Cotinine

Manufactured by Merck Group

Sourced in United States, Germany, United Kingdom, Japan

Cotinine is a laboratory equipment product manufactured by Merck Group. It is a chemical compound that is widely used as a biomarker to measure exposure to nicotine. Cotinine is the primary metabolite of nicotine and is present in the body for a longer period than nicotine, making it a reliable indicator of recent nicotine exposure.

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Lab products found in correlation

Nicotine, by Merck Group (20 mentions) Cotinine d3, by Merck Group (7 mentions) Nornicotine, by Merck Group (7 mentions) Myosmine, by Merck Group (7 mentions) Anabasine, by Merck Group (5 mentions) Formic acid, by Thermo Fisher Scientific (5 mentions) Nicotine d4, by Merck Group (4 mentions) Acetonitrile, by Thermo Fisher Scientific (3 mentions) Arecoline hydrobromide, by Merck Group (3 mentions) Guvacine hydrochloride, by Merck Group (3 mentions) Hydroxycotinine, by Merck Group (3 mentions) Anatabine, by Cayman Chemical (3 mentions) Methanol, by Merck Group (3 mentions) Methanol, by Thermo Fisher Scientific (2 mentions) Nicotyrine, by LGC (2 mentions) Nicotine hydrogen tartrate salt, by Merck Group (2 mentions) Acetonitrile, by Merck Group (2 mentions) Mecamylamine, by Merck Group (2 mentions) Varenicline, by Merck Group (2 mentions) Serotonin, by Merck Group (2 mentions)

Close spelling variants of this product

Cotinine-d3 (9 citations) (-)-cotinine free base (3 citations) [2H3]-cotinine (2 citations) Anti-cotinine-IgG (2 citations) Cotinine reagent (1 citations)

48 protocols using cotinine

Quantifying Nicotine and Cotinine in Mice

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The plasma concentration of nicotine and its principal metabolite cotinine were measured from the 14 days JUUL/TS-exposed mice by LCMS/MS analysis using cotinine-d3 (MilliporeSigma, St. Louis, MO, USA) as internal standard (IS) following a previously published method [23 (link)]. In brief, samples were prepared by protein precipitation of 25 µl mouse plasma using acetonitrile at a 1:8 ratio. The mass spectrometer was operated in positive polarity under the multiple reaction monitoring mode using the electrospray ionization technique. The transitions of m/z 163.2 → 132.1, 177.2 → 98.0, and 180.2 → 101.2 were used to measure the nicotine, cotinine, and IS, respectively. The elution of nicotine (MilliporeSigma), cotinine (MilliporeSigma), and IS were at 1.89, 1.77, and 1.76 min, respectively. This was achieved with a mobile gradient phase consisting of 5 mM ammonium bicarbonate, acetonitrile, and methanol (3:1, v/v) at a 0.3 ml/min flow rate on a Kinetex EVO C18 column (Phenomenex, Torrance, CA, USA).

Sifat A.E., Archie S.R., Nozohouri S., Villalba H., Zhang Y., Sharma S., Ghanwatkar Y., Vaidya B., Mara D., Cucullo L, & Abbruscato T.J. (2022). Short-term exposure to JUUL electronic cigarettes can worsen ischemic stroke outcome. Fluids and Barriers of the CNS, 19, 74.

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Quantification of Nicotine and Cotinine

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S(−)-nicotine (1 mg/mL in methanol), (−)-cotinine (1 mg/mL in methanol), (+/−)-nicotine-d₄ (0.1 mg/mL in acetonitrile), and (+/−)-cotinine-d₃ (1 mg/mL in methanol) were purchased from Millipore-Sigma (St. Louis, MO, USA). For the in vivo studies, an S(−)-nicotine base was purchased from MP Biomedicals, Inc. (Solon, OH, USA), dissolved in normal saline (sterilized 0.9% sodium chloride in deionized water) and injected subcutaneously (s.c.).

Nguyen K., Kanamori K., Shin C.S., Hamid A, & Lutfy K. (2020). The Impact of Sex on Changes in Plasma Corticosterone and Cotinine Levels Induced by Nicotine in C57BL/6J Mice. Brain Sciences, 10(10), 705.

