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2 protocols using dichlorofluorescin dcfh

1

HPLC-MS-QTOF Analysis of Polyphenols

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Standard polyphenols for HPLC-MS-QTOF analysis (phloroglucinol, gallic acid, catechin, (−)-epicatechin, quercetin, quercetin-3-O-glucoside and resveratrol were obtained from Sigma-Aldrich (Madrid, Spain). Delphinidin-3-glucoside, cyanidin-3-glucoside, peonidin-3-glucoside and malvidin-3-glucoside standards were obtained from Extrasynthese (Lyon, France), 6-OHDA, dichlorofluorescin (DCFH) and Dulbecco’s Phosphate Buffered Saline were purchased from Sigma-Aldrich Quımica S.A. (Madrid, Spain). HPLC-grade solvents were purchased from Merck (Darmstadt, Germany). Dulbecco Modified Eagle Medium‎ (DMEM), fetal bovine serum (FBS), penicillin/streptomycin, trypsin and l-glutamine were purchased from Lonza (Barcelona, Spain). Dimethyl sulfoxide (DMSO) was obtained from Panreac (Barcelona, Spain). Cell culture dishes were obtained from BD Falcon (Madrid, Spain).
The wine used was a commercial Cabernet Sauvignon, “Postales del Fin del Mundo” from Neuquén province, Argentina. Its alcohol content was 13.7% and its pH 3.6. Total polyphenol content was 2230 ± 160 mg GAE/L as determined by the Folin-Ciocalteau method [23 ]. The carbohydrates used for encapsulation were maltodextrin dextrose equivalent 10 (MD10) from Productos de Maíz, S.A., Argentina and gum arabic (from Gelfix, Buenos Aires, Argentina).
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2

Antioxidant Evaluation of Epicatechin

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Tert-butylhydroperoxide (t-BOOH), (−)-epicatechin (EC, >95% of purity), Glutathione reductase (GR), reduced glutathione (GSH) and oxidized glutathione, dichlorofluorescin (DCFH), o-phthaldialdehyde (OPT), nicotine adenine dinucleotide phosphate reduced salt (NADPH), gentamicin, penicillin G and streptomycin were purchased from Sigma-Aldrich Chemical (Madrid, Spain). Bradford reagent was from BioRad Laboratories S.A (Madrid, Spain). Methanol of HPLC grade, dimethyl sulfoxide (DMSO), light petroleum and phosphoric acid of analytical grade were purchased by Panreac (Barcelona, Spain). DMEM culture media and foetal bovine serum (FBS) were from Cultek (Madrid, Spain). All other reagents were of analytical quality.
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