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Determination of cotinine, 3'-hydroxycotinine, and their glucuronides in urine by ultra-high performance liquid chromatography


Shastri, M and Lu, W and Ferguson, SG and Narkowicz, CK and Davies, NW and Jacobson, GA, Determination of cotinine, 3'-hydroxycotinine, and their glucuronides in urine by ultra-high performance liquid chromatography, Analytical Letters, 48, (8) pp. 1217-1233. ISSN 0003-2719 (2014) [Refereed Article]

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Copyright 2015 Taylor & Francis Group, LLC

DOI: doi:10.1080/00032719.2014.979363


The measurement of the primary nicotine metabolites, cotinine and trans-3′-hydroxycotinine, is a useful biomarker of nicotine exposure and metabolism genetics for smoking cessation research. Herein is described an ultra-high performance liquid chromatography–tandem mass spectrometry method for the determination of these primary nicotine metabolites in urine. Urine samples were diluted one hundred-fold with water and introduced into an ultra-high performance liquid chromatography triple quadrupole mass spectrometer using positive ion electrospray ionization with multiple reaction monitoring. Levels of urinary nicotine metabolites: cotinine, trans-3′-hydroxycotinine, and their respective glucuronides were determined directly using deuterated internal standards and compared with indirect determination by enzymatic hydrolysis. The assay was applied to a community sample of smokers’ urine (n = 280). The assay demonstrated satisfactory performance (relative standard deviation of 1.6–6.5 percent at the 1000 nanograms per milliliter level and > 98 percent recovery) suitable for application to smoking studies with a run time less than five minutes. The mean (min-max) levels of cotinine and cotinine-glucuronide were 968 (31-3831) and 976 (9-5607) nanograms per milliliter. The mean (min-max) levels of trans-3′-hydroxycotinine and trans-3′-hydroxycotinine-glucuronide were 3529 (13-21337) and 722 (0-4633) nanograms per milliliter. Direct determination of glucuronide metabolites was superior to indirect measurement using enzymatic hydrolysis, where there was evidence of loss of metabolites during sample preparation. A sensitive and selective ultra-high performance liquid chromatography–tandem mass spectrometry assay was successfully developed for the determination of cotinine, trans-3′-hydroxycotinine, and their glucuronides in urine. The rapid and simple sample preparation makes this assay suitable for high throughput studies involving nicotine metabolism phenotype for both cytochrome P450 2A6 and uridine 5′-diphospho-glucuronosyltransferase, smoking prevalence, and cessation studies.

Item Details

Item Type:Refereed Article
Keywords:cotinine, nicotine, smoking cessation
Research Division:Biomedical and Clinical Sciences
Research Group:Pharmacology and pharmaceutical sciences
Research Field:Pharmaceutical sciences
Objective Division:Health
Objective Group:Other health
Objective Field:Other health not elsewhere classified
UTAS Author:Shastri, M (Mr Madhur Shastri)
UTAS Author:Lu, W (Dr Monica Lu)
UTAS Author:Ferguson, SG (Professor Stuart Ferguson)
UTAS Author:Narkowicz, CK (Dr Christian Narkowicz)
UTAS Author:Davies, NW (Associate Professor Noel Davies)
UTAS Author:Jacobson, GA (Professor Glenn Jacobson)
ID Code:97538
Year Published:2014
Web of Science® Times Cited:4
Deposited By:Pharmacy
Deposited On:2014-12-22
Last Modified:2017-11-02
Downloads:3 View Download Statistics

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