The aqueous extract of A. bisporus had a higher content of polyphenols. 50 g/ml), EESTG, ascorbic acid and butylated hydroxytoluene (BHT) had activity values corresponding to 0.0830 0.0101, 0.0247 0.0055, and 0.1750 0.0113, respectively. The total polyphenol content of dried samples was higher in aqueous extracts obtained both in room temperature and boiling. 8600 Rockville Pike The scavenging activity on hydroxyl radicals was expressed as. Antioxidant activity, phenolic and flavonoid contents in the leaves of different varieties of sweet potato (Ipomoea batatas). [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. [34] It is evident from our findings that the extract possesses antioxidant activity in a concentration-dependent manner, which may imply its relevance in attenuating oxidative damage to cellular components and thereby prevent oxidative stress. J. Agr. nikolajeremic. The flavonoid content was calculated in mg quercetin equivalents per g of dried sample (mg QE/g). and P. florida. Pearson's correlation test was used to assess correlations between means. 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%). Biomed. J. Rev. 968. DPPH radical scavenging activity of P. ostreatus. Free Radicals in Biology and Medicine; pp. The petroleum ether and methanolic extract of P. florida presented higher EC50 values compared with the aqueous extract (obtained by boiling) of dried and fresh fruiting bodies reported in this research (EC50 = 25.53 0.80 and 42.4 0.72 mg GAE/L, respectively). [28] The reaction mixture in a final volume of 1.0 ml contained 100 l of 2-deoxy-D-ribose (28 mM in 20 mM KH2PO4 buffer, pH 7.4), 500 l of the extract at various concentrations (50800 g/ml) in buffer, 200 l of [1.04 mM EDTA and 200 M FeCl3] (1:1v/v), 100 l of 1.0 mM hydrogen peroxide (H2O2) and 100 l of 1.0 mM ascorbic acid. All values are reported as means SD (n = 3). 40, 945948. The injector flow rate was 250C; carrier gas was He of 99.9995% purity, column flow rate 1.2926 mL/min. [29] The value of the TPC of the extract obtained confirms that the extract is very rich in phenolic contents. The result we obtained based on this assay confirms the antioxidant potency of EESTG in comparison to BHT as a standard antioxidant. The best extract in ferric reducing antioxidant power (FRAP) was the methanolic of fresh primordium obtained by boiling. Food Chem. Antioxidant activity and bioactive components of the aerial parts of Hypericum perforatum L. from Epirus, Greece. EC50 values of the extracts evaluated in this study are shown in Table 5. Halliwell B, Gutteridge JM. The extracts were quantified using gas chromatograph (Agilent Technologies, 7890A GC System) equipped with a column Agilent DB-WAX (30 m 320 m 0.25 m) coupled to a mass spectrometer Agilent Technologies (5975C V2-MSD with Triple-Axis Detector) with Autosampler and injector (G4513A), programmed at temperature 60C for 5 min, then 11C/min to 250C. The results were expressed as mean SD (standard deviation) and the EC50 values were obtained from the linear regression plots. In general, fruiting bodies showed the highest antioxidant activity and reducing power, while the mycelium showed the highest chelating activity. Values are the average of three replicates DS. [22] Briefly, 2.5 ml of each extract solution (800 g/ml) was mixed with 2.5 ml AlCl3 reagent in 90% ethanol and allowed to stand for 40 min at room temperature. Shimada et al. Antioxidant flavonoids: Structure, function and clinical usage. After the samples had cooled to room temperature, the absorbance was measured at 695 nm against the blank using a UV spectrophotometer. The extracts (0.050 mL) of the different concentrations were added with 0.2 mL of 0.2 M phosphate buffer (pH 6.6) and 0.2 mL of 1% potassium ferricyanide. Pleurotus ostreatus is a fungus very important because is industrially produced as food (oyster mushroom), a ligninolytic enzyme producer and as bioremediation agents in decontamination processes of materials rich in phenolic compounds, and recently as a biocompounds source (da Luz et al., 2012). Screening of plant extracts for antioxidant activity: A comparative study on three testing methods. Braz. The total polyphenol content of the fresh samples obtained at room temperature and boiling was higher in aqueous extract of mycelium and in the methanolic extract of the fruiting body. (1999) with some modifications. In addition, those organic acids may have a protective role against various diseases due to their antioxidant activity (Valentao et al., 2005). Reducing power assay of methanolic extracts of fresh P. ostreatus. Antioxidant properties of wild edible mushrooms. Figure 4 shows the DPPH radical scavenging activity of ascorbic acid, BHT and EESTG, while Figure 5 is a representation of the corresponding anti-oxidant activities based on DPPH assay. (2015) reported the profile of organic acids of the mushroom Agrocybe aegerita (Brig.). Invitro antioxidant activities, total phenolics and flavonoid of wild edible mushroom Macrolepiota mastoidea (FR.) (2009) reported in aqueous extracts of Pleurotus sp. 7, 16471654. Microbiol. Antioxidant properties and antioxidant compounds of various extracts from the edible basidiomycete Grifola frondosa (Maitake). Food Res. 122. (2009) reported radical scavenging activity of methanol extract of dried P. djamor (EC50 of 0.0293 mg sample/mL). Technol. 