Kaempferol 3 o rutinoside

Acetonitrile (hypergrade for LC-MS), formic acid (for LC-MS) and methanol (analytical grade) were purchased from Merck, Sofia, Bulgaria. The authentic standards used for compound identification were obtained from Extrasynthese (Genay, France) for protocatechuic, p-hydroxyphenylacetic, vanillic, gentisic, ferulic, p-coumaric, o-coumaric acid, rutin, hyperoside, luteolin 7-O-rutinoside, luteolin 7-O-glucoside, isoquercitrin, kaempferol 3-O-rutinoside, isorhamnetin 3-O-rutinoside, kaempferol 3-O-glucoside, isorhamnetin 3-O-glucoside, apigenin 7-O-glucoside, quercetin, apigenin, luteolin, chrysoeriolq eupatilin and genkwanin. Caffeic, 3,4-dicaffeoylquinic, 3,5-dicaffeoylquinic 1,5-dicaffeoylquinic, 4,5-dicaffeoylquinic and 3,4,5- tricaffeoylquinic acids were supplied by Phytolab (Vestenbergsgreuth, Germany). Chlorogenic acid and isorhamnetin were purchased from Sigma-Aldrich (St. Louis, MO, USA). Diclofenac, silymarin, collagenase, 1-chloro-2,4-dinitrobenzene, beta-nicotinamide adenine dinucleotide 2′-phosphate reduced tetrasodium salt (NADPH), ethylenediaminetetraacetic acid (EDTA), bovine serum albumin (fraction V) and 2,2′-dinitro-5,5′dithiodibenzoic acid (DTNB) were obtained from Merck (Darmstadt, Germany). Reduced glutathione (GSH), oxidized glutathione (GSSG), glutathionereductase (GR) and cumene hydroperoxide were purchased from Sigma Chemical Co. (Taufkirchen, Germany).

All solvents used (methanol, acetonitrile, acetic acid, water and ethyl acetate) were HPLC-grade and were purchased from Merck (Darmstadt, Germany). Protocatechuic acid, 4-hydroxybenzoic acid, vanillic acid, chlorogenic acid, trans-p-coumaric acid, eriodictyol-7-O-glucoside, eriodictyol, naringenin, naringenin-7-O-glucoside, kaempferol-3-O-rutinoside, kaempferol-3-O-glucoside, isorhamnetin-3-O-glucoside, quercetin, kaempferol, isorhamnetin, quercetin-3-O-rutinoside, quercetin-3-O-galactoside, quercetin-3-O-glucoside, isorhamnetin-3-O-rutinoside, catechin and epicatechin were purchased from Extrasynthese (Genay, France). Other chemicals were of analytical grade.

Chicoric acid, chlorogenic acid, caffeic acid, catechin, epicatechin, rosmarinic acid, ferulic acid, rutin, vitexin, quercetin-3-O-glucoside, lutein, and β-carotene were obtained from Sigma (St. Louis, MO, USA). Quercetin-3-O-galactoside, kaempferol-7-O-glucoside, kaempferol-3-O-rutinoside, and 3,5-di-O-caffeoyl quinic acid were obtained from Extrasynthese (Genay, France). Methanol and formic acid (LC–MS grade) were obtained from Merck KGaA (Darmstadt, Germany). Ultrapure water was produced by a Milli-Q Gradient A10 water purification system. The purity of the standards was 98%, and all were prepared as initial stock solutions of 1 mg ml⁻¹ in Methanol. lutein and β-carotene stocks of 1 mg ml⁻¹ were prepared in chloroform. Multiple standards stock solutions were prepared as combinations of individual standard stock solutions with further dilutions made with Methanol to obtain standard calibration curves in the range of 0.01–5.0 mg L^-1.

