Received 17 october 2012 received in revised form 30 march 20 accepted 8 may 20 available online 23 may. Dpph is a common abbreviation for the organic chemical compound 2,2diphenyl1picrylhydrazyl. Generally the radical scavenging activity of flavonoids depends on their structure and hydroxyl group arrangement. Hydroxyl scavenging activity of glucose, fructose, and. Most notably hydroxyl radicals are produced from the decomposition of hydroperoxides rooh or, in atmospheric chemistry, by the reaction of excited atomic. Hydroxyl and superoxide anion radical scavenging activities of. Hydroxyl radical scavenging activities of the spondias pinnata extract and the reference compound mannitol. Reducing power showed dose dependent increase in concentration absorption compared to standard, quercetin. Hydroxyl radical scavenging assay hydroxyl radical scavenging activity of fruit extracts was assayed by the method of14. The free radical scavenging activity of all the extracts was evaluated by 1, 1diphenyl2picrylhydrazyl dpph according to the previously reported method by shen et al.
Please share your experiences about the advantages and disadvantages of radical scavenging methods from medicinal plants. Structureradical scavenging activity relationships of flavonoids. The antioxidant potential and free radical scavenging activity were analysed using reducing power assay and hydrogen peroxide scavenging activity methods. This was assayed as described by elizabeth and rao with a slight modification. Amentoflavone protects against hydroxyl radical induced. Free radicals are generated by hydrogen peroxide h2o2. The results obtained from the study proved the free radical scavenging of the extract. Superoxide anion scavenging activity superoxide anion scavenging activity was. Novel fluorometric assay for hydroxyl radical scavenging. Presences of flavonoids generally possess higher antioxidant activity because of double bonds existing in cring.
The scavenging capacity for hydroxyl radical was measured according to the modified method of halliwell et al. The hydroxyl radicals thus generated quench the fluorescence of fluorescein over time. A new, simple assay has been optimized to generate and detect hydroxyl radical by fenton reaction. Hydroxy radical and dpph scavenging activity of crude protein. The extractives showed appreciable hydroxyl radical scavenging activity when compared with standard antioxidant, ca fig. Solvent effects and improvements in the deoxyribose. The developed method is less lengthy, more specific, and of a higher yield than the classical tbars assay. Evaluation of antioxidant and free radical scavenging. Materials and methods dpph free radical scavenging activity processing of plants for extract preparation.
Hydroxyl radical and hypochlorous acid scavenging activity of small centaury centaurium erythraea infusion. Assay of scavenging capacity against the hydroxyl radical the hydroxyl radical. The assay is the nitric oxide radical scavenging assay. Hydroxyl radicals are highly reactive easily becoming hydroxy groups and consequently shortlived. Methods used to evaluate the free radical scavenging. Somebody could please provide me hydroxyl radical scavenging. The antioxidant activities were determined by total antioxidant capacity, dpph 1,1diphenyl2picrylhydrazine radical scavenging assay, hydroxyl radical scavenging assay, ferrous reducing antioxidant capacity and lipid peroxidation inhibition assay methods. Hydroxyl radical scavenging assay of phenolics and flavonoids with. Taxifolin was also found to be the most effective antioxidant in the oxygen radical antioxidant capacity orac assay with a trolox equivalent of 2. Antioxidant and free radical scavenging potential have been reported to be rich in the fresh leaf than dry leaf 5. The total antioxidant activity, hydroxyl radical and hydrogen peroxide scavenging of a. Antioxidant capacity and radical scavenging effect of.
The hydroxyl radical footprinting methodology has wide applications to studying proteindna interactions, as well as structural perturbations e. The assay is based on quantification of the degradation product of 2deoxyribose by condensation with tba. The assay is based on oxidation of catechol, monitored at 360 nm following incubation at room temperature for ca. Mitochondrial hydroxyl radical detection assay kit. A comparative study on the antioxidant activity of. Hydroxyl radical antioxidant capacity horac assay kit 26 catalog number. Free radicals are generated by hydrogen peroxide h 2 o 2.