2014, 18. Biotech. (2015). DPPH radical scavenging activity was determined according to Moraes-de-Souza et al. L-ascorbic acid solution (50 l, at the concentrations of 50-800 g/ml in ethanol) was used as positive control [i.e. Influence of processing and storage on the phenolic composition of Thompson seedless grape juice. zgen U, Mavi A, Terzi Z, Yildirim A, Cokun M, Houghton PJ. Here is what I do, so just tell me if I`m doing it right: x-axis is log values of concentration, y is Inhibition(%) than I plot that, and perform DoseResp analysis(just by opening the dialog, selecting DoseResp under function and than I click fit). Could you (or anybody else) please further explain how to do that. Foods 25, 113. doi: 10.1016/j.foodchem.2006.07.038. J. Agric. (2013). We found the value of the TPC of the extract to be 596.57g GAE/mg extract. The methanolic extract obtained at room temperature had more radical scavenging activity than the methanolic extract obtaining by boiling of the dried fruiting body obtained in this investigation with EC50 de 32.26 and 136.98 mg sample/mL, respectively. It was observed that the DPPH radical scavenging activity was positively correlated to the concentration of the extract. (2009). J. The antioxidant activity of the extract was assayed based on the -carotene bleaching (BCB) method developed by Velioglu et al. [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. For extracts obtained by boiling, the Soxhlet apparatus was used for methanolic extracts (Association of Official Analytical Chemists [AOAC], 2000) and for the aqueous extracts were shaken in boiling for 5 min. doi: 10.1016/S1360-1385(00)01741-6, Pornariya, C., and Kanok-Orn, I. Free Radic. 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. T1 and T2 as Figure 1. The results were also expressed as AEAC (Ascorbic acid equivalent antioxidant capacity) i.e. Jan-Ying et al. Using the DoseResp or Logistic function will calculate EC50/IC50 values. The reducing power of all samples was concentration dependent (Figures 14). For fresh samples, methanolic extract of primordium and aqueous extract of the fruiting body, showed the highest polyphenols values, 12.06 0.02 and 9.92 0.05 mg GAE/g, respectively. Res. 21, 661668. Biol. No use, distribution or reproduction is permitted which does not comply with these terms. Total flavonoids content of P. ostreatus. Based on these facts, DPPH assay is one of the most widely employed methods for screening antioxidant activities of plant extracts. The ABTS+ solution was diluted with water to an absorbance of 0.70 (0.02) at 734 nm. doi: 10.1590/S1517-838220120004000035, Fatih, K., Mustafa, O., and Hsniye, K. (2010). Asample = Absorbance of sample after 45 min. Antioxidative properties of xanthan on the autoxidation of soybean oil in cyclodextrin emulsion. For statistical analysis, data were analyzed using Sigma Plot (version 11.0). Jung MJ, Heo SI, Wang MH. Non-site-specific hydroxyl radical-mediated 2-deoxy-D-ribose degradation: Hydroxyl radical scavenging activity was measured by the ability of the extract to scavenge the hydroxyl radicals generated by the Fe3+ -ascorbate-EDTA-H2O2 system (Fenton reaction). In pharmacology, a dose-response experiment determines the effects of a drug on cells grown in vitro. The aqueous extracts showed similar values about 45 mg GAE/g. J. Asample = Absorbance of sample after 6 min. The aim of this study was evaluated antioxidant activity of aqueous and methanolic extracts of P. ostreatus at different growth stages of developing on wheat straw. All tests were performed in triplicate and the graph was plotted with the average of the three determinations. Guzmn et al. Nasir E, Ali SI. 111. The reaction mixture was incubated at 37C for 10 min and the absorbance was measured at 593 nm. (2016), reported that the mushrooms do not contain flavonoids, and those found in the hyphae could be due to the facility of these organisms to absorb many nutrients and compounds from the substrate where they grow or from neighboring plants by spreading their hyphae or forming mycorrhizae. Antioxidant properties in the oyster mushrooms (Pleurotus spp.) (2009). Adv. Malic acid was the most abundant organic acid (1.82 g/100 g), followed by citric acid (0.88 g/100 g), then fumaric acid (0.26 g/100 g) and oxalic acid (0.09 g/100 g). 