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7 protocols using aluminum chloride

1

Antioxidant and Hepatoprotective Assays

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Adenine, aluminum chloride, potassium hydroxide, sodium nitrate, and Folin–Ciocalteu reagent were purchased from Carl-Roth (Germany). 2, 2-Diphenyl-1-picrylhydrazyl (DPPH), (2′, 2′-azobis (2-methylpropionamidine) dihydrochloride) (AAPH), gallic acid, quercetin, ascorbic acid, Tris, hydrogen peroxide, trichloroacetic acid, ferric chloride, and hydrogen chloride were purchased from Sigma–Aldrich (Germany). Captopril was purchased from Denk Pharma GmbH and Co., KG (Germany). Thiobarbituric acid was from Fluka. Sodium chloride and potassium chloride were purchased from BDH Chemicals Ltd. (England). Creatinine, urea, aspartate aminotransferase (AST), and alanine aminotransferase (ALT) kits were purchased at SGM Italia (Roma). Sodium thiopental was from Samarth Life Sciences Pvt Ltd. (Mumbai).
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2

Phytochemical Characterization of Extracts

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Acetone, Hydrochloric acid (37%), Folin–Ciocâlteu reagent, methanol and ethanol were purchased from Merck (Darmstadt, Germany). 2,2-diphenyl-1-(2,4,6-trinitro-phenyl) hydrazine (DPPH), 2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox) (97%), ferric chloride, sodium carbonate, 2,4,6-Tris(2-pyridyl)-S-triazine (TPTZ) (≥99%), dimethyl sulfoxide (DMSO) (≥99%), 3,4-Dihydroxy-L-phenylalanine (L-DOPA) (≥98%), 6-hydroxy-diammonium 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) (>98%), mushroom tyrosinase, phosphate buffer, and kojic acid were purchased from Sigma (Sigma Aldrich Chemie GmbH, Schnelldorf, Germany). Aluminum chloride (≥98%) was acquired from Carl Roth (Karlsruhe, Germany). Water was of Milli-Q-quality. All solvents were of LC grade and all reagents were of analytical grade.
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3

Comprehensive Chemical Inventory for Research

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Hexamethylenetetramine (≥99%), lactic acid (≥98%) and aluminum chloride (98%) were obtained from Carl Roth Gmbh & Co Kg (Karlsruhe, Germany). 2,2-diphenyl-1-picrylhydrazyl (DPPH) (99%), chlorogenic acid (≥95%), ferulic acid (≥99%), salicylic acid (≥99%), methanol (≥99.9%), hesperidin (≥90%), benzoic acid (≥97%), myricetin (≥96%), rutin (95%), naringenin (≥95%), coumarin (≥99%), caffeic acid (≥98%), vanillic acid (≥97%), gallic acid (≥98%), ellagic acid (≥97%), and syringic acid (≥97%), of analytical grade were obtained from Sigma-Aldrich Corp (Taufkirchen, Germany). Folin-Ciocalteu reagent, trifluoracetic acid (99%) and acetonitrile (≥99.8%) were supplied by Merck KGAA (Darmstadt, Germany). MRS with Tween 80 broth was obtained from Biolife Italiana S. r. l. (Milan, Italy). Coumaric acid and luteolin were obtained from: Chromadex, Inc. (Los Angeles, CA, USA). Sodium carbonate and acetic acid (99.9%) were bought from Reachem S.r. o. (Bratislava, Slovakia). Bidistilled water was prepared by means of distillation apparatus Thermo Fisher Scientific, Inc. (Fremont, CA, USA).
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4

Antioxidant Capacity Evaluation Protocol

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All chemicals utilized in this study were of analytical reagent grade. Quercetin, gallic acid, β-carotene, and DPPH (1,1-diphenyl-2-picrylhydrazyl) were acquired from Sigma-Aldrich (St. Louis, MA, USA). Folin–Ciocalteu reagent was supplied by AppliChem (Darmstadt, Germany). Sodium sulfate anhydrous, sodium carbonate, sodium hydroxide, and aluminum chloride were purchased from Carl Roth (Karlsruhe, Germany). Methanol, acetone, and petroleum ether were sourced from VWR Chemicals BDH (Fontenay-sous-Bois, France), while sodium nitrite and ascorbic acid were obtained from Centrohem (Stara Pazova, Serbia). Double-distilled water was used throughout all experiments.
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5

Multivariate Analytical Characterization

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Hexametiltetraamine (≥99%) and aluminum chloride (98%) were obtained from Carl Roth Gmbh & Co Kg (Karlsruhe, Germany). The 2,2-diphenyl-1-picrylhydrazyl (DPPH) (99%), methanol (≥99.9%), rutin (95%), hydrochloric (≥37%) and nitric (≥65%) acids of analytical grade were obtained from Sigma-Aldrich Corporation (Taufkirchen, Germany). Folin-Ciocalteu reagent was supplied by the Merck (Darmstadt, Germany). Acetonitrile (99.8%) was obtained from Avantor Performance Materials (Gliwice, Poland). Sodium carbonate and acetic acid (99.9%) were bought from Reachem S. r. o. (Bratislava, Slovakia). The standard mixture solution of multiple microelements (As, Ba, Ca, Cd, Co, Cr, Cu, Fe, K, Mg, Mn, Na, Ni, P, Pb, Se, Sr, V and Zn) in 2% nitric acid was obtained from CPAchem (Bulgaria). Bidistilled water was prepared by means of distillation apparatus Thermo Scientific (Fremont, CA, USA).
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6

Phytochemical Analytical Techniques

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All chemicals used in this study were of analytical reagent grade. Quercetin, gallic acid and DPPH (1,1-diphenyl-2-picrylhydrazyl) were purchased from Sigma-Aldrich (St. Louis, MA, USA). Hydrochloric acid, sulfuric acid, hydroquinone and potassium dihydrogen phosphate were obtained from Merck (Darmstadt, Germany). Folin–Ciocalteu’s reagent was supplied from AppliChem (Darmstadt, Germany). Sodium sulfite, sodium carbonate, sodium hydroxide, aluminum chloride and ammonium heptamolybdate were purchased from Carl Roth (Karlsruhe, Germany). Methanol and n-hexane were from VWR Chemicals BDH (Fontenay-sous-Bois, France), while sodium nitrite was from Centrohem (Stara Pazova, Serbia). In all experiments, doubly distilled water was used.
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7

Antioxidant and Tyrosinase Inhibition Assays

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The reagents used in this study were: sodium carbonate, ferric chloride, 6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox) (97%), diammonium 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) (>98%), 2,2-diphenyl-1-(2,4,6trinitro-phenyl) hydrazine (DPPH), tri-2-pyridyl-s-triazine (TPTZ) (≥99%), dimethyl sulfoxide (DMSO) (≥99%), phosphate buffer, mushroom tyrosinase, 3,4-dihydroxy-L-phenylalanine (L-DOPA) (≥98%), and kojic acid were purchased from Sigma (Sigma Aldrich Chemie GmbH, Schnelldorf, Germany). Folin-Ciocâlteu reagent, hydrochloric acid (37%), acetone, ethanol, and methanol were purchased from Merck (Darmstadt, Germany). Aluminum chloride (≥98%) was purchased from Carl Roth (Karlsruhe, Germany). All reagents were of analytical grade and all solvents were of LC grade. Water was of Milli-Q-quality.
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