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Cotinine

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Cotinine is a biochemical compound that serves as a biomarker for nicotine exposure. It is the primary metabolite of nicotine and can be detected and measured in various biological samples, such as blood, urine, or saliva. Cotinine analysis is widely used to assess tobacco and nicotine consumption levels.

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Lab products found in correlation

Acetonitrile, by Thermo Fisher Scientific (4 mentions) Lc ms grade ammonium acetate, by Thermo Fisher Scientific (2 mentions) Nicotine gluc, by LGC (2 mentions) Cotinine gluc, by LGC (2 mentions) 3hc gluc, by LGC (2 mentions) 3 hydroxycotinine, by LGC (1 mentions) Nicotine n oxide, by LGC (1 mentions) Methanol, by Thermo Fisher Scientific (1 mentions) Mecamylamine hcl, by Merck Group (1 mentions) Nicotine free base, by Merck Group (1 mentions) Nicotine hydrogen tartrate salt, by Merck Group (1 mentions) Ammonium acetate, by Merck Group (1 mentions) Silicone cap mats, by Phenomenex (1 mentions) Dulbecco s pbs, by Thermo Fisher Scientific (1 mentions) Recombinant β glucuronidase, by Merck Group (1 mentions) 13c6 nnal, by LGC (1 mentions) Cerex 96 well positive pressure processor, by Chromtech (1 mentions) Cotinine d3, by LGC (1 mentions) Oasis hlb cartridge, by Waters Corporation (1 mentions) Thc cooh, by Lipomed (1 mentions)

7 protocols using cotinine

1

In Vitro Micronucleus Assay Protocols

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For the in vitro MN assays carried out in the presence of chemicals that raise the luminal pH of acidic organelles, duplicate cultures were co‐exposed to the same range of nicotine concentrations and a fixed concentration of either ammonium chloride (NH4Cl; 5 mM), bafilomycin A1 (BafA1; 0.9 nM), or nigericin (100 nM) for 24 hr in the absence of S9 on three independent test occasions. Similarly, for experiments conducted on the major mammalian nicotine metabolites (e.g., cotinine, nornicotine (‐/L enantiomer), nicotine N′‐oxide, and 3′‐hydroxycotinine) (the latter three from Toronto Research Chemicals, Toronto, Canada), cells were exposed on two test occasions up to a maximum concentration of 10 mM, except for nicotine N′‐oxide (5 mM due to solubility limitations), prior to harvesting and enumerating the nuclei species.
Smart D.J., Helbling F.R., Verardo M., McHugh D, & Vanscheeuwijck P. (2019). Mode‐of‐action analysis of the effects induced by nicotine in the in vitro micronucleus assay. Environmental and Molecular Mutagenesis, 60(9), 778-791.
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2

Nicotine and Mecamylamine Pharmacology

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Mecamylamine HCl and (-)-nicotine hydrogen tartrate salt (Sigma-Aldrich, St. Louis, MO) were dissolved in physiological saline (Patterson Veterinary, Devens, MA), and the pH was adjusted to approximately neutral (pH ~ 7), as necessary. (-)-Nicotine free base (Sigma-Aldrich) was mixed with a 50:50 propylene glycol and glycerin solution (Sigma-Aldrich). Doses of nicotine for injection are expressed as mg/kg of the base. Nicotine and mecamylamine were injected subcutaneously (s.c.) at a volume of 10 ml/kg. Concentrations for aerosol administration are expressed as mg/ml in the e-cigarette tank, and may not be representative of the actual amount of nicotine inhaled.
Chemicals and reagents for the analysis of biological samples were purchased commercially and included nicotine (Sigma-Aldrich), cotinine (Toronto Research Chemicals, Toronto, ON), nicotine-d3 (Cambridge Isotope Laboratories, Tewksbury, MA), cotinine-d3 (Santa Cruz Biotechnology, Dallas, TX), ammonium acetate (Sigma-Aldrich), and formic acid and acetonitrile (Fisher Scientific, Fair Lawn, NJ). An internal standard solution was prepared in methanol (Fisher Scientific) containing 48 µg/mL nicotine-d3 and 38 μg/mL cotinine-d3. Working solutions containing both nicotine and cotinine were prepared in methanol at concentrations of 10,000 and 100 ng/mL.
Lefever T.W., Lee Y.O., Kovach A.L., Silinski M.A., Marusich J.A., Thomas B.F, & Wiley J.L. (2017). Delivery of nicotine aerosol to mice via a modified electronic cigarette device. Drug and alcohol dependence, 172, 80-87.
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3

