Extraction and determination of antioxidant activity of. Antioxidant activity of extract and subfraction were measured using a series of wellestablished assays including the 2, 2diphenyl1picrylhydrazyl dpph, the reducing power, the nitric oxide. Relatively high levels of phenolics were observed in s. Pdf dpph free radical scavenging activity of some bangladeshi. Molyneux, the use of the stable free radical diphenylpicrylhydrazyl dpph for estimating antioxidant activity, songklanakarin journal of science and technology, vol. The measurement of the dpph radical scavenging activity was performed according to methodology described by brandwilliams et al. Dpph 1,1diphenyl2picrylhydrazyl analysis is one of the bestknown, accurate, and frequently employed methods for evaluating antioxidant activity.
It has also been used to quantify antioxidants in complex biological systems in recent years. The most correct name, which described the mechanism of the reaction, is the first one. An examination of table 4 reveals that the total antioxidant activity, measured by dpph method, ranged from 0. Among them, thyme and oregano exhibited the highest antioxidant activity, with i dpph values of 98. With contributions from worldclass experts in the field, the text presents the general mechanisms underlying the various assessments, the types of. This method was developed by blois with the viewpoint to determine the antioxidant activity in a like manner by using a stable free radical. Antiradical scavenging activity was tested by the dpph model table 5. Kinetics and mechanisms of antioxidant activity using the dpph. Study of antioxidant activity of pyrimidinium betaines by dpph. Genesis and development of dpph method of antioxidant assay.
The percentage of antioxidant activity aa% of each substance was assessed by dpph free radical assay. Kinetics and mechanisms of antioxidant activity using the. The results from the antioxidant assay showed that extract of all plants can scavenge the radical to a certain extent. Antioxidant compounds act through several chemical mechanisms. In dpph radical scavenging method the free radicals, 2, 2 diphenyl 1 picrylhydroazyl dpph was used to find antioxidant scavenging activity of. Raquibul hasan and mokarram hossain and raushanara akter and mariam jamila and md. Total phenolic content in tea samples were determined.
Dpph is a stable free radical with an absorption band at 515 nm. Antioxidant activity and free radical scavenging capacity. The free radical scavenger ability of antioxidants can be predicted from standard oneelectron potentials. Rphplc method was developed on a phenomenex lunac18. The principle of this assay is based on the reduction of dpph, a free stable radical by an antioxidant according to the following reaction15. Comparative analysis of the antioxidant activity of cassia. Free radical scavenging activity of crude extracts and 4 bioline. In this work, we reported that pm displayed antioxidant activities in a series of in vitro tests such as total antioxidant activity, determination of total phenolics contents tpc, ferric reducing antioxidant power assay frap, dpph 1, 1diphenyl2. The methanolic extract and its four water, ethyl acetate, chloroform, and nhexane fractions were prepared and subjected to antioxidant evaluation. In vitro antioxidant activity of coumarin compounds by. Comparison of dpph and abts assays for determining. Pdf dpph antioxidant activity, total phenolic and total. The substantial differences are in sample preparation.
The lipid peroxidation radical activity of the oils were similar to vitamin c, weak. Pdf paperbased dpph assay for antioxidant activity analysis. Pdf in this study antioxidant activity was performed by dpph 1, 1diphenyl2 picryl hydrazyl. All experiments were done in threeelectrode electrochemical cell at. Antioxidant activity can be evaluated as total free radicalscavenging capacity, by spectrophotometrically measuring the disappearance of the free 2,2diphenyl1picrylhydrazyl dpph radical. An antioxidant is a substance that at low concentrations delays or prevents oxidation of a substrate. One of the quick methods to evaluate antioxidant activity is the scavenging activity on dpph, a stable free radical and widely used index. It loses this absorption when reduced by an antioxidant or a free radical species.
Improved dpph determination for antioxidant activity spectrophotometric assay article pdf available in chemical papers slovak academy of sciences 6. Pdf antioxidant activity by dpph radical scavenging method of. In the dpph free radical scavenging activity, three coumarin compounds i, ii and iii were evaluated for their free radical scavenging activity with ascorbic acid as standard compound. Dpph method is widely used to determine antiradical antioxidant activity of purified phenolic compounds as well as natural plant extracts7. The assay is based on the measurement of the scavenging capacity of antioxidants towards it.
Conversely, the essential oil of anise in which the percentage of monoterpenes was as low as 2. Dpph free radical scavenging activity of the extracts of. Is it possible to use the dpph and abts methods for. The plant essential oil and methanol extract were also subjected to screenings for the evaluation of their antioxidant activities using 2, 2. The use of the stable free radical diphenylpicrylhydrazyl dpph for estimating antioxidant activity songklanakarin j.
The antioxidant activity of the different pomegranate juices was compared to those. Dpph with an odd electron delocalized over the molecule shows. Ascorbic acid is used as a standard compound to validate the analytic method and served as a reference antioxidant to measure the relative antioxidation. Negative impact of radicals on humans and animals is responsible for growing research interest in antioxidant properties of substances, which protect living organisms from the damaging influence of these reactive species. Rhesta aerial parts were extracted with hexane, ethyl acetate. A series of antioxidant concentrations was tested to determine linear response. Dpph has two major applications, both in laboratory research. Evaluation of the antioxidant activity of syzygium cumini. Pdf improved dpph determination for antioxidant activity. The importance of antioxidant mechanisms is to understand the biological meaning of antioxidants, their possible uses. It is a darkcolored crystalline powder composed of stable freeradical molecules. Detection and activity evaluation of radical scavenging. Hatbased methods measure the classical ability of an antioxidant to quench free radicals by hydrogen donation ah any h donor hence, many scientists feel these are most relevant to reactions where antioxidants typically act. The free radical scavenging capacity rsc of antioxidants from sesame cake extract was studied using the 2,2diphenyl1picrylhydrazyl radical dpph on a kinetic model.