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Quantitative Analysis of Nicotine and Cotinine

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(−)-Nicotine (Cat # 36733); (−)-Cotinine, 1 mg/mL solution in methanol (Cat # C0430); deuterated internal standard (IS) (±)-Cotinine-d₃ (see Fig. 1a), 1 mg/mL solution in methanol (Cat # C-035) and LC/MS grade eluent additive ammonium bicarbonate (Cat # 40867) were purchased from Sigma-Aldrich, USA. LC/MS grade solvents water (Cat # W6-4), acetonitrile (Cat # A955-4) and methanol (Cat # A456-4) were obtained from Fisher Scientific, USA. Tobacco reference product, 3R4F equivalent to regular cigarettes were purchased from University of Kentucky.Fig. 1

Mass spectral data and representative peaks. Chemical structure of analytes and IS (a), mass spectral data of nicotine and cotinine (b). c Mass spectral data for the Internal Standards

Kaisar M.A., Kallem R.R., Sajja R.K., Sifat A.E, & Cucullo L. (2017). A convenient UHPLC-MS/MS method for routine monitoring of plasma and brain levels of nicotine and cotinine as a tool to validate newly developed preclinical smoking model in mouse. BMC Neuroscience, 18, 71.

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Nicotine and Metabolite Standards

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Nicotine, cotinine, and norNicotine were purchased from Sigma-Aldrich (St. Louis, MO) as standards. PseudooxyNicotine, nicotyrine and 6-hydroxyNicotine were obtained from Toronto Research Chemicals (Toronto, ON, Canada). All analytical and high-performance liquid chromatography (HPLC) grade reagents were bought from Merck China (Beijing, China).

Lei L., Xia Z., Liu X, & Wei H.L. (2015). Occurrence and variability of tobacco rhizosphere and phyllosphere bacterial communities associated with nicotine biodegradation. Annals of microbiology, 65(1).

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Cigarette Smoke Extract Preparation

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CSE was extracted and prepared as previously described (8). In brief, CSE was collected by drawing CS through filters and bubbled through PBS. The resultant extract represented 83.6 µg/ml of Nicotine and was stored in liquid nitrogen until further use. Reported levels of Nicotine in saliva and gingival crevicular fluid (GCF) after smoking one cigarette are 1.82 ± 0.61 µg/ml and 5.96 ± 0.77 µg/ml, respectively (mean ± SE) (15). CSE was diluted with PBS to final concentrations of 1%, 5% and 10%, which is equivalent to 0.84 µg/ml, 4.2 µg/ml and 8.4 µg/ml of Nicotine, respectively. Notably, neutrophils enter the oral cavity through the gingival crevice and are thus exposed to the CS component/metabolite concentrations found in GCF and saliva. Nicotine, cotinine and SCN-were obtained from Sigma Aldrich (Dorset, UK).

White P.C., Hirschfeld J., Milward M.R., Cooper P.R., Wright H.J., Matthews J.B, & Chapple I.L.C. (2018). Cigarette smoke modifies neutrophil chemotaxis, neutrophil extracellular trap formation and inflammatory response-related gene expression. Journal of periodontal research, 53(4).

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Platelet Activation and Signaling Pathways

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Thrombin was purchased from Chronolog Corporation (Havertown, PA), whereas ADP, nicotine and cotinine were from Sigma Aldrich (St. Louis, MO). Fluorescein isothiocyanate (FITC)-conjugated anti-P-selectin and FITC-conjugated Annexin V were purchased from Cell Signaling Technology, Inc (Danvers, MA). The JON/A antibody was obtained from Emfret analytics (Würzburg, Germany). The phycoerythrin-conjugated anti-CD69 and FITC-conjugated anti-CD45 were obtained from BD Biosciences (San Jose, CA). Stir bars and other disposables were purchased from Chrono-Log Corporation (Havertown, PA). Akt, phospho-Akt (Ser473), ERK and phospho-ERK antibodies were purchased from Cell signaling (Danvers, MA). The anti-PAR4 and anti-GPIIb-IIIa (αIIbβ3) antibodies were from Abcam (Cambridge, MA), whereas the anti-p85/PI3K antibody was from Proteintech (Rosemont, IL). The ELISA cotinine detection kit was purchased from Calbiotech (El Cajon, CA). The fibrinogen plasma detection/quantification ELISA kit was purchased form Innovative Research (Novi, MI).

