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4 protocols using naringenin 7 o glucoside

1

Antioxidant and Cytotoxic Evaluation

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BHA (butylated hydroxyanisole) and DPPH (2,2-diphenyl-2-picrylhydrazyl radical) were obtained from Sigma Chemical Co (St Louis, MO, USA). Porcine trypsin was purchased from Roche (Barcelona, Spain). Tripan blue, dimetilsulfoxide (DMSO), “Minimum Essential Medium Eagle (MEM)” and RPMI-1640 culture media, mix of antibiotics and antimycotic, sodium piruvate, sodium bicarbonate, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dichlorofluorescein diacetate, cisplatin, and potassium dichromate were purchased from Sigma-Aldrich (Madrid, Spain) and fetal bovine serum (FBS) was obtained from T.D.I. (Madrid, Spain). The phenolic standards eriodictyol-7-O-glucoside, naringenin-7-O-glucoside, luteolin-7-O-glucoside, quercetin-7-O-rutinoside, rosmarinic acid, and verbascoside were obtained from Extrasynthese (Genay Cedex, France). Ascorbic acid, formic acid and ethanol were purchased from Panreac (Barcelona). n-Hexane, methanol and acetonitrile, all with HPLC purity, were purchased from Lab-Scan (Lisbon, Portugal).
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2

Quantification of Phenolic Compounds

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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.
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3

Phytochemical Characterization of Blackthorn Flowers

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Hydrochloric acid (37%, AR grade), butan-1-ol, acetic acid glacial (AR grade), acetone (AR grade), acetonitrile (HPLC grade), dichloromethane (LR grade), hexane (GLC, pesticide residue grade) and methanol (HPLC grade) were obtained from ThermoFisher Scientific (Loughborough, UK); benzyl mercaptan (99%), catechin hydrate (≥98%), epicatechin (90%), epigallocatechin (95%), gallocatechin (≥97%), catechin gallate (≥98%), epicatechin gallate (≥98%), epigallocatechin gallate (≥95%), gallocatechin gallate (≥98%), quercetin (≥99%), isoquercitrin (≥90%), rutin hydrate (≥95%), naringin (≥95%), (±)-eriodictyol (≥90%) from Sigma-Aldrich (Poole, UK); naringenin (97%) from Alfa Aesar (Lancashire, UK); (±)-taxifolin (98%) from Apin Chemicals (Abingdon, UK); procyanidin A2 (PC A2, ≥99%) and naringenin-7-O-glucoside ( ≥99%) from Extrasynthese (Genay Cedex, France); afzelechin (96-98%) from Plantech UK (Reading, UK) and Sephadex LH-20 from GE Healthcare (Little Chalfont, UK). 1). Details of other samples are in Table A1 and Appendix A.1.1. Plant materials were purchased in 2012/2013 and ground to pass a 1 mm sieve (impeller SM1 cutting mill, Retsch, Haan, Germany). Leaves and stalks were removed from blackthorn flowers and only flowers were used.
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4

Quantification of Phenolic Compounds

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Standards of the following phenolic acids and flavonoids are available commercially and used in the present work: protocatechuic acid, caffeic acid, ferulic acid, (+) catechin, luteolin, and eriodictyol were purchased from Sigma-Aldrich (Sydney, Australia); apigenin, naringenin, naringenin-7-O-glucoside, quercetin dihydrate, isorhamnetin, and taxifolin were purchased from Extrasynthese (Genay Cedex, France). Organic solvents acetonitrile and methanol (HPLC grade) were supplied by Crown Scientific (Sydney, Australia), and Milli-Q water (Milli-Q plus 185, Sydney, Australia) was used to make all aqueous solutions and HPLC mobile phase.
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