6, 375380. The dried sample was grinded to powder, sieved and packed into polythene bags and stored at 4C. Mushrooms 10, 315324. 85, 231237. Food Chem. [4] IC 50 values can be used to compare the potency of two antagonists. The mixture was incubated in the water bath for 20 min at 50C. After incubation for 45 min, absorbance was determined in a spectrophotometer at 517 nm. Free radical scavenging and total phenolic contents from methanolic extracts of. Measurement of Total Phenolics and ABTS Assay for Antioxidant Activity (Personal Communication), Nickavar B, Kamalinejad M, Mohandesi S. Comparison of the components of the essential oils from leaves and fruits of. Antioxidant and antidiabetic activities of extracts from Cirsium japonicum roots. . Koudou J, Roblot G, Wylde R. Tannins constituents of Terminalia glaucescens. Specifically, EESTG displayed a significantly (P < 0.001) higher anti-oxidant activity when compared to BHT, and the activity of EESTG was higher than that of quercetin although not significant (P = 0. and P. ostreatus 9.01 and 7.23 mg GAE/g, respectively. The properties of the reducing power can be an indicator of antioxidant potential of compound evaluated (Meir et al., 1995). Values are the average of three replicates DS. Furthermore, treatment of boiling for extraction favored the DPPH radical scavenging activity, since these extracts showed higher activity than the extracts obtained at room temperature, except for the methanol extract of dry fruiting body and dry primordium obtained at room temperature, which they showed the lowest values of EC50 (30.89 1.83 and 26.99 0.47 mg GAE/L, respectively). This ability of an extract is a product of the presence of different antioxidants which can neutralize the linoleate-free radical and other free radicals formed in the system. The antioxidant values for the extracts were evaluated with the one-way ANOVA and Tukeys. Prieto P, Pineda M, Aguilar M. Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex: Specific application to the determination of vitamin E. Kulisic T, Radonic A, Katalinic V, Milos M. Use of different methods for testing antioxidative activity of oregano essential oil. Indian J. Nat. Pract. The antioxidant activity may be as a result of the presence of different molecules or substances no determined in this study which are present in the extracts. Two milliliter (2 ml) aliquots of the emulsion were pipetted into test tubes and immediately placed in a water bath at 50C. It was observed that aqueous extracts showed generally higher DPPH radical scavenging activity than methanol extract samples in both dry and fresh. B., and Asmah, R. (2013). This is the first report where the antioxidant activity of P. ostreatus in different growth stage was reported. Mycosphere 4, 661673. Here is what I do, so just tell me if I`m doing it right: x-axis is log values of concentration, y is Inhibition (%) than . 111, 370376. 139). (2011) found a DPPH radical scavenging activity, of methanol extract of A. bisporus, P. dryinus, Boletus edulis, and P. ostreatus with EC50 = 78.43, 58.06, 38.31, and 29.66 mg GAE/mL, respectively (EC50 calculated from the original values reported by the authors). The hydroxyl radical scavenging activity of EESTG and the positive controls (BHT and quercetin) at different concentrations (50, 100, 200, 400, 800 g/ml) are shown in Figure 6. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). In general, fruiting bodies showed the highest antioxidant activity and reducing power, while the mycelium showed the highest chelating activity. Accessibility [23] The Fe2+ can be monitored by measuring the formation of Perl's Prussian blue at 700 nm. Chye, F. Y., Wong, J. Y., and Lee, J.-S. (2008). For example, a dose-response experiment studies how well a drug decreases the growth of tumors grown in cell culture. The Review of Natural Products I, Facts and Comparison; p. 637. This method which is routinely employed to study phenolic antioxidants is fast, convenient, simple and most importantly reproducible. 