The solvents acetonitrile and formic acid were obtained from Tedia (HPLC grade, Radnor, PA, USA), and ultrapure water was obtained from the Milli-Q® Plus apparatus by Millipore (Billerica, MA, USA). Methanol, ethanol, isopropanol, polysorbate 80 (Tween® 80), dimethyl sulfoxide (DMSO), and sodium hydroxide were obtained from Vetec (Duque de Caxias, Brazil), and medium chain triglycerides (MCT) and egg lecithin (Lipoid® E80) were purchased from Lipoid GmbH (Ludwigshafen am Rhein, Germany). Monobasic potassium phosphate was obtained from Dinâmica (São Paulo, Brazil), and NBD-PE [N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine, triethylammonium salt] fluorescent-labelled phospholipid was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Roswell Park Memorial Institute 1640 broth medium (RPMI-1640), 3(N-morpholino) propane sulfonic acid (MOPS), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and itraconazole were obtained from Sigma-Aldrich (St. Louis, MA, USA), and potato dextrose agar (PDA) was purchased from Acumedia (San Bernardino, CA, USA). Tissue-Tek® O.C.T.™ was purchased from Sakura Finetechnical Co. (Tokyo, Japan). Log P (calculated log P) values of the coumarins were assigned using SwissADME http://www.swissadme.ch/ (accessed on 10 March 2023).
Isopropanol
Manufactured by Vetec
Sourced in Brazil
Isopropanol is a clear, colorless liquid chemical compound. It is a type of alcohol commonly used in various industrial and laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Itraconazole, by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Formic acid, by Merck Group (1 mentions)
Milli q plus, by Merck Group (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (1 mentions)
Monobasic potassium phosphate, by Dinâmica (1 mentions)
Nbd pe, by Thermo Fisher Scientific (1 mentions)
N 7 nitrobenz 2 oxa 1 3 diazol 4 yl 1 2 dihexadecanoyl sn glycero 3 phosphoethanolamine triethylammonium salt, by Thermo Fisher Scientific (1 mentions)
Potato dextrose agar (pda), by Acumedia (1 mentions)
3 n morpholino propanesulfonic acid mops, by Merck Group (1 mentions)
Sodium hydroxide (naoh), by Vetec (1 mentions)
Ethanol, by Vetec (1 mentions)
Methanol, by Vetec (1 mentions)
Rpmi mops, by Merck Group (1 mentions)
Mueller hinton broth 2, by Merck Group (1 mentions)
Acetonitrile, by Tedia (1 mentions)
Hexane, by Vetec (1 mentions)
Sodium sulfate na2so4, by Vetec (1 mentions)
Milli q system, by Merck Group (1 mentions)
Glc 603, by Nu-Chek Prep (1 mentions)
6 protocols using isopropanol
1
Characterization of Coumarins' Physicochemical Properties
2
Antimicrobial Polymer-Based Formulation
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Poly(epsilon-caprolactone) (PCL, Mn 80 kDa), chitosan of low molecular weight (50–190 kDa) and 75%–85% deacetylation degree, Mueller-Hinton broth 2, RPMI-MOPS, and MTT bromite of 3-(4,5-dimetiltiazol-2-il)-2, 5-dipheniltet razolium) were obtained from Sigma-Aldrich (St Louis, MO, USA). Medium-chain triglyceride (MCT) was purchased from Delaware (Porto Alegre, RS, Brazil), α-bisabolol and triclosan were acquired from Fragon (São Paulo, SP, Brazil), isopropanol and acetone were obtained from Vetec (São Paulo, SP, Brazil), and acetonitrile and ethanol HPLC standard were purchased from Tedia (São Paulo, SP, Brazil). Glacial acetic acid, Lipoid S75® (soybean lecithin), and polysorbate 80 were acquired from Fmaia (Belo Horizonte, MG, Brazil), Lipoid (Ludwigshafen, RP, Germany), and Henrifarma (São Paulo, SP, Brazil), respectively.
3
Triglyceride Quantification in Liver Tissue
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Triglyceride content was determined as previously described by Folch et al.
(27 (link)). Briefly, 50 mg of liver tissue
was homogenized in 1 mL of isopropanol (Vetec, Brazil) and centrifuged (740
g, 10 min, 4°C). The triglyceride content of the
supernatant was determined with a colorimetric kit following the manufacturer’s
instructions (Bioclin, Brazil). The absorbance was measured at 490–540 nm
(Hidex, Finland).
(27 (link)). Briefly, 50 mg of liver tissue
was homogenized in 1 mL of isopropanol (Vetec, Brazil) and centrifuged (740
g, 10 min, 4°C). The triglyceride content of the
supernatant was determined with a colorimetric kit following the manufacturer’s
instructions (Bioclin, Brazil). The absorbance was measured at 490–540 nm
(Hidex, Finland).
4
Chocolate Samples Fatty Acid Analysis
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Thirty-two chocolate samples (white, dark, and milk chocolates) from different national and multinational brands produced in Brazil were used to carry out this research. Samples were purchased in local markets. The addition of PHVO is still acceptable in this country, and some products had high trans fats contents declared. All reagents used in the experiments were of analytical grade, and water used in aqueous solutions was purified by deionization (Milli-Q system; Millipore, Bedford, MA, USA). Methanol, potassium hydroxide (KOH) and sodium bisulfate (NaHSO4) were purchased from Panreac Química S.A. (Madrid, Spain). Hexane, isopropanol, and sodium sulfate (Na2SO4) were obtained from Vetec (Rio de Janeiro, RJ, Brazil). Analytical FAME standards (GLC-603, GLC-611, GLC-643, GLC-80, GLC-409) were acquired from Nu-Chek Prep Inc. (Elysian, MN, USA).
5
Liver Lipid Quantification Protocol
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Liver samples (50 mg) were homogenized in 1 mL isopropanol (Vetec, Rio de Janeiro, Brazil) and centrifuged (2000× g, 10 min, 4 °C). We used n = 10 offspring per sex/group. The total triglyceride and cholesterol levels were measured using a colorimetric method with a commercial kit (Bioclin, Belo Horizonte, Brazil), as previously reported [62 (link)].
6
Liver Lipid and Glycogen Analysis
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Liver samples (50 mg) were homogenised in 1 ml isopropanol (Vetec) and centrifuged (5900 rpm, 10 min, 48C). Total TAG and cholesterol levels were measured by a colorimetric method using a commercial kit (Bioclin) (14) . The glucose produced by glycogen hydrolysis was measured using a commercial kit (Glucox; Doles). Liver was homogenised with 4 ml TCA (10 %). After centrifugation (1000 g, 48C, 10 min), 2 ml of supernatant were added to 5 ml of absolute ethanol and frozen. After 24 h, the mixture was centrifuged (1000 g, 48C, 10 min), and the supernatant was discarded. Glycogen was hydrolysed by boiling the pellet for 30 min with 1 M-HCl. After addition of 1 ml of 1 M-NaOH to neutralise the mixture, glucose was measured in 200 ml of supernatant as described previously (15) .
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