Quantitative Analysis of Nicotine and Metabolites

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Liquid chromatography mass spectrometry (LC/MS)-grade ammonium acetate and acetonitrile were purchased from Thermo Fisher Scientific (Waltham, MA), while the ACQUITY UPLC BEH-HILIC (1.7 m 2.1 x 100 mm) column was purchased from Waters (Milford, MA). Nicotine and eight of its major metabolites (cotinine, 3HC, NOX, COX, nicotine-Gluc, cotinine-Gluc, 3HC-Gluc, and HPBA) as well as their corresponding deuterium labeled (d3) internal standards were purchased from Toronto Research Chemicals (North York, ON, Canada).
Perez-Paramo Y.X., Watson C.J., Chen G., Thomas C.E., Adams-Haduch J., Wang R., Khor C.C., Koh W.P., Nelson H.H., Yuan J.M, & Lazarus P. (2022). Impact of Genetic Variants in the Nicotine Metabolism Pathway on Nicotine Metabolite Levels in Smokers. Cancer Epidemiology, Biomarkers & Prevention, 32(1), 54-65.
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4

HILIC-LC/MS Nicotine Metabolome Analysis

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LC/MS-grade ammonium acetate and acetonitrile were purchased from Thermo Fisher Scientific, whereas the ACQUITY UPLC BEH-HILIC (1.7 m 2.1 × 100 mm) column was purchased from Waters. Nicotine and eight of its major metabolites (cotinine, 3HC, NOX, COX, nicotine-Gluc, cotinine-Gluc, 3HC-Gluc, and HPBA) as well as their corresponding deuterium labeled (d3) internal standards were purchased from Toronto Research Chemicals.
Perez-Paramo Y.X., Watson C.J., Chen G., Thomas C.E., Adams-Haduch J., Wang R., Khor C.C., Koh W.P., Nelson H.H., Yuan J.M, & Lazarus P. (2022). Impact of Genetic Variants in the Nicotine Metabolism Pathway on Nicotine Metabolite Levels in Smokers. Cancer Epidemiology, Biomarkers & Prevention, 32(1), 54-65.
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5

Quantitative Analysis of Tobacco Biomarkers

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Cotinine, [13CD3]Cotinine, NNAL, and [13C6]NNAL (Figure 1), were procured from Toronto Research Chemicals (Ontario, Canada). Recombinant β-glucuronidase (catalog # G8295) was purchased from Sigma-Aldrich (Milwaukee, WI). True Taper® 96-well plates for sample processing and analysis were from Analytical Sales & Services (Pompton Plains, NJ) while silicone cap mats used to cover the 96-well plates were purchased from Phenomenex (Torrance, CA). Four hundred μL Isolute SLE+ diatomaceous earth solid-phase extraction 96-well plates were from Biotage (Charlotte, NC) while Oasis MCX 10 mg, 30 μm solid-phase extraction 96-well plates were from Waters (Milford, MA). A Cerex 96-well positive pressure processor (Chromtech, Apple Valley, MN) was utilized during sample processing. Dulbecco’s PBS was purchased from Invitrogen (Grand Island, NY) and the rest of the chemicals were obtained either from Fisher Scientific (Fairlawn, NJ), Sigma-Aldrich (Milwaukee, WI) or Alfa Aesar (WardHill, MA) and used without further purification unless otherwise noted. An Eppendorf multichannel pipettor was used during sample processing.
Kotandeniya D., Carmella S.G., Ming X., Murphy S.E, & Hecht S.S. (2014). Combined Analysis of the Tobacco Metabolites Cotinine and 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanol in Human Urine. Analytical Chemistry, 87(3), 1514-1517.
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6

Comprehensive Quantification of Illicit Drugs

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BE, AMPH, METH, MDMA, MOR, THC-COOH, and their deuterated analogues used as internal standards (BE-d3, MOR-d3, THC-COOH-d3, MDMA-d5) were purchased from Lipomed (Arlesheim, Switzerland). EtS and EtS-d5 were obtained from Restek (Bellefonte, PA, USA). Cotinine and Cotinine-d3 were purchased from Toronto Research Chemicals (TRC) (Ontario, Canada). Oasis HLB cartridges (6 mL/500 mg) were obtained from Waters Corporation (Milford, MA, USA). Glass-fiber filters (Whatman® glass microfiber filters, Grade GF/F, 0.7 μm) were purchased from Merck, (Darmstadt, Germany) as well as ammonium acetate, liquid chromatography grade methanol, ultrapure water, acetonitrile, and acetic acid (98 % purity).
, & Guzel E.Y. (2022). Monitoring of changes in illicit drugs, alcohol, and nicotine consumption during Ramadan via wastewater analysis. Environmental Science and Pollution Research International, 29(59), 89245-89254.
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7

Nicotine, Apocynin, and Cotinine Analysis

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Nicotine hydrogen tartrate (CAS No.: 65-31-6; purity 95%) was purchased from Nacalai Tesque, Inc. (Kyoto, Japan). Apocynin (CAS No.: 498-02-2; purity 98%) was purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan). Cotinine (CAS No.: 486-56-6; purity 98%) was purchased from Toronto Research Chemicals (North York, ON, Canada).
Suzuki S., Cohen S.M., Arnold L.L., Pennington K.L., Gi M., Kato H., Naiki T., Naiki-Ito A., Wanibuchi H, & Takahashi S. (2021). Cell proliferation of rat bladder urothelium induced by nicotine is suppressed by the NADPH oxidase inhibitor, apocynin. Toxicology letters, 336.
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