The Anzunbaengi wheat cultivar grown in Jinju, South Korea, was used to prepare GWB in this study. Gamma-Aminobutyric acid (GABA), catechin, gallic acid, 2,2-diphnyl-1-picrylhydrazyl (DPPH), potassium persulfate, trichloroacetic acid, potassium persulfate, aluminium chloride, and sodium hypochlorite were purchased from Sigma-Aldrich (St. Louis, MO, USA). Sodium hydroxide, methanol, and phenol were obtained from Dae Jung Co., Ltd. (Seoul, South Korea).
Potassium persulfate
Manufactured by Merck Group
Sourced in United States, Germany, Italy, Poland, Spain, Portugal, Australia, United Kingdom, Canada, India, Switzerland, France, China, Mexico, Singapore, Hungary, Slovenia, Israel, Brazil
Potassium persulfate is an oxidizing agent used in various laboratory and industrial applications. It is a white, crystalline solid that is soluble in water. Potassium persulfate is commonly used as an initiator in free-radical polymerization reactions, as an oxidizing agent, and as a bleaching agent.
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Lab products found in correlation
Gallic acid, by Merck Group (185 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (140 mentions)
Sodium carbonate, by Merck Group (125 mentions)
Trolox, by Merck Group (104 mentions)
Methanol, by Merck Group (102 mentions)
Hydrochloric acid (hcl), by Merck Group (93 mentions)
Folin ciocalteu reagent, by Merck Group (90 mentions)
Sodium hydroxide (naoh), by Merck Group (89 mentions)
2 2 azinobis 3 ethylbenzothiazoline 6 sulfonic acid (abts), by Merck Group (88 mentions)
Quercetin, by Merck Group (86 mentions)
Ascorbic acid, by Merck Group (72 mentions)
Ethanol, by Merck Group (70 mentions)
Acetic acid, by Merck Group (67 mentions)
Formic acid, by Merck Group (64 mentions)
Catechin, by Merck Group (56 mentions)
Sodium acetate, by Merck Group (53 mentions)
Acetonitrile, by Merck Group (46 mentions)
2 2 azino bis 3 ethylbenzothiazoline 6 sulfonic acid diammonium salt, by Merck Group (44 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (43 mentions)
2 2 azino bis, by Merck Group (40 mentions)
671 protocols using potassium persulfate
1
Antioxidant-rich Anzunbaengi Wheat GWB
2
Trolox Equivalent Antioxidant Capacity Assay for Microalgae
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The Trolox equivalent antioxidant capacity (TEAC) assay was used in this study with some modifications31 (link),36 (link). Briefly, lyophilized microalgal samples were extracted with 75% ethanol using a mini-beadbeater (BioSpec Products, USA) at the highest speed for 5 cycles, 20 s each, and cooled down on ice for 2 min between two cycles. Under dim light, ABTS (A-1888, Sigma-Aldrich, USA) and potassium persulfate (216,224, Sigma-Aldrich, USA) were dissolved in water to generate the blue-green ABTS·+ (7 mM ABTS and 2.45 mM potassium persulfate). This mix was allowed to react in the dark at room temperature for 16 h and its absorbance at 734 nm (OD734) was adjusted to 0.70 ± 0.05 before use. To build the standard curve of OD734 against Trolox quantity, different amounts of Trolox (238813, Sigma-Aldrich, USA) were mixed with the freshly prepared ABTS·+ solution to reduce the color, and the resulting OD734 readings were recorded. The antioxidation capacity of the samples was expressed as equivalent amount of Trolox per mg dry biomass.
3
Produced Water Treatment via Magnetite-Based Hydrogel
4
ABTS Radical Cation Antioxidant Assay
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The ABTS radical cation assay based on the method of Re et al. (1999) (link) was used to determine antioxidative capacity. Briefly, ABTS radical cation was prepared by reacting a 7 mM ABTS (Roche Diagnostics) stock solution with a known concentration of potassium persulfate (Sigma-Aldrich) so that the final concentration of potassium persulfate is 2.45 mM. The mixture was left to stand in the dark at room temperature for 16 h before use. A working ABTS•+ solution with an absorbance of 0.70 ± 0.02 at 734 nm was prepared by diluting with Phosphate-Buffered Saline (PBS) buffer. Ten microlitres of protein extract or standard was added to 190 µL of ABTS•+ and mixed thoroughly in a microplate well before being read at 734 nm (Varioskan LUX, Thermo Fisher Scientific). The antioxidant capacity was calculated as: , where Acontrol is the absorbance of the working ABTS•+ solution and Asample is the absorbance of the samples with ABTS•+ reagent. Gallic acid was used as a standard.
5
Antioxidant Capacity Determination
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ABTS (2,2’-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) di-ammonium salt) (Sigma-Aldrich, St. Louis, MO, USA), potassium persulfate (Sigma-Aldrich, St. Louis, MO, USA), L-ascorbic acid (Sigma-Aldrich, St. Louis, MO, USA), methanol (Merck, Darmstadt, Germany), and distilled water were used. All reagents were of analytical grade.
6
HPLC Analysis of Amino Acids and Antioxidants
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All the chemicals used in the experiments were of analytical grade. Acetonitrile (ACN) and methanol (MeOH) were HPLC gradient-grade (Merck KGaA (Darmstadt, Germany). Per-chloric (60%) and hydrochloric acid (37%) were from J.T. Baker (NJ, United States). Sigma-Aldrich Chemie (Sant Quentin Fallavier, France) provided Alcalase 2.4 L FG (CAS no. 9014-01-1), ortho-phthaldialdehyde reagent (OPA), l-glutathione, borate buffer, formic acid (for LC-MS LiChropur™), ammonium acetate, ammonium formate, phenyl isothiocyanate (PITC), triethylamine (TEA), pure standards for 19 amino acids, taurine, hypotaurine and homotaurine, trolox (6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid), 2,20-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), α-diphenyl-β- picrylhydrazyl (DPPH), potassium persulfate, Folin-Ciocalteu’s reagent, and gallic acid were purchased from Sigma-Aldrich (St. Louis, MO). The SIRT1 Direct Fluorescent Screening Assay Kit was from ABNOVA (Cat # KA1366, Taiwan).
