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4 protocols using cafeic acid

1

Analyzing Tea Plant Compounds

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Cammelia sinensis (tea plant), Symbiotic Cultures of Bacteria and Yeasts (SCOBYs) from German, American, Canadian and Jun SCOBY, and honey were obtained from local shops in Goiânia, Goiás, Brazil. All solutions were prepared using analytical grade reagents and double distilled Milli-Q water (conductivity 0.1 µS.cm−1) (Millipore S.A., Molsheim, France). Cafeic acid, catechin, catechol, gallic acid, epicatechin and rutin were purchased from Sigma Chemical Co. (St. Louis, MO, USA). All solution standards were prepared with a concentration of 100 µM from the dilution of stock solutions (1 mM). The carbon paste was prepared using graphite powder and mineral oil, both from Sigma-Aldrich (St. Louis, MO, USA).
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2

Bioactive Compound Evaluation in Cell Culture

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Caffeine, theobromine, teofiline, (±)-catechin, (−)-epigalocathechin gallate, (−)-epicatechin, (−)-epicathechin gallate, (−)-epigallocatechin, cafeic acid, p-coumaric acid, vanillic acid, rosmarinic acid, quercetin, naringenin, luteolin, kaempferol, ursolic acid, pinocembrine, carnosic acid, apigenin, ferulic acid, cyanidin-3-rutinosede, pelargonidin-3-glucoside, cyanidin, pelargonidin, quercetine 3-glucoside, and kaempferol 3-glucoside were from Sigma Aldrich (St. Louis, MO, USA). Dulbecco’s Modified Eagle Medium (DMEM, high glucose), qualified fetal bovine serum (FBS), MEM non-essential amino acid solution (NEAA), Hank’s balanced salt solution (HBSS), penicillin/streptomycin, and TRIzol reagent were purchased from Life Technologies (Grand Island, NY, USA). Interleukin 1 beta (IL-1β), tumor necrosis factor alpha (TNF-α), interferon gamma (IFN-γ), lipopolysaccharide (LPS), bovine serum albumin (BSA), and trypsin were obtained from Sigma Aldrich (St Louis, MO, USA). Transwell plates and inserts (24-well plates) were acquired from Corning, Inc. (Lowell, MA, USA).
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3

Granadilla Phytochemical Profiling and Bioactivity

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Fresh granadilla, a total of 10 kg, was purchased in a local market (Manizales -Colombia). Acetone, sodium carbonate, and Folin-Ciocalteu reagent were purchased from PanReac AppliChem, ITW Reagents, (Darmstadt -Germany); acetonitrile, ammonium formate, formic acid, methanol, sodium dodecyl sulphate, and hydrated sodium taurocholate were purchased from Sigma Aldrich (St. Louis, MO -USA). Sodium nitrite and aluminium chloride were obtained from LOBA Chemie (Mumbai -India), and sodium hydroxide was obtained from EMSURE Merck (Darmstadt -Germany). UHPLC reference standards apigenin, cafeic acid, caffeine, carnosic acid, (±)-catechin, cyanidin, cyanidin-3-rutinoside, (-)-epicatechin, (-)-epicathechin gallate, (-)-epigallocatechin, (-)-epigalocathechin gallate, ferulic acid, kaempferol, kaempferol 3-glucoside, luteolin, naringenin, p-coumaric acid, pelargonidin, pelargonidin-3-glucoside, pinocembrine, quercetin, quercetin 3-glucoside, kaempferol 3-glucoside, rosmarinic acid, teofiline, theobromine, ursolic acid, and vanillic acid were acquired from Sigma Aldrich (St. Louis, MO -USA). Human tumor necrosis factor alpha (TNF-α) and interleukin -6 (IL-6) kits were obtained from BioLegend (San Diego, CA -USA).
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4

Antioxidant Assays and Polyphenol Characterization

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(2,2ʹ-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), 6-hydroxy-2,5,7,8tetramethylchroman-2-carboxylic acid (Trolox), anthralin or dithranol (1,8-dihydroxy-9,10dihydroanthracen-9-one), NaCl, 2,5-dihydroxybenzoic acid, gallic acid, ellagic acid, phydroxybenzoic acid, vanillic acid, cafeic acid, p-coumaric acid, ferulic acid, punicalagin, procyanidin B2, quercetin, myricetin and kaempferol were all obtained from Sigma-Aldrich (St. Louis, MO). 2,4,6-Tris(2-pyridyl)-S-triazine (TPTZ) was from Fluka Chemicals (Madrid, Spain). All the reagents used for the preparation of phenolic compounds fractions and spectrophotometric determinations were of analytical grade; while for MS analysis, they were of MS grade.
Condensed tannin concentrate from Mediterranean carob pods (Ceratonia siliqua L) was supplied by Nestlé Ltd. (Vers-chez-les Blancs, Switzerland).
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