how to calculate ec50 value for antioxidant activity using

In general, fruiting bodies showed the highest antioxidant activity and reducing power, while the mycelium showed the highest chelating activity. Food Process Technol. Initial Choices for Modeling IC50 There are several initial choices to make when modeling IC50. The method was based on Hue et al. Phenolics have received much scientific attention because they are the most widely-spread secondary metabolites in the plant kingdom and aside this, they are also known as sources of potential natural antioxidants because of their abilities to act both as efficient radical scavengers and metal chelators.[30]. Slowly, 1 mL of sample solution was premixed with 0.05 mL of FeCl2 solution (2 mM) and 1.85 mL of double distilled water. Jayaprakasha GK, Jena BS, Negi PS, Sakariah KK. Study on in vitro antioxidant potential of some cultivated Pleurotus species (Oyster mushroom). All values are reported as means SD (n = 3). The blank was made up of 2.5 ml of 90% ethanol plus sample solution (2.5 ml). where ln is natural log, a is the initial absorbance (at 470 nm) at time 0, b is the absorbance (at 470 nm) at 20, 40, 60, 80, 100 or 120 min and t is the initial absorbance (470 nm) at time 0. Dermarderosian A. Missouri: Lippincott Williams and Wilkins; 2002. Reducing power assay was expressed as mg GAE/L. Technol. [41] In this assay, the incubation of ferric-EDTA with H2O2 and ascorbic acid at pH 7.4 led to the production of hydroxyl radicals. This mixture was incubated at room temperature for 30 min in darkness. In general, the dried samples had higher activity in the methanolic extract. Flavonoids are reported as antifungal compounds, because plants produce them as protection against fungal infections, then, these compounds might negatively affect fungal growth. The value reported was lower compared with that observed in this investigation in methanolic extract obtained at room temperature and boiling of the dried fruiting body with EC50 = 217.24 1.31 and 76.63 0.070 mg GAE/L, respectively. Int. (2009). Ellagic acid derivatives of agrostistachys hookeri. EC50 estimation of antioxidant activity in DPPH assay using several statistical programs J. *Correspondence: Jorge Soriano-Santos, jss@xanum.uam.mx Gerardo Daz-Godnez, diazgdo@hotmail.com, Association of Official Analytical Chemists [AOAC], 2000, Creative Commons Attribution License (CC BY). EESTG was found to have 358.33 5.77 g BHTE/mg dry extract. Reducing power assay of methanolic extracts of dry P. ostreatus. Food Res. doi: 10.1021/jf0201273, Meir, S., Kanner, J., Akiri, B., and Philosoph-Hadas, S. (1995). The methanol extract of the dried primordium obtained at room temperature showed the highest activity with an EC50 = 22.89 0.37 mg GAE/L. The mixture was then added to 0.2 ml of extract or standard (BHT and quercetin) or ethanol (as control). Finding an EC50 value using a semi-logarithmic plot - YouTube Oxford: Clarendon Press; 1989. Sudha et al. 5, 161166. The baseline is about 20%, and the maximum is 100%, so the EC50 is the concentration of agonist that evokes a response of about 60% (halfway between 20% and 100%). In general, the flavonoids content was reported with a low value, representing a very small percentage of the total polyphenol content, however, the majority of the extracts presented antioxidant activity. (2011) found in methanol extracts of P. ostreatus and P. dryinus high antioxidant 11600 and 2385.71 M de FeSO4/g. This analysis consists of plotting X values that represent the drug concentrations or function of concentrations (log . Both extracts of A. brasiliensis showed higher content of total flavonoids than extracts of A. bisporus. Evaluation of antioxidant activities and antimutagenicity of turmeric oil: A byproduct from curcumin production. The properties of the reducing power can be an indicator of antioxidant potential of compound evaluated (Meir et al., 1995). Agro. Association of Official Analytical Chemists [AOAC] (2000). Yim, H. S., Chye, F. Y., Ho, S. K., and Ho, C. W. (2009). Antioxidant activity (AA) is expressed as percent of inhibition relative to the control, using the following formula: The antiradical activity of the extract was estimated according to the procedure described by Nickavar et al. During this process, mycelium grown on wheat straw, primordia and fruiting bodies were obtained. EESTG showed an AEAC value of 1.43 mg Vitamin C equivalents/g dry wt of extract. In addition to the phytochemicals, fungi have also been considered as a source of biologically active substances that can be used to reduce oxidative damage in humans and useful in disease prevention (Chye et al., 2008; Fatih et al., 2010; Adebayo et al., 2012; Shirmila and Radhamany, 2012). T1 and T2 as Figure 1. 