Aliquots of extracts were made to total volume of 3ml using methanol. This method was developed by blois with the viewpoint to determine the antioxidant activity in a like manner by using a stable free radical. A change in colour from purple to yellow is observed. Among them, the 2,2diphenyl1picrylhydrazyl dpph spectrophotometric method is one of the most widely applied and is appreciated for its reliability. The derivatives showed reasonable free radical scavenging activities. The antioxidant activity of the extract was measured with the dpph method 18 with slight modifications. Dpph radical scavenging activity was expressed as milligrams of trolox equivalents mg teg sample. Estimation of antioxidant activity a dpph radical scavenging activity. 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.
These results suggest that tpl had the highest radical. Free radical scavenging and antibacterial activities of helichrysum caespititium dc harv. Kinetics and stoichiometry of reactions between the 2,2diphenyl1picrylhydrazyl dpph stable radical and 25 antioxidant compounds with. The measurement of the dpph radical scavenging activity was performed according to methodology described by brandwilliams et al. Free radical scavenging activity of crude extracts and 4. Antioxidant activity by dpph radical scavenging method of. 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. Evaluation of the antioxidant capacity of food samples. The method dpph is widely used for measurement of free radical scavenging ability of antioxidants perezjimenez and sauracalixto, 2008. Evaluation of the freeradical scavenging and antioxidant. A simple and a reliable method to quantify antioxidant activity in vivo.
Dpph assay for antioxidant activity acs publications american. Antioxidant activity was determined by dpph radical scavenging and reducing power assays. The result showed that the antioxidant activity in unheated oils are more compared to heated oils using abts radical scavenging activity ic 50 24. Free radical scavenging capacity and antioxidant activity. Comparison of dpph and abts assays for determining. Highthroughput relative dpph radical scavenging capacity assay. In the dpph radical scavenging assay, the activity of the positive control, ascorbic acid, was the highest.
Synthesis of salidroside analogues and their ability of. Assessment of free radical scavenging potential and. The total phenolic content of the extracts was found to be higher in a. Antioxidant activity by dpph assay of potential solutions. Book of abstracts of the institute of food technologists annual meeting. The highest dpph radical scavenging activity was detected in the methanolic extract of dried sample with 87.
Applicability of the dpph assay for evaluating the. In vitro antioxidant assay dpph radical scavenging assay. Detection and activity evaluation of radical scavenging. The 2,2diphenylpicrylhydrazyl dpph assay is widely used in plant biochemistry to evaluate the properties of plant constituents for scavenging free radicals. The thin layer coated on the sugar crystal was extracted by soxhlet extractor with dichloromethane. Invitro antioxidant and free radical scavenging activity. Determination of the effect of plant extract on 1,1diphenyl2picrylhydrazyl dpph radical. In the present study, the high dpph radical scavenging activity of the. Antioxidant capacity and radical scavenging effect of polyphenol. Issn phytochemical analysis and free radical scavenging.
The samples were reacted with the stable dpph radical in an ethanol solution. Determination of free radical scavenging activity of plant. Dr prietos dpph microplate protocol 020712 procedure. Antioxidant capacity and radical scavenging effect of. For validation of this method several well known antioxidants ascorbic acid6palmitate, gallic, chlorogenic, ferulic, caffeic, uric, gentisic and vanillic acids, catechin. The generation of pure hydroxyl radicals under the experimental conditions was evaluated and confirmed using electron spin. 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. All the synthetic derivatives 125 were submitted to in vitro dpph radical scavenging activity as per literature protocol 19, 20. Dpph scavenging activity was assessed using the method of hatano et al. Dpph 1,1diphenyl2picrylhydrazyl is considered as a stable radical because. Assessment of antioxidant activity of cane brown sugars by.
Structureradical scavenging activity relationships of. Dpph is converted to 1, 1diphenyl 2picryl hydrazine when it reacts with antioxidants. As positive controls, epicatechin and lascorbic acid were also examined for dpph radical scavenging activity. Superoxide, hydroxyl radical, lipid peroxidation, nitric oxide assay showed a. Bioactive constituents, in vitro radical scavenging and. Several methods have been developed to assess the radical scavenging activity. 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. Application of free radical diphenylpicrylhydrazyl dpph. Determination of free radical scavenging activity from. Dpph scavenging activity of outer bark extracts of f. 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. Synthesis of benzophenone hydrazone analogs and their dpph. Principle of dpph radical scavenging capacity assay.
Mpe exhibits significant strong scavenging activity on dpph and abts assay. Dpph 1,1diphenyl2picrylhydrazyl is a stable free radical. Dpph inhibition in mpe was determined by using the protocol of brandwilliams et al. 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. The methods for preparing each reagent were detailed in the analytical procedures. Dpph free radical scavenging assay free radical scavenging activity of the cell free extract was measured using the procedure described by 21. Science and technology, general antibacterial agents rites, ceremonies and celebrations antioxidants antioxidants nutrients biphenyl compound chlorinated solvents herbal medicine hydrogen peroxide medicinal plants. Free radical scavenging activity of ethanolic extracts from herbs and. The scavenging activity sa of extracts was defined as.
