Acetone, ascorbic acid, gallic acid, sodium hydroxide, potassium hydroxide, potassium sulphate, selenium metal powder, sodium carbonate and standard solutions for inductively coupled plasma optical emission spectrometry (ICP-OES) (boron, calcium, copper, iron, magnesium, manganese, phosphorus, potassium, sulphur and zinc) were from Panreac (Barcelona, Spain), 2,2-diphenyl-1-picrylhydrazyl (DPPH) was from Sigma-Aldrich (St. Louis, MO, USA) and boric acid were from Chem-lab (Zedelgem, Belgium), ethanol absolute and sulphuric acid were from Riedel-de Haën (Seelze, Germany), hydrochloric acid, nitric acid and Folin-Ciocalteu reagent were from Merck (Darmstadt, Germany). All reagents used in the analytical procedures were of analytical reagent grade. All the water used was purified using a Milli-Q water system (Millipore, Bedford, MA, USA).
Ascorbic acid
Manufactured by ITW Reagents
Sourced in Spain, United States
Ascorbic acid is a chemical compound commonly known as Vitamin C. It is a water-soluble vitamin that functions as a reducing agent and antioxidant. Ascorbic acid is a colorless, crystalline solid that is widely used in various laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (3 mentions)
Sodium hydroxide (naoh), by ITW Reagents (2 mentions)
Gallic acid, by ITW Reagents (2 mentions)
Formic acid, by ITW Reagents (2 mentions)
Luteolin 7 o glucoside, by Extrasynthese (2 mentions)
Sodium chloride (nacl), by ITW Reagents (2 mentions)
P coumaric acid, by Merck Group (2 mentions)
Folin ciocalteu reagent, by Merck Group (1 mentions)
Nitric acid, by Merck Group (1 mentions)
Milli q water system, by Merck Group (1 mentions)
Hydrochloric acid (hcl), by Merck Group (1 mentions)
Acetone, by ITW Reagents (1 mentions)
Boric acid, by Chem-Lab (1 mentions)
L cysteine, by ITW Reagents (1 mentions)
Glutathione (gsh), by Merck Group (1 mentions)
Uric acid, by Merck Group (1 mentions)
α tocopherol, by Merck Group (1 mentions)
Ionol, by Merck Group (1 mentions)
Caffeic acid, by ITW Reagents (1 mentions)
2 2 azobis 2 amidinopropane dihydrochloride, by Merck Group (1 mentions)
12 protocols using ascorbic acid
1
Analytical Procedures for Phytochemical Characterization
2
Potentiometric Measurements of Antioxidants
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Potentiometric measurements were carried out using an Expert-pH pH-meter (OOO Ekoniks-expert, Moscow, Russia) with the EMF measurement function and the RS-232 interface. A two-electrode electrochemical cell was used. The working electrode was the EPV-1 redox platinum electrode, and the reference electrode was the EVL-1M silver/silver chloride electrode (Ag/AgCl/3 mol/dm3 KCl) (Gomel ZIP, Gomel, Belarus). The studies were carried out in a thermostated cell at 37 °C using the LOIP LT-205a circulation thermostat (ZAO LOIP, Saint Petersburg, Russia).
The following reagents were used in the work: 2,2’-azobis(2-amidinopropane) dihydrochloride (AAPH), α-tocopherol, 2,6-ditretbutylphenol, ionol, uric acid, glutathione (Sigma-Aldrich, St. Louis, MO, USA); K4[Fe(CN)6], K3[Fe(CN)6], KH2PO4, Na2HPO4·12H2O (Reakhim, Moscow, Russia); ascorbic acid, L-cysteine, catechol, gallic acid, chlorogenic acid, caffeic acid (Panreac, Barcelona, Spain). All reagents were analytical grade. The studies were carried out in phosphate buffer solution (PS) pH = 7.4 KH2PO4/Na2HPO4·12H2O.
