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5 protocols using hplc grade ethanol

1

Extraction and Analysis of Bioactive Compounds

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HPLC grade methanol, acetone, and methyl tert-butyl ether were acquired from Fisher Scientific Scharlau Chemie (Loughborough, UK). Sodium chloride was purchased from POCH S.A. (Sowińskiego, Poland). Ultrapure water was obtained with a Milli-Q system (Millipore Ibérica, Madrid, Spain). Ammonium carbonate, acetonitrile, hexane, ethanol (HPLC grade), magnesium chloride hexahydrate, acetic acid, and ammonium acetate were acquired from Scharlab (Sentmenat, Spain). Butyl hydroxytoluene (BHT) was purchased from Scharlau Chemie S.A. (Barcelona, Spain). Calcium chloride dihydrate was obtained from Merck (Darmstadt, Germany). Sodium hydrogen carbonate and potassium dihydrogen phosphate were acquired from VWR (Llinars del Vallès, Spain). Potassium chloride was obtained from Panreac (Castellar del Vallès, Spain). Digestive enzymes (porcine pepsin, porcine bile extract, porcine pancreatin, and porcine lipase) were acquired from Sigma-Aldrich (Darmstadt, Germany).
Caffeic acid, ferulic acid, and p-coumaric acid commercial patterns were obtained from Sigma-Aldrich (St. Louis, MO, USA). B-carotene standard was acquired from Carote-Nature (Ostermundigen, Switzerland), and α-carotene was acquired from Supelco-Merck (Darmstadt, Germany).
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2

Standards for Volatile Compound Analysis

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Standards of 2-nonanone (99%, CAS 821-55-6), n-undecane (99%, CAS 1120-21-4), camphor (95%, CAS 76-22-2), trans-2-nonenal (97%, CAS 18829-56-6), 1,3,5-triethybenzene (97%, CAS 102-25-0), 1-decanol (99%, CAS 112-30-1), 2-6-ditetrbutyl-1,4-benzoquinone (95%, CAS 719-22-2), diisobutyl phthalate (DIBP) (99%, CAS 84-69-5), methyl palmitate (99%, CAS 112-39-0), 4-heptanone (CAS 123-19-3), octanal (99%, CAS 124-13-0), isomentol (98%, CAS 89-78-1), isobornyl acrylate (98.5%, CAS 5888-33-5), diphenyl ether (99%, CAS 101-84-8), 2,4-di-tert-butylphenol (99%, CAS 96-76-4), 3,5-Di-tert-4-butylhydroxytoluene (BHT) (99%, CAS 128-37-0), 3,5-di-tert-Butyl-4-hydroxybenzaldehyde (99%, CAS 37942-07-7), benzophenone (99%, CAS 119-61-9), ethyl 4-(dimethylamino)benzoate (99%, CAS 10287-53-3) and n-alkanes (C8 to C20) were purchased from Sigma (Madrid, Spain). Ethanol (HPLC grade) was from Scharlau (Barcelona, Spain). Ultrapure water was obtained from a Milli-Q system (Millipore, Billerica, MA, USA).
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3

Synthesis and Characterization of Compound 2

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Cesium cobalt(III) bis(dicarbollide) (COSAN) (Katchem Ltd., Prague, Czech Republic), Gold(III) chloride trihydrate (HAuCl4·3H2O, Aldrich, Madrid, Spain), Sodium citrate (Sigma-Aldrich, St. Louis, MO, USA) and poly (ethylene glycol) methyl ether thiol (MW 5000, Sigma-Aldrich) were used as purchased. All other reagents and anhydrous solvents, stored over 4 Å molecular sieves, were purchased from Aldrich Chemical Co. (Madrid, Spain) and used without further purification. HPLC grade ethanol, methanol, and acetonitrile were purchased from Scharlab (Sentmenat, Barcelona, Spain).
Experiments were carried out, except when noted, under a dry, oxygen-free dinitrogen atmosphere. Column chromatography was performed using silica gel 60 (Scharlab). Analytical thin layer chromatography (TLC) measurements were conducted with silica gel 60 F254 plates (Macherey-Nagel, Hoerdt, France); and the spots were visualized under UV lamp. Compound 2 was prepared and characterized according to the previously reported protocol [40 (link)].
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4

Analytical Protocol for Brimonidine

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Brimonidine and 2-bromoquinoxaline were obtained from Sigma-Aldrich (Madrid, Spain). LAPONITE®-RD (LAP) (surface density 370 m 2 g -1 , bulk density 1000 kg m -3 , chemical composition: SiO 2 59.5%, MgO 27.5%, Li 2 O 0.8%, Na 2 O 2.8%) was obtained from BYK Additives (Widnes, Cheshire, UK). Balanced 0.9% salt solution (9 mg ml -1 NaCl) (BSS) was obtained from Fresenius Kabi (Barcelona, Spain). HPLC-grade ethanol, acetonitrile, methanol, ammonium formate, formic acid, ammonia and phosphoric acid (85% w/w) were obtained from Scharlab (Barcelona, Spain). Oasis MCX Prime 96-well µElution plates were obtained from Waters Chromatography (Barcelona, Spain).
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5

Characterization and Purity Analysis of Cathinone Compounds

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Research chemicals containing cathinones were provided by Energy Control (ABD Foundation, Barcelona, Spain). All the compounds were characterized and puritytested by UHPLC-HRMS and nuclear magnetic resonance, following the same procedures already reported in literature 42, (link)43 (link) . Cathinone stock solutions were prepared at approximately 1 mg/mL in methanol (0.01 mg accuracy). HPLC-grade water was obtained by purifying demineralized water using a Milli-Q system from Millipore (Bedford, MA, USA). HPLC-grade methanol, HPLC-grade acetonitrile, HPLC-grade ethanol, formic acid and acetone were purchased from Scharlau (Scharlab, Barcelona, Spain). Physiological saline solution was purchased from Laboratorios ERN (Barcelona, Spain).
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