2 4 6 tris 2 pyridyl s triazine

Methyl jasmonate, 2,2'‐azino‐bis(3‐ethylbenzothiazoline‐6‐sulphonic acid) (ABTS), HCl 37%, 2,4,6‐Tris (2‐pyridyl)‐s‐triazine (TPTZ), DPPH radical (diphenyl‐1‐picrylhydrazyl), (+)‐catechin, (−)‐epicatechin, quercetin 3‐O‐glucoside, and transresveratrol were obtained from Sigma‐Aldrich (US). L‐phenylalanine, 2‐methoxyphenol (Guaiacol), pyrocatechol, iron (III) chloride hexahydrate (FeCl₃ (6H₂O)), potassium persulfate (K₂S₂O₈), iron (II) sulfate heptahydrate (FeSO₄ (7H₂O)), sodium nitrite, Tween‐80, hydrogen peroxide 30%, Folin–Ciocalteu reagent, and polyvinylpolypyrrolidone (PVP) were purchased from Merck. Acetonitrile and rutin trihydrate were purchased from CHEMSOLUTE and Fluka, respectively. Trifluoroacetic acid (TFA) and methanol (all of HPLC grade) were obtained from VWR CHEMICALS. Milli‐Q water was acquired by SG water apparatus.

Antioxidant activity and NO concentration measuring reagents such as 2,2-Diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azobis(2-methylpropionamidine) dihydrochloride (AAPH), 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), Trolox, Folin–Ciocalteu reagent, 2,4,6-Tris(2-pyridyl)-s-triazine (TPTZ), gallic acid, quercetin, trichloroacetic acid, aluminum chloride (AlCl₃), thiazolyl blue tetrazolium bromide (MTT), and lipopolysaccharide (LPS) were purchased from Sigma-Aldrich (St. Louis, MO, USA). The mouse macrophage cell line, Raw 264.7, were obtained from the American Type Culture Collection (Manassas, VA, USA). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum, and antibiotics used Raw 264.7 cell culture were purchased from Wellgene (Daegu, Korea). All antibodies used in Western blotting were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Fu brick tea (Camellia sinensis L.) was obtained from a dark tea factory in Yiyang city, Hunan province, China. The glutinous rice (Oryza sativa var.) was bought from a local supermarket.
The compounds 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,4,6-tris (2-pyridyl)-S-triazine (TPTZ), 2,2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS), ferrozine, ascorbic acid (vitamin C), 1,10-phenanthroline, and pUC18 plasmid DNA were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). The standards: gallic acid (GA), (−)-gallocatechin (GC), theobromine (Tb), 3,4-dihydroxybenzoic acid (DbA), theophylline (Tp), (−)-epigallocatechin (EGC), 3,4-dihydroxybenzaldehyde (Dbd), caffeine, catechin (C), caffeic acid (CA), (−)-epicatechin (EC), (−)-epigallocatechin gallate (EGCG), (−)-gallocatechin gallate (GCG), and (−)-epicatechin gallate (ECG) were purchased from Yuan-ye Biotech Co. (Shanghai, China). LC-grade methanol, acetonitrile, and trifluoroacetic acid were purchased from Merck (Darmstadt, Germany). Folin–Ciocalteu reagent and all other chemicals were analytical grade and purchased from Sinopharm Chemical Reagent Co. (Shanghai, China).

The Folin-Ciocalteu reagent, isobutanol, quercetin (95% purity), sodium hydroxide (NaOH), 2,2-diphenyl-1-picryl-hydrazyl radical (DPPH), pyrocatechol violet (3,3′ acid, 4-trihydroxyfuchsone-2′-sulfonic acid), 2,4,6-tris (2-pyridyl)-S-triazine (TPTZ), hydrogen peroxide, heneicosanoic acid (C:21), cholesterol, dimethyl sulfoxide (DMSO), 2′,7′-dichlorofluorescein diacetate (DCFH-DA), dehydrated ferric chloride, ascorbic acid, and ferrozine 3-(2-Pyridyl)-5,6-di(2-furyl)-1,2,4-triazine-5′,5″-disulfonic acid disodium salt) were obtained from Sigma-Aldrich (São Paulo, Brazil). Anhydrous sodium sulfate, n-hexane, chloroform, and methyl alcohol were obtained from Anidrol (São Paulo, Brazil). Ethyl alcohol was obtained from Neon (São Paulo, Brazil). A549, IMR90, HepG2, and Caco-2 were obtained from the cell bank of Rio de Janeiro/Rio de Janeiro, Brazil.

Potassium hydroxide (KOH) was supplied from Fisher Chemicals (Fisher Scientific, Loughborough, Leicestershire, UK), while yeast extract and bactopeptone were ordered from Biolife (Milano, Italy) and sodium hydrogen phosphate heptahydrate (Na₂HPO₄·7 H₂O) was product of Acros Organics (Verona, Spain). Acetonitrile was purchased from Merck (Darmstadt, Germany), formic acid from Sigma Aldrich (Munich, Germany), cyanidin-3-O-rutinoside from Extrasynthese (Genay, France), ethanol and sodium carbonate anhydrous (Na₂CO₃) from Centrohem (Stara Pazova, Serbia), and hydrochloric acid (HCl; 36.2%) from Zorka Pharma (Sabac, Serbia). Folin–Ciocalteu reagent was purchased from Carlo Erba Reagents (Emmendingen, Germany), sodium carbonate anhydrous (Na₂CO₃) from Centrohem (Stara Pazova, Serbia), and gallic acid, 2,2-dyphenyl-1-picrylhydrazyl (DPPH), ascorbic acid, sodium acetate, 2,4,6-tris(2-pyridyl)-(S)-triazine (TPTZ), sodium acetate trihydrate, iron(III) chloride hexahydrate (FeCl₃∙6H₂O), iron(II) sulfate heptahydrate (FeSO₄∙7H₂O), and citric acid were purchased from Sigma Aldrich (Munich, Germany).

Methanol, sodium hydroxide, formic acid, acetonitrile, hydrochloride acid, sodium carbonate, ferric chloride hexahydrate, potassium chloride, sodium bicarbonate, sodium chloride, and Folin–Ciocalteu reagent were supplied by Panreac Química SLU (Barcelona, Spain). Pure phytochemical standards (≥96%), including gallic, caffeic, chlorogenic, p-coumaric, protocatechuic, and vanillic acids, rutin and quercetin, and caffeine, were purchased from Sigma-Aldrich (Sigma-Aldrich, St. Louis, MO, USA), and extrasynthese (Genay, France). 6-Hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox), 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), potassium persulfate, 2,4,6-tris(2-pyridyl)-s-triazine (TPTZ), ammonium carbonate, calcium chloride dihydrate, magnesium chloride hexahydrate, potassium phosphate monobasic, porcine pepsin, porcine pancreatin, pronase E, and Viscozyme ® were purchased from Sigma-Aldrich (Sigma-Aldrich, St. Louis, MO, USA).

Analytical reagents; acetic acid, acetone, acetone and hydrochloric acid were purchased from R & M Marketing (Essex, UK). The standard reagents and solutions; sodium acetate, pyridine, ascorbic acid, 2,2-diphenyl-1-picrylhydrazyl (DPPH) of 95% potency, 2,4,6-tris (2-pyridyl)-s-triazine (TPTZ) of 99% potency, iron (III) chloride hexahydrate; N-methyl-N- (trimethylsilyl) trifluoroacetamide (MSTFA) of 97% potency and methoxyamine hydrochloride of 98% potency brand were Sigma-Aldrich (St. Louis, MO., USA).