2 2 azobis 2 amidinopropane dihydrochloride

The compounds 2,2-diphenyl-1-picrylhydrazyl, 2,2′-azobis-(2-amidinopropane) dihydrochloride, 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid), Folin–Ciocalteu reagent, carbazole, galacturonic acid, gallic acid, and catechin were purchased from Sigma-Aldrich (St. Louis, MO, USA). Phosphate-buffered saline (PBS) was purchased from Welgene (Gyeongsan, Korea). Ascorbic acid was purchased from Reagent Duksan (Ansan-si, Republic of Korea). HPLC ginsenoside standards were purchased from Ambo Institute (Daejeon, Republic of Korea).

Soluble starch, D-glucose, α-glucosidase (type V from rice), p-nitrophenyl α-D-glucopyranoside, acarbose, 3,5-dinitrosalicylic acid (DNS), Folin-Ciocalteu reagent, gallic acid, quercetin, 6-hydroxy-2-5-7-8-tetramethylchroman-2-carboxylic acid (Trolox), 1,1-diphenyl-2-picryl-hydrazyl (DPPH), 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), potassium persulphate, 2,2′-azobis (2-amidinopropane) dihydrochloride (AAPH), sodium fluorescein, 2,4,6-tripyridyl-s-triazine (TPTZ), and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich, USA. α-amylase (Bacillus amyloliquefaciens) was purchased from Roche Diagnostics, USA. All the other chemicals and reagents were of analytical grade. All the analyses were carried out using high-throughput 96-well microplate readers (SpectraMax Plus384, Molecular Devices, USA and SPECTRAmax-Gemini EM, Molecular Devices Inc, USA).

Potentiometric measurements were carried out using an Expert-pH pH-meter (OOO Ekoniks-expert, Moscow, Russia) with the EMF measurement function and the RS-232 interface. A two-electrode electrochemical cell was used. The working electrode was the EPV-1 redox platinum electrode, and the reference electrode was the EVL-1M silver/silver chloride electrode (Ag/AgCl/3 mol/dm³ KCl) (Gomel ZIP, Gomel, Belarus). The studies were carried out in a thermostated cell at 37 °C using the LOIP LT-205a circulation thermostat (ZAO LOIP, Saint Petersburg, Russia).
The following reagents were used in the work: 2,2’-azobis(2-amidinopropane) dihydrochloride (AAPH), α-tocopherol, 2,6-ditretbutylphenol, ionol, uric acid, glutathione (Sigma-Aldrich, St. Louis, MO, USA); K₄[Fe(CN)₆], K₃[Fe(CN)₆], KH₂PO₄, Na₂HPO₄·12H₂O (Reakhim, Moscow, Russia); ascorbic acid, L-cysteine, catechol, gallic acid, chlorogenic acid, caffeic acid (Panreac, Barcelona, Spain). All reagents were analytical grade. The studies were carried out in phosphate buffer solution (PS) pH = 7.4 KH₂PO₄/Na₂HPO₄·12H₂O.

Resveratrol (1) with a purity of >98% was purchased from Career Henan Chemical Co., Ltd. (Zhengzhou, China). HPLC solvents were purchased from ChemLab (Zedelgem, Belgium), and organic solvents and reagents (bis(trifluoroacetoxy)iodo)benze (PIFA), (diacetoxyiodo)benzene (PIDA), 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH), oxone, anhydrous, monobasic, potassium phosphate (P5379), and xanthine (0626) were purchased from Sigma Aldrich (Budapest, Hungary). Periodic acid, iron (III) chloride hexahydrate and sodium periodate were purchased from Reanal Laboratory Chemicals (Budapest, Hungary).

Absolute ethanol, sodium phosphate monobasic (NaH₂PO₄), fluorescein, 2,2′-azobis(2-amidinopropane)dihydrochloride (AAPH), Dulbecco’s Modified Eagle Medium (DMEM), dimethyl sulfoxide (DMSO), [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide] (MTT), 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA), N-acetyl-l-cysteine (NAC), and tert-butyl hydroperoxide (t-BuOOH) were purchased from Sigma Aldrich S.p.A. (Milan, Italy). Trypsin-EDTA solution, fetal bovin serum (FBS), glutamine, penicillin-streptomycin, and phosphate saline buffer (PBS) were purchased from Euroclone (Milan, Italy). Reagents used for RT-PCR were purchased from Euroclone (Milan, Italy). Solvents used for LC-ESI/LTQOrbitrap/MS extraction and water were purchased from VWR (Milan, Italy), while acetonitrile and formic acid were purchased from Merck (Merck KGaF, Darmstadt, Germany).
Phospholipon 90G (>90% phosphatidylcholine; P90G) was purchased from Lipoid GmbH (Ludwigshafen, Germany).

Benitaka grape (Vitis vinifera L. fruits) was purchased in single batch in a local supermarket. Dimorphandra mollis Beth fava beans were harvested in Private Biological Reserve (22°18′S/47°11′W), in Mogi Guaçu (São Paulo, Brazil), in May 2015. Ruta graveolens leaves were harvested in the Chemical, Biological and Agricultural Pluridisciplinary Research Center—CPQBA (22°48′S/47°0′W) at the University of Campinas (UNICAMP) (Campinas, São Paulo, Brazil) in April 2015. Ginkgo biloba L. dry extract was purchased at Galena (Campinas, São Paulo, Brazil). Ethyl alcohol, acetic acid, hydrochloric acid, boric acid, oxalic acid, acetone, ether, metallic magnesium, metallic zinc, ferric chloride, and aluminum chloride were provided by Synth (São Paulo, Brazil), and 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2-2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS), 2,2′-azobis (2-amidinopropane) dihydrochloride (AAPH), and fluorescein by Sigma-Aldrich (São Paulo, Brazil). Quercetin (93.3% purity) and rutin (97.3% purity) standard by Acros (Sao Paulo, Brazil). RPMI-1640 and fetal bovine serum were provided by Gibco (Sao Paulo, Brazil) and Tris [hydroxymethyl] aminomethane and neutral red dye was provided by Sigma-Aldrich (São Paulo, Brazil).

P. domestica fruits of the Methley, Pisardii and Satsuma cultivars were acquired in a ripe state from Frutalcoop local producers in Los Santos, Costa Rica. The cultivars were confirmed with the support of the Costa Rican National Herbarium, and vouchers were deposited there. The reagents, such as fluorescein, 2,2-azobis(2-amidinopropane) dihydrochloride (AAPH), 2,2-diphenyl-1-picrylhidrazyl (DPPH), Trolox, gallic acid, Amberlite XAD-7 resin, fetal bovine serum, glutamine, penicillin, streptomycin, amphotericin B, and trypsin–EDTA, were provided by Sigma-Aldrich (St. Louis, MO, USA). The human gastric adenocarcinoma cell line AGS, human colorectal adenocarcinoma SW 620 and monkey normal epithelial kidney cells Vero were obtained from the American Type Culture Collection (ATCC, Rockville, MD), while the solvents—such as acetone, chloroform and methanol—were purchased from Baker (Center Valley, PA, USA), and the DMSO was acquired from Sigma-Aldrich (St. Louis, MO, USA).