Protocol - Biomarker of exposure to nicotine-containing products - Urine
Description
A biospecimen is collected from the respondent to measure cotinine, a marker of either smoking or of environmental tobacco smoke exposure. One of the most common methods is via a urine sample obtained from the respondent. Cotinine in urine is best measured by liquid chromatography - tandem mass spectrometry (LC-MS/MS).
Specific Instructions
Cotinine is excreted as both “free” cotinine and as its glucuronide conjugate. Most methods measure free cotinine. If total (free + glucuronide) concentrations are needed, the urine sample can be treated with a glucuronidase enzyme to deconjugate the glucuronides prior to analysis.
Because of the sensitive nature of this assay, analysts cannot be actively using tobacco products, and measurements must be performed in a smoke-free environment.
Safety Precautions: Personal protective equipment (PPE) including eye protection, gloves and suitable protective clothing when used to extract and process samples.
The Content Expert Panel acknowledges that a Total Nicotine Equivalents (TNE) might also be valuable to collect.
Availability
Protocol
The respondent is asked to urinate in a sterile 90-mL urine specimen container. Children above the age of 10 and adults can self-collect the urine. The specimen should be refrigerated immediately after collection. Aliquot 3mLs into a 4 mL cryovial. Freeze the cryovial at -20?C until analysis. If the samples need to be stored greater than 1 year freeze at -80?C.
Liquid chromatography – tandem mass spectrometry (LC-MS/MS) is the preferred method to accurately measure cotinine in urine samples, especially when used to assess secondhand smoke exposure. (See source references for details.) The limit of quantitation is 0.05 ng/mL.
The urinary cotinine cutpoint to distinguish people who smoke from those who do not is 30-50 ng/ml (see source reference 3). Note that this range relates to background population exposure to secondhand smoke and to some degree of metabolic differences. When background secondhand exposure is low, the cutpoint is low.
Personnel and Training Required
Urine samples can be self-collected by children aged 10 or older with adequate instructions. A laboratory technician trained to process and analyze biological specimens is necessary to analyze urine samples. Laboratory training in the use of liquid chromatography and tandem mass spectrometry is required, as well as general laboratory training on the safe use of chemicals and solvents.
Equipment Needs
Standard urine collection supplies that have been sterilized. Laboratory supplies and instruments are needed to measure cotinine in biological fluids. Biological samples may be shipped using appropriate shipping procedures to laboratories that specialize in these types of analysis. This method requires high-performance liquid chromatography and tandem mass spectrometry on a triple quadrupole mass spectrometer.
Requirements
| Requirement Category | Required |
|---|---|
| Major equipment | No |
| Specialized training | Yes |
| Specialized requirements for biospecimen collection | No |
| Average time of greater than 15 minutes in an unaffected individual | No |
Mode of Administration
Bioassay
Lifestage
Child, Adolescent, Adult, Senior, Pregnancy
Participants
Ages 3 and up
Selection Rationale
Urine is easily collected and with the proper laboratory equipment, cotinine can be measured accurately from a sterile urine sample. Urinary cotinine is a frequently measured biomarker and produces more quantifiable results than other metabolites.
There are other assays (e.g., gas chromatography (GC), gas chromatography mass spectrometry (GC-MS), radioimmunoassay and enzyme-linked immunosorbent assays) used to measure cotinine but liquid chromatography – tandem mass spectrometry (LC-MS/MS) is preferred, especially when used to assess secondhand smoke exposure.
Determination of cotinine in urine using liquid chromatography – tandem mass spectrometry (LC-MS/MS) is the preferred method for determination of cotinine in urine (see source reference 1) and other biological fluids, especially for measuring concentrations in non-smokers for exposure to SHS and THS (see source reference 2).
Language
English
Standards
| Standard | Name | ID | Source |
|---|
Derived Variables
None
Process and Review
The Tobacco Regulatory Research (TRR) Content Expert Panel (CEP) reviewed the measures in the Tobacco Regulatory Research collection in February 2024.