246350. [41] It is generated mainly through Fenton reaction; and other routes such as the reaction between hypochlorous acid and superoxide anion as well as the decomposition of peroxynitrous acid. If your Inhibition is indeed % then it is reasonable to set A1 and A2 as 0 and 100 and set both to fixed. Food Chem. Degradation rate (DR) was calculated according to first order kinetics, using the following equation based on Al-Saikhan, Howard, and Miller:[26]. (2014) investigated the antioxidant activity and the total polyphenol amount, of methanol extracts of mycelium and fruiting body of P. sajor-caju, P. ostreatus and P. sapidus. Washington, DC: Association of Official Analytical Chemists. Could you (or anybody else) please further explain how to do that. 3 Posts. J. Both values are lower than those reported in this study for the aqueous extract of fresh fruiting body obtained by boiling (9.92 0.05 mg GAE/g) and the aqueous extract of the dry fruiting body obtained at room temperature (11.36 mg GAE/g). Keles et al. B., and Asmah, R. (2013). FIGURE 3. The higher the AEAC value, the greater is the antioxidant activity. The GC Mass analysis revealed in some samples, mainly of the fruiting body the presence of methyltartronic acid (RT 20.185) which could be the responsiblity of the antioxidant activity. Overall, the fruiting body of P. ostreatus showed the best results and the possibility of continuing to investigate its functional properties of this fungus is opened. Antioxidant properties of several tropical fruits: A comparative study. djamor with an absorbance of 0.874, followed by P. djamor var. Terashima S, Schimizu M, Nakayama H, Ishikura M, Ueda Y, Imai K, et al. How Do I Perform a Dose-Response Experiment? - GraphPad Reducing power assay of aqueous extracts of fresh P. ostreatus. Methanolic extract obtained at room temperature of the fruiting body and primordium, both dried showed the higher radical scavenging activity of DPPH and ABTS. Figure 2 shows the degradation rates of EESTG in comparison with the negative and positive controls while a comparison of anti-oxidant activities is displayed in Figure 3. Phenolic profiles of selected edible wild mushrooms as affected by extraction solvent, time and temperature. Concentration changes of malondialdehyde across the cerebral vascular bed and shedding of L-selectin during carotid endarterectomy. J. Med. (2002). Trichloroacetic acid (0.25 mL) was added to the mixture and was centrifuged at 1000 rpm for 10 min at room temperature. 90% ethanol (1 ml) plus each sample solution (50 l) was used as blank. Chye, F. Y., Wong, J. Y., and Lee, J.-S. (2008). Summed up together, the EC 50 and AEAC values obtained for EESTG imply that the extract contains phytochemicals with antioxidant properties. 122. Biol. Trends Plant. The reaction mixture consisted of 0.07 mL of extract and 3 mL of the ABTS radical. Chye et al. J. Afterward, 0.1 mL of ferrozine solution (5 mM) was added and mixed vigorously. Food Chem. The results of the metal ion chelating activity indicate that all extracts tested had acted. In the case of the phenolics compounds, the best extracts were the methanolic of fresh primordium obtained by boiling and aqueous of the dry fruiting body obtained at room temperature. (2012) reported 32.21 mg GAE/g in the methanol extract of P. eryngii. 8600 Rockville Pike Chia-Chi, C., Ming-Hua, Y., Hwei-Mei, W., and Jiing-Chuan, C. (2002). (2012) reported high chelating activity of methanol extract of the mushroom P. eryngii collected from different regions of the Tunceli province of Turkey (EC50 = 470.23, 218.31, and 292.99 mg GAE/mL for Ovacik, Pulumur and City center, respectively). Food Chem. Our results show that the hydroxyl radical scavenging activities of the standard compounds (BHT and quercetin) significantly (P < 0.001) exceeded those of the extract at both the lowest (50 g/ml) and highest (800 g/ml) concentrations used in this study. Choi YH, Pezzuto JM, Kinghorn AD, Farnsworth NR. DPPH assay allows the evaluation of antioxidant capacity through the determination of EC50. Antioxidant properties in the oyster mushrooms (Pleurotus spp.) EC 50 represents the concentration at which a substance exerts half of its maximal response. The reaction mixtures were incubated for 60 min in the dark at room temperature and then, the absorbances were measured at 725 nm. (2009) reported in aqueous extracts of Pleurotus sp. Technol. Antioxidative properties of xanthan on the autoxidation of soybean oil in cyclodextrin emulsion. [21] To 0.50 ml of each sample (800 g/ml), 2.5 ml of 1/10 dilution of Folin-Ciocalteau's reagent and 2 ml of Na2CO3 (7.5% w/v) were added and incubated at 45C for 15 min. The flavonoid content was determined by the colorimetric method of aluminum chloride according to methodology previously described by Chia-Chi et al. We can