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. Not only the name but the conditions of the described in the literature methods are very different. Measurement of antioxidant activity and capacity offers a muchneeded resource for assessing the antioxidant potential of food and includes proven approaches for creating healthy food products. Antioxidant activity was evaluated by free radical scavenging activity using 2,2diphenylpicryl1picrylhydrazyl dpph assay. Flavonoid content and antioxidant activity of vegetables.
No report on the antioxidant activities of pm was presently available. The antiradical activity of crude extracts 80% methanol, 20% water of s. It has been mentioned that antioxidant activity of plants might be due to their phenolic compounds cook and samman, 1996. Herein, we have demonstrated that excised leaf disc has. Kinetics and stoichiometry of reactions between the 2,2diphenyl1picrylhydrazyl dpph stable radical and 25 antioxidant compounds with different structure, molecular weight, number of. Antioxidant activity by dpph radical scavenging method of. All the essential oils showed antioxidant activity. It is a stable free radical because of its spare electron delocalization over the whole molecule. The following assay procedure was modified from those described by blois 1958 and yamasaki, et al. When the free radical is completely quenched, the color changes to yellow. The most used names of the method dpph are the free radical scavenging activity and antioxidant activity.
Caution dpph is a harmful radical species and all contact with skin or eyes should be avoided. Reevaluation of the 2,2diphenyl1picrylhydrazyl free. Standardized methods for the determination of antioxidant. For validation of this method several well known antioxidants ascorbic acid6palmitate, gallic, chlorogenic, ferulic, caffeic, uric, gentisic and vanillic acids, catechin. In the present study, antioxidant activities of the phenolic extracts from h. Principle of dpph radical scavenging capacity assay. Dpph is a common abbreviation for the organic chemical compound 2,2diphenyl1picrylhydrazyl. Pdf a novel amperometric method for antioxidant activity. Dpph and abts radical scavenging activities were discovered. 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 antioxidant activity of the aerial part extract of m. The aim of the present study was to determine the invitro antioxidant activity by dpph and abts free radical scavenging methods. Dpph free radical scavenging activity of two extracts from. Oh groups, and redox potential were investigated by recording the loss of dpph absorbance at 515 nm continuously for 10 min.
The samples were reacted with the stable dpph radical in an ethanol solution. Four methods were used to test the antioxidant activity of pomegranate juices including three based on the evaluation of the freeradical scavenging capacity of the juices, and one based on measuring their ironreducing capacity. Free radical scavenging behavior of antioxidant compounds. Estimation of phytochemical content and antioxidant. Use of a free radical method to evaluate antioxidant activity. Ehsanul hoque mazumder and shafiqur rahman, year2009. Dpph free radical scavenging activity of some bangladeshi. Leaf disc assays for rapid measurement of antioxidant activity. Dpph free radical scavenging activity of some bangladeshi medicinal plants.
Relevance and standardization of in vitro antioxidant. Invitro antioxidant and free radical scavenging activity. Antioxidant activity by dpph assay of potential solutions. Antioxidant and free radical scavenging capacity of seed and. Antioxidant compounds and their antioxidant mechanism. The dpph test has previously been successfully employed in assessing the antioxidant activity 14. Antioxidant and free radical scavenging activities of. Pdf hydromethanol extracts of 15 bangladeshi medicinal plants, traditionally used in different ailments, were evaluated for antioxidant potential. 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. Trolox equivalent antioxidant capacity teac, ferric reducing antioxidant power frap, and oxygen radical absorbance capacity orac assays were used less. Pure lignans and lignan glycosides isolated from methanolic extract by preparative hplc were used in the study.
Determination of dpph free radical scavenging activity by rp. Page 21 dpph free radical scavenging activity of some leafy vegetables used by tribals of odisha, india rajani kanta sahu 1, manoranjan kar 2, rasmirani routray 3, 1. The antioxidant activity of syzygium cumini leaf extracts was investigated using the 2,2diphenyl1picrylhydrazyl dpph free radicalscavenging and ferricreducing antioxidant power frap assays. This table indicates that the 2,2diphenyl1picrylhydrazylhydrate free radical assay method dpph assay is the most often used assay for estimation of antioxidant activity of sprouts. Antioxidant levels are key parameters for studies of food quality, stress responses, and plant health.
To understand the kinetic behavior better and to determine the rsc of sesame antioxidants, the secondorder. Among numerous methods for antioxidant activity estimation, dpph and abts are the most popular and commonly used ones due to their ease, speed, sensitivity and the. The crude aqueous extract of the plant contains the phenolics and flavonoids which are said to be the potent antioxidants 11. Dpph is widely used to test the ability of compounds to act as free radical scavengers or hydrogen donors, and to evaluate antioxidant activity of foods. While all extracts had antioxidant activity in the dpph assay, s. Antiradical activity was defined as the amount of anti. The dpph paperbased assay has also been developed to allow for fast screening of the radical scavenging activity of antioxidants 27, 28. A new method for the determination of antioxidant activity based on the amperometric reduction of 2,2diphenyl1picrylhydrazyl dpph at the glassy carbon electrode is proposed.