Guidance from the TRR CEP includes:
· Replaced protocol
· New Data Dictionary
Previous version in Toolkit archive (link)
Protocol Name from Source
Wang L, et al. Collaborative Method Performance Study of the Measurement of Nicotine, Its Metabolites, and Total Nicotine Equivalents in Human Urine. CANCER EPIDEM BIOMAR, 2018
Source
Wang, L., Bernert, J. T., Benowitz, N. L., Feng, J., Jacob, P., 3rd, McGahee, E., Caudill, S. P., Scherer, G., Scherer, M., Pluym, N., Doig, M. V., Newland, K., Murphy, S. E., Caron, N. J., Sander, L. C., Shimizu, M., Yamazaki, H., Kim, S., Langman, L. J., Pritchett, J. S., ? Pirkle, J. L. (2018). Collaborative Method Performance Study of the Measurement of Nicotine, Its Metabolites, and Total Nicotine Equivalents in Human Urine. Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology, 27(9), 1083?1090. https://doi.org/10.1158/1055-9965.EPI-17-1127
Jacob, P., 3rd, Yu, L., Duan, M., Ramos, L., Yturralde, O., & Benowitz, N. L. (2011). Determination of the nicotine metabolites cotinine and trans-3-hydroxycotinine in biologic fluids of smokers and non-smokers using liquid chromatography-tandem mass spectrometry: biomarkers for tobacco smoke exposure and for phenotyping cytochrome P450 2A6 activity. Journal of chromatography. B, Analytical technologies in the biomedical and life sciences, 879(3-4), 267?276. https://doi.org/10.1016/j.jchromb.2010.12.012
Benowitz, N. L., Bernert, J. T., Foulds, J., Hecht, S. S., Jacob, P., Jarvis, M. J., Joseph, A., Oncken, C., & Piper, M. E. (2020). Biochemical Verification of Tobacco Use and Abstinence: 2019 Update. Nicotine & tobacco research : official journal of the Society for Research on Nicotine and Tobacco, 22(7), 1086?1097. https://doi.org/10.1093/ntr/ntz132
General References
Avila-Tang, E., Al-Delaimy, W. K., Ashley, D. L., Benowitz, N., Bernert, J. T., Kim, S., Samet, J. M., & Hecht, S. S. (2013). Assessing secondhand smoke using biological markers. Tobacco control, 22(3), 164?171. https://doi.org/10.1136/tobaccocontrol-2011-050298
Benowitz N. L. (1999). Biomarkers of environmental tobacco smoke exposure. Environmental health perspectives, 107 Suppl 2(Suppl 2), 349?355. https://doi.org/10.1289/ehp.99107s2349
Benowitz, N. L. (1996). Cotinine as a biomarker of environmental tobacco smoke exposure. Epidemiologic Reviews, 18(2), 188-204.
Dhar P. (2004). Measuring tobacco smoke exposure: quantifying nicotine/cotinine concentration in biological samples by colorimetry, chromatography and immunoassay methods. Journal of pharmaceutical and biomedical analysis, 35(1), 155?168. https://doi.org/10.1016/j.jpba.2004.01.009
Kuo, H. W., Yang, J. S., & Chiu, M. C. (2002). Determination of urinary and salivary cotinine using gas and liquid chromatography and enzyme-linked immunosorbent assay. Journal of chromatography. B, Analytical technologies in the biomedical and life sciences, 768(2), 297?303. https://doi.org/10.1016/s1570-0232(01)00613-4
Protocol ID
91707
Variables
Export Variables| Variable Name | Variable ID | Variable Description | dbGaP Mapping | |
|---|---|---|---|---|
| PX091707_Biomarker_Of_Exposure_To_Nicotine_Containing_Products_Urine_Concentration | ||||
| PX091707010000 | Concentration of analyte | N/A | ||
| PX091707_Biomarker_Of_Exposure_To_Nicotine_Containing_Products_Urine_Make_Manufacturer_Equipment | ||||
| PX091707020000 | Make and manufacturer of equipment | N/A | ||
| PX091707_Biomarker_Of_Exposure_To_Nicotine_Containing_Products_Urine_Repeatability_Coefficients_Variation | ||||
| PX091707030000 | What is the repeatability and coefficients more | N/A | ||
Measure Name
Biomarker of exposure to nicotine-containing products
Release Date
October 31, 2024
Definition
Cotinine is a major metabolite of nicotine and is an indicator of exposure to nicotine from tobacco or other nicotine containing products.
Purpose
To assess smoking and environmental tobacco smoke (ETS) exposure by measuring cotinine, a metabolite of nicotine. To screen for tobacco use and quantity and to estimate exposure to secondhand smoke (SHS) and all tobacco exposure. Also used as an outcome measure in smoking cessation trials to determine if an individual has quit smoking.
Keywords
Biomarker of exposure to nicotine-containing products - urine, cotinine, Respiratory, Liquid Chromatography, Tandem mass spectrometry (LC-MS/MS), LC-MS/MS, Cigaette smoking, secondhand smoke, thirdhand smoke
Measure Protocols
| Protocol ID | Protocol Name |
|---|---|
| 91707 | Biomarker of exposure to nicotine-containing products - Urine |
| 91708 | Biomarker of exposure to nicotine-containing products - Saliva |
| 91709 | Biomarker of exposure to nicotine-containing products - Serum |
Publications
There are no publications listed for this protocol.