Dpph is a stable free radical that reacts with compounds able to donate a hydrogen atom. A 70% methanol extract of spondias pinnata stem bark was studied in vitro for total antioxidant activity, for scavenging of hydroxyl radicals. The ability of the extractives to quench hydroxyl radicals might directly relate to the prevention of lipid peroxidation. Novel hydroxyl radical scavenging antioxidant activity. Hydroxyl radical scavenging activity assay the scavenging activity for hydroxyl radicals was measured with fenton reaction 12. It is a darkcolored crystalline powder composed of stable free radical molecules. The extracts were prepared from a 10 mgml ethanol crude extract. Comparative analysis of the antioxidant activity of cassia. The horac assay is based on the oxidation of fluorescein by hydroxyl radicals via a classic hydrogen atom transfer hat mechanism. For determination of radical scavenging activity of different foods, beverages and substrates were elaborated a. Please share you experiences about the best method to determine radical scavenging on natural product samples.
Introduction it is now well established that oxidative stress is a major risk factor for the development of. The antioxidant activity of the extracts was measured on the basis of the scavenging activity of the stable 1, 1 diphenyl 2picrylhyorazyl dpph free radical according to the method described by brandwilliams et al22 with slight modifications. Antioxidant and free radical scavenging activity of. The generation of pure hydroxyl radicals under the experimental conditions was evaluated and confirmed using electron spin resonance. Evaluation of antioxidant capacity and free radical. Evaluation of free radical scavenging activity of various leaf extracts from. In the present study, the ability of scavenging free radicals of the ethanolic extract of mollugo nudicaulis was determined by using 1,1diphenyl2picrylhydrazyl dpph, reducing power, ferric reducing antioxidant power frap, hydroxyl radical scavenging assay, superoxide radical. Jan 19, 20 the antioxidant activities were determined by total antioxidant capacity, dpph 1,1diphenyl2picrylhydrazine radical scavenging assay, hydroxyl radical scavenging assay, ferrous reducing antioxidant capacity and lipid peroxidation inhibition assay methods. Scavenging capacity assay kit chekine hydroxyl free radical. The antioxidant activity of the memq was evaluated by the phosphomolybdenum method according to the procedure of prieto et al. In the hydroxyl radical scavenging assay, the activity of the leaf fraction was greater than that of the other fractions but lower than that of ascorbic acid. Ros, which include free radicals such as superoxide anion. Development and validation of a radical scavenging. Antioxidant activity and free radical scavenging capacity.
Solvent effects and improvements in the deoxyribose degradation assay for hydroxyl radicalscavenging xican li. Based on dpph and hydroxyl radical scavenging activity, tpl showed. Therefore, the search for methods to determine free radical scavenging is important. Oxiselect hydroxyl radical antioxidant capacity horac. The hydroxyl radical scavenging rate constants of ascorbic acid, formate, and hexacyanoferrateii that caused interference in other assays could be easily found with the proposed procedure. Is there any easy method for the estimation of hydroxyl radical scavenging activity.
Amentoflavone protects against hydroxyl radicalinduced. In vitro antioxidant and free radical scavenging activity. Radical scavenging and antioxidant activity of ethanolic. Unlike other antioxidant activity assays, the fluorescent horac assay provides a direct measurement of antioxidant capacity against hydrophilic chainbreaking hydroxyl radicals.
Dpph radical scavenging assay an overview sciencedirect. Antioxidant and free radical scavenging potential have been reported to. This effect can be a result of the protien exsisted in cuttlfish tissue, mechanism of direct radical scavenging activity, or absorbtion of heavy metals. Mpe exhibits significant strong scavenging activity on dpph and abts assay. The method dpph is widely used for measurement of free radical scavenging ability of antioxidants perezjimenez and sauracalixto, 2008. Antioxidant, free radical, hydroxyl radical, dpph, frap, sepia pharaonis. These were then serially diluted with distilled water to make concentrations from 100. In vitro free radical scavenging activity dpph radical scavenging assay the scavenging activity for dpph free radicals was measured according to the procedure described by blios 20.
