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Methadone

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Methadone is a laboratory reagent used as an internal standard in the analysis of opioid compounds. It is a synthetic opioid that can be utilized to quantify the presence and concentration of related substances in analytical samples.

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Lab products found in correlation

Morphine, by Merck Group (2 mentions) Codeine, by Merck Group (2 mentions) Ammonium molybdate, by Merck Group (2 mentions) Sodium formate, by Merck Group (2 mentions) Haloperidol, by Merck Group (2 mentions) Pethidine, by Merck Group (2 mentions) 2 nitrophenyl octyl ether npoe, by Merck Group (2 mentions) Tributyl phosphate tbp acetonitrile, by Merck Group (2 mentions) Nortriptyline, by Merck Group (2 mentions) Loperamide, by Merck Group (2 mentions) Acetic acid, by Merck Group (2 mentions) Nitric acid, by Merck Group (2 mentions) Sodium dihydrogen phosphate, by Merck Group (2 mentions) Formic acid, by Merck Group (2 mentions) Phenolphthalein, by Merck Group (2 mentions) Phosphoric acid, by Merck Group (2 mentions) Disodium hydrogen phosphate, by Merck Group (2 mentions) Sodium acetate, by Merck Group (2 mentions) Sulphuric acid, by Merck Group (2 mentions) Imipramine, by Merck Group (2 mentions)

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Methadone hydrochloride (18 citations)

13 protocols using methadone

1

Methadone Dose Selection for Morphine Withdrawal

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The selection of Methadone doses was based on pharmacological range and below LD50 value which might be effective and safe for the use in this experiment (47 (link), 48 ). Methadone (0, 0.5, 1.0, 2.0 mg/kg was dissolved in vehicle (20% Tween 80; Sigma Aldrich, UK) and injected intraperitoneally 10 min before withdrawal testing, i.e. 23 h 50 min after the last morphine dose (47 (link)). Thereafter, Methadone was repeatedly administered every 8 h (47 (link)) for testing the duration of withdrawal inhibiting effects. This design is a modified version of a substitution routine described by Ruiz et al., 1996 (49 (link)).
Hassan R., Pike See C., Sreenivasan S., Mansor S.M., Müller C.P, & Hassan Z. (2020). Mitragynine Attenuates Morphine Withdrawal Effects in Rats—A Comparison With Methadone and Buprenorphine. Frontiers in Psychiatry, 11, 411.
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2

Heroin and Methadone Pharmacokinetics

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Heroin (diamorphine HCl) was administered by s.c. injection and by vapor inhalation (see below). methadone was administered by s.c. injection and naloxone by i.p. injection. Inhalation/vapor doses are varied, and are therefore described, by altering the concentration in the propylene glycol (PG) vehicle, the puff schedule and/or the duration of inhalation sessions in this approach (Javadi-Paydar et al., 2019b (link); Nguyen et al., 2016a (link); Nguyen et al., 2016b (link)). Heroin was dissolved in PG to achieve target concentrations and then loaded in e-cigarette tanks in a volume of ~ 0.5 ml per session. Fresh solutions were used for each session. naloxone, methadone and heroin were dissolved in saline (0.9% NaCl), for injection. The heroin was provided by the U.S. National Institute on Drug Abuse and the PG, methadone and naloxone were obtained from Sigma-Aldrich Corporation (St. Louis, MO, USA).
Gutierrez A., Nguyen J.D., Creehan K.M, & Taffe M.A. (2020). Female rats self-administer heroin by vapor inhalation. Pharmacology, biochemistry, and behavior, 199, 173061.
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3

Reconstitution of Membrane Transporters

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Loperamide was purchased from MP Biomedicals (Santa Ana, CA). Digoxin, ethylene glycol tetraacetic acid (EGTA), and imidazole were purchased from Alfa Aesar (Tewksbury, MA). The detergent n-dodecyl-β-D-maltoside (DDM) was purchased from EMD Millipore Corporation (San Diego, CA). Cholesterol, disodium ATP, and Tris-HCl were purchased from Amresco (Solon, OH). Escherichia (E.) coli total lipid extract powder was purchased from Avanti Polar Lipids Inc (Alabaster, AL). Dithiothreitol (DTT) was purchased from Gold Biotechnology (Olivette, MO). Deuterated dithiothreitol was purchased from CDN Isotopes (Quebec, Canada). HEPES and acrylamide were purchased from Calbiochem (San Diego, CA). Deuterium oxide was purchased from Cambridge Isotope Laboratories, Inc (Tewksbury, MA). Sodium orthovanadate (Na3VO4) was purchased from Enzo Life Sciences (Farmingdale, NY). Ammonium chloride (NH4Cl), deuterated dimethyl sulfoxide, and methadone were purchased from Sigma-Aldrich (Milwaukee, WI). All remaining chemicals used in this study were purchased from Thermo Fischer Scientific (Waltham, MA).
Gibbs M.E., Wilt L.A., Ledwitch K.V, & Roberts A.G. (2018). A conformationally-gated model of methadone and loperamide transport by P-glycoprotein. Journal of pharmaceutical sciences, 107(7), 1937-1947.
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4

