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8 protocols using sodium carbonate anhydrous

1

Spectrophotometric Analysis of Antioxidant Compounds

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Acetic acid (99.5%), aluminum chloride hexahydrate, copper(II) chloride dihydrate, disodium hydrogen phosphate dihydrate, 96% ethanol, 36% hydrochloric acid, iron(III) chloride hexahydrate, methanol, phosphoric acid, potassium persulfate, potassium dihydrogen phosphate, potassium hexacyanoferrate(III), sodium acetate anhydrous, sodium carbonate anhydrous, sodium hydroxide, sodium nitrite, and trichloroAcetic acid were purchased from Chempur, Poland. Neocuproine was delivered by J&K Scientific, Germany. Folin–Ciocalteu reagent, acetonitrile, iron(II) sulfate heptahydrate, gallic acid were supplied by Merck, Germany, whereas rutin trihydrate by Roth, Germany. ABTS (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), DPPH (2,2-diphenyl-1-picrylhydrazyl), TPTZ (2,4,6-tris(2-pyridyl)-s-triazine), Trolox (6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid), caffeine were purchased from Sigma-Aldrich, USA. All the chemicals were of analytical grade.
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2

Antioxidant Potential of Medicinal Plants

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The following amino acids were used for the synthesis: l-threonine (Thr, >99 wt%, pure), l-methionine (Met, >99 wt%, pure), l-arginine (Arg, >99 wt%, pure), l-isoleucine (Ile, >99 wt%, pure), purchased from Roth, and triethanolamine (TEA) (>99 wt%, pure) from Acros Organics. DPPH (2,2-diphenyl-1-picrylhydrazyl), TPTZ (2,4,6-tris(2-pyridyl)-s-triazine) and Trolox (6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid) were purchased from Sigma-Aldrich (USA). Folin–Ciocalteu phenol reagent, iron(ii) sulfate heptahydrate and gallic acid were obtained from Merck (Germany), whereas ethanol, 99.5% acetic acid, sodium acetate anhydrous, 36% hydrochloric acid, iron(iii) chloride hexahydrate, copper(ii) chloride dihydrate, and sodium carbonate anhydrous were obtained from Chempur (Poland). Neocuproine was delivered by J&K Scientific (Germany).
The research plant material consisted of dry and cut L. clavatum herbs (Dary Natury, Poland), C. islandica (Kawon, Poland) and D. fullonum leaves (NatVita, Poland).
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3

Antioxidant Capacity Assays Protocol

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DPPH (2,2-diphenyl-1-picrylhydrazyl), TPTZ (2,4,6--tris(2-pyridyl)-s-triazine), ABTS (2,2′-azino-bis(3--ethylbenzothiazoline-6-sulfonic acid) and Trolox (6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxy-lic acid) were purchased from Sigma-Aldrich, USA; gallic acid, Folin-Ciocalteu reagent and iron(II) sulfate heptahydrate from Merck, Germany; rutin trihydrate from Roth, Germany; ferrozine from Serva Electrophoresis, Germany; neocuproine from J&K Scientific, Germany. Methanol, potassium persulfate, 36% hydrochloric acid, iron(III) chloride hexahydrate, 99.5% acetic acid, sodium acetate anhydrous, sodium carbonate anhydrous, copper(II) chloride dihydrate, iron(II) chloride tetrahydrate, potassium chloride, sodium nitrite, aluminum chloride hexahydrate and sodium hydroxide were obtained from Chempur, Poland; ethanol from Linegal Chemicals, Poland. All reagents were of analytical grade.
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4

