Preview

Dpph Method

Powerful Essays
Open Document
Open Document
4901 Words
Grammar
Grammar
Plagiarism
Plagiarism
Writing
Writing
Score
Score
Dpph Method
ORIGINAL ARTICLE

The use of the stable free radical diphenylpicrylhydrazyl (DPPH) for estimating antioxidant activity
Philip Molyneux

Abstract
Molyneux, P.

The use of the stable free radical diphenylpicrylhydrazyl (DPPH) for estimating antioxidant activity
Songklanakarin J. Sci. Technol., 2004, 26(2) : 211-219
The use of the stable free radical diphenylpicrylhydrazyl (DPPH) to estimate the activity of antioxidants is reviewed. Current applications of the method are examined, particularly the use of the parameter EC50 (substrate concentration to produce 50% reduction of the DPPH). Some recommendations are made as to the most suitable ways of carrying out this assay and evaluating the data produced.

Key words : DPPH, diphenylpicrylhydrazyl, free radical, antioxidant activity
Ph.D.(Polymer Chemistry), Macrophile Associates, 9 Brewery Lane, Salisbury, Wiltshire, SP1 2LJ, U.K. E-mail: molyneux@easynet.co.uk Received, 8 June 2003 Accepted, 15 December 2003

Songklanakarin J. Sci. Technol. Vol. 26 No. 2 Mar.-Apr. 2004

212

Use of DPPH to estimate antioxidant activity Molyneux, P.

There is an increasing interest in antioxidants, particularly in those intended to prevent the presumed deleterious effects of free radicals in the human body, and to prevent the deterioration of fats and other constituents of foodstuffs. In both cases, there is a preference for antioxidants from natural rather than from synthetic sources (Abdalla and Roozen, 1999). There is therefore a parallel increase in the use of methods for estimating the efficiency of such substances as antioxidants ′ (S a nchez-Moreno, 2002; Schwarz, et al., 2001). One such method that is currently popular is based upon the use of the stable free radical diphenylpicrylhydrazyl (DPPH). The purpose of this paper is to examine the basis of this method, and to further examine the use of the parameter “EC50” (equivalent concentration to give 50% effect) which is currently used in the interpretation



References: Abdalla, A.E. and Roozen, J.P. 1999. Effect of plant extracts on the oxidative stability of sunflower oil and emulsion, Food Chemistry, 64: 323-329. Blois, M.S. 1958. Antioxidant determinations by the use of a stable free radical, Nature, 181: 11991200. Bondet, V., Brand-Williams, W. and Berset, C. 1997. Kinetics and mechanisms of antioxidant activity using the DPPH• free radical method, Lebensmittel-Wissenschaft und -Technologie/Food Science and Technology, 30: 609-615. Brand-Williams, W., Cuvelier, M.E. and Berset, C. 1995. Use of a free radical method to evaluate antioxidant activity, Lebensmittel-Wissenschaft und -Technologie/Food Science and Technology, 28: 25-30. Songklanakarin J. Sci. Technol. Vol. 26 No. 2 Mar.-Apr. 2004 ′ 219 Use of DPPH to estimate antioxidant activity Molyneux, P. G O mez-Alonso, S., Fregapane, G., Salvador, M.D. and Gordon, M.H. 2003. Changes in phenolic composition and antioxidant activity of virgin olive oil during frying, J. Agric. Food Chem., 51: 667672. Guo, J.-T., Lee, H.-L., Chiang, S.-H., Lin, F.I. and Chang, C.-Y. 2001. Antioxidant properties of the extracts from different parts of broccoli in Taiwan, J. Food Drug Anal., 9(2): 96-101. Kim, J.-K., Noh, J.H., Lee, S., Choi, J.S., Suh, H., Chung, H.Y., Song, Y.-O. and Choi, W.C. 2002. The first total synthesis of 2,3,6-tribromo-4,5dihydroxybenzyl methyl ether (TDB) and its antioxidant activity, Bull. Korean Chem. Soc., 23(5): 661-662. Lebeau, J., Furman, C., Bernier, J.-L., Duriez, P., Teissier, E. and Cotelle, N. 2000. Antioxidant properties of di-tert-butylhydroxylated flavonoids, Free Radic. Biol. Med., 29(9): 900-912. ˜ o, G.G., Leit a o, S.G. and Vilegas, W. 2002. ˜ Leit a Quick preparative separation of natural naphthoquinones with antioxidant activity by high-speed counter-current chromatography, Z.Naturforsch., 57c: 1051-1055. Lu, Y. and Foo, L.Y. 2000. Antioxidant and radical scavenging activities of polyphenols from apple pomace, Food Chemistry, 68: 81-85. ′ S a nchez-Moreno, C. 2002. Review: methods used to evaluate the free radical scavenging activity in foods and biological systems, Food Sci. Tech. Int., 8(3): 121-137. ′ S a nchez-Moreno, C., Larrauri, J.A. and Saura-Calixto, F. 1998. New parameter for evaluation of free radical scavenging capacity of polyphenols, 2nd International Electronic Conference on Synthetic Organic Chemistry (ESCOC-2), http://www. mdpi.org/escoc/, September 1-30, 1998 [dp130]; http://ecsoc2.hcc.ru/DP_TOP1/dp130/ dp130. htm. ′ S a nchez-Moreno, C., Larrauri, J.A. and Saura-Calixto, F. 1999. Free radical scavenging capacity and inhibition of lipid oxidation of wines, grape juices and related polyphenolic constituents, Food Res. Int., 32: 407-412. Schwarz, K., Bertelsen, G., Nissen, L.R., Gardner, P.T., Heinonen, M.I., Hopia, A., Huynh-Ba, T., Lambelet, P., McPhail, D., Skibsted, L.H. and Tijburg, L. 2001. Investigation of plant extracts for the protection of processed foods against lipid oxidation. Comparison of antioxidant assays based on radical scavenging, lipid oxidation and analysis of the principal antioxidant compounds, Eur. Food Res. Technol., 212: 319-328. Wanjek, C. 2001. Mixed messages: antioxidants may in some cases do more harm than good. The Washington Post, 7 Aug, p HE01. ′ ˜ Yepez, B., Espinosa, M., L O pez, S. and Bola n os, G. 2002. Producing antioxidant fractions from herbaceous matrices by supercritical fluid extraction, Fluid Phase Equil., 194-197: 879-884. Zhu, Q.Y., Hackman, R.M., Ensunsa, J.L., Holt, R.R. and Keen, C.L. 2002. Antioxidative activities of oolong tea, J. Agric. Food Chem., 50: 6929-6934.