Patch-Clamp Recording with Methadone

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The extracellular solution for patch-clamp experiments contained (in mM): 130 NaCl, 3 KCl, 1 CaCl2, 1 MgCl2, 10 HEPES, and 10 glucose; pH 7.4 with 1 M NaOH (∼4 mM Na+ added). The internal solution for patch electrodes contained (in mM): 138 K-gluconate, 4 KCl, 10 Na2-phosphocreatine, 0.2 CaCl2, 10 HEPES (Na+ salt), 1 EGTA, 4 Mg-ATP, 0.3 Na-GTP; pH 7.4 with 1 M KOH (∼3 mM K+ added). In some whole-cell recordings, the internal solution also includes 1 mg/ml Lucifer yellow dye. The osmolarity of all solutions was adjusted to 290 mOsm. Methadone (Sigma-Aldrich, St. Louis, MO, United States) was dissolved in water to make 1 mg/mL stock solutions. Methadone was diluted in Media2 to working concentrations (1 and 10 μM). The cell culture media (Media2 + Methadone) was filtered with Tube-Top Vacuum Filter (Corning, Tewksbury, MA, United States) to ensure sterility.
Wu W., Yao H., Dwivedi I., Negraes P.D., Zhao H.W., Wang J., Trujillo C.A., Muotri A.R, & Haddad G.G. (2020). Methadone Suppresses Neuronal Function and Maturation in Human Cortical Organoids. Frontiers in Neuroscience, 14, 593248.
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5

Quantification of Drugs in Biological Samples

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Cotinine (COT), trans-3-hydroxycotinine (OH-COT), cotinine-d3 and trans-3-hydroxycotinine-d3 were obtained from LGC Standards (Molsheim, France). Caffeine, ibuprofen, methadone, morphine, pseudoephedrine, codeine, cocaine, clomipramine, propranolol and Δ-9-tetrahydrocannabinol analytical reference standards presented in Methanol were obtained from Cerilliant (Sigma-Aldrich, Saint-Quentin-Fallavier, France). Potassium dihydrogen phosphate and dipotassium hydrogen phosphate were obtained from Sigma-Aldrich (St Louis, MO, USA). Methanol, hexane, isopropanol were of analytical grade and were obtained from Merck (Darmstadt, Germany). Sterile water was obtained from Fresenius (Sevres, France). Ammonia solution 25% was purchased from VWR (Fontenay-sous-Bois, France). Hydrochloric acid 0.2 m was obtained from VWR (Fontenay-sous-Bois, France). Clean Screen SPE columns 200 mg were purchased from UCT (Bristol, PA, USA). The derivatizing agent utilized was bis(trimethylsilyl)trifluoroacetamide (BSTFA) + 1% trimethylchlorosilane (TMS) (Sigma-Aldrich, Saint-Quentin-Fallavier, France).
Chiadmi F, & Schlatter J. (2014). Simultaneous determination of cotinine and trans-3-hydroxycotinine in urine by automated solid-phase extraction using gas chromatography–mass spectrometry. Biomedical Chromatography, 28(4), 453-458.
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6

Methadone and Doxorubicin Combination Protocol

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D,L-Methadone hydrochloride (Methadone, Sigma), doxorubicin (Sigma), and naloxone (Fagron GmbH&Co) were freshly dissolved in sterile distilled water prior to each experiment to ensure constant quality of the preparations. 3-Isobutyl-1-methylxanthine (IBMX, Sigma) was freshly dissolved in 0.01N NaOH.
For in vivo application, we used D,L-Methadone (Methaddict, Hexal) as 5 mg tablets purchased from the local pharmacy. The tablets were pulverized and solubilized freshly before use in 10% Tween 80 in saline.
Friesen C., Hormann I., Roscher M., Fichtner I., Alt A., Hilger R., Debatin K.M, & Miltner E. (2014). Opioid receptor activation triggering downregulation of cAMP improves effectiveness of anti-cancer drugs in treatment of glioblastoma. Cell Cycle, 13(10), 1560-1570.
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7

