Amiodarone hydrochloride

Manufactured by Merck Group

Sourced in United Kingdom, United States

Amiodarone hydrochloride is a white to off-white crystalline powder. It is a class III antiarrhythmic agent used to treat and prevent certain types of abnormal heart rhythms.

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Lab products found in correlation

Dimethyl sulfoxide (dmso), by Merck Group (3 mentions) Lc ms grade methanol, by Thermo Fisher Scientific (2 mentions) Doxorubicin hydrochloride, by Merck Group (2 mentions) Acetaminophen, by Merck Group (2 mentions) Chloroquine diphosphate, by Merck Group (2 mentions) Blasticidin s hydrochloride, by Thermo Fisher Scientific (1 mentions) Water and acetonitrile, by Thermo Fisher Scientific (1 mentions) Dulbecco s modified eagle s medium, by Merck Group (1 mentions) Ketoconazole, by Merck Group (1 mentions) Amiodarone d4 hydrochloride, by LGC (1 mentions) Desethylamiodarone d4 hydrochloride, by LGC (1 mentions) α naphthoflavone, by Merck Group (1 mentions) Formic acid, by Merck Group (1 mentions) Desethylamiodarone hydrochloride, by LGC (1 mentions) Hanks balanced salt solution (hbss), by Thermo Fisher Scientific (1 mentions) Nigericin, by Merck Group (1 mentions) Chloroquine, by Merck Group (1 mentions) Nh4cl, by Merck Group (1 mentions) Monensin, by Merck Group (1 mentions) Anti gapdh, by Santa Cruz Biotechnology (1 mentions)

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Amiodarone (42 citations) Hydrochlorides (2 citations)

18 protocols using amiodarone hydrochloride

Amiodarone Metabolism Profiling

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Dulbecco’s Modified Eagle’s Medium (DMEM), amiodarone hydrochloride, formic acid, dimethyl sulfoxide (DMSO), α-naphthoflavone, and ketoconazole were purchased from Sigma-Aldrich (St. Louis, MO). Desethylamiodarone hydrochloride, amiodarone-d4 hydrochloride, and desethylamiodarone-d4 hydrochloride were obtained from Toronto Research Chemicals Inc. (Toronto, ON, Canada). Blasticidin S hydrochloride was purchased from Life Technologies (Grand Island, NY). Human microsomes were obtained from BioreclamationIVT (Baltimore, MD). Water and acetonitrile were LC–MS grade and obtained from Fisher Scientific (Pittsburgh, PA).

Wu Q., Ning B., Xuan J., Ren Z., Guo L, & Bryant M.S. (2016). The role of CYP 3A4 and 1A1 in amiodarone-induced hepatocellular toxicity. Toxicology letters, 253, 55-62.

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Autophagy Protein Detection and Inhibition

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The following primary antibodies were used at the indicated dilutions: anti-LC3A/B (1:1000 for western blotting and 1:100 for immunofluorescence; Cell Signaling Technology, 4108), anti-LC3A (1:100 for immunohistofluorescence; Abgent, 1805a), anti-ATG12 (1:100; Cell Signaling Technology, 2010), anti-ATG13 (1:1000; Cell Signaling Technology, 6940), anti-ATG16L1 (1:1000; Cell Signaling Technology, 8089), anti-human LAMP1 and anti-mouse LAMP1 (1:100; Becton Dickinson, 555798 and 553792), anti-WIPI2 (1:100; Bio-Rad, MCA5780GA), anti-CDSN/corneodesmosin (R&D Systems, AF5725), anti-GAPDH (1:2000; Santa Cruz Biotechnology, sc-25778), and anti-ATP6V0D1 (1:50; ABCAM, ab56441). The following inhibitors and reagents were used: Amiodarone hydrochloride (Sigma, A8423), bafilomycin A₁ (Tocris Biosciences, 1334), betahistine dihydrochloride (Sigma, B4638), chloroquine (Sigma, C6628), CCCP (Sigma, C2759), HBSS (Gibco, 14025–092), hydroxychloroquine (Sigma, H0915), lidocaine (Sigma, L7757), lidocaine hydrochloride monohydrate (Sigma, L5647), monensin (Sigma, M5273), NH₄Cl (Sigma, 213330), nigericin (Sigma, N7143), PP242 (Tocris, 4257), procainamide hydrochloride (Sigma, P9391). Mito-TMRE (abcam, 113852), PIK3C3/VPS34 inhibitor (IN-1) was kindly provided by Dr I. Ganley, University of Dundee.

