Sunflower oil was supplied from a local market. Butyric anhydride (>98%), acetic anhydride (>98%), ferrous sulfate, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, barium chloride, sesamol (>98%), and ammonium thiocyanate were supplied from Sigma-Aldrich Company (St. Louis, MO). Methanol, hydrochloric acid, chloroform, and n-heptane were supplied from Merck Company (Darmstadt, Germany). Hexanoic anhydride (>97%) was supplied from Acros Company (New Jersey, USA).
2 2 diphenyl 1 picrylhydrazyl radical dpph
Manufactured by Merck Group
Sourced in United States, Germany, Italy, Spain
2,2-diphenyl-1-picrylhydrazyl radical (DPPH•) is a stable free radical compound commonly used in biochemical research. Its core function is to serve as a reagent for evaluating the antioxidant activity of various substances.
Automatically generated - may contain errors
Lab products found in correlation
Gallic acid, by Merck Group (34 mentions)
Folin ciocalteu reagent, by Merck Group (17 mentions)
Quercetin, by Merck Group (12 mentions)
Ascorbic acid, by Merck Group (11 mentions)
Caffeic acid, by Merck Group (9 mentions)
Sinapic acid, by Merck Group (9 mentions)
Methanol, by Merck Group (9 mentions)
Catechin, by Merck Group (9 mentions)
6 hydroxy 2 5 7 8 tetramethylchroman 2 carboxylic acid trolox, by Merck Group (8 mentions)
Trolox, by Merck Group (8 mentions)
Chlorogenic acid, by Merck Group (7 mentions)
Acetic acid, by Merck Group (7 mentions)
Folin ciocalteu s phenol reagent, by Merck Group (6 mentions)
Acetonitrile, by Merck Group (6 mentions)
Potassium persulfate, by Merck Group (6 mentions)
Ferulic acid, by Merck Group (6 mentions)
Rutin, by Merck Group (6 mentions)
P coumaric acid, by Merck Group (5 mentions)
L ascorbic acid, by Merck Group (5 mentions)
Formic acid, by Merck Group (5 mentions)
93 protocols using 2 2 diphenyl 1 picrylhydrazyl radical dpph
1
Sunflower Oil Modification and Antioxidant Assay
2
Radical Scavenging Activity of Enterobactin
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In vitro radical scavenging activity of enterobactin was determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical (Sigma-Aldrich) following the method described by Brand-Williams et al. (1995) [47 ]. An aliquot of 900 μL of pure enterobactin or ascorbic acid at different concentrations (1–50 μM) were mixed with 100 μL of 1 mM DPPH in methanol. Methanol was used as control for zero scavenging activity. Solutions were incubated in dark at room temperature for 30 min. As an indication of DPPH reduction, absorbance at 517 nm was measured. The percentage of reduced DPPH was calculated using the following equation:
To estimate the ability of enterobactin to scavenge radicals when chelating iron, 5 μM enterobactin aliquots with or without 5 μM FeCl3 were mixed with 100 μL of 1 mM DPPH. Absorbance was measured at 517 nm after 30 min of incubation at room temperature. FeCl3 was used as a negative control to confirm that iron did not react with DPPH.
To estimate the ability of enterobactin to scavenge radicals when chelating iron, 5 μM enterobactin aliquots with or without 5 μM FeCl3 were mixed with 100 μL of 1 mM DPPH. Absorbance was measured at 517 nm after 30 min of incubation at room temperature. FeCl3 was used as a negative control to confirm that iron did not react with DPPH.
3
Evaluating Antioxidant and Antibacterial Potential
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Ciprofloxacin hydrochloride, Folin–Ciocalteu’s phenol reagent, gallic acid, sodium carbonate, formic acid (HPLC), acetonitrile (HPLC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), (R)-(+)-6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), silver nitrate (AgNO3), and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (Steinheim, Germany). 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) was obtained from Merck (Darmstadt, Germany). Dulbecco’s Modified Eagle’s Medium/Dulbecco’s Minimum Essential Media (DMEM) and penicillin-streptomycin solution were purchased from Biological Industries Israel Beit Haemek Ltd. (Beit Haemek, Israel). Mueller-Hinton broth was supplied by Oxoid (Basingstoke, UK). Fetal bovine serum and trypsin/ethylenediaminetetraacetic acid (EDTA) solution were provided by Biochrom (Berlin, Germany). All other chemicals and reagents were of analytical grade. Ultra-pure water was obtained from SG Water Ultra Clear TWF water purification system (Barsbüttel, Germany).