Hydroxyl radical scavenging assay of phenolics and flavonoids with a modified cupric reducing antioxidant capacity cuprac method using catalase for hydrogen peroxide degradation. Dpph free radical scavenging activity of the extracts of the. The highest radical scavenging activity is exhibited by compounds that have an ortho 3. Among the extracts, tsb showed the highest antioxidant activity followed by trb, tf.
A new fenton assay for hydroxyl radical scavengers by. The structural elements of the flavonoid molecule most important for hydroxyl radical scavenging are hydroxylation of ring b and a c2c3 double bond connected with a c. The hydroxyl radical antioxidant capacity horac assay is a classic tool for measuring the antioxidant capacity of biomolecules from a variety of samples. Cpll extract showed in vitro antioxidant activity in both hydroxyl radical scavenging and as well as in dpph scavenging. Antioxidant and free radical scavenging activity of spondias pinnata. Dpph radical scavenging assay an overview sciencedirect topics. Protocol used in this study for the use of rat as animal model for lipid.
Stock solutions of edta 1 mm, fecl3 10 mm, ascorbic acid 1 mm, h2o2 10 mm and deoxyribose 10 mm, were prepared in distilled deionized water. Mitochondrial hydroxyl radical detection assay kit ab219931 is a sensitive fluorometric onestep assay to detect intracellular hydroxyl radical oh in live cells. Flavonoids as potent scavengers of hydroxyl radicals treml. The dpph and abts free radical scavenging assay are the commonly used methods generally used evaluation of proton. Free radical scavenging and antioxidant activities of. The following assay procedure was modified from those described by blois 1958 and yamasaki, et al. Is there any easy method for the estimation of hydroxyl radical.
There are several assays to measure the antioxidant potential of compounds, the 1,1diphenyl2picrylhydrazine dpph radical scavenging assay being the most extensively used antioxidant assay for plant samples. Jun 02, 2008 hydroxyl radical scavenging assay of phenolics and flavonoids with a modified cupric reducing antioxidant capacity cuprac method using catalase for hydrogen peroxide degradation. The relationship between hydroxyl radical scavenging and the structure of the flavonoids is discussed. What is the best method for radical scavenging assay. The hydroxyl radical is generated by a coiimediated fentonlike reaction, and the hydroxyl radical formation under the experimental condition is indirectly confirmed by the hydroxylation of phydroxybenzoic acid. The order of greatest hydroxyl radical scavenging activity was green tea. Hydroxyl radical and hypochlorous acid scavenging activity. Superoxide, hydroxyl radical, lipid peroxidation, nitric oxide assay showed a comparable scavenging activity compared to its standard. Horac assay hydroxyl radical assay oxford biomedical research. Ftir spectral studies clearly indicate the presence of various functional groups which may be attributed to the antioxidant and free radical scavenging properties of these extracts. Is there any easy method for the estimation of hydroxyl. In vitro antioxidant and free radical scavenging activity of different. Hydroxy radical and dpph scavenging activity of crude. Hydroxyl radical scavenging assay of phenolics and.
A comparative study with green tea camellia sinensis. The successive antioxidant activity were done for dpph radical scavenging activity, hydroxyl radical scavenging activity, nitric oxide radical scavenging activity, reducing power, lipid peroxidation inhibiting activity and total antioxidant assay using standard procedure. Frap assay, hydrogen peroxide scavenging activity, nitric oxide scavenging activity, superoxide radical scavenging activity, hydroxyl radical scavenging activity, phosphomolybdate assay, reducing power, metal ion chelating activity, and carotene. The amount of nadph utilized in a direct measure of enzyme activity. Superoxide radical scavenging activity the superoxide radical scavenging activity of annona squamosa seeds extracts were studied and compared with gallic acid. A comparative study on the antioxidant activity of methanolic. As cpll extract, scavenge hydroxyl radical more than the dpph radical, it might have dna protectant activity also. Antioxidant capacity and radical scavenging effect of polyphenol. What is the best method for radical scavenging assay antioxidant assay. A novel fluorometric method was developed and validated for hydroxyl radical scavenging capacity hosc estimation using fluorescein as the probe. All mrps exhibited detectable, but variable, non site. Free radical scavenging properties of annona squamosa.
