(−)-Nicotine bitartrate, (+/−) nornicotine, (+/−) anabasine, (+/−) myosmine, and (−)-cotinine were obtained from Sigma (St Louis, MO). (+/−) Anatabine was obtained from Toronto Research Chemicals, Inc. (Ontario, Canada). All drugs were prepared in sterile saline, adjusted to a pH of 7.4 using dilute NaOH, and administered s.c. in a volume of 1 ml/kg. All drug doses are expressed as the base.
Myosmine
Manufactured by Merck Group
Sourced in United States
Myosmine is a laboratory equipment product manufactured by Merck Group. It is designed for use in scientific research and analysis. The core function of Myosmine is to facilitate the measurement and detection of specific molecules or compounds in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Cotinine, by Merck Group (7 mentions)
Anabasine, by Merck Group (4 mentions)
Nicotine, by Merck Group (3 mentions)
Formic acid, by Thermo Fisher Scientific (3 mentions)
Nornicotine, by Merck Group (2 mentions)
Nornicotine, by LGC (2 mentions)
Anatabine, by Cayman Chemical (2 mentions)
Anatabine, by LGC (1 mentions)
Nicotine bitartrate, by Merck Group (1 mentions)
Harmane, by Merck Group (1 mentions)
Norharmane, by Merck Group (1 mentions)
Nicotine hydrogen tartrate, by Merck Group (1 mentions)
Tyramine hcl, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Blue dye, by Molecular Devices (1 mentions)
Blue dye, by Merck Group (1 mentions)
Methamphetamine, by Merck Group (1 mentions)
Menthone, by Merck Group (1 mentions)
Piperonal, by Merck Group (1 mentions)
6 methylcoumarin, by Merck Group (1 mentions)
9 protocols using myosmine
1
Nicotinic Alkaloid Administration Protocol
2
Tobacco Constituents Bioavailability Protocol
3
GABA receptor assay in WSS1 cells
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WSS1 cells from ATCC (Manassas, VA, USA) which express functional GABAA receptors were cultured as previously described by Wong et al. [57 (link)]. The cells were grown to near confluence and then transferred to black well, clear bottom 96-well assay plates (Corning Incorporated, Corning, NY, USA) 12–18 h prior to the assay. Molecular Devices blue dye (Sunnyvale, CA, USA) was reconstituted as previously described [58 (link)]. The WSS1 cells were allowed to come to room temperature, the culture medium was then aspirated, replaced with the blue dye and equilibrated for 30 min before being placed on a FlexStation 3 plate reader (Molecular Devices, Sunnyvale, CA, USA). Varying concentrations of γ-aminobutyric acid (GABA, Tocris Bioscience, Bristol, United Kingdom), myosmine (Sigma, St. Louis, MO, USA) and anabaseine [31 (link),32 ] in blue dye solution were added to the cells by the FlexStation. Programmed readings and data analysis were performed as previously described [58 (link),59 (link)]. The cellular responses to GABA were normalized to the maximum dye response generated by 1 mM GABA. The fifty percent effective concentration for GABA was determined using a sigmoidal dose-response equation (log(agonist) vs. normalized response—Variable slope) with Prism version 6.03 (GraphPad Software, Company, San Diego, CA, USA, www.graphpad.com ).
4
Nicotine and Methamphetamine Pharmacology
5
Compound Purchasing and Preparation
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The following compounds were purchased from Sigma-Aldrich (St. Louis, MO): piperonal (99.0%), 2,3,5-trimethylpyrazine (99.0%), menthone (99.0%), (E)-beta-damascone (99.0%), cinnamic acid methyl ester (99.0%), myosmine (99.0%), cotinine (98.5%), and the internal standard (ISTD) 6-methyl coumarin (99.0%). Hydroxyacetone (96.0%) was purchased from Santacruz Biotechnology (Dallas, TX). 200 proof ethanol was purchased from Pharmco-Aaper. Propylene glycol (PG) and vegetable glycerol (VG) were purchased from Spectrum (New Brunswick, NJ).
6
Quantitative Analysis of Tobacco Alkaloids and Polymerization Compounds
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Cotinine, nicotine, myosmine, N-nitrosonorniCotinine, methacrylic acid (MAA), ethylene glycol dimethacrylate (EGDMA), azobisisobutyronitrile (AIBN), potassium phosphate monobasic (KH2PO4), sodium phosphate dibasic (Na2HPO4), and agarose powder were purchased from Sigma-Aldrich (St. Louis, MO, USA). MAS® DOA TOTAL Control liquid assayed drugs of abuse control was purchased from Thermo Fisher Scientific (Waltham, MA, USA). High-performance liquid chromatography (HPLC) grade chemical solvents including acetonitrile, dichloromethane, acetic acid, 2-propanol, and methanol were purchased from Sigma-Aldrich and Merck (Kenilworth, NJ, USA). Ammonia (30%) was purchased from Panreac Quimica SAU (Barcelona, Spain). Glass fiber (GF) membrane filter was purchased from a local manufacturer in Thailand (Pacific Biotech CO. LTD, Phetchabun, Thailand). Filter paper (Whatman, Grade 1) was purchased from GE Healthcare Life Science (Chicago, IL, USA). CHROMABOND® column C18ec was purchased from Macherey-Nagel GmbH & Co. KG (Düren, Germany). Deionized water was purified by a Milli-Q system (Merck Millipore, Billerica, MA, USA) with resistivity of 18.2 Ω cm.
