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4 protocols using ferric chloride

1

Cell Culture and Cytotoxicity Assay

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Roswell Park Memorial Institute 1640 (RPMI 1640) culture medium and foetal bovine serum (FBS) were purchased from Gibco Invitrogen (Karlsruhe, Germany). Penicillin/streptomycin, dimethyl sulphoxide (DMSO) and salt 3-(4,5-dimethyl-2-thiazole)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) were purchased from Sigma Chemical (St. Louis, MO, USA). Folin-Ciocalteu reagent and 2,2-diphenyl-1-picryhydrazyl (DPPH) were purchased from Merck KGaA (Darmstadt, Germany). Methanol, hydrochloric acid, ethanol, ferric chloride, sodium hydroxide, chloroform, anhydrous sodium sulphate, sulphuric acid, sodium carbonate, gallic acid and ethyl acetate were purchased from Vetec Química Fina Ltda. (Duque de Caxias, RJ, Brazil).
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2

Antioxidant Capacity Evaluation Protocol

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Gallic acid, 2, 2′-azinobis-3-ethylbenzotiazoline-6-sulfonic acid (ABTS), 2, 2-diphenyl-1-picrylhydrazyl (DDPH), phosphate buffer, sodium carbonate and potassium persulfate were from Sigma-Aldrich (USA). Aluminum chloride, potassium ferricyanide and ferric chloride were from Vetec (Brazil). Calcium chloride was from Anidrol (Brazil), sodium alginate from Dinamica (Brazil) and EOs were from Ferquima (Brazil).
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3

Phytochemical and Antioxidant Assays

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We purchased 2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), acetone, acetonitrile, citric acid, DPPH, ferrous-ion-o-dianisidine, Folin–Ciocalteu phenol reagent, formic acid, gallic acid, hydrochloric acid, methanol, β-nicotinamide adenine dinucleotide 2-phosphate reduced tetrasodium salt (NADPH), ultrapure phenolic standards, 4-amino-3-hydrazino-5-mercapto-1,2,4-triazole (Purpald), sodium acetate, superoxide dismutase determination kit, 2,4,6-tris(2-pyridyl)-s-triazine (TPTZ), and 6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox) from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). Ascorbic acid, boric acid, ferric chloride, ferrous sulfate, hydrogen peroxide, potassium chloride, sodium carbonate, sodium hydroxide, and sulfuric acid were obtained from Vetec (Rio de Janeiro, RJ, Brazil). Cyanidin-3-O-glucoside, delphinidin-3-O-glucoside, delphinidin, quercetin-3-O-glucoside, 3-O-caffeoyl-quinic acid, and myricetin were purchased from Extrasynthese (Genay, France). HA, PCA, VA, isovanillic acid (iVA), ellagic acid, FA, and kaempferolwere purchased from Sigma-Aldrich (St Louis, MO, USA). The kit to analyze urine creatinine was purchased from Labtest Diagnóstica S.A. (Lagoa Santa, MG, Brazil).
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4

Quantification of Polyphenolic Compounds

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Folin Ciocalteu's reagent and arabic gum were purchased from Dinâmica Química Contemporanea (Diadema, Brazil). Vanillin, ferric chloride, ferric ammonium sulfate were purchased from from Vetec Química (São Paulo, Brazil). Fluorescein, ammonium sulfate, sodium acetate and sodium chloride were purchased from Ecibra (São Paulo, Brazil). 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid was purchased from Acros Organics (Belgium). Monobasic and dibasic sodium phosphate, sodium carbonate, hydrochloric acid, and formic acid were purchased from LabSynth (Diadema, Brazil). LC grade methanol was purchased from JT Baker (Center valley, PA, USA). Tannic acid was purchased from Ajinomoto OmniChem Division (Wetteren, Belgium). The enzymes cellulase and pectinase were obtained from Prozyn (São Paulo, Brazil). Gallic acid, catechin, caffeic acid, procyanidin B2, transresveratrol, quercetin, rutin, Tannic acid, 2,2-diphenyl-1-picrylhydrazyl, 2,2′-Azobis(2-amidinopropane) dihydrochloride, 4,6-tripryridyl-s-triazine were purchased from Sigma-Aldrich, (St. Louis, MO, USA). The water used was produced from an ultrapure Milli-Q system (Merck Millipore, Darmstadt, Germany).
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