Petunidin 3 o glucoside chloride

Bilberry anthocyanin-rich dry extract, Anthocyan^®, was supplied by Ferlux SA (Cournon d’Auvergne, France). Methanol was of analytical grade and purchased from Carlo Erba Reagents SAS (Val-de-Reuil, France). Acetonitrile (MeCN) was of chromatographic grade (Carlo Erba Reagents SAS, Val-de-Reuil, France). All aqueous solutions were prepared with pure water produced by Milli-Q water (18.2 MΩ) system (Merck, Darmstadt, Germany). Phosphoric acid (purity 85%) was purchased from VWR Prolabo (Fontenay-sous-Bois, France). Delphinidin 3-O-glucoside chloride, cyanidin 3-O-glucoside chloride, cyanidin 3-O-galactoside chloride, petunidin 3-O-glucoside chloride, peonidin 3-O-glucoside chloride, malvidin 3-O-glucoside chloride, and malvidin 3-O-galactoside chloride were purchased from Extrasynthese (Genay, France). Aminoguanidine chloride, MGO (40% in water), bovine serum albumin (BSA), D-ribose, 6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox) and DPPH were bought from Sigma–Aldrich Chemical (Saint-Quentin Fallavier, France). MGO and DPPH radical solutions were freshly prepared every day and kept protected from light.

Graphite powder, mineral oil, hexacyanoferrate (II)/(III) (K₄[Fe(CN)₆]/K₃[Fe(CN)₆]), potassium chloride (KCl), hydroquinone, catechol, resorcinol, gallic acid, vanillin, sodium dihydrogen phosphate (NaH₂PO₄ · 2H₂O), sodium hydrogen phosphate (Na₂HPO₄ · 2H₂O), acetic acid (CH₃COOH), and sodium acetate anhydrous (CH₃COONa) were analytical grade and were purchased from Sigma-Aldrich (Milan, Italy). BioDea acidic solution (a liquid by-product of the gasification of woody waste) was provided by Romana Maceri Centro Italia S.r.l. (Civitella in Val di Chiana, Italy). The anthocyanins delphinidin-3-O-glucoside chloride (myrtillin chloride), peonidin-3-O-arabinoside chloride, peonidin-3-O-glucoside chloride, cyanidin-3-O-galactoside chloride, cyanidin-3-O-glucoside chloride (kuromanin chloride), petunidin-3-O-glucoside chloride, malvidin-3,5-di-O-glucoside chloride, malvidin-3-O-glucoside chloride (oenin chloride), and pelargonidin-3-O-rutinoside chloride were from Extrasynthese (Geneve, France).
Water solutions were prepared with deionized water obtained with a Milli-Q water purification system (Milan, Italy).

Lutein (LUT) and six anthocyanidin glucosides: cyanidin-3-O-glucoside chloride (CG) (or kuromanin chloride), delphinidin-3-O-glucoside cholride (DG) (or myrtillin chloride), pelargonidin-3-O-glucoside chloride (PLG) (or callistephin chloride), malvidin-3-O-glucoside chloride (MG) (or oenin chloride), peonidin-3-O-glucoside chloride (PNG), and petunidin-3-O-glucoside chloride (PTG) were ordered from Extrasynthese (Lyon, France). Other chemical reagents were ordered from Sigma Aldrich (Sydney, NSW, Australia). Cell culture medium components were purchased from Gibco^TM (Life Technologies, Mulgrave, VIC, Australia).

Methanol and acetone (HPLC grade) were purchased from Merck (Zedelgem, Belgium). Methanol (HPLC-Ultra LC-MS) from HiPerSolv CHROMANORM, VWR Chemicals BDH (The Netherlands), was also purchased for the production of standard solutions. Formic acid (99%) and hydrochloric acid (37%) for analysis were purchased from Carlo Erba (Chaussée du Vexin, France), and trifluoroacetic acid for LC-MS was obtained from Fluka (Buchs, Switzerland). Standards of delphinidin-3-O-glucoside chloride, cyanidin-3-O-glucosidechloride, petunidin-3-O-glucoside chloride, peonidin-3-O-glucoside chloride, and malvidin-3-O-glucoside chloride were obtained from Extrasynthese (Genay Cedex, France). The standards of delphinidin-3-O-glucoside (Dlp), cyanidin-3-O-glucoside (Cyn), petunidin-3-O-glucoside (Pt), peonidin-3-O-glucoside (Pn), and malvidin-3-O-glucoside (Mlv) were diluted separately in methanol LC-MS with 0.1% HCl (concentration 1000 mg/L).

Streptozotocin, 5,5′-dithiobis (2-nitrobenzoic acid), thiobarbituric acid, trichloroacetic acid, 2,4-dinitrophenylhydrazine, and chlorogenic acid were delivered from Sigma–Aldrich (St. Louis, MO, USA). Acetonitrile for liquid chromatography–mass spectrometry was purchased from POCh (Gliwice, Poland). Petunidin 3-O-glucoside chloride was purchased from Extrasynthese (Lyon Nord, France). All other reagents were of analytical grade.