calculate the IC50 values in EXCEL program by plotting the curve of inhibitions and corresponding concentrations following the . Oxidative stress, caused by endogenous factors such as reactive oxygen species (ROS) including the hydroxyl radical, superoxide anion radical, hydrogen peroxide, singlet oxygen, nitric oxide radical, hypochlorite radical, etc., and exogenous factors such as smoking, ionizing radiation, pollution, organic solvents, pesticides, etc., are able to attack nucleic acids, proteins, enzymes, and other small molecules causing loss of structure and function. IC 50 values are very dependent on conditions under which they are measured. IC50 Calculator | AAT Bioquest The reaction mixture consisted of 0.5 mL of extract, 3 mL of methanol, and 0.3 mL of 0.5 mM 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical solution in methanol. Ferric reducing antioxidant power assay was measured according to the procedure described by Sudha et al. Whereas at the highest concentration (i.e. Frontiers | Evaluation of the Antioxidant Activity of Aqueous and The percentage yield of the extraction was 9.39% w/w. A thorough examination of the various in vitro antioxidant and free radical scavenging assays carried out on ethanol extract of stem bark of Terminalia glaucescens points to the fact that the extract contains some phytocomponents with potent antioxidant activity as evident most emphatically from -carotene-linoleate bleaching assay and reducing power activity. The absorbance was determined at 562 nm after the mixture stood for 10 min at the room temperature. 5, 5661. Bio. Finally, absorbance at 415 nm was read. HHS Vulnerability Disclosure, Help 43, 18131819. Comparing the previous results with those reported in our study, the value obtained from first region was higher than the reported for aqueous extract (obtained at room temperature and boiling) of fresh fruiting body, as well as of the methanolic extract (obtained at room temperature) of fresh fruiting body and the aqueous extract (obtained by boiling) of the dried fruiting body. Determination and involvement of aqueous reducing compounds in oxidative defense systems of various senescing leaves. mg Vitamin C equivalents/mg dry wt, which was calculated as follows:[27]. The methanol extracts of dried samples showed low values (between 0.62 0.012.39 0.02 mg GAE/g). In general, the extracts of dried samples showed higher reducing power than the extracts of fresh samples and tend to show greater reducing power by aqueous than methanolic extracts. Aruoma OI, Laughton MJ, Halliwell B. Carnosine, homocarnosine and anserine: Could they act as antioxidants. It was observed that the DPPH and ABTS radical scavenging activities were positively correlated to the concentration of the extract. Vol. Total polyphenols and flavonoids contents, as well as ferric reducing antioxidant power (FRAP), metal ions chelating activity, reducing power assay and scavenging activity of DPPH and ABTS radicals in aqueous and methanolic extracts obtained from mycelium, primordium, and fruiting body of Pleurotus ostreatus in both fresh as dry, were evaluated. Antioxidant properties of several commercial mushrooms. For obtaining extracts, mycelium, primordia, and fruiting bodies, both fresh and dried samples were used. They also found in their methanol extracts a low antioxidant activity of mushroom Hydnum repandum with 145.50 M de FeSO4/g. Nutritional quality and antioxidant activity of selected edible wild mushrooms. (2013). and P. ostreatus 9.01 and 7.23 mg GAE/g, respectively. This calculator generates an EC 50 value typically thought of as the relative EC 50. Values are the average of three replicates DS. Mushrooms 10, 315324. The antioxidant activity measured by FRAP method was observed in all extracts (Table 3). No use, distribution or reproduction is permitted which does not comply with these terms. doi: 10.1080/11358120809487626. Cite Penn State Hershey Medical Center and Penn State College of. J. Agr. Measurement of antioxidant activity - ScienceDirect It was freshly prepared and warmed at 37C. Fejes S, Blzovics A, Lugasi A, Lemberkovics E, Petri G, Kry A. Meir S, Kanner J, Akiri B, Philosoph-Hadas S. Determination and involvement of aqueous reducing compounds in oxidative defense systems of various senescing leaves. EC50 value, defined as the concentration of the sample leading to 50% reduction in the initial DPPH concentration, was obtained from a calibration curve for the extract. The absolute EC50/IC50 is the response corresponding to the 50% control (the mean of the 0% and 100% assay controls). doi: 10.1080/21501203.2010.511292. Int. that publishes the EC 50 prediction of antioxidant activity for honey, bee pollen and propolis using comparative different regression models. Antioxidant activity and total phenolics