ROS scavenging potential of Ful-containing cements and CMC/Gel were determined using a 1,1-Diphenyl-2-picrylhydrazine (DPPH) assay.[98 ,99 ] DPPH solutions was prepared by dissolving DPPH (31.62 mg, Sigma-Aldrich, United States) in 80:20 v/v% solution of ethanol (400 mL, Thermo Fisher Scientific, United States) and deionized water. Respective cement samples (10 mg) were placed into DPPH solution (2 mL) and shook at 37 °C in the dark. At 1, 12, and 24 h, samples of DPPH (100 μL) were removed and absorbance was measured on a Tecan MPlex microplate reader at 517 nm. Absorbance values of DPPH solutions incubated with Ful containing cements ( ) were compared to absorbance readings of DPPH solution incubated with CMC/gel cement as shown in Equation (1) below.
2 2 diphenyl 1 picrylhydrazyl (dpph)
Manufactured by Tecan
Sourced in Switzerland
The DPPH is a compact and versatile instrument designed for the analysis of antioxidant activity. It utilizes the DPPH (2,2-diphenyl-1-picrylhydrazyl) radical assay to quantify the free radical scavenging capacity of various samples, such as plant extracts, food products, or pharmaceutical compounds. The DPPH instrument provides accurate and reliable results, making it a valuable tool for researchers and laboratories focused on investigating antioxidant properties.
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2 protocols using 2 2 diphenyl 1 picrylhydrazyl (dpph)
1
DPPH Assay for ROS Scavenging Potential
2
DPPH Radical Scavenging Assay for Lettuce
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The chain-breaking activity was measured following the bleaching rate of a stable free radical, 2,2-diphenyl-1-picrylhydrazyl (DPPH ) in the presence of the sample (Manzocco, Mastrocola, & Nicoli, 1998) . A volume of 150 μL of 6.1 × 10
-5 mol/L DPPH (Sigma-Aldrich, St. Louis, Missouri, USA) methanol solution was used. The reaction was started by the addition of 10 μL of lettuce waste extract. DPPH bleaching was followed at 515 nm (Sunrise™, Tecan, Männedorf, Switzerland) at 25 °C for 10 min. DPPH bleaching rate was proportional to sample concentration. Trolox (Sigma-Aldrich, St. Louis, Missouri, USA) was used as the standard for the calibration curve in the assay (5-150 μg/mL, R 2 = 0.999), and the antioxidant capacity was expressed as μg of Trolox equivalents (TE) per mL of extract.
-5 mol/L DPPH (Sigma-Aldrich, St. Louis, Missouri, USA) methanol solution was used. The reaction was started by the addition of 10 μL of lettuce waste extract. DPPH bleaching was followed at 515 nm (Sunrise™, Tecan, Männedorf, Switzerland) at 25 °C for 10 min. DPPH bleaching rate was proportional to sample concentration. Trolox (Sigma-Aldrich, St. Louis, Missouri, USA) was used as the standard for the calibration curve in the assay (5-150 μg/mL, R 2 = 0.999), and the antioxidant capacity was expressed as μg of Trolox equivalents (TE) per mL of extract.
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