Jacquin E., Leclerc-Mercier S., Judon C., Blanchard E., Fraitag S, & Florey O. (2017). Pharmacological modulators of autophagy activate a parallel noncanonical pathway driving unconventional LC3 lipidation. Autophagy, 13(5), 854-867.

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MALDI-TOF Mass Spectrometry Protocols

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Methanol (LC-MS grade) was purchased from Fisher Scientific (Leicestershire, UK); purified water, ELGA Purelab Option (Marlow, UK); MALDI matrices and analytes, 2,5-dihydroxybenzoic acid (2,5-DHB, >99%), 9-aminoacrinid (9AA, ≥99.5%), d-(+)-raffinose pentahydrate (≥98%) were purchased from Sigma-Aldrich (Dorset, UK) and used as supplied. Amiodarone hydrochloride (≥98%) was purchased from Sigma-Aldrich (Dorset, UK) and used as supplied or blended with lactose monohydrate, Lactohale (GMS 15284, Grade 4), EP and USP and JP, bovine (DFE Pharma, Goch, Germany) for use by inhalation. All other materials were of analytical grade.

Patel A., Hoffman E., Ball D., Klapwijk J., Steven R.T., Dexter A., Bunch J., Baker D., Murnane D., Hutter V., Page C., Dailey L.A, & Forbes B. (2019). Comparison of Oral, Intranasal and Aerosol Administration of Amiodarone in Rats as a Model of Pulmonary Phospholipidosis. Pharmaceutics, 11(7), 345.

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Preparation and Characterization of Samples

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Methanol (LC-MS grade) was purchased from Fisher Scientific (Leicestershire, UK); purified water from was purchased from ELGA Purelab Option (Marlow, UK); and raffinose pentahydrate (≥ 98%) and amiodarone hydrochloride (≥ 98%) were purchased from Sigma-Aldrich (Dorset, UK) and used as supplied. Superfrost Plus (Thermo Scientific, Waltham, MA, USA) glass slides were used for all experiments.

Dexter A., Steven R.T., Patel A., Dailey L.A., Taylor A.J., Ball D., Klapwijk J., Forbes B., Page C.P, & Bunch J. (2019). Imaging drugs, metabolites and biomarkers in rodent lung: a DESI MS strategy for the evaluation of drug-induced lipidosis. Analytical and Bioanalytical Chemistry, 411(30), 8023-8032.

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HepG2 Cell Cytotoxicity Assay

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HepG2 cells were seeded at a density of 1 × 10⁵ cells/cm² on culture dishes in Dulbecco’s modified Eagle’s medium (Wako) supplemented with 10% (v/v) fetal bovine serum (Biowest), and then cultured in 5% CO₂ at 37°C. Reagents were purchased from the indicated suppliers: ampicillin, DMSO, and amiodarone hydrochloride (Sigma-Aldrich); sodium arsenite (Merck-Millipore); G418 (Calbiochem); and MG132, CCl₄, N-nitrosodiethylamine, APAP, and methotrexate (Wako). All reagents used in cytotoxicity tests were dissolved in 1% DMSO.

Tsuji S., Ohbayashi T., Yamakage K., Oshimura M, & Tada M. (2016). A Cytoplasmic Form of Gaussia luciferase Provides a Highly Sensitive Test for Cytotoxicity. PLoS ONE, 11(5), e0156202.