You May Also Find These Documents Helpful

  • Better Essays

    Abstract The structure of a major component of star anise oil was derived by analyzing its IR spectroscopy melting point of after oxidation and purification. From the evidence obtained, it was concluded that the exact structured should be p-methoxybenzoic acid. Introduction The purpose of this experiment is to determine the structure of the major component of star anise oil. It is already known that this component has a molecular formula C10H12O. This molecular formula equates to an IHD of 5. It is evident, by calculating the IHD, that this compound contains a benzene ring. By heating anisene with hydroiodic acid the following reaction occurs: [rxn 1] The resultant of a phenol indicates that anisene is, in fact, an aryl methyl ether with the formula C9H9OCH3. Since all these facts have been established about the structure of anisene, what is to be determined from this experiment is the structure of the unsaturated side chain and its location. In order to do this, anisene will first be oxidized with KMNO4 so as to then identify the oxidation product and establish the positioning of the side chain. The reaction is as follows: Finally purification of the structure of the unknown oxidized component, its structure shall be determined by obtaining the melting point and by means of IR spectroscopy of the original substance. Experimental The experiment was carried out following the procedure described in Experiment 24 of the assigned lab book (Lehman 188-195) with some deviation: When it was time to use the condenser the first time around, the tubes for the water to circulate were not attached, causing the solvent to evaporate. Consequently, more solvent had to be added and the experiment continued normally. Calculations The weight of the total final product is equal to the weight of the watch glass and the final product minus the weight of the watch glass. Wt of Final product = (wt of watch glass and product)-(wt…

    • 1271 Words
    • 6 Pages
    Better Essays
  • Satisfactory Essays

    Extraction from nutmeg

    • 341 Words
    • 2 Pages

    PURPOSE: The purpose of this experiment was to illustrate the extraction of a solid natural product from its natural source by partition with the aid of an organic solvent. Extraction is the process by which a compound or mixtures of, is transferred by separation from one phase into another. In this case, the major triglyceride contains a single fatty acid identified as myristic acid. It is a substituent of the triglyceride called trimyristin, which embodies 20-25% of the dried weight of ground nutmeg we will eventually use in this experiment. The purpose of this lab is to extract trimyristin from nutmeg with the organic substance, diethyl ether, to evidently produce trimyristin with a small portion of myristicin (solid-liquid extraction). Also, it is good to know that due to impurity the best way purify the product of trimyristin is by recrystallization through hot acetone.…

    • 341 Words
    • 2 Pages
    Satisfactory Essays
  • Good Essays

    Antioxidants can neutralize free radicals. Antioxidants acts as scavengers looking for free radicals, helping to prevent…

