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4 protocols using sodium bicarbonate

1

Vasoactive Compound Preparation Protocol

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Magnesium sulfate (MgSO4), potassium chloride (KCl), calcium chloride (CaCl2), sodium chloride (NaCl) and formaldehyde were purchased from Vetec Química Fina Ltda. (Brazil). Glucose (C6H12O6) and sodium bicarbonate (NaHCO3) were purchased from Dinâmica (Brazil). Potassium monobasic phosphate (KH2PO4), sodium hydroxide (NaOH) and hydrochloric acid (HCl) were purchased from Nuclear (Brazil). These substances, except glucose, NaCl and NaHCO3 were diluted in distilled water to obtain each solution, which were maintained under refrigeration.
Phenylephrine (Phe) was purchased from Pfizer (USA). Acetylcholine (ACh), sodium nitroprusside (SNP), L-Nω-nitroarginine methyl ester (L-NAME), indomethacin, tempol, apocynin, Cremophor® and R-(-)-apomorphine were obtained from Sigma-Aldrich (Brazil). All substances were diluted in distilled water as needed for each experimental protocol. The carbogen mixture (95% O2 and 5% CO2) was acquired from White Martins (Brazil).
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2

Characterization of Pharmacological Agents

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Magnesium sulphate (MgSO4), potassium chloride (KCl), calcium chloride (CaCl2), and sodium chloride (NaCl) were purchased from Vetec Química Fina Ltda. (Brazil). Glucose (C6H12O6) and sodium bicarbonate (NaHCO3) were purchased from Dinâmica (Brazil). Sodium dihydrogen phosphate (NaH2PO4) was purchased from Nuclear (Brazil). Histamine, atropine, Cremophor®, Triton-X 100®, apamin, cesium chloride (CsCl), tetraethylammonium chloride (TEA+), 4–aminopyridine (4–AP), and glibenclamide were obtained from Sigma–Aldrich (Brazil). Carboxymethylcellulose and castor oil were obtained from Fórmula (Brazil). Loperamide was obtained from Janssen Cilag Farmacêutica Ltda. (Brazil) and the activated charcoal was obtained from Proquímios (Brazil).
All substances were diluted in distilled water and the extract was solubilized in Cremophor®, dissolved in distilled water as needed for each experimental protocol. The final concentration of Cremophor® did not show any interference in the in vivo experiments, according to data from previous experiments.
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3

Synthesis and Characterization of Acetylated Carvacrol

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A solution containing 1 g of carvacrol (purity ≥ 98%; Sigma-Aldrich ® , St. Louis, USA), 15 mL of acetic anhydride P.A. (Dinâmica ® , São Paulo, Brazil) and 1.5 g of sodium acetate P.A. (Dinâmica ® , São Paulo, Brazil) was refluxed for 1 h at room temperature. Cold water was added (20 mL), and the solution was neutralized to pH 7.0 with 5% sodium bicarbonate P.A. (Dinâmica ® , São Paulo, Brazil). The solution was transferred to a separating funnel and washed three times with chloroform P.A. (100 mL) (Dinâmica ® , São Paulo, Brazil). The chloroform phase, containing the acetylated material, was washed with water and dried with sodium sulfate P.A. (Dinâmica ® , São Paulo, Brazil). The solvent was rotoevaporated (Matos, 1997) . CVA was subjected to thin layer chromatography and characterized by gas chromatography mass spectrometry (GC-MS). The yield of CVA was 83%.
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4

TNBS-Induced Colitis Model Evaluation

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TNBS, non-selective COX inhibitor, acetylsalicylic acid (ASA), selective COX-1 inhibitor (SC-560), ethanol (etOH), fluorescein, hexadecyltrimethylammonium bromide (HTAB), hydrogen peroxide (H2O2), and O-dianisidine were purchased from Sigma® (Brazil). The selective COX-2 inhibitor (celecoxib) was purchased from Pfizer (Brazil). The salts used for the Krebs solution (sodium chloride, potassium chloride, calcium chloride, magnesium sulfate, monobasic potassium phosphate, sodium bicarbonate, and glucose) were obtained from Dinâmica® (Brazil).
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