Jan 25, 2018 non sitespecific hydroxyl radical scavenging activity. Moreover, bht, which is a synthetic antioxidant, was evaluated as a positive control and compared with acetone and methanol extracts of the plant. Hydroxyl radical scavenging assay of phenolics and. Antioxidant activity of polyphenolic compounds isolated from. Superoxide anion radical scavenging activity biology essay.
Most notably hydroxyl radicals are produced from the decomposition of hydroperoxides rooh or, in. Structureradical scavenging activity relationships of flavonoids. Free radicals are generated by hydrogen peroxide h2o2. The free radical scavenging activity of the extract and fractions of t cordifolia was evaluated using the stable free radical i.
It reacts with polyunsaturated fatty acid moieties of cell membrane. The scavenging capacity may change with time, and it would be useful to continuously measure this change to be able to optimize aop doses. The hydroxyl radical producing compound of the root extract showed 53%, whereas the leaf extract showed only 28% scavenging activita. In addition, hydroxyl radicals can reduce disulfide bonds in proteins. The hydroxyl radical is often referred to as the detergent of the troposphere because it reacts with many pollutants, decomposing them through cracking, often acting as the first step to their removal. Various concentrations 20, 40, 60, 80 and 100 gml in methanol of extracts were taken in different test tubes and evaporated to dryness. Is there any easy method for the estimation of hydroxyl radical. Place 20 l of sample or standard into the appropriate wells of the provided microplate. Free radicals, natural antioxidants, and their reaction. Evaluation of free radical scavenging activity of various extracts of. The ic 50 values table 1 of the extract and standard in this assay were 112.
For determination of radical scavenging activity of different foods, beverages and substrates were elaborated a. Evaluation of free radical scavenging activity of an. Abbkine chekine hydroxyl free radical scavenging capacity assay kit is designed to assess the hydroxyl free radical scavenging capacity in the samples. 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. The method dpph is widely used for measurement of free radical scavenging ability of antioxidants perezjimenez and sauracalixto, 2008. The fenton reaction mixture consisted of 200 l 10 mm. The reaction mixture contained 50 mm phosphate buffer ph 7.
Cpll extract showed in vitro antioxidant activity in both hydroxyl radical scavenging and as well as in dpph scavenging. Flavonoids as potent scavengers of hydroxyl radicals treml. Hydroxyl radical scavenging activity of the extracts was determined according to. On the direct scavenging activity of melatonin towards hydroxyl and a series of peroxyl radicals. The relationship between hydroxyl radical scavenging and the structure of the flavonoids is discussed. Chrysin, rutin and quercetin were run to explore the effect of. The hydroxyl radical scavenging activity of the microparticle s suspension was slightly less than bht but more.
According to principles of electrochemistry antioxidant must be endowed with a. In vitro free radical scavenging activities of the extracts were assessed against dpph and hydroxyl radicals. Hydroxyl radical scavenging assay of phenolics and flavonoids. Horac microplate assay kit hydroxyl radical antioxidant. 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. Free radical scavenging and antioxidant activities of. 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. In vitro antioxidant and free radical scavenging activity. Deoxyribose assay is used to determine the hydroxyl radical scavenging activity in an aqueous medium. In vitro and in vivo determination of hydroxyl radical scavenging activity hrsa of fractions of aqueous extract of moringa oleifera leaves aemol address for correspondence. The % hydroxyl radical scavenging activity of test sample was determined accordingly in comparison with negative control.
It also has an important role in eliminating some greenhouse gases like methane and ozone. The present study was aimed to investigate the in vitro free radical scavenging activity of various leaf extracts aqueous, methanol, acetone chloroform and petroleum ether of kedrostis foetidissima. In vitro free radical scavenging activities of the extracts were assessed against. Our finding further provides evidence that mallotus fruit extract is a potential natural source of antioxidants which have a protective role on human erythrocytes exhibiting minimum hemolytic activity and this. Unlike other antioxidant activity assays, the fluorescent hydroxyl radical antioxidant capacity horac assay provides a direct measurement of antioxidant capacity against hydrophilic chainbreaking hydroxyl radicals. Scavenging activity of extracts of root and leaf is inversely proportional to the free radical production. Hydroxy radical and dpph scavenging activity of crude protein. Hydroxy radical and dpph scavenging activity of crude. Hydroxyl radical antioxidant capacity horac the horac activity assay is based on the oxidation of a fluorescent probe fluorescein by hydroxyl radical via hydrogen atom transfer hat process 22. The objective of the present study was to evaluate the antioxidant potential and free radical scavenging activity of a 70% methanol extract of s. If so, it would have relevance to the free radical scavenging activity of ndga.