Commercial extracts of RO and BC were obtained from PK Components (Warsaw, Poland). Cyanidin-3-O-glucoside, a 3-O-rutinoside, delphinidin-3-O-glucoside, delphinidin-3-O-rutinoside, quercetin-3-O-glucoside, quercetin-3-O-rutinoside, myricetin-3-O-rutinoside, myricetin-3-O-galactoside, kaempferol-3-O-rutinoside, (+)-catechin, and (−)-epicatechin were purchased from Extrasynthese (Lyon, France). Neochlorogenic acid was obtained from TRANS MIT GmbH (Giessen, Germany). Acetonitrile was purchased from Merck (Darmstadt, Germany). The extracts were preliminarily tested for 1,1-diphenyl-2-picrylhydrazyl free radical (DPPH) to assess the synthetic capacity to neutralize the reactive oxygen species that are the main mediator of peroxidation process [25 ,26 (link)].

The standards caffeic acid, caftaric acid, coutaric acid, fertaric acid, kaempferol-3-O-rutinoside, quercetin-3-rutinoside, kaempferol 3-O-galactoside, quercetin-3-glucuronide, quercetin-3-galactoside, quercetin-3-glucoside, quercetin-3-rhamnoside, kaempferol-3-O-glucoside, isorhamnetin 3-glucoside, dihydroquercetin-3,5-rhamnoside, dihydrokaempferol-3-glucoside, procyanidin trimer, procyanidin B2, procyanidin tetramer, (−)-epicatechin, (−)-epicatechin 3-gallate, petunidyn-3-O-glucoside, malvidyn-3-O-glucoside, delphinidyn-3-O-(6′-acetyl)-glucoside, petunidyn-3-O-(6″-p-coumaroyl)-glucoside and malvidyn-3-O-(6″-caffeoyl)-glucoside were purchased from Extrasynthese (Lyon Nord, France). 2,2′-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid, ABTS), gallic acid, FeCl₃, Trolox and all solvents were purchased from Sigma-Aldrich (Steinheim, Germany). Pectinex^® Ultra SP-L was obtained from Novozymes (Bagsvaerd, Denmark).

HPLC grade acetonitrile, ethyl acetate, dichloromethane, hexane, and methanol were purchased from VWR International (Radnor, PA, USA). Kaempferol-3-O-rutinoside (35) and 3,5-di-O-caffeoylquinic acid (41) were purchased from Extrasynthèse (Genay, France). N-trans-p-coumaroyltyramine (48) was purchased from Phytolab GmbH & Co. KG (Vestenbergsgreuth, Germany). Chlorogenic acid (3-O-caffeoylquinic acid) (13), rutin (29), N-trans-feruloyltyramine (moupinamide) (49), dimethylsulfoxide (DMSO), camptothecin, (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), fetal bovine serum (FBS), and lipopolysaccharides (LPS from E. coli serotype O55:B5) were purchased from Sigma-Aldrich (Bornem, Belgium).

Reagents for antioxidant activities and cholinesterase inhibition assays were from Sigma-Aldrich (St. Louis, MO, USA and Steinheim, Germany), except magnesium chloride hexahydrate, which was bought from VWR (Leuven, Belgium), sodium chloride from Fisher Scientific (Fair Lawn, NJ, USA), sodium nitroprusside dihydrate from Fluka, and sodium carbonate from Merck. Hydrogen peroxide 35.5% w/w was purchased from Labchem (Zelienople, PA, USA). Methanol Chromasolv for HPLC was from Riedel-de Haën (Seelze, Germany), and formic acid was from Carlo Erba (Val de Reuil, France). The standards used for HPLC analyses were acquired from Sigma-Aldrich (4-O-caffeoylquinic acid, quercetin-3-O-rutinoside hydrate, protocatechuic acid, ferulic acid, and p-coumaric acid), Fluka (caffeic acid), Alfa Aesar (5-O-caffeoylquinic acid), and Extrasynthèse (Genay, France): 3-O-caffeoylquinic acid, apigenin-8-C-glucoside, apigenin-6-C-glucoside, quercetin-3-O-glucoside, kaempferol-3-O-glucoside, kaempferol-3-O-rutinoside, and tiliroside. Ultrapure water (with resistivities of 18.2 MΩ/cm) was obtained using a Milli-Q water purification system from Millipore (Molsheim, France). Standard solutions of Pb and Cd, Atomic Absorption standard with 4% HNO₃, were purchased from SPC Science (Quebec, Canada). Suprapur nitric acid 65% (v/v) and NH₄H₂PO₄ (Suprapur) were from Merck (Darmstad, Germany).