    • 1036 Words
    • 5 Pages
    Good Essays
  • Good Essays

    Due to the necessity of keeping the food's natural characteristics and appearance for longer, to meet the requirements of consumers, and because of the short life that natural food have, certain chemical compound have discovered and developed certain ways to preserve food and extend their life for longer. Furthermore, there are many different types of chemical compounds were added to our daily diet, yet certain chemical compounds can be listed on the nutrition label, and even they cannot be tested or seen on the products. Butylated hydroxyanisole and Butylated hydroxytoluene and Ethanol are among of that chemical compounds, which were used in certain food and beverages in order to preserve them. The aim of this essay is to discuss the use of BHA (butylated hydroxyanisole), BHT (butylated hydroxytoluene) and Ethanol in the food industry, also this essay will discuss and define the side effects of them on our daily diet.…

    • 784 Words
    • 4 Pages
    Good Essays
  • Better Essays

    Cited: "Antioxidants: MedlinePlus." U.S National Library of Medicine. U.S. National Library of Medicine. Web. 30 Oct. 2013.…

    • 946 Words
    • 4 Pages
    Better Essays
  • Powerful Essays

    Free Radicals

    • 1078 Words
    • 5 Pages

    I had previously been looking into the effects of free radicals on the body, and the types of supplements that would work to inactivate these dangerous toxins. Webster’s Dictionary defines a free radical as, “An atom or group of atoms that has at least one unpaired electron, and is therefore unstable and highly reactive. In animal tissues, free radicals can damage cells and are believed to accelerate the progression of cancer, cardiovascular disease, and age-related diseases” (Merriam-Webster, 2010). In this paper, I will discuss: exposure to free radicals, their effects on the body, “The Free Radical Theory”, both supplements and foods which will help to inactivate free radicals, and finally; my take on the dangerous effects of free radicals as they pertain to the elderly population.…

    • 1078 Words
    • 5 Pages
    Powerful Essays
  • Good Essays

    It has been estimated that the average person has around 10,000–20,000 free radicals attacking each body cell each day. Although free radicals appear to be the main factor in many diseases, cells have evolved defense mechanisms for protection against ROS -mediated oxidative damage by producing antioxidants, which are either naturally produced in situ, or externally supplied through foods and supplements.…

    • 330 Words
    • 2 Pages
    Good Essays
  • Powerful Essays

    Cancer Prevention

    • 2112 Words
    • 9 Pages

    Kaur, C. & Kapoor, H.C. (2001). Antioxidants in fruits and vegetables – the millenium’s health. International Journal of Food Science and Technology, 36, 703-725.…

    • 2112 Words
    • 9 Pages
    Powerful Essays
  • Good Essays

    In our daily diets, carbohydrates, fats, proteins and water are the main components in our food. However, if the food is left untreated, oxidative deterioration of the flavor and odor of fats and fatty constituents in it will be resulted. Hence, antioxidant are used widely in the production of food which fats and oils are used as raw materials and in the marketing of foods containing fats under modern conditions.…

    • 914 Words
    • 3 Pages
    Good Essays
  • Good Essays

    The ABTS values of the antioxidant extracts are presented in Table 2. Mahua extracts from different extraction solvents differed significantly (P < 0.05) in their ABTS values. ABTS radical scavenging ability of samples can be ranked as acetone > methenol > meth – ethanol > water > ethenol fraction. The ABTS values of mahua flower ranged from 12.243±0.745 to 74.18±1.155 µmol Trolox/g fw ; unripe fruit from 20.196±0.332 to 59.1±0.1 µmol Trolox/g fw ; and ripe fruit from 13.64± 0.529 to 45.22 ± 1.105 µmol Trolox/g fw . The results suggested that 50 % acetone was the best solvent for extracting ABTS antioxidant assay for mahua flower, unripe and ripe…