4
Phytochemical Analysis of Medicinal Plants
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All the standards, reagents, and solvents used through experiments were of analytical grade. Apigenin, diosmetin, hyperoside, rutin, quercitrin, luteolin-7-O-glucoside, Apigenin-7-O-glucoside, p-coumaric acid, caffeic acid, rosmarinic acid, and chlorogenic acid standards were purchased from Extrasynthese (Genay, France), and avicularin from Chromadex (Santa Ana, CA, USA). Acetic acid and acetonitrile were purchased from Sigma-Aldrich GmbH (Buchs, Switzerland), and ethanol from Vilniaus degtine, AB (Vilnius, Lithuania). Potassium persulfate, sodium acetate trihydrate, iron (III) chloride hexahydrate, and 2,4,6-tri(2-pyridyl)-S-triazine (TPTZ), 2,2-diphenyl-1-picrylhydrazyl (DPPH·) radical, and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) were obtained from Sigma-Aldrich (Steinheim, Germany) and Trolox from Fluka Chemika (Buchs, Switzerland). Folin-Ciocalteu reagent, gallic acid monohydrate, aluminum chloride hexahydrate, sodium carbonate, hexamethylenetetramine, copper chloride, and neocuproine were purchased from Sigma-Aldrich GmbH (Buchs, Switzerland). Deionized water used in HPLC and for the samples preparation produced by the Crystal E high-performance liquid chromatography (HPLC, Adrona SIA, Riga, Latvia) water purification system.
5
Antioxidant Screening of Biomolecules
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λ-DNA was purchased from Bangalore Genei, India. Agarose, H2O2, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), dimethyl sulfoxide (DMSO) were purchased from Sisco Research Laboratories. FeSO4·7H2O, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, ascorbic acid, Tris base, ethidium bromide (EtBr), ethylenediaminetetraacetic acid (EDTA), Enhanced Chemiluminescence solution (ECL), vitexin, isovitexin and quercetin were purchased from Sigma-Aldrich Chemicals Co. (St. Louis, MO, USA). Polyvinylidene fluoride membrane (PVDF) was purchased from pall corporation, (New York, USA). BCA protein assay kit was purchased from Thermo Fisher Scientific (Massachusetts, USA). All other chemicals used were of analytical grade.
6
Antioxidant and Elemental Analysis
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All chemicals and reagents were of analytical grade and were used without further purification. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical, ascorbic acid, boric acid, gallic acid, D-glucose, phenol, hydrochloric acid (HCl), nitric acid (HNO3), sodium carbonate (Na2SO4), aluminum chloride (AlCl3), sulfuric acid (H2SO4), and sodium hydroxide (NaOH) were purchased from Sigma-Aldrich (USA), while ethanol (96%) and multielement standard solution (ICP TraceCERT) were purchased from Merck (Darmstadt-Germany). Ultrapure water was used throughout the sample preparations and measurements. Standard solutions of each gallic acid, ascorbic acid, and quercetin were prepared in ethanol.
7
Synthesis and Radical Scavenging Evaluation
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10-Undecenoic acid, 1,1’-azobis(cyclohexanecarbonitrile) (ABCN), hydroxybenzotriazole (HOBt) and 1-ethyl-3-(3’-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl), cysteamine hydrochloride, cinnamic acid, sinapic acid, ferulic acid, p-coumaric acid, caffeic acid, α-tocopherol (α-TP), tert-butylhydroquinone (TBHQ), linoleic acid (LA) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical were purchased from Sigma–Aldrich (St.Louis, USA), and pre-coated TLC plates (silica gel 60 F254) were purchased from Merck (Darmstadt, Germany). All solvents were purchased from Sd Fine Chemicals (Mumbai, India) and were of the highest grade of purity available.
