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6 protocols using vitamin b6

1

Bentonite Characterization with Reagents

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Vitamin B6 (pyridoxine hydrochloride), hydrochloric acid (HCl, 37% w/w, Merck), sodium hydroxide (NaOH, analytical reagent; Merck), sodium chloride (NaCl, analytical reagent; Merck), and calcium chloride, (CaCl2, analytical reagent; Merck) was used as received. Deionized water (the water was purified to a resistivity of ≥18.2 MΩ cm) was used in all experiments. The bentonite sample was provided by Salafchegan bentonite mine (Iran).
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2

Electrochemical Analysis of Biomolecules

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The measurements were performed on an Autolab potentiostat/ galvanostat (PGSTAT 302 N, Eco Chemie, the Netherlands), while the experimental parameters were controlled using General Purpose Electrochemical System (GPES) software. In addition to the working electrode, which was the modified GCE, the three-electrode cell further included an Ag/AgCl/KCl (3.0 M) reference electrode and a platinum wire as the auxiliary. The pH measurements were performed on a Metrohm 710 pH meter.
The solutions used for the experiments were freshly prepared. Levodopa, vitamin B6, theophylline, guaifenesin and the other chemicals used were of analytical grade and were procured from Merck chemical company (Darmstadt, Germany). Orthophosphoric acid and its salts were used to prepare the buffer solutions with pH values in the range of 2.0 -9.0. Hematoxylin was obtained from Sigma Aldrich Co. (USA) and graphene/ZnO nanorods were prepared as described elsewhere. 40
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3

Comprehensive Antioxidant Profiling

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All the reagents used were of analytical or HPLC grade. Methanol and ethanol (96%) were purchased from Fisher Chemicals (Fair Lawn, NJ, USA). DPPH (2,2-diphenyl-1-picrylhydrazyl), Trolox (6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid), acetic acid, sodium acetate, TPTZ (2,4,6-tris(2-pyridyl)-s-triazine), hydrochloric acid, iron (III) chloride hexahydrate, Folin-Ciocalteau reagent, sodium carbonate, gallic acid, MES monohydrate, tris(hydroxymethyl)aminomethane, L(+)-ascorbic acid, DL-malic acid, L(+) tartaric acid, β-carotene, vitamin B6, and vitamin E were purchased from Sigma-Aldrich (St. Louis, MO, USA). D(+)-Glucose and D(−)-Fructose were purchased from VWR International (Radnor, PA, USA). Lutein was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Celite was purchased from AppliChem (Darmstadt, Germany). Sodium hydroxide and Acetone were purchased from LabChem (Zelienople, PA, USA). Ultrapure water was obtained from a Synergy® Water Purification System (MilliporeSigma, Burlington, MA, USA).
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4

Analytical Workflow for Bifonazole and CTZ

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Bifonazole and CTZ were obtained from Sigma-Aldrich Corp. (St. Louis, MO, USA). HPLC-grade methanol, acetonitrile, and formic acid (98%) were obtained from JT Baker. HPLC-grade dichloromethane was obtained from Merck (Darmstadt, Germany).
Malt extract, glucose, casein hydrolyzate, l-asparagine, adenine, vitamin B6, and yeast extract were obtained from Sigma-Aldrich Corp. (St. Louis, MO, USA). Chemicals such as KH2PO4, MgSO4·7H2O, FeCl3, MnSO4·H2O, NH4Cl, CaCl2·6H2O, and ZnSO4·7H2O were obtained from PPH Golpharm (Kraków, Poland). Water (quadruple-distilled) with a conductivity of less than 1 µS cm−1 was prepared using S2-97A2 distillation apparatus (ChemLand, Stargard Szczecin, Poland).
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5

Quantification of essential vitamins

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The standards used during the analyses, thiamin (≥ 99.9% purity), riboflavin (≥ 99.9%), niacin (≥ 99.9%), pantothenic acid (≥ 99.9%), vitamin B6 (≥ 99.9%), vitamin C (≥ 98.7%), vitamin A (≥ 99.2%), vitamin E (≥ 99.9%) and β-carotene (≥ 97.6%) were purchased from Sigma–Aldrich (St. Louis, MO, USA). Acetonitrile, formic acid and ammonium formate of LC–MS grade were purchased from Thermo Fisher Scientific (Fair Lawn, NJ, USA). The 0.22 μm syringe filters were purchased from Agela Technologies (Tianjin, China). All other chemicals were of analytical grade and purchased locally. Ultra–pure water was provided by a Siemens Water Technologies system (Warrendale, PA, USA).
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6

Nutrient-mediated Modulation of Cellular Signaling

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The following substances were purchased from Sigma Aldrich and used without further purification: lipopolysaccharide (LPS, from Escherichia coli 055:B5, cat #L6529), vitamin B 6 (cat #P5669-5G), B 9 (folic acid, cat #F7878), vitamin B 12 (cat #V6629), vitamin A (cat #95,144), all-trans retinoic acid (RA, cat #R2625), vitamin D 3 in a form of 7-dehydrocholesterol/vitamin 25(OH)D 3 (100 mg/ml in ethanol, cat #47,763), cis-4,7,10,13,16,19-docosahexaenoic acid (DHA, cat #D2534), cis-5,8,11,14,17-eicosapentaenoic acid (EPA, cat #E7006) , valproic acid (VPA, cat #P4543), S-(5 0 -adenosyl)-L-homocysteine (SAH, cat # A9384), anacardic acid (ANA, cat # A7236). Inhibitors of retinoic acid receptors (RAR) were purchased from Tocris Bioscience: BMS 195614 (4-[[[5,6-dihydro-5,5-dimethyl-8-(3-quinolinyl) -2-naphthalenyl]carbonyl]amino]benzoic acid, cat # 3660), 8, 9, 7, 7, 10, naphtho [2,3-b][1,4] diazepin-13-yl)benzoic acid, cat # 2021), MM 11253 (6-[2-(5,6,7,8-tetrahydro-5,5,8,8-
). The concentrations of nutrient stock solutions used in the experiments are listed in Table 1.
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