The following reagents were used in the work: 2,2’-azobis(2-amidinopropane) dihydrochloride (AAPH), α-tocopherol, 2,6-ditretbutylphenol, ionol, uric acid, glutathione (Sigma-Aldrich, St. Louis, MO, USA); K4[Fe(CN)6], K3[Fe(CN)6], KH2PO4, Na2HPO4·12H2O (Reakhim, Moscow, Russia); ascorbic acid, L-cysteine, catechol, gallic acid, chlorogenic acid, caffeic acid (Panreac, Barcelona, Spain). All reagents were analytical grade. The studies were carried out in phosphate buffer solution (PS) pH = 7.4 KH2PO4/Na2HPO4·12H2O.
3
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).
4
Bioactive Compounds Extraction and Analysis
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Ethyl acetate, petroleum ether (boiling point 40–60 °C), HCl (37%), FeCl2·4H2O, FeCl3·6H2O, butanol, methanol, and 2,2-diphenyl-1-picrylhydrazyl (DPPH•) were obtained from Sigma-Aldrich (Munich, Germany). Folin–Ciocalteau reagent, vanillin and hydrogen peroxide solution (30% w/w) were purchased from Sigma (Steinheim, Germany), and potassium phosphate (monobasic and dibasic), sodium carbonate and quercetin from Fluka (Buchi, Switzerland). Trichloroacetic acid (TCA), gallic acid and catechin were obtained from Riedel-de Haen (Seelze, Germany), and ascorbic acid from Panreac (Barcelona, Spain).
5
Antioxidant Biochemical Assay Protocol
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Dithiothreitol (DTT), nitroblue tetrazolium (NBT), H2O2, titanium dioxide, dichloromethane, pyridine, N,O-bis(trimethylsilyl) trifluoroacetamide, trimethylsilyl chloride, sodium azide, thiobarbituric acid (TBA), methionine, catalase, peroxidase, cholesterol, maltose, octadecane, phenol, palmitic acid, sorbitol, ammonia and potassium phosphate buffer were purchased from Sigma-Aldrich (Saint Louis, MO, USA). Tetracosane and 4-aminoantipyrine were purchased from Fluka (Bucharest, Romania). Sulfuric acid was purchased from Fisher Chemical (New Hampshire, EUA). TrichloroAcetic acid (TCA) was purchased from BIOCHEM Chemopharma (Cosne-Cours-sur-Loire, France). Guaiacol and polyvinylpyrrolidone (PVP) were purchased from Acrós Organics. Triton-X and ascorbic acid were purchased from Panreac. Acetic acid, ethylenediaminetetraAcetic acid (EDTA) and phenylmethylsulfonyl fluoride (PMSF) were purchased from VWR (Pennsylvania, EUA). Riboflavin, n-hexane p.a., methanol p.a., sodium carbonate (Na2CO3), dimethyl sulfoxide (DMSO), potassium sulfate and quercetin were purchased from Merck (Darmstadt, Germany). Luteolin and oleuropein were purchased from Extrasynthese (Genay, France).
6
Synthesis of Rhodamine 6G-labeled BSA
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All chemicals were of reagent grade and have been used without further purification: tetraethylrhodamine hydrochloride (Rhodamine 6G) and BSA from Sigma Aldrich (Madrid, Spain), silver nitrate, sodium citrate, ascorbic acid and hydroxylamine hydrochloride from Panreac (Barcelona, Spain). Water was purified using a Milli-Q (18.2 MΩ) water system from Millipore (Madrid, Spain).
7
Culturing Aggressive Tongue Carcinoma Cell Line
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A HPV-negative, highly aggressive metastatic human tongue squamous cell carcinoma cell line (HSC-3, Japan Health Sciences Foundation, Japan) was cultured with Dulbecco’s Modified Eagle’s Medium (DMEM)/F-12 (Gibco Paisley, UK) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Gibco), 100 µg/ml streptomycin (Gibco), 100 U/ml penicillin (Gibco), 5 µg/ml amphotericin b (Sigma-Aldrich, St. Louis, Missouri, USA), and 50 µg/ml ascorbic acid (PanReac AppliChem, Darmstadt, Germany) in 75 m2 flasks. Cells were divided after reaching 80% confluence with 10% trypsin/EDTA (Gibco). The HSC-3 cells were tested negative for mycoplasma using a PromoKine PCR Mycoplasma Test Kit I/C (PromoCell, Heidelberg, Germany). The cell line was authenticated by FIMM Technology Center (Helsinki, Finland).