in selected fruits, vegetables, and grain products. Tecnol. Lpez-Vlez M, Martnez-Martnez F, Del Valle-Ribes C. The study of phenolic compounds as natural antioxidants in wine. (2014). Sing. doi: 10.1021/jf00055a012, Miliauskas, G., Venskutonis, P. R., and van Beek, T. A. (2012). These results show that EESTG displayed significantly (P < 0.001) higher activity than BHT but lower activity than ascorbic acid at 50 g/ml. Jung MJ, Heo SI, Wang MH. doi: 10.3390/molecules16043197, Jeena, G. S., Punetha, H., Prakash, O., Chandra, M., and Kushwaha, K. P. S. (2014). Arbaayah and Umi (2013) reported a high content of flavonoids from ethanol extracts of various species of Pleurotus, with values of 1.40 to 29.80 mg QE/g. In methanol extracts of Pleurotus sp. Several research had proven the antioxidant activity of different mushrooms. It was identified by a plant taxonomist in the Department of Botany, University of Ibadan, Nigeria and a voucher specimen was deposited in the Herbarium of same Department with the Herbarium number UIH-22404. Fr) Kummer stored in packaging materials. Vinson JA, Su X, Zubik L, Bose P. Phenol antioxidant quantity and quality in foods: Fruits. The higher reducing power (at 10 mg sample/mL) was shown for P. djamor var. For technical assistance on using this calculator, please contact websupport@aatbio.com. In general, values of antioxidant activity of extracts of fresh samples were higher with respect to the dried samples. [25] An aliquot of 0.1 ml of the fractions (50-800 g/ml) was combined with 1.0 ml of reagent (0.6 M sulfuric acid, 28 mM sodium phosphate and 4 mM ammonium molybdate). Metal ion chelating activity was expressed as mg GAE/L. Values are mean SD for triplicate assay. the contents by NLM or the National Institutes of Health. doi: 10.1016/j.fct.2012.02.013, Joan-Hwa, Y., Hsiu-Ching, L., and Jeng-Leun, M. (2002). Antioxidants in fruits and vegetables-the millennium's health. The absorbance was measured at 700 nm. The EC50 values of the root, leaf, whole plant and stem of Sida rhombifolia have been reported by . If polar compounds such as ascorbic acid, rosmarinic acid, etc., are tested by the BCB method, they would be considered as weak antioxidants;[37] as a result of this factor, the reference compounds that we used in this study were quercetin and BHT. B., and Alencar, S. M. (2008). The total antioxidant capacity is quantitatively expressed as microgram BHT equivalents per mg (g BHTE/mg) of dry extract. T1 and T2 as Figure 1. 2014, 18. All tests were performed in triplicate and the graph was plotted with the average of the three determinations. doi: 10.5943/mycosphere/4/4/2. The different results also suggested that antioxidant activity couldnt be by polyphenols. Methods Enzymol. Results are expressed as micrograms of gallic acid equivalents per milligram of dry weight (g GAE/mg) of extract. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The values observed with this method were generally low compared with those reported in other studies, Keles et al. Four hundred grams (400 g) of the powered plant sample was soaked in 70% ethanol (1600 ml) for 72 h with intermittent stirring/shaking. Reducing power of EESTG and the positive controls [BHT and ascorbic acid]. FIGURE 4. For statistical analysis, data were analyzed using Sigma Plot (version 11.0). Al-Saikhan MS, Howard LR, Miller JC., Jr Antioxidant activity and total phenolics in different genotypes of potato (. All water and methanolic extracts possess phenolics compounds and flavonoids. Antioxidant activity, phenolic and flavonoid contents in the leaves of different varieties of sweet potato (Ipomoea batatas). Pearson's correlation test was used to assess correlations between means. Our result confirms the presence of flavonoids in the extract based on quercetin as the reference compound and the TFC expressed in microgram quercetin equivalents per milligram (g QE/mg) of dry extract. Antioxidant activity applying an improved ABTS radical cation decolorization assay. This fact still promisingly indicates the presence of potent phytoconstituents in EESTG that have the capability to scavenge hydroxyl radicals although the activities of such components may have been shielded by the presence of other components in the heterogenous extract. 6, 4147. Food Chem. Degradation rate of the ethanol extract of the stem bark of Terminalia glaucescens [EESTG] assayed by -carotene bleaching method (n = 3), Antioxidant activity (%) of ethanol extract of the stem bark of Terminalia glaucescens [EESTG] assayed by -carotenelinoleate bleaching. The relative EC50/IC50 is the parameter c in the 4-parameter logistic model and is the concentration corresponding to a response midway between the estimates of the lower and upper plateaus.

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