    • 1312 Words
    • 6 Pages
    Good Essays
  • Good Essays

    The kinetic equation, half-life values, correlation coefficient and pre-harvest interval (PHI) of the trifloxystrobin and tebuconazole residue dissipation were calculated from the experimental data and summarized in Table 4. The dissipation dynamics of trifloxystrobin on green tea leaves followed a first-order rate equation as follows C = 2.5472e-0.245x (r=0.9797) and C = 6.5457e-0.207x (r= 0.9804) with the half-life of 2.83 and 3.34 days for trifloxystrobin at RD and DRD, respectively. The half-life of trifloxystrobin in processed tea was 2.74 and 3.63 days respectively for 31.25 and 62.50 g a.i./ha and its dissipation dynamics were C = 5.5593e-0.253x and C = 9.8395e-0.191x. The calculated half-lives for tebuconazole were in a range of 2.98 to 3.36 and 3.01 to 3.07 days from the treatment at 62.50 and 125.0 g a.i./ha, respectively for green tea leaves and processed tea. The half-lives of mixed formulation consisting of trifloxystrobin and tebuconazole in the present study was similar to those of gherkin reported by Paramasivam, Selvi, Deepa, Jayaprakash and Chandrasekaran (2015) who observed half-lives were in the range of 3.31-3.38 and 3.0-3.04 days at different doses and slight variation was found the results studied in ginseng (5.92-9.76 and 4.59-7.53 days) reported by Wang, Gao, Liu, Cui and Li (2015). The half-lives of trifloxystrobin and tebuconazole on grape…

    • 767 Words
    • 4 Pages
    Good Essays
  • Better Essays

    Adhesives Out of a Betel Nut

    • 3971 Words
    • 16 Pages

    Soon, it was realized to be an invasive species due to their adaptability to a wide type of fresh water ecosystems and interference with human activities. Thus a huge amount of money, and efforts have been invested since then for their management. However, it was recently realized that they could be substainably managed in their natural ecosystem and used in biofuel production, generating ample avenues of research, development and marketing of their end product (i.e. bioethanol and biogas). As the search for alternatives to fossil fuel intensifies in this age of modernization and industrialization, fuelled by increasing energy costs, water hyacinth holds a strong promise in the 21st century biofuel industry. The antoxidative properties of water hyacinth leaves were investigated by evaluating the scavenging capacity of liquid extracts in a competitive protective process against oxygenated free radicals (OFRs) released via electrolysis in a phosphate buffer with a Pt electrode. Colorimetric measurements carried out at 515 nm, through a N,N-diethyl-1,4-phenylenediamine (DPD) assay, showed a decreased absorbance of the sample, as compared to the blank obtained by electrolysis of the buffer without plant extract, revealing, thereby, the presence of antioxidizing agents in the liquid extracts. The antioxidative activity was estimated in terms of equivalent-glutathione (EG, in nmoles equivalent-glutathione per gram of dry plant material selected (eg/gdp)), and compared to those of soya beans and garlic bulbs. The EG value increased with decreasing dilution factors, regardless to the plant type, suggesting a strong influence of the medium pH on the…

    • 3971 Words
    • 16 Pages
    Better Essays
  • Good Essays

    Previously, Yeoh, & Wong (1993) reported the nutritional values of A. gangetica. Ogle et al., 2001 reported the folate composition of A. gangetica. However, not much information on the utility of this leafy vegetable has been reported. To the best of our knowledge, there is no report on the optimization of polyphenols and their radical scavenging activities from the edible leaves of A. gangetica. Therefore, the main aim of this study is to optimize extraction conditions such as EtOH concentration, liquid-to-solid ratio and extraction time to obtain the maximum extraction yield, phenolic content, flavonoid content and DPPH and ABTS radical scavenging activity from the A. gangetica leaves. Furthermore, the polyphenols present in the optimized…

    • 920 Words
    • 4 Pages
    Good Essays
  • Powerful Essays

    rancidity fat and oil

    • 1970 Words
    • 8 Pages

    However, due to its high melting point, it is not gaining due status in spite of being rich in natural antioxidants, vitamins, high oxidative stability and having a long self life (Edem, 2002). Palm oil is also rich in β-carotene which helps to prevent liver and lung cancer (Murakoshi et al., 1992). The food value of the edible lipids also depends on chemical properties like iodine value, peroxide value, p-anisidine value, acidity etc, as well as on some…

    • 1970 Words
    • 8 Pages
    Powerful Essays
  • Better Essays

    Flavonols are found in spinach with its prominent food flavonoids such as Myricetin, Quercetin and Kaempferol. Most flavonoids are commonly concentrated in leafy vegetables or the skin and peels of a certain fruit. It has an aromatic chromophores that in nature, it is in colorful pigments of vegetable plants that attract pollinating insects and assist in propagation. And it also serves as phytoalexins which acts as antimicrobial compound (Corcoran, McKay, et. al., 2012). A lot of studies show that a regular intake of foods with flavonoids may reduce the risk of several chronic conditions such as the neurodegenerative diseases, atherosclerosis and other types of cancer. These researches contain considerable interest in flavonoids as it is associated to a positive health outcomes. (Corcoran, McKay, et. al.,…

    • 1200 Words
    • 5 Pages
    Better Essays