Dpph free radical scavenging activity of two extracts from. Dodecane is a preferred solvent for the extraction of hydrophobic compounds from live cultures due to its low toxicity and good phase separation2022. First of all, grids with 10mm spacing were made on a thin layer chromatography plate. Determination of total phenolic, flavonoid content and. Free radical scavenging ability of the extracts was tested by dpph radical scavenging assay as described by blois and desmarchelier et al. Antioxidant activity by dpph assay of potential solutions. Phytochemical screening, chemical composition and antioxidant activity of spearmint aerial parts methanolic. Dpph is a stable free radical in a methanolic solution. The dpph assay provides an easy and rapid way to evaluate potential antioxidants. Evaluation of free radical scavenging activity of butea monosperma lam. Pdf the crude protein hydrolysates from aqueous extract of velvet antler aeva were prepared by simulated gastrointestinal digestion sgi. Antiradical scavenging activity was tested by the dpph model table 5.
Dpph scavenging activity of outer bark extracts of f. Abts free radical scavenging assay, determination of total phenolics contents tpc, ferric reducing antioxidant power assay frap, rapid screening of antioxidant by dotblot dpph 1, 1diphenyl2picrylhydrazyl staining, dpph radical. Scavenging activity dpph assay the free radical scavenging activities of the extracts were determined by using 2, 2 diphenyl1picrylhydrazyl dpph free radical scavenging method 10. A highthroughput relative 2,2diphenyl1picryhydrazyl dpph radical scavenging capacity rdsc assay was developed and validated in the present study. A number of protocols have been followed for this assay resulting in variation in the results of different laboratories.
The odd electron of nitrogen atom in dpph is reduced by receiving a hydrogen. Au515 a0a1 a0ka1k where au515 is the antiradical activity of the extract, a0 the absorbance of the sample at the beginning of the reaction 0 min, a1 the absorbance of the sample after incubation times 20120 sec of the reaction. Among them, the 2,2diphenyl1picrylhydrazyl dpph spectrophotometric method is one of the most widely applied and is appreciated for its reliability. This assay provides useful information on the reactivity of the compounds with the stable free radical, because of the odd number of electrons. Sem suggests that gallic acid was dispersed into lecithin while dsc and xrd results for the complex mainly showed the. Inhibitory effect of dpph radical scavenging activity and. Dpph radical scavenging activity the free radical scavenging activity of the extracts of a. Antioxidant activity, nitric oxide scavenging activity and phenolic. Dpph free radical scavenging activity of the extracts of. Dpph 1,1diphenyl2picrylhydrazyl radical scavenging. The ethanolic extract exhibited higher free radical scavenging effect than the water extract at all tested concentrations.
Antioxidant activity and total phenolic compound content. The measurement of the dpph radical scavenging activity was performed according to methodology described by brandwilliams et al. All the essential oils showed antioxidant activity. Characterization of multifunctional, biodegradable sodium. Among them, thyme and oregano exhibited the highest antioxidant activity, with i dpph values of 98. Free radical scavenging activity of crude extracts and 4. Dpph is a stable free radical at room temperature which accepts an electron or hydrogen radical to form a stable diamagnetic molecule.
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. Structureradical scavenging activity relationships of. Several studies have revealed that a major part of the antioxidant activity may be from. This assay uses this character to show herbs free radical scavenging activity.
Applicability of the dpph assay for evaluating the antioxidant. 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. Box 719, hr31107 osijek, croatia bthe rugjer bokovi institute, p. The free radical of dpph has an odd electron, which gives a maximum absorption at 517 nm purple colour. The assay was carried out in buffered medium methanol. Keywords dpph assay, interlaboratory study, antioxidant, food additive. In vitro antioxidant and free radical scavenging activity. Several methods have been developed to assess the radical scavenging activity. In conclusion, the antioxidant assay based on scavenging of dpph radical at a dpph concentration of 50 lm in methanol or buffered methanol, depending upon the solubility of the compound under investigation, is. Standardized methods for the determination of antioxidant. This method was developed by blois with the viewpoint to determine the antioxidant activity in a like manner by using a stable free radical. Scavenging of dpph free radical is the basis of a common antioxidant assay. The assay is based on the measurement of the scavenging capacity of antioxidants towards it.
Dpph radical scavenging assay an overview sciencedirect topics. Comparison of dpph and abts assays for determining. Dpph radical scavenging activity was measured using a film extract solution according to the method described by lee, yang, lee, and song. Mpe exhibits significant strong scavenging activity on dpph and abts assay. Invitro antioxidant and free radical scavenging activity. A perusal of the publications in the recent past table 1 shows that various research groups have used widely different protocols which differed in. In 96wells plate, the test samples were allowed to react w ith 2,2di4 tertoctylphenyl1picrylhydrazyl stable free radical dpph.
