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2 protocols using 1 1 diphenyl 2 picrylhydrazyl dpph butylated hydroxyanisole

1

Antioxidant Compound Characterization

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Methanol (HPLC gradient grade) and tetrachloroethylene (synthesis grade) were purchased from Scharlab (Barcelona, Spain). Triethylamine (analysis quality) and phenylisothiocyanate (PITC) of reagent grade were supplied by Panreac (Barcelona, Spain); 1,1-diphenyl-2-picrylhydrazyl (DPPH), butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), and anthrone (reagent grade) were supplied by Sigma-Aldrich (St. Louis, MO, USA). Formic acid (synthesis grade) and amino acids (aspartic acid, glutamic acid, histidine, arginine, proline, valine, lysine, methionine, isoleucine, and phenylalanine) were provided by Merck (Darmstadt, Germany). Phenolic compounds were supplied as follows: gallic acid, protocatechuic acid, (−) epicatechin, ferulic acid, p-coumaric acid, vanillic acid, syringic acid, and (+) catechin by Sigma–Aldrich Chemie (Steinheim, Germany), and rutin and gentisic acid by Merck (Darmstadt, Germany). Ultrapure water was obtained from a Milli-Q system from Millipore (Bedford, MA, USA).
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2

Antioxidant and ACE Inhibitory Assays

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The chemicals and solvents used in the present study were purchased at the analytical grade or highest level of purity available. Alcalase ® 2.4 L serine-protease from Bacillus licheniformis was obtained from Novozymes ® (Bagsvaerd, Denmark). The chemicals used in the experimental assays, including 1,1-diphenyl-2-picrylhydrazyl (DPPH), butylated hydroxyanisole (BHA), angiotensin I-converting enzyme from rabbit lung, the ACE synthetic substrate hippuryl-A c c e p t e d M a n u s c r i p t 5 l-histidyl-l-leucine (HHL), β-carotene, and L-ascorbic acid, were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Diethylaminoethyl (DEAE)-cellulose were supplied by Pharmacia (Uppsala, Sweden). All other chemicals and reagents were used at the analytical grade, and all solutions were freshly prepared in distilled water.
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