Betanin

3T3-L1 cell (BCRC Number: 60159) was obtained from the Bioresource Collection and Research Center (BCRC, Food Industry Research and Development Institute, Hsinchu, Taiwan). Oil red O solution, 3-isobutyl-1-methylxanthine, dexamethasone, insulin, betanin, rutin and kaempferol were obtained from Sigma (St. Louis, MO, USA). The dye MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) was purchased from PanReac AppliChem (Darmstadt, Germany). Dulbecco’s modified Eagle’s medium (DMEM), Trypsin-EDTA, penicillin-streptomycin, bovine calf serum, and fetal bovine serum were purchased from Gibco (Grand Island, NY, USA). Anti-PPARγ and anti-β-actin were purchased from Cell Signaling Technology (Danvers, MA, USA). All other chemicals were reagent grade.

The used red beetroots were from Dezful (in the south of Iran) and purchased from the local market in the city of Dezful, Khoozestan, Iran. PBS (Phosphate buffered saline), DMEM (Dulbecco’s Modified Eagle Medium) medium, FBS (fetal bovine serum) were purchased from GIBCO, UK. MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide], LPS (lipopolysaccharide), Betanin (CAS 7659-95-2), sulphanilamide, TLC (Thin-layer chromatography sheets), silica gel (high-purity grade pore size 60 Å, 60–100 mesh) and NED (N-1-naphthyl ethylenediamine dihydrochloride) were from Sigma-Aldrich, USA. methanol, and acetonitrile were HPLC-grade, citric acid, ascorbic acid, phosphoric acid and DMSO (Dimethyl sulfoxide) were obtained from Merck, Germany. https://www.biocompare.com. Anti-CD11b antibody (ab128797, Cambridge, UK, https://www.abcam.com), Alexa-488 Cross-adsorbed anti-rabbit IgG (H+L) (A11008, Invitrogen, https://www.thermofisher.com).

Gallic acid, rutin, betanin, 1,6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), potassium persulfate, Folin–Ciocalteu reagent, 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis (3-ethylbenzothiazoline)-6-sulfonic acid (ABTS) were purchased from Sigma-Aldrich Canada Ltd. (Oakville, ON, Canada). HPLC grade methanol was obtained from Karal (León; Guanajuato, Mexico). The water used was obtained from a Millipore Milli-Q water system with a resistivity of 18.2 MΩ cm (25 °C).

Dimethyl sulfoxide (DMSO) was provided by Glentham Life Sciences (Corsham, UK). PCR-grade nano-pure water was purchased from Jena Bioscience (Munich, Germany). Vivid CYP1A2, CYP2B6, CYP2C19, CYP2D6, and CYP3A4 screening kits, EOMCC, and BOMCC were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Betanin, bupropion, furafylline, omeprazole, quinidine, rifampicin, and Krebs-Henseleit buffer were supplied by Sigma-Aldrich (St. Louis, MO, USA). Sulfaphenazole, fluvoxamine, and ketoconazole were obtained from Toronto Research Chemicals (Toronto, ON, Canada). P450-Glo CYP1A2, P450-Glo CYP2B6, and P450-Glo CYP2C9 assay kits were provided by Promega (Wisconsin, MD, USA).

Standard ochratoxin A (OTA), aflatoxin B₁ (AFB₁) and zearalenone (ZEA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Individual stock solutions of mycotoxins were prepared in AcN at 500 µg/mL and formed solutions in MeOH at 100 µg/mL. The solutions were maintained at −20 °C in the dark. Betanin was purchased from Sigma-Aldrich and diluted to prepare solution in EtOH.
To prepare the gastrointestinal solutions, KCl 89.6 g/L, KSCN 20 g/L, NaH₂PO₄ 88.8 g/L, NaSO₄ 57 g/L, NaCl 175.3 g/L, NaHCO₃ 84.7 g/L, urea 20 g/L and Milli-Q water (Milli-Q water purification system (Millipore, Bedford, MA, USA)) were used. First, 145 mg α-amylase in 100 mL of Milli-Q water was prepared. A Pepsin solution with 0.5 mg Pepsin (1 g in 25 mL HCl (0.1 N) (Pepsin from porcine gastric mucosa, powder, ≥250 units/mg solid, P-7000, Sigma-Aldrich, St. Louis, MO, USA) was used. A solution of pancreatin: 1.10 mg (0,1 g Pancreatin (Pancreatin, from Porcine Pancreas, P1750, Sigma-Aldrich, St Louis, MO, USA) and 0.625 g bile salts (Bile extract porcine, B8631, Sigma-Aldrich, St. Louis, MO, USA) in 25 mL of NaHCO₃ (0.1 N) was prepared. The saliva solution was prepared mixing: 10 mL of KCl 89.6 g/L, 10 mL of KSCN 20 g/L, 10 mL of NaH₂PO₄ 88.8 g/L, 10 mL of NaSO4 57 g/L, 1.7 mL of NaCl 175.3 g/L, 20 mL of NaHCO₃ 84.7 g/L, 8 mL of urea (20g/L) and was completed to 500 mL with Milli-Q water.

Betanin (red beet extract diluted with dextrin) (Product number, P.N. CDS000584) was purchased from Sigma-Aldrich (St. Louis, MO, USA). 3,4-Dihydroxyphenethylamine hydrochloride (P.N. 62317), sucrose (P.N. 57501), sodium carbonate (P.N. 497198), methanol (P.N. 67561), acetonitrile (P.N. 75058), formic acid (P.N. 64186), and ultrapure water (P.N. 7732185) were purchased from Wako Pure Chemical Industries (Osaka, Japan). Calcium lactate for the Ca²⁺-added treatments (P.N. 814802) and trifluoroacetic acid (TFA) (P.N. 350709) were purchased from Nacalai Tesque Inc. (Kyoto, Japan). Folin-Ciocalteu phenol reagent (P.N. 02195186) was purchased from MP Biomedicals (Santa Ana, CA, USA). High-TEMPO Ar and High-TEMPO Cu were developed for liquid fertilizer by Mitsubishi Plastics Agri Dream Co. (Tokyo, Japan), as described previously [16 (link)].

Thioacetamide, betanin, N-acetylcysteine, GSH/GSSG kit, sodium chloride, procaine, heparin, paraFormaldehyde, sodium hydroxide, gallic acid, Folin–Ciocalteu reagent, catechin, ascorbic acid, dithiothreitol (DTT), DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate), Trolox^TM ((±)-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid), sodium acetate, TPTZ (2,4,6-tris(2-pyridyl)-s-triazine), ABTS^•+ (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt), potassium persulfate, AAPH (2,2′-azobis (2-methylpropionamidine) dihydrochloride), sodium nitroprusside, sulphanilic acid, N-1-naphthyl ethylenediamine were all obtained from Sigma Aldrich (St. Louis, MO, USA). Acetonitrile (nanograde) was from Mallinckrodt. Formaldehyde, absolute methanol and glacial acetic acid were from J.T. Baker (Madrid, Spain). Hydrochloric acid and ferric chloride were from Fermont (Monterrey, México) and sodium pentobarbital was from Pisabental, Pisa Agropecuaria, México.