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Cardiac and Bone Tumor Cell Assays

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Cardiac tissues were studied using caffeine (50 mM in water; Sigma-Aldrich), amiodarone hydrochloride (2.418 μM in DMSO; Sigma-Aldrich), isoproterenol hydrochloride (a series of drug concentrations in water; Sigma-Aldrich), or doxorubicin hydrochloride (1 μM in DMSO; Sigma-Aldrich), all diluted in RPMI Medium 1640 supplemented with B-27™. Response to isoproterenol was analyzed 10 minutes after exposure to 1 mM isoproterenol hydrochloride, diluted in RPMI Medium 1630 supplemented with B27™.
ES bone tumor cell lines and tissues were studied using either doxorubicin hydrochloride (10 mM in water; Sigma-Aldrich), linsitinib (OSI-906) (various concentrations in DMSO; Santa Cruz Biotechnology), all diluted in either non-metastastic media (RPMI Medium 1640, 10% FBS, 1% PenStrep) or metastatic media (EMEM, 10% FBS, 1% PenStrep).
Both tissues were treated with linsitinib, dissolved at a 10 mM concentration in DMSO (Corning) and mixed in with the respective cell medium at a 12 mM concentration unless otherwise noted. Vehicle treatments involved just the addition of DMSO at identical volumes as a control. Tissues were randomly assigned to experimental groups. Medium was changed every day.

Chramiec A., Teles D., Yeager K., Marturano-Kruik A., Pak J., Chen T., Hao L., Wang M., Lock R., Tavakol D.N., Lee M.B., Kim J., Ronaldson-Bouchard K, & Vunjak-Novakovic G. (2020). Integrated human organ-on-a-chip model for predictive studies of anti-tumor drug efficacy and cardiac safety. Lab on a chip, 20(23), 4357-4372.

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Amiodarone Biophysical Effects Protocol

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amiodarone hydrochloride (Sigma-Aldrich, St. Louis, MO) was dissolved in
dimethyl sulfoxide (DMSO; Sigma-Aldrich) to create a stock concentration of 30
mM. For experiments, amiodarone stock was diluted into bath solution for use at
3 μM concentration. Fresh dilutions were made on the day of experiments.
amiodarone or DMSO was continually present in the superfusate during experiments
examining the biophysical effects of amiodarone. DMSO concentration never
exceeded 0.01% in control or test conditions.

Beckermann T.M., McLeod K., Murday V., Potet F, & George AL J.r. (2014). Novel SCN5A Mutation in Amiodarone-responsive Multifocal Ventricular Ectopy-associated Cardiomyopathy. Heart rhythm : the official journal of the Heart Rhythm Society, 11(8), 1446-1453.

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HPLC Characterization of Pharmaceutical Compounds

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Pharmaceutical-grade standards of terfenadine (TER), fexofenadine hydrochloride (FEX), amiodarone hydrochloride, papaverine hydrochloride, and starch purchased from Sigma-Aldrich, St. Louis, MO, USA; acetonitrile, methanol, sodium hexanesulfonate, and triethylamine (TEA) from Merck, Darmstadt, Germany; acetic acid (CH₃COOH), sodium acetate (CH₃COONa), hydrochloric acid, sodium chloride (NaCl), sodium tetraborate (Na₂B₄O₇), sodium hydrogen phosphate (NaHPO₄), sodium hydroxide (NaOH), kalium dihydrogen phosphate (KH₂PO₄), and kalium hydroxide (KOH) from POCh, Gliwice, Poland were used. Buffers for LC methods (acetate buffers of pH 3.1 and 4.8) as well as buffers for degradation (acetate buffer of pH 4.0, phosphate buffer of pH 7.0 and borate buffer of pH 10.0) were prepared as described in European Pharmacopoeia [26 ]. The buffers used for degradation have the same ionic strength of 1 M, which was attained with 4 M NaCl.