The scavenging or inhibiting activity, expressed as mole hydroxyl radical scavenged or reduced per mole of test agent, was calculated as follows. Hydroxyl radical scavenging activity a0 a1a0100 where, a0 is the absorbance of the control reaction, and a1 is the absorbance in presence of all of the extract samples and reference. Scavenging activity on dpph radical quantitative fig. Scavenging ability of hydride antioxidants results the reaction scheme for the direct measurement of the hydroxyl radical scavenging capacity 78 is based on the principle that any molecule introduced into the analytical solution that is able to react with the oh will compete with the terephthalate for oh and therefore will reduce. There was no significant difference in the scavengingfree radical activity of methanol and water extracts from fmof compared with fmos group. Mustard varieties, thyme, oregano, clove, and allspice all exhibited strong ohscavenging activity. This was assayed as described by elizabeth and rao. Free radical scavenging activity and inhibition of oxidative stress. Scavenging of hydroxyl radical was determined using dmpo. Non sitespecific hydroxyl radical scavenging activity. Increasing the concentration of the extracts did not result in an increase in the nitrite radical scavenging activity. The contents of free radical scavenging activity in the mycelia of rcef0868 were soluble in.
Antioxidant and free radical scavenging activity of spondias. The effects of methanol extracts of 51 spices on oh scavenging were studied in detail. A new fenton assay for hydroxyl radical scavengers by. Activity was calibrated using a standard material, l. Determination of hydroxyl radical scavenging activity. Evaluation of free radical scavenging activity of various. Chekine hydroxyl free radical scavenging capacity assay.
Hydroxyl radical scavenging assay of phenolics and flavonoids with. Free radical scavenging activity of crude extracts and 4. Structureradical scavenging activity relationships of. Hydroxyl radicals are highly reactive easily becoming hydroxyl groups and consequently shortlived. The assay is based on quantification of the degradation product of 2deoxyribose by condensation with tba. Hydroxyl radical scavenging activity klein, 1995 17. Fentons reaction, scavenging of oh, and luminol cl reaction. The hydroxyl radical footprinting methodology has wide applications to studying proteindna interactions, as well as structural perturbations e. The extract was examined for different reactive oxygen species ros scavenging activities including hydroxyl, superoxide, nitric oxide, hydrogen peroxide, peroxynitrite, singlet oxygen and. K1, an analog of vitamin c and vitamin e which is water soluble and.
Hydroxyl radical scavenging capacities of the powder suspension t obtained from hypsizygus. Hydroxyl radical scavenging activity hrsa of the extractives was determined by the method of klein et al. Extraction and determination of antioxidant activity of. Development of an evaluation method for hydroxyl radical. Box 180, hr2 zagreb, croatia received march 26, 2002. Calculations the data were subjected to routine statistical analysis. External standard calibration method to measure the hydroxyl. The metal chelating activity and reducing power ability of the extracts were also. Hydroxyl radicals cleave dna by abstracting a hydrogen atom from c4 of the sugar in the minor groove. There was no significant difference in the scavenging free radical activity of methanol and water extracts from fmof compared with fmos group. The hydroxyl radical oh scavenging capacity is a useful parameter for the design and operation of an advanced oxidation process aop in water treatment. The hydroxyl radical, oh, is the neutral form of the hydroxide ion oh. Bioactive compounds, radical scavenging, antioxidant.
The compound sodium nitroprusside is known to decompose in aqueous solution at physiological ph 7. Hydroxyl radical scavenging activity fenton reaction based. However, they form an important part of radical chemistry. Evaluation of hydroxyl radicalscavenging abilities of two. Hydroxyl radical and its scavengers in health and disease hindawi.
Hydroxyl radical antioxidant capacity horac assay kit. In this study we determined the free radical scavenging activity, total phenolic content, total oxidant level, and total antioxidant level in methanol and acetone extracts of t. The calculated results with trolox standard curve, the teac value explain the antioxidant potential of the ge overhaul ble. Assay of nitric oxide scavenging activity nitric oxide scavenging activity can be estimated by the use of griess iliosvoy reaction garrat, 1964. Assessment of free radical scavenging and antiproliferative. Structureradical scavenging activity relationships of flavonoids dragan ami,a, duanka davidoviami,a drago belo,a and nenad trinajstib afaculty of agriculture, the josip juraj strossmayer university, p. The antioxidant activity increased with an increase in concentration of the extracts reaching a plateau. The antioxidant principles present in the extracts of k. The hydroxyl radicalscavenging activity was monitored using 2deoxyribose method of halliwell et al. As positive controls, epicatechin and lascorbic acid were also examined for dpph radical scavenging activity. In vitro antioxidant and free radical scavenging activity of different. Free radical scavenging activity, total phenolic content. Antioxidant capacity and radical scavenging effect of polyphenol. Box 719, hr31107 osijek, croatia bthe rugjer bokovi institute, p.
