Phenoxodiol

Phenoxodiol (Sigma-Aldrich Cat#D7446) was made as a stock solution of 10 mg/ml in DMSO and stored at -20°C, protected from light. The stock was used within 7 days. The stock solution was diluted 100-fold in growth medium as a working dilution. This was then further diluted in growth medium to a final concentration of 10 μM or 30 μM. Vehicle control and 10 μM Phenoxodiol treatment were augmented accordingly with DMSO to ensure uniform concentration.

Phenoxodiol, equol-4'-O-α-glucoside, equol-7-O-α-glucoside and genistin β-glucoside were purchased from Sigma-Aldrich (USA), whereas daidzin β-glucoside was from TCI Research Chemicals (Japan). Isopropyl β-D-1-thiogalactopyranoside (IPTG), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), L-ascorbic acid, pepsin, pancreatin and bile salts were obtained from Sigma-Aldrich, while cisplatin was from Roche Diagnostics (Germany). The others were of reagent grade.

All chemicals used were of analytical reagent grade. All reagents were purchased from Sigma, Aldrich: Ascorbic acid, Acetic acid, acetylcholinesterase (AChE), acetylthiocholine (ACTHI) acetonitrile (ACN), N, O-bis (trimethylsilyl) trifluoroacetamide (BSTFA), cyclohexane (CYH), dichloromethane (DCM), Dulbecco's modified eagle medium (DMEM), dimethyl sulfoxide (DMSO), 1-1-diphenyl-2-picryl hydrazyl (DPPH), 5, 5-dithiobis-2-nitrobenzoic acid (DTNB), Folin-Ciocalteu reagent (2 N), methanol (MeOH), human colon cancer cells (HCT116), human breast cancer cells (MCF-7), 3- [4, 5-dimethylthiazol-2yl]-2, 5-diphenyl tetrazolium bromide (MTT), Roswell Park Memorial Institute (RPMI), Chlorotrimethylsilane (TMCS), and tetrahydrofuran (THF).
The analytical standards used for the identification and quantification of the principal phenolic compounds found in the extracts: 3-amino-4-hydroxybenzoic acid, Gallic acid, 3, 4-dihydroxy-5 methoxybenzoic acid, 7-hydroxycoumarin-3-carboxylic acid, rutin hydrate, Butyl gallate, 4-hydroxytamoxifen, cardamonin, phenoxodiol, pinostilbene hydrate, 3-benzyloxy-4, 5-dihydroxy-benzoic acid methyl ester, ethyl trans-2-hydroxycinnamate, 4′, 5-dihydroxy-7-methoxyflavone, pinosylvin monomethyl ether, 3, 6, 3′-trimethoxyflavone, shikonin, 5-hydroxy-3′-methoxyflavone, and 3′-hydroxy-b-naphthoflavone were also purchased from Sigma, Aldrich.