Pelargonidin 3 glucoside

The chemicals 1,1-Diphenyl-2-picrylhydrazyl (DPPH), 2,4,6-Tris(2-pyridyl)-s-triazine (TPTZ), (±)6-Hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (TROLOX), 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), diammonium salt (98%TLC), ABTS, gallic acid, sodium carbonate monohydrate ACS reagents, and ethanol (final ethanol grade) were purchased from Sigma-Aldrich (Steinheim, Germany); Manganese (IV) oxidise activated (˃90%), and Folin–Ciocalteu’s phenol reagents were purchased from Fluka (Buchs, Switzerland). Analytical grade solvents and reagents were used for all of the work. Ellagic acid, gallic acid, punicalagin A and B, punicalin, cyanidin 3-glucoside, cyanidin 3,5-diglucoside, pelargonidin 3,5-diglucoside, and pelargonidin 3-glucoside were obtained from Sigma-Aldrich (Stenheim, Germany). Formic acid and acetonitrile for LC/MS were purchased from Carlo Erba Reagents (Cornaredo, MI, Italy). Sigma Aldrich provided all the other chemicals (Steinheim, Germany). The ultrapure water was produced from the Millipore system (Millipore Sigma, Darmstadt, Germany), and filtered with a 0.20 μm Sartolon polyamide filter (Sartorius Stedim Biotech, Göttingen, Germany).

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).

For the determination of sugars and organic acids, we used the following standards: sucrose, glucose, fructose, tartaric, quinic, malic, citric, oxalic, shikimic and fumaric acid from Sigma-Aldrich (Steinheim, Germany). For phenolic compounds’ identification and calculation, the following standards were purchased: procyanidin B1, p-coumaric acid, epicatechin ferulic acid, catechin, quercetin-3-O-rutinoside, naringenin, quercetin-3-O-galactoside, quercetin-3-O-glucoside, quercetin-3-O-xyloside, kaempferol-3-O-glucoside and phloridzin from Fluka Chemie (Seelze, Germany), quercetin-3-O-arabinofuranoside from Apin Chemicals LTD (Compton, UK), isorhamnetin-3-O-glucoside from Extrasynthèse (Genay, France), and chlorogenic and caffeic acid, quercetin-3-O-rhamnoside, 3-O-caffeoylquinic acid, cyanidin-3-O-glucoside, pelargonidin-3-glucoside, cyanidin-3-O-rutinoside and 4-O-caffeoylquinic acid from Sigma-Aldrich. For the extraction of phenolic compounds, methanol from Sigma-Aldrich, double-distilled water purified with a Milli-Q system (Millipore, Bedford, MA, USA) and 3% formic acid from Fluka Chemie were used. The chemicals for the mobile phases were acetonitrile (HPLC-MS), formic acid, sulphuric acid and double-distilled water.

Phenolic compounds were extracted from whole fruit samples as previously described (Terry et al., 2007b (link)). Concentrations of total anthocyanins, pelargonidin-3-glucoside, pelargonidin-3-glucoside derivative, cyanidin-3-glucoside, quercetin-3-glucoside, catechin, epicatechin, ellagic and chlorogenic acids (Sigma-Aldrich, Dorset, UK or Extrasynthese, Lyon, France) were calculated against authentic standards and expressed as mg kg⁻¹ of dry weight.

Available comparable reference phenolics were caffeine, theobromine, theophylline, (±)-catechin, (-)-epigallocatechin gallate, (-)-epicatechin, (-)-epicatechin gallate, (-)-epigallocatechin, caffeic acid, p-coumaric acid, vanillic acid, rosmarinic acid, quercetin, naringenin, luteolin, kaempferol, ursolic acid, pinocembrin, carnosic acid, apigenin, ferulic acid, cyanidin-3-rutinosede, pelargonidin-3-glucoside, cyanidin, pelargonidin, quercetin 3-glucoside, and kaempferol 3-glucoside from Sigma Aldrich (St. Louis, MO, USA) [20 (link)].

Acetonitrile, methanol, formic acid, hydrochloric acid, water (HPLC grade), MK-571, Amberlite XAD7-HP, anthocyanin standards (pelargonidin-3 glucoside, pelargonidin-3-rhamnosylglucoside, cyanidin-3-glucoside, delphinidin-3- glucoside, malvidin-3-glucoside, petunidin-3-glucoside, peonidin-3-glucoside), 4’,6-diamidino-2-phenylindole (DAPI) were purchased from Sigma-Aldrich, Gillingham, UK. Caco-2 cells were purchased from European Collection of Authenticated Cell Cultures (ECACC), Salisbury, UK. Concentrate of Strawberry was purchased from Cobell, Exeter, UK and red grape concentrate from Homebrew shop, Aldershot, UK. The other reagents were from Lonza, Basle, Switzerland.