The principle of this assay is based on the reduction of dpph, a free stable radical by an antioxidant according to the following reaction15. We optimized the dpph assay for use with dodecane, as previouslypublished dpph radicalscavenging assays used methanol or ethanol as the solvent. In the antiradical scavenging property test the extract showed at 64. Dpph rapid assay highthroughput assay scavenging capacity index abstract a new microplateadapted dpph rapid assay was developed to assess the antioxidant capacity of pure compounds and foods. Superoxide, hydroxyl radical, lipid peroxidation, nitric oxide assay showed a. Thereafter, the absorbance of the assay mixture was measured at. Dpph wako pure chemical industries, osaka, japan of the same lot was distributed to the participating laboratories. The percentage of antioxidant activity aa% of each substance was assessed by dpph free radical assay. Box 180, hr2 zagreb, croatia received march 26, 2002. Free radical scavenging capacity and antioxidant activity of. An online nphplcdpph method for the determination of. Study of antioxidant activity of pyrimidinium betaines by.
Dpph radical scavenging capacity of phenolic extracts from. Hence, it is commonly used in dpph assay for measuring the antioxidant activity of different natural samples such as wine, fruits, herbal tea etc. 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. Determination of dpph radicals scavenging activity was estimated with the method used by kato 5. Dpph radical caused by antioxidants is due to the reaction between antioxidant molecules and radical. Conversely, the essential oil of anise in which the percentage of monoterpenes was as low as 2.
Dpph has two major applications, both in laboratory research. The dpph assay method is based on the reduction of dpph, a stable free radical. In its oxidized form, the dpph radical has an absorbance maximum centered at about 520 nm molyneux, 2004. Flavonoids are reported to exhibit various biological activities, including antioxidative and free radical scavenging activities. Dpph in oxidized form gives a deep violet color in methanol. Department of agriculture, arkansas childrens nutrition center, 1120 marshall street. Materials and methods dpph free radical scavenging activity processing of plants for extract preparation.
Dpph 1,1diphenyl2picrylhydrazyl is considered as a stable radical because. The samples were reacted with the stable dpph radical in an ethanol solution. Genesis and development of dpph method of antioxidant assay. Available on line journal of chemical and pharmaceutical. Ocimum gratissimum extract, antioxidant activity, no. Evaluation of the methods for determination of the free radical scavenging activity by dpph 11 bulgarian journal of agricultural science, 17 no 1 2011, 1124. The dpph radical scavenging assay was determined according to the method of shimada et al. Freshly prepared dpph solution was taken in test tubes and extracts were added followed by serial dilutions 15. Dpph is listed in the worlds largest and most authoritative dictionary database of abbreviations and acronyms. It is a darkcolored crystalline powder composed of stable freeradical molecules.
Dpph radical scavenging assay in this study, the dpph assay was conducted according to the following procedure. Dpph radical scavenging assay the free radical scavenging capacity of the extracts was determined using dpph 8. Dpph scavenging assay of eighty four bangladeshi medicinal. Calibration curve was prepared by adding 0, 1, 2, 5, 8 and 10 ml of the ascorbic acid stock solution into 100 ml volumetric flasks, and then dilute to volume with water. 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. The uv and ir spectra of the complex showed an additive effect of gallic acid and lecithin, in which the characteristic absorption peaks of gallic acid and lecithin were retained. Absorbance of the resulting solution was measured spectrophotometrically at 517nm. Pdf genesis and development of dpph method of antioxidant assay.
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. Smpcl films 25 mg were cut into small pieces and dissolved in 5 ml distilled water with shaking at 20 c for 30 min. 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. The free radical scavenging activity of the extracts, based on the scavenging activity of the stable 1, 1 diphenyl2 picryl hydrazyl dpph free radical was determined by the method described7. Evaluation of free radical scavenging activity of butea. Scavenging of dpph radical is the basis of the popular dpph antioxidant assay alma et al. This rdsc assay is easy to perform and has acceptable accuracy 90. The radical scavenging activity of films was determined using a dpph and abts assay. Oxide scavenging methods using uv vis spectrophotometer were employed. Dpphfree radical scavenging capacity of legume extracts was evaluated according to the method of chen and ho 11 as modified by xu and chang 10.
The hydrogen atom donating ability of the plant extractives was determined by the decolorization of methanol solution of 2,2diphenyl1picrylhydrazyl dpph. Dpph is a common abbreviation for the organic chemical compound 2,2diphenyl1picrylhydrazyl. Dpph scavenging activity was assessed using the method of hatano et al. Applicability of the dpph assay for evaluating the. Extraction and determination of antioxidant activity of. Antioxidant and cytotoxic activities of sterculia setigera. Pdf antioxidant activity of protein hydrolysates from. Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements ronald l. Reducing power showed dose dependent increase in concentration absorption compared to standard, quercetin. Recent automated versions combine the dpph test with an hlpc assay.
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