In the determination of total phenolics contents tpc and ferric. Determination of the effect of plant extract on 1,1diphenyl2picrylhydrazyl dpph radical. Abts and 2,2diphenyl1picrylhydrazyl dpph radical method. In addition, the same order as found in the present work was reported by dorman et al. Radical scavenging and antioxidant activities of methanolic. The samples were reacted with the stable dpph radical in an ethanol solution. The reaction mixture was vortexed thoroughly and left in the dark. Dependence of free radicals scavenging activity on concentration. The same ranking order for the free radical scavenging activity measured by the dpph test was found by kosar et al. The antiradical activity of crude extracts 80% methanol, 20% water of s. As seen from the graph 2, dpph free radicals scavenging activity of these plants equals to 843. Feb 25, 2011 dpph assay is considered a valid accurate, easy and economic method to evaluate radical scavenging activity of antioxidants, since the radical compound is stable and need not be generated.
Therefore, rate reduction of a chemical reaction upon addition of dpph is used as an indicator of the radical nature of that reaction. To show values directly dependent on antioxidant activity, antiradical activity ara was calculated as 1ic50. Good antioxidant activity for dpph radical scavenging activity ic 50 0. Antioxidant activity by dpph assay of potential solutions. Methanolic extracts of rice bran from four varieties. Dpph scavenging activity of outer bark extracts of f. The method is based on the spectrophotometric measurement of the dpph concentration. The uvvis absorption spectrum of kaempferol and dpph was investigated, the optimum determination wavelength and reaction time for determining the dpph radical scavenging activity of kaempferol was 517 nm and 30 min, respectively. Antioxidant activity by dpph assay of potential solutions to. Detection and activity evaluation of radical scavenging.
In the abts free radical scavenging assay, the nbuoh fraction displayed the highest total antioxidant activity 17. A change in colour from purple to yellow is observed. Total phenol and total flavonoid content in extracts, which reason antioxidant and free radical scavenging activity, was. In vitro antioxidant activity of coumarin compounds by dpph. Dpph is a deeppurple colored stable free radical, color of which changes from. Estimation of antioxidant activity a dpph radical scavenging activity. Lower absorbance of the reaction mixture indicated higher free radical activity. Free radical scavenging activity by tlc dpph method semiquantitative assay thin layer chromatography was used to determine the capacity of scavenging of the free radical dpph by the plant extracts as described by kwape and chaturvedi 2011. Determination of anti oxidant activity dpph radical scavenging activity the dpph assay method is based on the reduction of dpph, a stable free radical. Dpph scavenging effect % or percent inhibition a 0 a 1 a 0. Comparison of dpph and abts assays for determining. Dpph is a common abbreviation for the organic chemical compound 2,2diphenyl1picrylhydrazyl. The antioxidant activity of the memq was evaluated by the.
Dpph assay is considered a valid accurate, easy and economic method to evaluate radical scavenging activity of antioxidants, since the radical compound is stable and need not be generated. Preparation of dpph radical, and antioxidant scavenging assay dr jose m. Principle of dpph radical scavenging capacity assay. Genesis and development of dpph method of antioxidant assay. The method dpph is widely used for measurement of free radical scavenging ability of antioxidants perezjimenez and sauracalixto, 2008. The scavenging activity percentage aa% was determined according to mensor et al.
First of all, grids with 10mm spacing were made on a thin layer chromatography plate. Dpph is a wellknown radical and a trap scavenger for other radicals. Structureradical scavenging activity relationships of. Dpph radical scavenging activity % abs c o n t r o labs s a m p l e abs c o n t r o l, where abs control is the absorbance of dpph radical in methanol.
Structureradical scavenging activity relationships of flavonoids. For validation of this method several well known antioxidants ascorbic acid6palmitate, gallic, chlorogenic, ferulic, caffeic, uric, gentisic and vanillic acids, catechin. Among them, the 2,2diphenyl1picrylhydrazyl dpph spectrophotometric method is one of the most widely applied and is appreciated for its reliability. Dependence of free radical scavenging activity on concentration 0. This rdsc assay is easy to perform and has acceptable accuracy 90. May 05, 2010 the ability to scavenge dpph radical was calculated by the following equation. Dpph has two major applications, both in laboratory research. Free radical scavenging activity of ethanolic extracts from.
The scavenging activity is also studied using dpph. Antioxidant and free radical scavenging activities of. The ability to scavenge dpph radical was calculated by the following equation. Abs sample is the absorbance of dpph radical solution mixed with sample extract standard. Dpph radical scavenging activity pph radical is a stable organic free radical with adsorption band at 517 nm. It is a darkcolored crystalline powder composed of stable free radical molecules. Dpph free radical scavenging activity of the extracts of. These compounds have been described as chainbreaking antioxidants acting through radical scavenging activity, that is related to their hydrogen or electron donating capacity and to the ability to delocalizestabilize the resulting phenoxyl radical within their structure. Evaluation of the radical scavenging activity of a series of. A highthroughput relative 2,2diphenyl1picryhydrazyl dpph radical scavenging capacity rdsc assay was developed and validated in the present study. Free radical scavenging activity of crude extracts and 4.