Antioxidant Capacity Assays Protocol

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DPPH (2,2-diphenyl-1-picrylhydrazyl), TPTZ (2,4,6--tris(2-pyridyl)-s-triazine), ABTS (2,2′-azino-bis(3--ethylbenzothiazoline-6-sulfonic acid) and Trolox (6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxy-lic acid) were purchased from Sigma-Aldrich, USA; gallic acid, Folin-Ciocalteu reagent and iron(II) sulfate heptahydrate from Merck, Germany; rutin trihydrate from Roth, Germany; ferrozine from Serva Electrophoresis, Germany; neocuproine from J&K Scientific, Germany. Methanol, potassium persulfate, 36% hydrochloric acid, iron(III) chloride hexahydrate, 99.5% acetic acid, sodium acetate anhydrous, sodium carbonate anhydrous, copper(II) chloride dihydrate, iron(II) chloride tetrahydrate, potassium chloride, sodium nitrite, aluminum chloride hexahydrate and sodium hydroxide were obtained from Chempur, Poland; ethanol from Linegal Chemicals, Poland. All reagents were of analytical grade.
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5

Comprehensive Antioxidant Analysis Protocol

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2,4,6-tripyridyl-s-triazine (TPTZ), 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2-azino-bis(3-ethylbenzo--thiazoline-6-sulphonic acid) (ABTS) were purchased from Sigma-Aldrich, USA; iron(III) chloride hexahydrate, Folin-Ciocalteu phenol reagent -from Merck, Darmstadt, Germany; 99.5% acetic acid, sodium acetate anhydrous, potassium persulfate, 36% hydrochloric acid, sodium carbonate anhydrous, ascorbic acid and applied solvents, i.e. ethanol, methanol, acetone and petroleum ether, were from Chempur, Piekary Śląskie, Poland. All reagents were of analytical grade.
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6

Antioxidant Capacity Assay Protocol

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Hydrochloric acid, anhydrous sodium carbonate, and Folin–Ciocalteu reagent were purchased from Chempur (Piekary Śląskie, Poland). Gallic acid anhydrous, 2,2-diphenyl-1-picrylhydrazyl (DPPH), L-ascorbic acid, oxalic acid, m-phosphoric acid, 6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox), and methanol were purchased from Sigma–Aldrich (Poznan, Poland). All reagents were of analytical grade.
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7

Antioxidant Adsorption on Functionalized Silica

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The mesoporous silica materials, MCM-41 (<150 μm particle size, 2.1–2.7 nm pore size, hexagonal pore morphology), SBA-15 (<150 μm particle size, 6 nm pore size, hexagonal pore morphology), SBA-NH2 (<150 μm particle size, 6 nm pore size, amine functionalized), and SBA-SH (<150 μm particle size, 6 nm pore size, thiol functionalized), were purchased from Merck and used as received. All antioxidants (i.e., GA, PCA, CGA, and 4-HBA) were of analytical grade (HPLC purity > 97.0%) and purchased from Sigma-Aldrich. All aqueous solutions were prepared using ultrapure water (18.2 MΩ·cm, Milipore Simplicity System, Billerica, MA, USA). HPLC grade chemicals were obtained from J.T. Baker (Mallinckrodt Baker, Phillipsburg, NJ, USA). Reagents for the Folin–Ciocalteu method were as follows: ethanol 96% (v/v) (Emsure, Darmstadt, Germany), anhydrous sodium carbonate (purity > 99.5%, Chempur, Piekary Śląskie, Poland), Folin–Ciocalteu reagent (analytical grade, Chempur, Piekary Śląskie, Poland).
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8

Antioxidant and Reducing Capacity Assays

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pH buffer solutions, hydrochloric acid, 0.1 M sodium hydroxide, anhydrous sodium carbonate, and Folin–Ciocalteu reagent were all analytical grade and purchased from Chempur (Piekary Śląskie, Poland). Disodium fluorescein of reagent grade, m-phosphoric acid, acetonitrile, and formic acid of HPLC gradient grade were purchased from Avantor Performance Materials Poland S.A. (Gliwice, Poland). Gallic acid anhydrous, phosphate-buffered saline (PBS), 2,2′-azobis(2-methylpropionamidine) dihydrochloride (AAPH), (±)-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox), methanol, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) were reagent or analytical grade, purchased from Sigma-Aldrich Sp. z.o.o (Poznań, Poland). Pure water (resistivity of 18.0 MΩ·cm) was also used in the research.
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