Gumieniczek A., Berecka-Rycerz A., Pietraś R., Kozak I., Lejwoda K, & Kozyra P. (2019). Comparative Study of Chemical Stability of Two H1 Antihistaminic Drugs, Terfenadine and Its In Vivo Metabolite Fexofenadine, Using LC-UV Methods. Journal of Analytical Methods in Chemistry, 2019, 5790404.

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Evaluation of Cardiac and Hepatic in vitro Models

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iCell Cardiomyocytes (Catalog No. CMC-100-010-001; Lot No. 1031999) and Hepatocytes (Catalog No. PHC-100-020-001; Lot No. 1636 and 1208), including plating and maintenance media, were purchased from Cellular Dynamics International (Madison, WI). EarlyTox Cardiotoxicity Kits, including reference standards isoproterenol, propranolol, and sotalol, and CatchPoint cAMP GPCR assay kits were purchased from Molecular Devices, LLC (Sunnyvale, CA). B-27 medium supplement, CellROX Deep Red reagent, gentamicin (50 mg/mL), Hank's Balanced Salt Solution, HCS LipidTOX Deep Red reagent, Hoechst 33342, MitoTracker Orange CMTMRos reagent, penicillin/streptomycin solution, and RPMI 1640 medium were obtained from Life Technologies (Grand Island, NY). 3-Isobutyl-1-methylxanthine, amiodarone hydrochloride, cisapride monohydrate, crizotinib, doxorubicin hydrochloride, formaldehyde solution, forskolin, Krebs-Ringer bicarbonate buffer (KRBG), sodium bicarbonate, and tetraoctylammonium bromide (TAB) were all obtained from Sigma-Aldrich (St. Louis, MO). Dimethyl sulfoxide (DMSO), dexamethazone, hydrogen peroxide (3%), menadione, recombinant oncostatin M, and sunitinib were purchased from Fisher Scientific (Waltham, MA).

Grimm F.A., Iwata Y., Sirenko O., Bittner M, & Rusyn I. (2015). High-Content Assay Multiplexing for Toxicity Screening in Induced Pluripotent Stem Cell-Derived Cardiomyocytes and Hepatocytes. Assay and Drug Development Technologies, 13(9), 529-546.

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Biorelevant Intestinal Media for Drug Dissolution

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Amiodarone hydrochloride, celecoxib, clopidogrel bisulfate, danazol, fluoxetine hydrochloride, furosemide, nefazodone hydrochloride, nifedipine, zidovudine and isoniazid were purchased from Sigma Aldrich Inc. (Milwaukee, WI, USA). Griseofulvin was provided by Pion. The gastrointestinal tract lipid (GIT-0 PN 110669), the acceptor sink-buffer (ASB PN 110139) and the PVDF (polyvinylidene difluoride) filter support (PN 120875) were purchased from Pion Inc. (Billerica, MA, USA). FaSSIF buffer concentrate, FeSSIF buffer concentrate and the simulated intestinal powder version I FaSSIF/FeSSIF/FaSSGF, were purchased from Biorelevant.com (London U.K.). DMSO (dimethyl sulfoxide) and methanol (HPLC Plus grade) used to prepare the stock solution for standards were purchased from Sigma Aldrich (Milwaukee, WI, USA).
Media preparation
Dissolution/flux tests were performed using the FaSSIF, FeSSIF version 1 to simulate the fasted state and fed state intestinal fluids (Table-1 for media composition). FaSSIF and FeSSIF were prepared using the buffer concentrate method with FaSGF/FaSSIF/FeSSIF powder as described in Biorelevant Media preparation tool on their website (Biorelevant.com). Table-1 summarizes the composition of the intestinal media used in this study.

Jankovsky C., Tsinman O, & Thakral N.K. (2022). Food effect risk assessment in preformulation stage using material sparing μFLUX methodology. ADMET & DMPK, 10(4), 299-314.

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