Dpph inhibition in mpe was determined by using the protocol of brandwilliams et al. Dpph 1,1diphenyl2picrylhydrazyl is considered as a stable radical because. The method is based on the spectrophotometric measurement of the dpph concentration change resulting from the reaction with an antioxidant. In vitro nitric oxide scavenging activity of methanol. In vitro antioxidant and free radical scavenging activity 39 roots are the main portions of the whole plant as they possess wide number of the therapeutic agents. To compare the antioxdant activity we used abts 2,2azinobis 3ethylbenzothiazoline6sulfonic acid diammonium radical scavenging assay which is applicable for both lipophilic and hydrophilic antioxidants. Chrysin, rutin and quercetin were run to explore the effect of. The antiradical activity of caffeic acid 1, dihydrocaffeic acid 5, and their corresponding nalkyl esters was evaluated by using the 2,2diphenyl1picrylhydrazyl radical dpph method.
Highthroughput relative dpph radical scavenging capacity. Dpph is converted to 1, 1diphenyl 2picryl hydrazine when it reacts with antioxidants. Characterization and dpph radical scavenging activity of. Dpph free radical scavenging assay free radical scavenging activity of the cell free extract was measured using the procedure described by 21. Prieto this is an assay for scavenging activity against free radicals. The antioxidant activity of the aerial part extract of m. Several methods have been developed to assess the radical scavenging activity. Evaluation of dpph free radical scavenging activity of. The methods for preparing each reagent were detailed in the analytical procedures. The result showed that the antioxidant activity in unheated oils are more compared to heated oils using abts radical scavenging activity ic 50 24. The dpph radical himedia is stable due to the delocalization of a spare electron over the molecule, thus preventing dimer formation. Based on dpph and hydroxyl radical scavenging activity, tpl showed strong scavenging activity 91.
The crude aqueous extract of the plant contains the phenolics and flavonoids which are said to be the potent antioxidants 11. Dpph free radical scavenging activity of the extracts of the. Aliquots of extracts were made to total volume of 3ml using methanol. Antioxidant activity by dpph radical scavenging method of. The free radical scavenger ability of antioxidants can be predicted from standard oneelectron potentials. Determination of free radical scavenging activity of plant.
Scavenging activity on dpph radical quantitative fig. Selective abts and dpph radical scavenging activity of. The control solution was prepared by mixing ethanol 3. Dpph radical scavenging activity of kaempferol scientific. This method was developed by blois with the viewpoint to determine the antioxidant activity in a like manner by using a stable free radical. Dpph scavenging activity was assessed using the method of hatano et al. Scavenging activity of hypericum extracts against dpph radical was assessed according to the method of blois with some modifications.
Statistical analysis the experiment was done in triplicate for each substance. Antioxidant capacity and radical scavenging effect of. The assay is based on the measurement of the scavenging capacity of antioxidants towards it. Invitro antioxidant and free radical scavenging activity of. These results suggest that tpl had the highest radical. Dpph radical scavenging assay the radicalscavenging activity rsa % of the chalcones 116 was determined using the dpph radical in ethanol 0. Dpph radical scavenging assay an overview sciencedirect.
Application of free radical diphenylpicrylhydrazyl dpph to. The 2,2diphenylpicrylhydrazyl dpph assay is widely used in plant biochemistry to evaluate the properties of plant constituents for scavenging free radicals. Applicability of the dpph assay for evaluating the. We show that increased cat, apx, gr, and dpphradical scavenging activity and protection of cat activity during chilling are correlated with heat shock. Synthesis of salidroside analogues and their ability of dpph. The odd electron of nitrogen atom in dpph is reduced by receiving a hydrogen. Vasumathi, yamini, jyothi and njavara were used to study their total phenolic and flavonoid contents, in vitro antioxidant activities including total antioxidant activity, scavenging of nitric oxide and 1,1diphenyl2picrylhydrazyl dpph radical, reducing power and cytotoxic. 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. For determination of radical scavenging activity of different foods, beverages and substrates were elaborated a. Dpph free radical scavenging capacity of legume extracts was evaluated according to the method of chen and ho 11 as modified by xu and chang 10.
Antioxidant enzymes and dpphradical scavenging activity in. Dpph, known formally as 2,2diphenyl1picrylhydrazyl, is a cellpermeable, stable free radical that is commonly used to evaluate the ability of compounds to act as free radical scavengers or hydrogen donors and to measure the antioxidant activity of tissue extracts. Available on line journal of chemical and pharmaceutical research. In vitro nitric oxide scavenging activity of methanol extracts of three bangladeshi medicinal plants rozina parul1, sukalayan kumar kundu 2 and pijush saha2 1. Dpph free radical, was calculated using log dose inhibition curve. Available on line journal of chemical and pharmaceutical. Dpph radical scavenging capacity of phenolic extracts from. Dpph free radical scavenging activity of methanolic extracts of magnolia coco flowers the dpph radical scavenging assay is a convenient and fast technique to evaluate antioxidative activity 10. In order to obtain information about the real antioxidant activity with respect to lipids or food stabilization, it is. It loses this adsorption when accepting an electron or a free radical species, which results in a visually noticeable discoloration from purple to yellow 9. As positive controls, epicatechin and lascorbic acid were also examined for dpph radical scavenging activity. Dr prietos dpph microplate protocol 020712 procedure. In vitro antioxidant and free radical scavenging activity of.
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