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2 protocols using aluminum chloride

1

Dulbecco's PBS Buffer Preparation

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Dulbecco’s
PBS (Sigma-Aldrich
Co., St. Louis), Tween-20 (0.0003% Arsenic, 0.0005% Copper, Fisher
BioReagents, Fair Lawn), hydrochloric acid (Honeywell GmbH, Seelze,
Germany), sodium hydroxide (potassium ≤0.02%, ≥98%,
Sigma-Aldrich Co., St. Louis), sodium chloride (Merck KGaA, Darmstadt,
Germany), potassium chloride (≥99%, Fisher Chemical, Loughborough,
U.K.), calcium chloride (≥93%, Sigma-Aldrich Co., St. Louis),
and aluminum chloride (≥99%, Chem-Supply Pty Ltd, Gillman,
Australia) were used as received without any further purification.
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2

Probiotic Bacteria and Cocoa Compounds Interaction

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The L. delbrueckii subsp. bulgaricus probiotic bacteria were used from the stock culture collections in the Food Microbiology Laboratory at the University of Melbourne. Fructooligosaccharides (FOS), Na-alginate, enzymes (salivary α-amylase, porcine pepsin, pancreatin), HCl, acetone, acetic acid, epicatechin, catechin, procyanidin B1, procyanidin B2, quercetin 3-O-galactoside, quercetin 3-O-glucoside were purchased from Sigma Aldrich (Castle Hill, NSW, Australia). A standard mixture of short chain fatty acids (SCFAs) was purchased from Cayman Chemical (Ann Arbor, MI, USA). The selective media DeMan, Rogosa and Sharpe (MRS), nutrient agar & broth, AnaeroGen sachets, yeast extract, beef extract, protease peptone, n-hexadecane, L-cysteine hydrochloride, bile salts, and trichloroacetic acid were purchased from Thermo Fisher Scientific Pty Ltd (Melbourne, VIC, Australia). NaOH, phosphate-buffered saline, CaCl2, dextrose, K2HPO4, (NH4)2SO4, MgSO4·7H2O, NaCl, KCl, NaHCO3, MgCl2(H2O)6, (NH4)2CO3, potassium persulfate, potassium acetate, aluminum chloride were ordered from the Chem-Supply Pty Ltd. (Melbourne, VIC, Australia). Chocolates with 45% and 70% cocoa mass, cocoa powder (pure), cocoa liquor, cocoa butter, sugar, soy lecithin were purchased from local supermarket (VIC, Australia).
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