Methadone-based Organoid Neural Differentiation

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Methadone (Sigma-Aldrich, St. Louis, MO) was dissolved in sterile nuclease-free water (Invitrogen, Waltham, MA) and diluted to a working concentration of 1 μM in fresh stage-specific medium each day of media change. Treatment began on Day 9 of organoid culture, the first day of the neural proliferation stage, and concluded at Day 60. nuclease-free water was used as a vehicular control.
Dwivedi I., Caldwell A.B., Zhou D., Wu W., Subramaniam S, & Haddad G.G. (2023). Methadone alters transcriptional programs associated with synapse formation in human cortical organoids. Translational Psychiatry, 13, 151.
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8

Analytical Reagent Procurement Protocol

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All chemicals were of high analytical grade (>95%) Haloperidol, loperamide, methadone, nortriptyline, pethidine and ammonium molybdate were all purchased from Sigma Aldrich (St Louis, Missouri, USA). Phosphoric acid, sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium acetate, acetic acid, sodium formate, formic acid, phenolphthalein, 2-nitrophenyl octyl ether (NPOE), tributyl phosphate (TBP) acetonitrile, sulphuric acid and nitric acid were purchased from Merck (Darmstadt, Germany). Ethanol was obtained from Arcus (Oslo, Norway) and water (18 MU cm) was obtained from a Milli-Q water purification system (Molsheim, France).
Restan M.S., Ramsrud S.B., Jensen H, & Pedersen-Bjergaard S. (2020). Influence of acid-base dissociation equilibria during electromembrane extraction. Journal of separation science, 43(15).
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9

Quantitative Analysis of Opioids

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Morphine, methadone, codeine, 6-MAM, EDDP standards, and Morphine-d3 and methadone-d3 internal standards (IS) were purchased from Sigma Aldrich (Milan, Italy). For the extraction procedure, distilled water (H2O) for analysis, sodium hydroxide (NaOH), methanol (MeOH) for analysis (≥99.9%), hydrochloric acid for analysis (37%), dichloromethane for analysis stabilized with amylin (≥99.9%), 2-propanol (99.8%) and ammonium hydroxide (32%) of reagent grade were purchased from Carlo Erba (Cornaredo, Italy). For the UHPLC analysis, methanol for UHPLC-MS (99.99%), ultrapure water for UHPLC-MS and formic acid (FA) for LC-MS (≥99%) were purchased from Carlo Erba (Cornaredo, Italy). All reagents and laboratory consumables were stored according to the manufacturer’s instructions.
Buratti E., Mietti G., Cippitelli M., Cerioni A., Froldi R., Cingolani M, & Scendoni R. (2023). Detection of Three Opioids (Morphine, Codeine and Methadone) and Their Metabolites (6-Monoacetylmorphine and 2-Ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine) in Larvae of Lucilia sericata Species by UHPLC-TF-MS and Validation. Molecules, 28(12), 4649.
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10

Characterization of Drug Metabolism Assays

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Uridine 59-diphosphoglucuronic acid trisodium salt (UDPGA), NADPH, alamethicin from Trichoderma viride, adenosine 3-phosphate 5-phosphosulfate, diclofenac sodium salt, midazolam hydrochloride, felodipine, S-(+)-mephenytoin, raloxifene hydrochloride, lovastatin, nisoldipine, nifedipine, methadone, quinidine, testosterone, benzydamine hydrochloride, cisapride, cyclosporin, terbutaline, sildenafil, verapamil, trazodone, atorvastatin, simvastatin, buspirone, rifabutin, saquinavir, terfenadine, zolpidem, repaglinide, indinavir, alprazolam, carbazeran, enalapril, ramipril , and dabigatran etexilate were purchased from Sigma-Aldrich (St. Louis, MO). NADPH-regenerating system containing NADP + , glucose-6phosphate, and glucose-6-phosphate dehydrogenase was purchased from Corning (Wiesbaden, Germany). Organic solvents were of LC-mass spectrometry (MS) or higher quality grade and were acquired from VWR International (Radnor, PA) or Fisher Scientific UK Ltd (Loughborough, UK). Purified water was obtained from Milli-Q Integral 5 Water Purification System (Merck KGaA, Darmstadt, Germany). Phosphate buffer, HQM Hepatocyte/Enterocyte Incubation Medium, and Co-Factor N for MetMax were purchased from In Vitro ADMET Laboratories (Columbia, MD).
Davies M., Peramuhendige P., King L., Golding M., Kotian A., Penney M., Shah S, & Manevski N. (2020). Evaluation of In Vitro Models for Assessment of Human Intestinal Metabolism in Drug Discovery. Drug metabolism and disposition: the biological fate of chemicals, 48(11).
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