Alarabi A.B., Karim Z.A., Montes Ramirez J.E., Hernandez K.R., Lozano P.A., Rivera J.O., Alshbool F.Z, & Khasawneh F.T. (2020). Short-Term Exposure to Waterpipe/Hookah Smoke Triggers a Hyperactive Platelet Activation State and Increases the Risk of Thrombogenesis. Arteriosclerosis, thrombosis, and vascular biology, 40(2), 335-349.

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Quantification of Blood Alkaline Phosphatase, Nicotine, and Cotinine

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Blood samples (10 ml) were obtained from jugular vein and used to dose alkaline phosphatase, nicotine and Cotinine serum levels, after eight weeks of surgery. Alkaline phosphatase activity was colorimetrically obtained by automated laboratory techniques using p-nitrophenilphosphate as substrate.
The major nicotine metaboliteCotinine -was detected and quantified by Gas Chromatography -Flame Ionization detector (CG/FID, Chromatographer HP 5890 II, Capillary Column BP 624 -30 m) after solid phase extraction (SPE, Varian HF Bond Elut LRC Certify).
Cotinine (98 % purity (-) -Cotinine, Sigma) was used as analytical standard. The method sensitivity was 0.1 ng/mL.

Linden M.S.S., Bittencourt M.E., Carli J.P., Miyagaki D.C., Santos P.L.D., Paranhos L.R., Groppo F.C, & Ramacciato J.C. (2018). The effects of subcutaneous injection of nicotine on osseointegration of machined and anodized implants in rabbits. Acta cirurgica brasileira, 33(1).

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ARPE-19 Cells Exposed to Nicotine and Cotinine

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(−)-Nicotine (N3876; Sigma-Aldrich, St. Louis, MO) and (−)-cotinine (C5923; Sigma-Aldrich) were dissolved in dimethyl sulfoxide (DMSO). ARPE-19 cells or RPE explants were treated with 1 or 2 μM nicotine, cotinine or nicotine-cotinine mixture in DF-12 culture medium (final concentration of DMSO was 0.1%) for 5–7 days. The control group was the cells treated with 0.1% DMSO in DF-12 culture medium. The medium was changed daily in order to maintain a constant level of nicotine and cotinine.

Zhang X.Y., Ng T.K., Brelén M.E., Chan K.P., Wu D., Yung J.S., Cao D., Wang Y., Zhang S., Chan S.O, & Pang C.P. (2017). Disruption of retinal pigment epithelial cell properties under the exposure of cotinine. Scientific Reports, 7, 3139.

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Quantification of Nicotine and Metabolites

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All reagents were of analytical grade unless otherwise indicated. (-)-Nicotine, (±)nornicotine, (±)-anabasine, (-)-cotinine and methanol CHROMASOLV® for HPLC were purchased from Sigma-Aldrich (St. Louis, MO, USA). Radiolabeled nicotine, DL-[Nmethyl-14 C] nicotine (50-60 mCi/mmol or 1.85-2.22 Gbq/mmol), was obtained from American Radiolabeled Chemicals, Inc. (MO, USA). All solutions and dilutions were prepared with double distilled de-ionised water, produced by an ELGA PURELAB Ultra water purification system from Veolia Water Systems Ltd. (High Wycombe, UK) unless otherwise stated.

du Rand E.E., Pirk C.W.W., Nicolson S.W, & Apostolides Z. (2017). The metabolic fate of nectar nicotine in worker honey bees. Journal of insect physiology, 98.

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Nicotine and Cotinine Standardization

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Nicotine and cotinine were purchased from Sigma-Aldrich Japan (Tokyo, Japan). Nicotine-d₃ (isotopic purity 98.4%), and cotinine-d₃ (isotopic purity 99.9%) purchased from Toronto Research Chemicals Inc. (Toronto, Ontario, Canada) were used as an internal standard (IS). Stock solutions of 1 mg mL⁻¹ of each compound were prepared by dissolving in methanol. The working solutions were prepared by diluting these stock solutions with distilled water to the required concentration. These prepared solutions were stored at 4 °C in refrigerator until use. Methanol and distilled water were of LC–MS grade, while all other chemicals were of analytical reagent grade.

Kataoka H., Kaji S, & Moai M. (2021). Risk Assessment of Passive Smoking Based on Analysis of Hair Nicotine and Cotinine as Exposure Biomarkers by In-Tube Solid-Phase Microextraction Coupled On-Line to LC-MS/MS. Molecules, 26(23), 7356.

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