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6 protocols using gallocatechin

1

Characterization of Catechin Standards

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All chemicals were at least of analytical grade. Toluene, 1-propanol, formic acid, acetic acid, hydrochloric acid (37%) and 4-dimethylaminocinnamaldehyde (DMACA) were from Merck (Darmstadt, Germany). Ethanol, acetone, as well as HPLC grade acetonitrile and mEthanol were purchased from Sigma-Aldrich (St. Louis, MO, USA). LC-MS grade acetonitrile and mEthanol used for MS analyses were from Fluka (Buchs, Switzerland). MilliQ 18.2 MΩ water (Millipore, Bedford, MA, USA) was also used.
Standards of (−)-epicatechin gallate, (−)-epigallocatechin, (−)-epigallocatechin gallate, (−)-catechin gallate, (−)-gallocatechin, (−)-gallocatechin gallate, procyanidin B1, procyanidin B2, procyanidin B3 and procyanidin C1 were obtained from Extrasynthèse (Genay, France), while (−)-epicatechin was from Sigma-Aldrich and (+)-catechin from Carl Roth (Karlsruhe, Germany).
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2

Quantitative Analysis of Polyphenols

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All chemicals were at least of analytical grade. Acetic acid, formic acid, hydrochloric acid (37%), toluene, 1-propanol, and 4-dimethylaminocinnamaldehyde (DMACA) were from Merck (Darmstadt, Germany). Acetone, ethanol as well as HPLC grade acetonitrile and methanol were purchased from Sigma-Aldrich (St. Louis, MO, USA). LC–MS grade acetonitrile and methanol used for MS analyses were from Fluka (Buchs, Switzerland). MilliQ 18.2 MΩ water (Millipore, Bedford, MA, USA) was also used.
Standard of (−)-epicatechin was obtained from Sigma-Aldrich and (+)-catechin from Carl Roth (Karlsruhe, Germany). Standards of procyanidin B1, procyanidin B2, procyanidin B3, procyanidin C1, (−)-catechin gallate, (−)-gallocatechin, (−)-gallocatechin gallate, (−)-epicatechin gallate, (−)-epigallocatechin and (−)-epigallocatechin gallate were obtained from Extrasynthèse (Genay, France).
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3

Intestinal Epithelium Differentiation Protocol

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MEM with Earle's salts, DMEM, fetal bovine serum (FBS), trypsin‐EDTA, and antibiotics were purchased from Biochrom GmbH (Berlin, Germany). Entero‐STIM Intestinal Epithelium Differentiation Medium and MITO+ Serum Extender were purchased from Corning (Wiesbaden, Germany). Hesperetin, peltatoside, hyperoside, kaempferol, ellagic acid, isoquercitrin, quercitrin, phloretin, (−)‐epicatechin, epigallocatechin, epicatechin gallate, procyanidin B1, procyanidin B2, epigallocatechin gallate, gallocatechin, and morine were obtained from Extrasynthese (Genay Cedex, France). Phloridzin, chlorogenic acid, caffeic acid, quercetin, (+)‐catechin, gallic acid, 2‐mercaptoethanol, NaCl, MgSO4, KCl, CaCl2, HEPES, and xylitol were purchased from Sigma‐Aldrich (Schnelldorf, Germany). Guaijaverin and avicularin were obtained from Glentham Life Sciences (Corsham, United Kingdom). Purified and concentrated GLE (cGLE) was provided by Belan GmbH (Wels, Austria).30 Guava fruit puree for preparation of fruit extracts was obtained from Tradework BV (Rotterdam, The Netherlands), commercially available GLE and apple extract (AE) were purchased from Pfannenschmidt (Hamburg, Germany). Transwell inserts (12 mm, collagen‐treated, 0.4 μm pore size) and 12‐well plates were obtained from Corning (Wiesbaden, Germany).
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4

Quantification of Polyphenolic Compounds

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Folin–Ciocalteau reagent, gallic acid, quercetin, methanol, trifluoroacetic acid (TFA), Na2CO3, and acetonitrile (HPLC grade) were purchased from Sigma-Aldrich Chemie GmbH (Schnelldorf, Germany). Isorhamnetin, myricetin, and epicatechin were from TCI Europe N.V. (Zwijndrecht, Belgium). Isorhamnetin-3-glucoside, rutin, and gallocatechin were from Extrasynthese (Lyon, France). Epigallocatechin and kaempferol were from Alfa Aesar Thermo Fisher Scientific, (Haverhill, MA, USA). Polyvinylpolypyrrolidone (PVPP) and thiabendazole (2-(4-Thiazoly) benzimidazole) were purchased from Sigma-Aldrich S.r.l. (Milan, Italy).
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5

Purification and Characterization of Polyphenol Compounds

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Gallocatechin gallate (GCG; HPLC purity > 98%), catechin gallate (CG; HPLC purity > 98%), epicatechin gallate (ECG; HPLC purity > 97.5%), catechin (C; HPLC purity > 99%), gallocatechin (GC; HPLC purity > 98%), and epicatechin (EC; HPLC purity > 99%) were from Extrasynthese (Genay Cedex, France); NADPH and L-idose were from Carbosynth (Compton, England). Bovine serum albumin (BSA), D,L-dithiothreitol (DTT), D,L-glyceraldehyde (GAL), EDTA, glycerol and glutathione (GSH) were purchased from Sigma-Aldrich (Saint Louis, MO, USA); ammonium sulfate, sodium phosphate monobasic were from J.T. Baker (Phillipsburg, NJ, USA); YM10 ultrafiltration membranes were from Merck-Millipore (Darmstadt, Germany). All other chemicals were of a reagent grade.
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6

Bioactive Compound Acquisition and Preparation

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Kaempferol (K) and luteolin (L) were purchased from Indofine (Hillsborough, New Jersey, USA). Tamarixetin (T), (-)-epicatechin (EC), (-)-gallocatechin (GC), (-)-epicatechin gallate (ECG) and (-)-epigallocatechin gallate (EGCG) were purchased from Extrasynthese (Genay, France). Quercetin (Q), isorhamnetin (I), Dimethyl sulfoxide (DMSO), Mc Coy's 5A medium, poly-DL-alanine and green tea catechins (GTC) mix (containing (+)-catechin (C), (-)-catechin gallate (CG), EC, ECG, EGCG, GC and (-)-gallocatechin gallate (GCG), 100 µg/mL of each component delivered in acetonitrile: water (8:2) with 5% 1 M HCl) were purchased from Sigma-Aldrich (Schnelldorf, Germany). Penicillin, streptomycin, Dulbecco's phosphate buffered saline (DPBS) and GlutaMax were purchased from Gibco TM (Life Technologies, Paisley, United Kingdom). Foetal bovine serum (FBS) was purchased from Lonza TM (VWR, Verviers, Belgium). Ascorbic acid was purchased from Prolabo TM (VWR, Leuven, Belgium). Acetonitrile (ACN) and formic acid (UPLC-MS grade) were purchased from BioSolve BV (Valkenswaard, The Netherlands). High-purity water was produced with a Synergy UV water purification system from Merck Millipore (Amsterdam, The Netherlands).
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