Petunidin 3 o glucoside

Black-seeded common beans (Phaseolus vulgaris L. cv Negro de vaina morada) were cultivated in November 2018 and collected in March 2019 in Chiapas, Mexico. Seeds were stored at 4 °C until use. Ultrapure water, ethanol (≥99%), sodium carbonate, 2 N Folin–Ciocalteu reagent, gallic acid, chlorohydric acid, potassium chloride, sodium acetate, Amberlite^® XAD-7HP, formic acid, n-hexane (≥95%), ethyl acetate (≥99.7%), methanol (≥99%), trifluoracetic acid (TFA), acetonitrile (≥99.9%), 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS), 6-hydroxy-2,5,7,8-tetramethylchrome-2-carboxylic acid (Trolox^®), potassium persulfate, sodium nitroprusside, Griess reagent, gallic acid (≥99%), genistein (≥98%), myricetin (≥96%), apigenin-7-glucoside (≥90%), epigallocatechin gallate (≥80%), kaempferol-3-glucoside (≥95%), quercetin-3-glucoside (≥90%), quercetin-3-D-galactoside (≥98%), delphinidin 3-O-glucoside (≥98%), malvidin 3-O-glucoside (≥98%), and petunidin-3-O-glucoside (≥98%) were purchased from Sigma-Aldrich (St. Louis, MO, USA). iNOS inhibitor screening kit (Fluorometric) was purchased from Biovision (Milpitas, CA, USA). COX-1 (ovine) and COX-2 (human) inhibitor screening assay kits were purchased from Cayman Co. (St. Louis, MO, USA). CO₂ gases were obtained from Grupo Infra (Guadalajara, Jal., México).

Folin–Ciocalteau reagent, 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,4,6-Tris(2-pyridyl)-1,3,5-triazine (TPTZ), cyanidin-3,5-di-O-glucoside, malvidin-3,5-di-O-glucoside, pelargonidin-3-O-glucoside, delfinidin-3-O-glucoside, cyanidin-3-O-glucoside, malvidin-3-O-glucoside, peonidina-3-O-glucoside, petunidin-3-O-glucoside, kaempferol-3-O-glucoside, rutin, isorhamnetin-3-O-glucoside, myricetin, trans-resveratrol, quercetin-3-β-d-glucoside, gallic acid, caftaric acid, caffeic acid, ferulic acid, chlorogenic acid, p-coumaric acid, (–)-epicatechin, (–)-epicatechin gallate, (–)-epigalatocatechin gallate, (+)-catechin, procyanidin A2, procyanidin B1, and procyanidin B2 compounds were purchased from Sigma-Aldrich^® (St. Louis, MO, USA). The solvents (methanol, acetonitrile, and ortho-phosphoric acid) used in the extraction and HPLC procedures were analytical/HPLC grade and purchased from Merck^® (São Paulo, Brazil).

Chromatography solvents (acetonitrile, methanol, water and formic acid) and phenolic standards (cyanidin 3-O-glucoside, delphinidin 3-O-glucoside, peonidin 3-O-glucoside, petunidin 3-O-glucoside, malvidin 3-O-glucoside, catechin, epicatechin, caffeic acid, vanillic acid, chlorogenic acid, syringetin 3-O-glucoside and quercetin 3-O-glucoside) were obtained from Sigma Aldrich (St. Louis, MO, USA). Internal standards, ethyl gallate and taxifolin, were obtained from TCI (Portland, OR, USA) and Selleckchem (Houston, TX, USA), respectively. Materials required for the in vitro digestion including mucin (M2378), α-amylase (A3176), pepsin (P7125), lipase (L3126), pancreatin (P7545), and bile (B8631) were obtained from Sigma Aldrich, while urea (U15-500), and uric acid (A13346-14) were purchased from Fisher Chemical (Fair Lawn, NJ, USA). The oral phase solution for the in vitro digestions was prepared as described^{40 (link)} by combination of KCl, Na₃PO₄, Na₂SO₄, NaCl, and NaHCO₃ in distilled water at concentrations of 242, 11, 8, 10, and 40 mmol/L, respectively.