Dpph assay was performed by spectrophotometry as previously reported chua et al. Dpph free radical scavenging activity of the extracts of. Box 180, hr2 zagreb, croatia received march 26, 2002. Dpph radical scavenging assay an overview sciencedirect topics. The antiradical activity of crude extracts 80% methanol, 20% water of s. Phytochemical screening and free radical scavenging. Box 719, hr31107 osijek, croatia bthe rugjer bokovi institute, p. Antioxidant and tyrosinase inhibitory in vitro activity. Abts and 2,2diphenyl1picrylhydrazyl dpph radical method. 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. First of all, grids with 10mm spacing were made on a thin layer chromatography plate.
Methanolic extracts from the hypericum species were analyzed for radical scavenging and antioxidant activities using dpph, abts free radicals, total antioxidant activity and inhibition of lipid peroxidation by ferric. Preparation of dpph radical, and antioxidant scavenging assay dr jose m. Dependence of free radicals scavenging activity on concentration. Scavenging activity on dpph radical quantitative fig. Antioxidants scavenge these free radicals, thereby protecting the cell from damage. As seen from the graph 2, dpph free radicals scavenging activity of these plants equals to 843. The dpph radical himedia is stable due to the delocalization of a spare electron over the molecule, thus preventing dimer formation. In order to obtain information about the real antioxidant activity with respect to lipids or food stabilization, it is. Selective abts and dpph radical scavenging activity of. 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. Free radical scavenging and antibacterial activities of. Good antioxidant activity for dpph radical scavenging activity ic 50 0.
Kaempferol exhibited strong dpph radical scavenging activity with a ic50 value of 0. Evaluation of the radical scavenging activity of a series. A novel fluorometric method was developed and validated for hydroxyl radical scavenging capacity hosc estimation using fluorescein as the probe. Based on dpph and hydroxyl radical scavenging activity, tpl showed strong scavenging activity 91. Characterization and dpph radical scavenging activity of. To show values directly dependent on antioxidant activity, antiradical activity ara was calculated as 1ic50. Dpph radical scavenging assay the radicalscavenging activity rsa % of the chalcones 116 was determined using the dpph radical in ethanol 0. In vitro antioxidant and free radical scavenging activity. The odd electron of nitrogen atom in dpph is reduced by receiving a hydrogen. Antioxidant and free radical scavenging activity of.
The results suggest that mentha spicata has promising antioxidant activity and could serve as potential source of natural. The method is based on the spectrophotometric measurement of the dpph concentration change resulting from the reaction with an antioxidant. Available on line journal of chemical and pharmaceutical. The crude extracts were diluted using ethanol according to the assay needs. The antioxidant activity of the aerial part extract of m. Dpph wako pure chemical industries, osaka, japan of the same lot was distributed to the participating laboratories. The scavenging activity is also studied using dpph. The crude aqueous extract of the plant contains the phenolics and flavonoids which are said to be the potent antioxidants 11. Prieto this is an assay for scavenging activity against free radicals. The present study was designed to examine the free radical scavenging potential and oxidative dna damage. Dpph radical scavenging assay in this study, the dpph assay was conducted according to the following procedure. The dpph radical scavenging activity of the chloroform extract of sida acuta was analyzed by the standard method. The dpph method is described as a simple, rapid and convenient method independent of sample polarity for screening of many samples for radical scavenging activity koleva et al.
Dependence of free radical scavenging activity on concentration 0. Dpph, no, h 2 o 2, and o 2radicals inhibition percentages were measured to assay the antiradical activity of extracts table 2. Thirteen hypericum species growing in bulgaria were investigated for free radicalscavenging activity, antioxidant activity, total tannins and total flavonoids contents. Genesis and development of dpph method of antioxidant assay. For determination of radical scavenging activity of different foods, beverages and substrates were elaborated a. The assay is based on the measurement of the scavenging capacity of antioxidants towards it. The characterization of compounds with antioxidant activity is of great interest due.
The antioxidant and free radical scavenging activity of the ethanol extracts of melia dubia hiern. Oxidation of biomolecules such as carbohydrates, proteins, lipids, and nucleic acids results in generation of free radicals in an organism which is the major cause of onset of various degenerative diseases. Carvacrol, pcymene, linalool, and borneol in oil from the errich population. Dpph is a stable free radical in a methanolic solution. Analysis of in vitro radical scavenging activity dpph radical scavenging activity. Novel fluorometric assay for hydroxyl radical scavenging. Dpph radical scavenging capacity of phenolic extracts from. 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. Dpph radical scavenging activity pph radical is a stable organic free radical with adsorption band at 517 nm.
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