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. [31,32] The antioxidant activities of phenolic compounds and flavonoids in biological systems have already been established based on their abilities to act as scavengers of singlet oxygen and free radicals;[33] thus validating the presence of antioxidants and free radical scavengers in the extract. This parameter is typically employed not only to express the antioxidant capacity but also to compare the activity of different compounds with each other. [22] Briefly, a 0.1 mM solution of DPPH radical solution in 90% ethanol was prepared and 1 ml of this solution was mixed vigorously with 50 l of different concentrations (50-800 g/ml in ethanol) of each extract. J. Pharm. Our result on total flavonoid content shows that the flavonoid content of EESTG is lower in quantity as compared to phenolic content just in consonance with the results earlier obtained by some other researchers. Antioxidant activities of three Indian commercially available Nagakesar: An. The highest values were 166.5 0.10 and 113.9 0.24 M de FeSO4/g in methanolic extracts of fresh primordium and in aqueous extracts of fresh fruiting body, respectively, both obtained by boiling. One peculiarity of this method is that it allows testing of both lipophilic and hydrophilic compounds[37,38] in comparison to other methods that are restricted in the nature of antioxidants that they can be used to quantify. Sorry, I forgot to ask, also how do I calculate standard deviation for IC50 given the SE?? Food Ag-Ind. The values of DPPH radical scavenging activity were higher compared with the obtained in this investigation in the methanolic extract (obtained at room temperature) of the dried fruiting body (EC50 = 26.99 0.47 mg GAE/L). 299, 152178. The antioxidant activity was calculated by using the following equation. Tecnol. IC 50 values are very dependent on conditions under which they are measured. Jeena et al. Identification and determination of antioxidant constituents of bioluminescent mushroom. The antioxidant capacities of Kyoho skin, seed, and flesh extracts were determined using DPPH and ABTS assays and a suitable statistical program was tested for the prediction of EC 50 values of Kyoho skin, seed, and flesh extracts obtained by DPPH and ABTS assays. Halliwell B, Gutteridge JM, Aruoma OI. Ferric reducing antioxidant power (FRAP) of P. ostreatus. FIGURE 3. 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. The EC50 values of the root, leaf, whole plant and stem of Sida rhombifolia have been reported by . [40] to be 546.1, 852.8, 983.8 and 1,222.5 g/ml respectively, as compared to 145.54 g/ml and 33.72 g/ml which we obtained for EESTG and ascorbic acid respectively. The overall effects of hydroxyl radicals have the inclination of causing mutagenesis, carcinogenesis and aging. 43, 18131819. 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. Halliwell B. Oxidative stress, nutrition and health. Biotechnology Research Center- CRBt Constantine ALGERIA. Studies on aldose reductase inhibitors from medicinal plant of sinfito, Potentilla candicans, and further synthesis of their related compounds. (2005). Technol. The EC50 is often used to express the amount or concentration of extracts needed to scavenge 50% of the free radicals. Fr) Kummer stored in packaging materials. (2012) with some modifications. Med. These results show that EESTG displayed significantly (P < 0.001) higher activity than BHT but lower activity than ascorbic acid at 50 g/ml. Food Chem. 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). Your choice of modeling IC50 or EC50 is simply based on whether you're modeling an inhibitor (antagonist) or a stimulant (agonist). EESTG showed an AEAC value of 1.43 mg Vitamin C equivalents/g dry wt of extract. Pharm. This calculator generates an IC 50 value typically thought of as the relative IC 50. Based on DPPH assay, we also expressed the antioxidant activities as the 50% effective concentration (EC50) and ascorbic acid equivalent antioxidant capacity (AEAC). Food Chem. Cienc. T1 = extract obtained at room temperature, T2 = extract obtained by boiling. J. Trop. In this study, the EC(50) estimation was performed using a comparative approach based on various regression models implemented in six specialized computer programs: GraphPad Prism version 5.01, BLeSq, OriginPro 8.5.1, SigmaPlot 12, BioDataFit 1.02, and IBM SPSS Statistics Desktop 19.0. In order to ascertain whether there is any link between the ethnomedicinal applications of Terminalia glaucescens and its antioxidant activities, different methods were employed to evaluate the free radical scavenging and antioxidant activities of ethanol extract of stem bark of Terminalia glaucescens (EESTG). ABTS radical scavenging activity of P. ostreatus. Arbaayah, H. H., and Umi, K. Y. In general, extracts of the fruiting body had a high amount of polyphenols, followed by primordium and by the mycelium (Table 1). 1 (Whatman Ltd., England). Lignocellulolytic enzyme production of Pleurotus ostreatus growth in agroindustrial wastes. 7, 6369. The antioxidant activity measured by the FRAP method of extracts from fresh samples were higher with respect to the dried samples. Vinson JA, Su X, Zubik L, Bose P. Phenol antioxidant quantity and quality in foods: Fruits. The results suggested that antioxidant activity could be due to polyphenols, but mainly by different molecules or substances present in the extracts. Vol. Rice-Evans C, Miller N, Paganga G. Antioxidant properties of phenolic compounds. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. Halliwell B, Gutteridge JM. In this assay, -carotene, a biologically oxidizable substrate, gives direct information on the ability of an extract to prevent oxidation. Antioxidant properties of wild edible mushroom Pleurotus eryngii collected from Tunceli province of Turkey. where EC50 Vit C and EC50 sample are the effective concentrations of vitamin C and sample respectively. In general, values of antioxidant activity of extracts of fresh samples were higher with respect to the dried samples. **P < 0.001; significantly different from quercetin and extract. FOIA 117, 398404. Antioxidant activity of various extracts from an edible mushroom Pleurotus eous. [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. Jacob RA. For aqueous extracts obtained by boiling showed very similar values of polyphenols content (around 4 mg GAE/g) and in these samples, the smallest value was reported for mycelium extract obtained at room temperature. Antioxidants in fruits and vegetables-the millennium's health. Whereas at the highest concentration (i.e. Cite Penn State Hershey Medical Center and Penn State College of. doi: 10.1016/S2221-1691(12)60194-4. J. Med. Statistical programs: OriginPro version 8, GraphPad Prism version 7, GraphPad Prism version 7 five-parameter (5P) logistic . Mustofa, Valentin A, Benoit-Vical F, Plissier Y, Kon-Bamba D, Malli M. Antiplasmodial activity of plant extracts used in west African traditional medicine. doi: 10.4162/nrp.2008.2.4.247, Zhengjun, X., Junrong, H., Xueming, X., and Zhengyu, J. Reducing power assay was expressed as mg GAE/L. The absorbance was determined at 562 nm after the mixture stood for 10 min at the room temperature. All water and methanolic extracts possess phenolics compounds and flavonoids. 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. They also found in their methanol extracts a low antioxidant activity of mushroom Hydnum repandum with 145.50 M de FeSO4/g. Dermarderosian A. Missouri: Lippincott Williams and Wilkins; 2002. Methanolic extract obtained at room temperature of the fruiting body and primordium, both dried showed the higher radical scavenging activity of DPPH and ABTS. (1999). Authors JS-S and GD-G designed the study, contributed reagents/materials, and supervised the work in all its aspects. In May 2023, Frontiers adopted a new reporting platform to be Counter 5 compliant, in line with industry standards. Reducing power assay of aqueous extracts of dry Pleurotus ostreatus. FIGURE 4. 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. The determination of the total flavonoid content (TFC) was carried out as described by Nickavar et al. 7:1099. doi: 10.3389/fmicb.2016.01099. A 900 L FRAP reagent was mixed with 90 L water and 30 L of the extract. J. 26, 12311237. The lower the EC50 value the higher the antioxidant activity of a sample. The amount of total phenolics in the plant extracts was determined with the Folin-Ciocalteau reagent using the method of Spanos and Wrolstad,[20] as modified by Lister and Wilson. Choi YH, Pezzuto JM, Kinghorn AD, Farnsworth NR. You do this on the Parameters tab. Food Sci. Apart from the advantage of being employed for the spectrophotometric quantitation of total antioxidant capacity, the determination of the total antioxidant capacity by phosphomolybdate method also employs cost-effective reagents. Total polyphenol content of Pleurotus ostreatus. These results seem to validate the basis for the therapeutic use of the extract in traditional medicine because correlations are known to exist between the reductive ability of a compound and its antioxidant activity, although it should be of note that a reductant is not necessarily an antioxidant, but an antioxidant is commonly a reductant.[35]. Only, the aqueous extracts of dry samples were not positively correlated to the concentration of the polyphenols. Food Chem. doi: 10.1016/j.foodchem.2009.04.016. The absorbance was measured at 700 nm. Although all extracts showed chelating activity, some of them had higher values, for example EC50 = 1932.06 0.95 mg GAE/L for the methanol extract obtained by boiling of dried primordium. The best extract with the reducing power assay was aqueous extract of dry fruiting body obtained by boiling. Estimation of total flavonoid content in propolis by two complementary colorimetric methods. Paste experimental data into the box on the right.
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how to calculate ec50 value for antioxidant activity using
