To evaluate the scavenging activity of DPPH free radicals, Govarthanan et al. (2015) [51 (link)] method was used with a slight modification. Initially, a methanolic extract of the sample (diluted 1:10) or gallic acid at various concentrations (ranging from 6.25 to 200 μg/mL) was mixed with 100 μL of 0.1 mM methanolic DPPH solution (Sigma-Aldrich, USA), followed by incubation for 20 min at room temperature in the dark. The resulting mixture was then measured at a wavelength of 517 nm using a microplate reader (BioTek Synergy H1 Multimode Reader, Agilent Technologies, USA). The negative control was sterile distilled water. The scavenging activity of the extracts was determined using the following formula: inhibition (%) = [(absorbance of control – absorbance of sample) x 100]/absorbance of control, where absorbance control is the absorbance of DPPH solution without extract and Abs sample is the absorbance of the sample with DPPH solution.
Methanolic dpph solution
Manufactured by Merck Group
Sourced in United States
Methanolic DPPH solution is a laboratory reagent used as a free radical scavenging assay. It is a solution of the stable free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) in methanol. The DPPH solution exhibits a deep purple color that can be spectrophotometrically measured at 517 nm. The degree of discoloration indicates the free radical scavenging ability of a sample.
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Lab products found in correlation
4 protocols using methanolic dpph solution
1
Evaluation of DPPH Radical Scavenging Activity
2
DPPH Radical Scavenging Assay for Kombucha
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The 2,2-diphenyl-1-picrylhydrazyl (DPPH) method described by Von Gadow et al. (22 (link)) was used to measure free radical scavenging activity (RSA) of the beverages. Kombucha samples were centrifuged at 1370×g for 10 min (Mistral 2000; MSE) and filtered through a filter with a pore diameter of 0.45 µm (GVS Filter Technology). A volume of 4 mL of methanolic DPPH solution (Sigma Aldrich, Merck) was added to 100 µL of the prepared samples and the reaction mixture was kept in the dark for 30 min. The absorbance of the samples was measured with a spectrophotometer (UV-5100; SOIF, Shanghai, PR China) at 516 nm and the results were expressed in ascorbic acid equivalents (AAE) in µmol/mL.
3
Antioxidant Activity of P. acutifolium Essential Oil
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The EO of P. acutifolium (150 µL) at different concentrations (0.1; 0.5; 1.0; 5.0 and 10.0%) reacted with 850 µL of 0.01 mM DPPH methanolic solution (Sigma Aldrich St Louis, MO, USA). The reaction lasted 30 min, and the absorbances were read at 517 nm. Each test was carried out in triplicate [40 (link)].
4
DPPH Assay for Antioxidant Evaluation
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A DPPH assay was used to detect free radical scavenging activity in different concentrations of the samples. Samples of 40 µL of FER at concentrations of 20, 10, 5, 2.5, and 1.25 (mM), or 625, 313, 156, 78, and 39 (µM), or DFER at concentrations of 20, 10, 5, 2.5, or 1.25 (mM), blank and standard, were added to 200 µL of DPPH methanolic solution (Sigma, Burlington, VT, USA) in 96-well plates. After being incubated in the dark for 1 h, the absorbance under 517 nm was read. The absorbance of the control solution was in the range of 0.8 ± 0.1. The free radical scavenging activity was calculated using the following equation:
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