8
Synthesis and Characterization of Functional Nanomaterials
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Titanium (IV) oxide (TiO2, anatase, 20–30 nm particle size), (3-Aminopropyl) triethoxysilane (KH-550, >98%), Rhodamine B, Rhodamine 6G, Rhodamine 123, acrylamide (AM, >99%), ethylene glycol dimethacrylate (EDMA), 2,2’-Azobisisobutyronile (AIBN), styrene (≥99%), divinylbenzene (DVB) (80%), magnesium sulfate (99%), copper (II) sulfate (>99%), iron (II) chloride tetrahydrate (>98%), iron (III) chloride hexahydrate (>97%), ethylene glycol (98%), ammonium acetate (98%), ammonium phosphate (>99%), phosphoric acid (>85%) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Acetic acid (>99.7%), nitric acid (HNO3, >69%), ascorbic acid (>98%), citric acid (>99%), glucose (>99%), maltose monohydrate (>99%) were obtained from Panreac (Barcelona, Spain). Methanol (MeOH), ethanol (EtOH), acetonitrile (ACN) and acetone were delivered by Fisher Scientific (Geel, Belgium). MWCNTs of 30 ± 15 nm diameter (95% purity) were obtained from NanoLab Inc. (Waltham, MA, USA). Purified water with a Milli-Q system (Millipore, Bedford, MA, USA) was used for all the preparations.
9
Synthesis of Nitrohydroxytyrosol Derivatives
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Acetonitrile, formic acid, sodium chloride, disodium hydrogen phosphate anhydrous, potassium dihydrogen phosphate, phenolsulfonphthalein (phenol red), and ascorbic acid were acquired from Panreac (Barcelona, Spain). Antibiotics (gentamicin, penicillin, and streptomycin), enzymes (catechol-O-methyltransferase, b-glucuronidase/sulfatase), Sadenosyl-L-methionine chloride, UDP-glucuronic acid, non-essential amino acids and dimethyl sulfoxide were purchased from Sigma Chemical Co. (Madrid, Spain). Media, trypsine and fetal bovine serum (FBS) were from Biowhitaker (Innogenetics, Madrid, Spain). All reagents were of analytical or chromatographic grade. Hydroxytyrosol (HT) was recovered with 95% purity from olive oil wastewaters. 21 Nitrohydroxytyrosol (NO 2 HT), nitrohydroxytyrosyl acetate (NO 2 HT-A) and ethyl nitrohydroxytyrosyl ether (NO 2 HT-E) were prepared from free HT, isolated from olive oil wastewaters, as described elsewhere. 9, 10
10
Comprehensive Analysis of Phenolic Compounds
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Water, methanol, acetonitrile, and formic acid (Fisher Scientific, Fair Lawn, NJ, USA) were of Optima® LC/MS grade; ascorbic acid (Panreac, Barcelona, Spain), analytical grade; and glacial acetic acid (Merck, Darmstadt, Germany), Suprapur® quality. Leucine Enkephalin acetate hydrate and sodium formate solution were provided by Sigma-Aldrich Chemie (Steinheim, Germany). Luteolin-7-O-glucoside, kaempferol-3-O-glucoside, quercetin-3-O-galactoside, quercetin-3-Orhamnoside were purchased from Extrasynthèse (Genay, France); caffeoyltartaric acid and quercetin-3-O-glucoside, from Chromadex (Irvine, CA, USA); 5-O-caffeoylquinic acid, p-coumaric acid, 1,5-dicaffeoylquinic acid, 1,3-dicaffeoylquinic acid, and quercetin-3-O-rutinoside, from Sigma-Aldrich Chemie (Steinheim, Germany); and ferulic acid, caffeic acid, and 3,4-dihydroxybenzoic acid, from Fluka Chemie (Steinheim, Germany). Standard stock solutions of phenolic compounds were prepared in methanol; and a reference solution of these compounds (5 μg/mL), in methanol-water-acetic acid (30:65:5, v/v/v).
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