrogen donor [46]. Arnao M.B. 2015 Oct 14;63(40):8765-76. doi: 10.1021/acs.jafc.5b03839. The range of linearity between the DPPH inhibition percentage and In Table 1, we provide an overview of methods for antioxidant capacity evaluation and some of them are briefly described [12,42,43,44]. (Elements & Compounds) any substance that retards deterioration by oxidation, esp of fats, oils, foods, petroleum products, or rubber. Antioxidants capable of hydrogen atom transfer decolorize the solution, absorbance decrease at 505 nm [12]. The main disadvantage of using this assay is that DMPD radical is soluble only in water, so there are limitations in using it for hydrophobic antioxidants determination [54]. Cao G., Prior R.L. Oliveira G.K.F., Tormin T.F., Sousa R.M.F., De Oliveira A., De Morais S.A.L., Richter E.M., Munoz R.A.A. Nanoparticles have been shown to have a more complicated mechanism of action in biological systems using in vivo activity-based methods. Elemike E.E., Fayemi O.E., Ekennia A.C., Onwudiwe D.C., Ebenso E.E. Comparison of ABTS/DPPH assays to measure antioxidant capacity in popular antioxidant-rich US foods. The scavenging capacity is usually determined in organic solvents, not in aqueous media [12,39,41,42,45,46]. Selvan D.A., Mahendiran D., Kumar R.S., Rahiman A.K. Floegel A., Kim D., Chung S., Song W.O.N.O., Fernandez M.L.U.Z., Bruno R.S., Koo S.I., Chun O.C.K.K. Several studies described the cytotoxic effect of AgNPs through ROS generation [31,32,33]. This method is popular mainly in the determination of antioxidant ability of foods and is limited by low reactivity of fluorescein and with peroxyl radicals [61]. For these reasons, various physical techniques were developed. However, the ABTS assay has some reservations in the overall applications, such as specificity for reaction of different antioxidants, storage of the radical, or processing method conditions [53]. Mini Rev Med Chem. Determination of antioxidant activity (or capacity) of samples of various origin is based on different methodologies and assays. Pluym et al. Nanotechnology solutions for alzheimer s disease: Advances in research tools, diagnostic methods and therapeutic agents. Silver nanoparticles mediated by Costus afer leaf electrochemical properties. In vivo methods use the whole cell for AgNPs biosynthesis, so nanoparticles are synthesized intra- or extracellularly, while during the in vitro process, the reduction of Ag+ ions takes places outside of a living organism (the most common are plant extracts containing the compounds with antioxidant and reducing propertiespolyphenols, flavonoids, terpenes, aldehydes, carbohydrates, etc.,) [100,101,102]. The two main problems observed are: first, the absolute determination of the fifty percent inhibitory concentration (IC 50) despite the initial concentration of DPPH used, when a relative value would be more coherent and could be compared with findings of other studies [6,7]; and second, the mistake in the determination of the percentage of scav. Arteaga J.F., Ruiz-Montoya M., Palma A., Alonso-Garrido G., Pintado S., Rodriguez-Mellado J.M. Chromatography encompasses a group of broadly applicable methods in detection and separation of various compounds present in the analyte, but can also be used for antioxidant capacity measurement. Authors explained this phenomenon by the role of phytochemicals. Do J.-L., Frii T. Mechanochemistry: A force of synthesis. Denev P., Ciz M., Ambrozova G., Lojek A., Yanakieva I., Kratchanova M. Solid-phase extraction of berries anthocyanins and evaluation of their antioxidative properties. Classification, physicochemical principles, mechanisms, and electron transfer (ET)-based assays. Generally, there are three methods for AgNPs synthesisbiological, physical, and chemical. The oxidation potential is usually scanned linearly in time from an initial to a final value and back as a triangular waveform. [127], who prepared Ag nanoparticles by Costus afer. Ilyasov I.R., Beloborodov V.L., Selivanova I.A., Terekhov R.P. Wang L., Wu Y., Xie J., Wu S., Wu Z. The higher degree of hydroxylation in chemical structures of phenolic compounds demonstrated the higher radical scavenging capacity and higher tendency to reduce Ag+ to AgNPs [161]. All tested nanoparticles have exhibited lower scavenging activity than ascorbic acid standard. ) radical scavenging activity is generally quantified in terms of inhibition percentage of the pre-formed free radical by antioxidants, and the EC(50) (concentration required to obtain a 50% The DPPH assay is one of the most commonly employed methods for measuring antioxidant activity. Otunola G.A., Afolayan A.J., Ajayi E.O., Odeyemi S.W. These findings make the silver nanoparticles useful for neurodegenerative diseases, cancer, or AIDS treatment [155,162]. http://creativecommons.org/licenses/by/4.0/, Reaction with peroxyl radical initiated by AAPH, Reaction with OH radicals generated by Co, Reaction with luminol-derived radicals, generated during the AAPH decomposition, Emission of light by a substance that has absorbed the light or other electromagnetic radiation of a different wavelength, The potential of working electrode is varying from initial to final value and back, current intensity is recorded, Measurement of the intensity of cathode or anode peak, The potential of working electrode is fixed to a reference electrode, Measurement of the intensity currently produced by oxidation or reduction of a sample, The reaction of an antioxidant with the oxidized form of a reversible redox couple, Measurement of the current flow between two identical working electrodes, Separation of compounds in a reaction mixture at a stationary phase in a liquid mobile phase, Enzyme-based biosensors measuring total phenolic content, Reaction of noble metal (Au, Ag) salt with antioxidant compound. The formation of nanoparticles from noble metals, such as gold and silver, by the reaction between metallic salt (Au3+, Ag1+) and antioxidant leads to the possibility of determination of antioxidant properties of studied samples [12]. John T., Odunayo O., Benjakul S., Rujiralai T. Synthesis and characterization of novel poly (3-aminophenyl boronic acid- co-vinyl alcohol) nanocomposite polymer stabilized silver nanoparticles with antibacterial and antioxidant applications. Some current insights into oxidative stress. Jing M., Ah K., Kyung I., Sun H., Kim S., Yun J., Choi J., Won J. 2.8. ABTS is usually first oxidized by potassium persulphate [49], manganese dioxide [50], or AAPH (2,2-azobis-(2-amidino-propane) dihydrochloride) [51] which gives rise to the cation radical ABTS+. Kaysen G.A., Eiserich J.P. They observed that the alfalfa root is able to absorb silver from agar medium to produce AgNPs.
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