7
Substrate Specificity Analysis of NicA2
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Substrate specificity of NicA2 was analyzed by the Amplex Red assay described above, using 10 μM of test compound and 160 nM NicA2 enzyme. Compounds tested were: (2’S)-nicotine-1’-N-oxide (Toronto Research Chemicals; TRC), (±)-nornicotine, nicotinamide, β-nicotinamide adenine dinucleotide (NAD), acetylcholine, choline, (−)-cotinine, varenicline, bupropion, (−)-cytisine, mecamylamine (TRC), dopamine, serotonin, (±)-norepinephrine, L-glutamate, γ-amino-N-butyric acid (GABA), (R,S)-anatabine (TRC), (R,S)-anabasine (TRC), and myosmine (compounds obtained from Sigma-Aldrich unless otherwise stated). Activities were expressed relative to the activity found for S-(−)-nicotine run in parallel.
8
Quantification of Tobacco Alkaloids and Metabolites
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Nicotine, anabasine, myosmine, nicotinamide, cotinine, norNicotine, norcotinine, nicotinic acid, and their corresponding deuterated or 13C internal standards were purchased from Sigma-Aldrich (St. Louis, MO, USA) unless otherwise stated below. Anatabine was purchased from Cayman Chemicals (Ann Arbor, MI, USA), and Anatabine-d4 and myosmine-d4 were obtained from Toronto Research Chemicals (Ontario, ON, Canada). Solvents used for LC-MS analysis were from Honeywell (Charlotte, NC, USA). Ammonium acetate was from Sigma-Aldrich, and formic acid was from Thermo Fisher Scientific (Waltham, MA, USA).
The following solvents used for microfractionation were purchased from Shanghai Titan Scientific Co. Ltd. (Shanghai, China): butanol (≥99.5%, certified analytical reagent (AR)), dichloromethane (≥99.5%, AR), ethyl acetate (≥99.5%, AR), 95% ethanol (95%, AR), ethanol (≥99.7%, AR), hexane (≥97.0%, AR), and methanol (≥99.5%, AR). Ultrapure water was produced by Bicoll (Planegg, Germany). The pipettes for 96-well microtiter plates were produced by Eppendorf (Hamburg, Germany). Polytetrafuoroethylene membrane filters (0.22 µm) and syringes were purchased from Anpel technologies (Shanghai, China).
The following solvents used for microfractionation were purchased from Shanghai Titan Scientific Co. Ltd. (Shanghai, China): butanol (≥99.5%, certified analytical reagent (AR)), dichloromethane (≥99.5%, AR), ethyl acetate (≥99.5%, AR), 95% ethanol (95%, AR), ethanol (≥99.7%, AR), hexane (≥97.0%, AR), and methanol (≥99.5%, AR). Ultrapure water was produced by Bicoll (Planegg, Germany). The pipettes for 96-well microtiter plates were produced by Eppendorf (Hamburg, Germany). Polytetrafuoroethylene membrane filters (0.22 µm) and syringes were purchased from Anpel technologies (Shanghai, China).
9
Comprehensive E-Liquid Chemical Analysis
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The standards of nicotine, cotinine, anabasine and myosmine and boric acid were purchased from Sigma Aldrich (St. Louis, USA). Nornicotine and -nicotyrine standards were bought from Toronto Research Chemicals Inc. (Toronto, USA). Nicotine-N-oxide and anatabine came from Cayman Chemical (Michigan, USA). The matrix components propylene glycol and glycerol were purchased from Merck (Schuchardt, Germany). The components used to reconstitute a flavoured e-liquid sample were all purchased from Sigma Aldrich (St. Louis, USA). These include diacetyl, acetylpropionyl, maltol, ethylmaltol, vanillin, ethylvanillin, citral, limonene, cinnamaldehyde, transcinnamaldehyde, benzaldehyde, carvone, pulgeone, damascenone, methylcyclopentone, methylcinnamate, methylaminobenzoaat and methylmyristate. The solvents acetonitrile, methanol, acetone and ethylactetate were HPLC-grade and were purchased from Biosolve (Valkenswaard, The Netherlands). Concentrated ammonia 28-30% was obtained from Merck (Darmstadt, Germany). Water was obtained using a milliQ-Gradient A10 system (Millipore, Billerica, USA).
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