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9 protocols using sodium sulphide

1

Synthesis and Characterization of Silver Nanoparticles

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Trisodium citrate dihydrate, silver nitrate, potassium chloride, ethylene glycol and acetone were purchased from POCH S.A. (Gliwice, Poland). Sodium sulphide was acquired from Sigma-Aldrich (St. Louis, MO, USA). Polyvinylpyrrolidone (PVP) with an average molar mass of ca. 4 × 104 g mol−1 was purchased from Fluka (Seelze, Germany). Glycerin and ammonium fluoride were purchased from Chempur (Piekary Slaskie, Poland). Pyridine was purchased from Ubichem (Eastleigh, UK). All of the chemicals were used without further purification or treatment. The water was purified by a Millipore Milli-Q system and had a resistivity of ca. 18 MΩ cm−1. The silver used as a material for electrodes was purchased from the Polish Mint (Warsaw, Poland). Titanium foil of 0.25 mm thick and 99, 5% purity was purchased from Alfa Aesar (Kandel, Germany). It was cut into 1 cm2 round plates. The plates were cleaned ultrasonically with acetone and ethanol, rinsed with deionized (DI) water, and dried in air.
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2

Synthesis and Characterization of Colloidal Nanoparticles

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Silver nitrate (Honeywell, 99.8%), indium(III) chloride (Sigma-Aldrich, 98%), L-reduced glutathione (Sigma-Aldrich), 3-mercaptopropionic acid (AppliChem), sodium sulphide (Abcr, 98%), ammonium hydroxide 5 M (Sigma-Aldrich), citric acid (Roth, 99.5%), zinc acetate (Chemsolute, 99.5%), ethanol (Merck, absolute), sodium 3-mercaptopro-1-propanesulfonic acid (ChemPUR), thioglycolic acid (Merck) were used in the present work without additional purification.
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3

Nanoparticle Synthesis and Characterization

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Copper chloride (CuCl2), indium chloride (InCl3), sodium citrate dihydrate (Na3C6H5O7), L-glutathione (GSH), sodium sulphide (Na2S), zinc acetate dihydrate (Zn(O2CCH3)2(H2O)2), thiourea (CH4N2S), sodium hydroxide (NaOH), hydrochloric acid (HCl), ethanol (CH3CH2OH), dichloromethane (CH2Cl2), methanol (CH3OH), pyrrole (C4H4NH), n-hexane (C6H14), 3-hydroxybenzaldehyde (C7H6O2), propionic acid (CH3CH2CO2H), chloroform (CHCl3), sodium bicarbonate (NaHCO3), acetone (CH3)2CO), and ethyl acetate (CH3COOCH2CH3), were purchased from Sigma Aldrich, South Africa and used as received. RPMI medium, fetal bovine serum, and Mueller Hinton (MH) agar plates. Deionized water was used for the synthesis and analysis.
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4

Porcine Pancreatin Digestion Protocol

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The pancreatin (from Alpha Aesar, Lancester, UK) was from porcine pancreas. The sodium di-hydrogen phosphate, tryptone, pepsin, porcine bile acids, cysteine, resazurin, sodium sulphide and salivary alpha-amylase were from Sigma-Aldrich (Darmstadt, Germany).
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5

Enzyme-Mediated Biomolecule Extraction

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Cysteine, sodium di-hydrogen phosphate, sodium sulphide, resazurin, salivary α-amylase, and pepsin from porcine bile acids (porcine bile extract) were provided by Sigma-Aldrich (Darmstadt, Germany). Pancreatin from porcine pancreas was provided by Alpha Aesar (Lancaster, UK).
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6

Synthesis and Characterization of Copper-Indium Nanoparticles

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Copper chloride (CuCl2), indium chloride (InCl3), sodium citrate (Na3C6H5O7), L-glutathione (GSH), sodium sulphide (Na2S), zinc acetate dihydrate (Zn(O2CCH3)2(H2O)2), thiourea (CH4N2S), sodium hydroxide (NaOH), hydrogen chloride (HCl), ethanol (CH3CH2OH), ethyl acetate (CH3COOCH2CH3), methanol (MeOH) pyrrole, n-hexane, 3-hydroxybenzaldehyde, propionic acid, sodium bicarbonate (NaHCO3), methylene blue (MB), 1,3-diphenylbenzofuran (DPBF) and dimethyl sulfoxide (DMSO) were all purchased from Sigma Aldrich, South Africa. The 4-L electric pressure cooker was bought from www.takealot.com. Amphotéricine B-1, DMEM - high glucose, glutamine, SVF, trypsin, Pénicilline streptomycin were purchased from Sigma Life Sciences, France. Isopropanol and chloroform were purchased from Carlo Erba reagents, France. Trizol extract-ALL Rna extraction reagent was purchased from OMEGA bio-tek and WST-1 cell proliferation reagent was purchased from Roche. iScript™ cDNA Synthesis Kit was purchased from Bio-Rad (Marnes-la Coquette, France). All chemicals were used without further purification except for pyrrole which was distilled before it was used.
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7

In Vitro Digestion and Fermentation Protocol

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The reagents used for the in vitro digestion and fermentation were: potassium di-hydrogen phosphate, potassium chloride, magnesium chloride hexahydrate, sodium chloride, calcium chloride dihydrate, sodium mono-hydrogen carbonate, ammonium carbonate, and hydrochloric acid, all purchased from Sigma-Aldrich (Germany). The enzymes (salivary alpha-amylase and porcine pepsin) and bile acids (porcine bile extract) were obtained from Sigma-Aldrich, and porcine pancreatin was from Alfa Aesar (United Kingdom). The fermentation reagents (sodium di-hydrogen phosphate, sodium sulphide, tryptone, cysteine, and resazurin) were obtained from Sigma-Aldrich (Germany).
Short chain fatty acids standards (acetic acid, propionic acid, and butyric acid) were purchased from Sigma-Aldrich (Germany).
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8

Biomass Preparation and Chemical Characterization

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For the study, eucalyptus and casuarina trees were felled from the research plots established in Forest College and Research Institute, Mettupalayam (Lat. 11°19′25.37’’ and Long. 76°56′8.84’’), Tamil Nadu, India. The logs collected from the felled trees were chipped using a pilot chipper and then graded using vibrating chip classifier to get uniform sized (about 2–3 cm long) wood chips. Then the wood chips were subjected to washing to remove impurities. After washing the wood chips were dried under atmospheric conditions for 2 weeks and then stored in air tight container for further use. Chemicals such as Sodium hydroxide, Sodium carbonate, Sodium sulphide, Oxalic acid, Acetic acid and Sodium chlorite were obtained from Sigma-Aldrich, India. All the chemicals used were of Analytical Reagent grade (AR grade) and used as received.
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9

Biochemical Analysis of Antioxidant Compounds

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Trolox ((±)-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid), 2,4,6-tri(2-pyridyl)-s-triazine (TPTZ), gallic acid, acetic acid, propionic acid, butyric acid, Folin-Ciocalteu reagent, sodium hydroxide, hydrochloric acid, ferric chloride hexahydrate, sodium acetate, potassium chloride, potassium dihydrogen phosphate, sodium monohydrogen carbonate, sodium chloride, magnesium chloride hexahydrate, ammonium carbonate, calcium chloride dihydrate, sodium dihydrogen phosphate, tryptone, cysteine, sodium sulphide, resazurin, salivary α-amylase, porcine pepsin, bile acids ( porcine bile extract), pancreatin and ethanol were from Sigma-Aldrich (Darmstadt, Germany).
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