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3 protocols using sodium acetate

1

Horseradish Peroxidase-Magnetic Nanoparticle Assay

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Horseradish peroxidase Type VI (EC 1.11.1.7), polyethylenimine (PEI) (MW 1300, 2000, 25000 and 60000), 2,2′-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) and hydrogen peroxide were from Sigma Aldrich (St. Louis, MO). Tetramethyl orthosilicate (TMOS), trehalose and potassium phosphate monobasic were from MERCK (Whitehouse Station, NJ). Dibasic sodium phosphate and sodium acetate were from Biopack (Buenos Aires, Argentina). Gel filtration PD10-Columns were from GE Healthcare (Buckimghamshire, UK). Magnetic nanoparticles (MNPs) fluidMag-PAA (200 nm of aggregate size) were from Chemicell (Berlin, Germany). All other chemicals used were analytical grade reagents.
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2

Spectroscopic Analyses of Bioactive Compounds

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Colour measurements were made with a hand-held tristimulus reflectance spectrocolorimeter (Minolta Co., Model CM-508-d, Japan). For total anthocyanin and phenolic compound contents, and antioxidant capacity (TEAC assay), a UV- VIS spectrophotometer (model V-630, JASCO, Japan) was used. For the determination of antioxidant capacity evaluated by ORAC assay, fluorescence measurements were made by means of a Multilabel Microplate Reader (Perkin Elmer, Victor3 model, USA). Fluorescence measurements in Caenorhabditis elegans assays were performed in a Spark 20M Multimode Microplate Reader (Tecan, NC, USA).
AAPH (2,2′-Azobis (2-methylpropionamidine) dihydrochloride, ABTS (2,2 -azinobis (3-ethylbenzothiazoline)-6-sulfonate), fluorescein sodium salt, gallic acid (GA), Trolox (C14H18O4) and 2,7 – dichlorodihydrofluorescein diacetate (H2DCF-DA) were purchased from Sigma-Aldrich (St. Louis, USA). Ethyl alcohol, sodium acetate, sodium carbonate, and Tween 80 were from Biopack (Buenos Aires, Argentina). Folin Ciocalteau reagent, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, disodium hydrogen phosphate, hydrochloric acid, potassium chloride, sodium chloride, sodium hydroxide, manganese sulphate, and malt extract agar (MEA) were purchased from Merck (Darmstadt, Germany).
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3

Brewing Coffee Roast Diversity

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BSGs with different roasting levels (Pilsen, Caramel 60, Caramel 120 and Chocolate) were obtained in the laboratory under equivalent industrial conditions (Palmer, 2006) . Sodium carbonate, sodium acetate and acetic acid were supplied by Biopack. DPPH reagent, gallic acid, Folin-Ciocalteu reagent, 6-Hydroxy-2,5,7,8-tetramethylchromane-2carboxylic acid (Trolox standard), ABTS reagent and potassium persulphate were purchased from Sigma-Aldrich, while TPTZ reagent was supplied by Fluka Chemicals. All these chemicals were of analytical grade.
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