8
Phytochemical Evaluation of Plant Extracts
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Acetone, methanol, ethyl acetate, hydrochloric acid, sodium hydroxide, sodium carbonate, sodium molybdate and sodium nitrite were obtained from POCh (Gliwice, Poland). Caffeic and chlorogenic acids were purchased from Carl Roth GmbH Co, Germany, Folin-Ciocalteu reagent and aluminum chloride from Merck (Darmstadt, Germany), dimethylosulfoxide (DMSO) from Ubichem Ltd. (Hampshire, England), 2,2-diphenyl-1-picrylhydrazyl (DPPH radical) from Sigma-Aldrich, USA, and butylhydroxyanisole (BHA) from Fluka, France. Absorbance was measured with a UV/VIS Lambda 35 spectrophotometer (Perkin-Elmer, USA).
9
In Vitro Digestion Protocol
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Cellulose membrane for dialyzing (avg. flat width 25 mm, molecular weight cut-off = 14,000) and digestive enzymes such as α-amylase (TDF-100 A, 24,975 U/mL), mucin from the porcine stomach—type II, pepsin from the porcine gastric mucosa (250 U/mg solid), pancreatin from the porcine pancreas (8 × USP specifications), porcine bile extract, and reagents such as sodium dodecyl sulfate—ACS reagent, sodium bicarbonate ≥ 99.5%, (±)-6-hydroxy-2,5,7,8-tetramethyl-chromane-2-carboxylic acid (Trolox), 2,2-diphenyl-1-picrylhydrazyl (DPPH• radical), 2,2′-Azino-bis (3-ethylbenzothiazoline-6-sulfonic) acid, diammonium salt (ABTS+• radical), acetonitrile (HPLC) and sodium hydroxide pellets ≥ 98.0% (NaOH) were provided by Sigma-Aldrich (St. Louis, MO, USA).
Reagents such as disodium hydrogen phosphate anhydrous pure p.a. ≥ 99.0% (Na2HPO4), dipotassium hydrogen phosphate (K2HPO4), sodium chloride pure p.a. ≥ 99.9% (NaCl), and di-sodium wersenate standard solution 0.01 mol/L were supplied from Chempur (Piekary Śląskie, Poland). In addition, Honeywell Fluka (Seelze, Germany) provided pure hydrochloric acid p.a. ACS reagent 37% (HCl) and potassium peroxodisulfate ≥ 99.0%, and Avantor (Gliwice, Poland) provided formic acid 98–100% CZDA, ethanol 96% CZDA and methanol (HPLC grade).
Reagents such as disodium hydrogen phosphate anhydrous pure p.a. ≥ 99.0% (Na2HPO4), dipotassium hydrogen phosphate (K2HPO4), sodium chloride pure p.a. ≥ 99.9% (NaCl), and di-sodium wersenate standard solution 0.01 mol/L were supplied from Chempur (Piekary Śląskie, Poland). In addition, Honeywell Fluka (Seelze, Germany) provided pure hydrochloric acid p.a. ACS reagent 37% (HCl) and potassium peroxodisulfate ≥ 99.0%, and Avantor (Gliwice, Poland) provided formic acid 98–100% CZDA, ethanol 96% CZDA and methanol (HPLC grade).
10
Antioxidant Capacity and Phenolic Analysis
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All reagents or solvents used were of analytical, HPLC or HPLC-MS grade. Absolute ethanol, acetone, aluminum nitrate nonahydrate (Al(NO3)3·9H2O), hydrochloric acid (HCl), L-ascorbic acid, sodium hydroxide (NaOH), and sodium nitrite (NaNO2) were obtained from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, formic acid, leucine enkephalin and methanol, as well as two standards, rutin and cyanidin 3-O-glucoside, were purchased from Sigma-Aldrich (St. Louis, MO, USA). The iTaqTM Universal SYBR® Green Supermix was purchased from Bio-Rad Laboratories (Hercules, CA, USA).
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