The maximum free radical scavenging activity and potency were interpolated from figure 1 to give results as shown in table 1. In vitro antioxidant and free radical scavenging activity of. However, 18 pure organic solvents replaced the sample solutions and they were not. 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. Determination of hydroxyl radical scavenging activity this was assayed as described by elizabeth and rao. In vitro nitric oxide scavenging activity of methanol. Hydroxyl radical scavenging assay of phenolics and flavonoids with a modified cupric reducing antioxidant capacity cuprac method using catalase for hydrogen peroxide degradation.
The objective of the present study were to determine the antioxidant activity, total phenolic content, reducing power activity, hydroxyl group reducing activity, estimation of ascorbic acid. Relative standard deviations n 3 of scavenging activity with 700. Mechanism of the oh radical scavenging activity of. Superoxide, hydroxyl radical, lipid peroxidation, nitric oxide assay showed a comparable scavenging activity compared to its standard. In dpph radical scavenging method the free radicals, 2, 2 diphenyl 1 picrylhydroazyl dpph was used to find antioxidant scavenging activity of. Hydroxyl radicalscavenging activity of 18 pure organic solvents the hydroxyl oh radicalscavenging activities of 18 pure organic solvents were determined, based on the original deoxyribose method, as described in the former section. The scavenging effect % was calculated by using the formulae given by duan et al. The extractives showed appreciable hydroxyl radical scavenging activity when compared with standard antioxidant, ca fig. What is the best method for radical scavenging assay. Hydroxyl radical scavenging activity of the extracts was determined according to the method reported by klein et al. Based on dpph and hydroxyl radical scavenging activity, tpl showed.
Analysis of radical scavenging active components in the. Hydroxyl radicals were produced by hydroxyl radical initiator h2o2 and fenton reagent, which quenches the fluorescent probe over time. 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. Flavonoids as potent scavengers of hydroxyl radicals. The total antioxidant activity, hydroxyl radical and hydrogen peroxide scavenging of a. Antioxidant capacity and radical scavenging effect of. Hydroxylation of ring a also enhances the activity, as does the presence of gallate and galactouronate moieties as substituents on the. Free radical scavenging activities of solutions of the plant extracts and synthetic antioxidant substances used in the study prepared in methanol at concentrations of 50, 100 and 200.
The hydroxyl radical scavenging activity of the extract is reported as % inhibition of deoxyribose degradation is calculated by using the following equation. In fba, c, bn and tea, the hydroxyl radical scavenging activity was decreased by over 50% by ascorbate oxidase treatment, indicating that ascorbate or ascorbatelike activity has a major effect in the hydroxyl scavenging of these preparations. A comparative study on the antioxidant activity of. Curcumin 38, a lipid soluble active principle of turmeric is a bis. Is there any easy method for the estimation of hydroxyl. The hydroxyl radical scavenging activity was monitored using 2deoxyribose method of halliwell et al. Thus, the hydroxyl radical scavenging activity of natural products is a crucial aspect of their cancerpreventive activity and hence, it is the emphasis in the evaluation of antioxidant activity. The ability of the extractives to quench hydroxyl radicals might directly relate to the prevention of lipid peroxidation. Solvent effects and improvements in the deoxyribose. Hydroxyl radical scavenging activity of microparticles.
The evaluation of scavenging activities in one sample n 3 was completed within 3. Department of anatomy, faculty of basic medical sciences, college of medicine, ekiti state university, adoekiti, nigeria phone. The super oxide anion scavenging activity was calculated according to the following equation. Purification and structural characterization of sulfated. Antioxidant activity of polyphenolic compounds isolated. Evaluation of hydroxyl radical scavenging activity and. The structural elements of the flavonoid molecule most important for hydroxyl radical scavenging are hydroxylation of ring b and a c2c3 double bond connected. The principle of the antioxidant assay is the formation.
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. The reaction mixture containing fecl3 100 m, edta 104 m, h2o2 1 mm and 2deoxy d. Evaluation of nitrite radical scavenging properties of. This was assayed as described by elizabeth and rao with a slight modification. To conclude, cpll showed a good antioxidant activity and might be an alternate to synthetic antioxidants available in the market. A comparative study on the antioxidant activity of methanolic. Leucas, antioxidant, protein extract, hydroxyl radical dpph. Antioxidant and free radical scavenging activity of.
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