Deoxyribose assay is used to determine the hydroxyl radical scavenging activity in an aqueous medium. The horac activity assay is based on the oxidation of a fluorescent probe by hydroxyl radicals by way of a hydrogen atom transfer hat process. Hydroxyl radical scavenging assay of phenolics and flavonoids. Structurefunction relationships of hydroxyl radical. Superoxide anion radical scavenging activity biology essay. Hydroxyl radicals cleave dna by abstracting a hydrogen atom from c4 of the sugar in the minor groove. Reactive oxygen species like hydroxyl radicals oh o are produced as the byproduct of normal cell metabolism and environmental stress 4. In vitro antioxidant activity and free radical scavenging. Free radicalscavenging capacity, antioxidant activity and. Hydroxyl radical scavenging o assay of phenolics and. Determination of the radical scavenging activity 1, 1diphenyl2picrylhydrazyl dpph assay introduction the importance of free radicals and reactive oxygen species ros has attracted increasing attention over the past decade.
Bioactive compounds, radical scavenging, antioxidant. Methods and applications of antioxidant activity assays. Deoxyribose assay for hydroxyl radical scavenging activity. Dna nicking assay for hydroxyl radical scavenging activity. Fe assay principle colorimetric hydroxyl free radical scavenging capacity assay kit provides a simple and easy assay for the study of hydroxyl radical scavenging capacity in various sample. The hydroxyl radical scavenging rate constants of ascorbic acid, formate, and hexacyanoferrateii that caused interference in other assays could be. Health, general biological sciences science and technology, general antioxidants analysis antioxidants nutrients comparative. Radical scavenging dpph assay radical scavenging dpph assay were conducted using the standard protocols. The data represent the percentage inhibition of deoxyribose degradation. Antioxidant and free radical scavenging activity of spondias. Novel fluorometric assay for hydroxyl radical prevention. The green fruit had the highest ic 50 value 119 mgml, followed by the leaf 167 mgml, the stem 267 mgml, and the ripe fruit 173 mgml 37.
Several methods have been developed to assess the radical scavenging activity. School of chinese herbal medicine, guangzhou university of chinese medicine, guangzhou, china article info article history. Deoxyribose assay was used to determine the hydroxyl radical scavenging activity in an aqueous medium. This was assayed as described by elizabeth and rao. Phytochemical analysis was performed on the plant extract to detect the presence of phytoconstituents.
Standard antioxidant assays like dpph, abts, hydroxyl, superoxide radical, nitric. Hydroxyl radical scavenging activity by pnda pbutrisidunethyl aniline. Evaluation of nitrite radical scavenging properties of. The hydroxyl radical, oh, is the neutral form of the hydroxide ion oh. Principles of procedure the hydroxyl radical antioxidant capacity horac assay is based on the oxidation of fluorescein by hydroxyl radicals via a classic hydrogen atom transfer hat mechanism.
Dpph has two major applications, both in laboratory research. On the other hand, hydrogen peroxide is used in the hrs assay to determine the scavenging effecincy of hydroxyl radicals by the chemlcals described in the working procedure depicted in my previous. Therefore for assessment of different antioxidants we need different assays. Development and validation of a radical scavenging antioxidant assay using potassium permanganate isaac kingsley amponsah, emmanuel orman, abraham yeboah mensah, francis mainoo sarpong, francis ackah armah, linda mensah sarpong the main objective of the study was to develop and validate a novel radical scavenging antioxidant assay using. Hydroxyl radical scavenging assay of annonasquamosa seed extracts aspe, asch, asea and asme at different concentrations. At 500 gml concentration of cpll, it showed 78 and 63% inhibition, with an ic50 value of 150 and 175 gml in hydroxyl radical and dpph assay respectively. Somebody could please provide me hydroxyl radical scavenging activity protocol. Hydroxyl radical and hypochlorous acid scavenging activity of. Determination of hydroxyl radical scavenging activity. Among them, the 2,2diphenyl1picrylhydrazyl dpph spectrophotometric method is one of the most widely applied and is appreciated for its reliability. Phytochemical analysis and hydroxyl radical scavenging. A novel fluorometricmethod has been developed to evaluate hydroxyl radical prevention capacity using fluorescein fl as the probe.
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