7
Olive Pomace Bioactive Extraction
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Olive pomace (OP), obtained after a three-phase process (Taggiasca cultivar), was supplied by a local producer (Liguria, Italy). The fresh OP was collected and dried in a laboratory oven at 50 °C until there was constant moisture, and then stored in dark conditions at room temperature.
Ethanol, mEthanol, acetonitrile, acetic acid, and sodium carbonate were purchased from Carlo Erba Reagents (Cornaredo, Milan, Italy), while Folin–Ciocalteu’s reagents included caffeic acid, 2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), potassium persulfate, 6-hydroxy-2,5,7,8-tetramethychroman-2-carboxylic acid (Trolox), ionophore A23187, NMDA, and neutral red solution from Merck-Sigma-Aldrich (Milan, Italy). Calcium Green™-1AM, Neurobasal™ medium, B-27 supplement, Glutamax®, penicillin, and streptomycin solution were purchased from Life Technologies Italia (Milan, Italy). DMEM, FBS, MEM non-essential amino acids 100×, and trypsin-EDTA 1× solutions were from Euroclone (Milan, Italy).
Ethanol, mEthanol, acetonitrile, acetic acid, and sodium carbonate were purchased from Carlo Erba Reagents (Cornaredo, Milan, Italy), while Folin–Ciocalteu’s reagents included caffeic acid, 2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), potassium persulfate, 6-hydroxy-2,5,7,8-tetramethychroman-2-carboxylic acid (Trolox), ionophore A23187, NMDA, and neutral red solution from Merck-Sigma-Aldrich (Milan, Italy). Calcium Green™-1AM, Neurobasal™ medium, B-27 supplement, Glutamax®, penicillin, and streptomycin solution were purchased from Life Technologies Italia (Milan, Italy). DMEM, FBS, MEM non-essential amino acids 100×, and trypsin-EDTA 1× solutions were from Euroclone (Milan, Italy).
8
Preparation and Characterization of Nanoparticles
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Zinc nitrate hexahydrate (98%), dimethylsulfoxide (DMSO, 96%), ascorbic acid (99%), osmium tetroxide (99.8%), glutaraldhyde (50% in water), phosphate-buffered saline (pH 7.4), ethanol (98%), methanol (99.85%), potassium persulfate (98.5%), 2,2-diphenyl-1-picrylhydrazyl (DPPH, 95%) and 2,20 -azino-bis [3-ethyl benzo thiazoline-6-sulphonic acid] (ABST, 98%) were purchased from Sigma-Aldrich (Hamburg, Germany).
9
Comprehensive Analytical Methods for Plant Phenolics
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Ethanol, hydrochloric acid (37%), and acetone of HPLC grade were purchased from Scharlab Chemie (Barcelona, Spain). Gallic acid, Folin-Ciocalteu reagent, epicatechin, vanillin, polyethylene glycol (8000 Da), polyethylene glycol (4000 Da), twin20 (1228 Da), ethylene glycol (62 Da), dextran (50,000 Da), iron(III) chloride, sodium carbonate, hydrogen peroxide, 4-dimethylaminocinnamaldehyde (DMAC), 6-hydroxy-2,5,7,8-tetramethyl-chromane-2-carboxylic acid (Trolox), potassium persulfate, 2,2’-azinobis(3-ethylbenzothiazoline-6-sulphonic acid) and diammonium salt (ABTS)were purchased from Sigma-Aldrich (Saint Louis, MO, USA). Sodium dihydrogen phosphate dihydrate and dipotassium hydrogen phosphate were obtained from Merck (Hesse, Darmstadt, Germany).
MEthanol (99.99%), formic acid, acetonitrile, and butanol of HPLC grade were supplied by Fisher Scientific (Midlands, Leicestershire, UK). Ultrapure water (18.2 MΩ/cm) was obtained from a Millipore system (Millipore, Billerica, MA, USA). Promod 439 LTM enzyme was a kind donation of Biocatalysts Ltd. (Wales, Cardiff, UK).
MEthanol (99.99%), formic acid, acetonitrile, and butanol of HPLC grade were supplied by Fisher Scientific (Midlands, Leicestershire, UK). Ultrapure water (18.2 MΩ/cm) was obtained from a Millipore system (Millipore, Billerica, MA, USA). Promod 439 LTM enzyme was a kind donation of Biocatalysts Ltd. (Wales, Cardiff, UK).
10
Antioxidant Potential of P. yezoensis
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The free radical scavenging abilities of P. yezoensis extract and porphyra 334 were investigated using the ABTS 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation decolorization test. The interaction of 2.45 mM potassium persulfate (7727-21-1, Sigma-Aldrich, St. Louis, MO, USA) and 7 mM ABTS (30931-67-0, Sigma-Aldrich, St. Louis, MO, USA) in ultra-purified water (UPW) formed the ABTS+ cation radicals, which were then kept at 20 °C in the dark for 12 to 16 h before use. UPW was then added to the ABTS+ solution for dilution, resulting in an absorbance of 0.700 at 734 nm. P. yezoensis extract and porphyra 334 were added to the ABTS+ solution at various concentrations. The positive control was 50 μm vitamin C. Samples were tested at an absorbance of 734 nm. The following equation was used to determine the radical scavenging activity:
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