Are there tests available to detect nicotine (nicotine) use?

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Last updated: December 22, 2025View editorial policy

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Nicotine Drug Testing

Yes, cotinine measurement is the gold standard laboratory test for detecting nicotine use, with urine cotinine (cutoff 30 ng/mL) being the recommended first-line test due to its optimal balance of sensitivity, 7-day detection window, and ease of collection. 1

Primary Testing Method: Cotinine

Cotinine is the major metabolite of nicotine and provides superior sensitivity compared to other nicotine biomarkers. 1 The test can be performed on multiple specimen types with specific cutoff values 2, 1:

  • Urine cotinine: 30 ng/mL (recommended first-line test)
  • Serum/plasma cotinine: 3-5 ng/mL
  • Saliva cotinine: 3-5 ng/mL

The half-life of cotinine is 16-18 hours, providing a biochemically verifiable detection window of up to 7 days after nicotine use. 2, 1 This extended window makes cotinine far more practical than other markers for clinical screening purposes.

Rapid point-of-care saliva cotinine dipstick tests are available and demonstrate 99% sensitivity and 96% specificity when compared to laboratory-based methods, making them suitable for immediate verification of smoking status in clinical settings. 3

Alternative Testing Methods

Carbon Monoxide (CO)

Exhaled CO can be measured with a cutoff of 5-6 ppm, but has significant limitations 2:

  • Half-life of only 2-8 hours with a 1-day detection window
  • Only detects combustible tobacco use (cigarettes)
  • Does not detect smokeless tobacco or nicotine replacement therapy (NRT)
  • Cannot distinguish tobacco smoking from marijuana smoking or other combustible exposures 2

A positive CO level alone is insufficient to confirm tobacco use in the era of vaping and cannabis use; combining CO with urine cotinine provides more robust assessment. 2

NNAL (Tobacco-Specific Nitrosamine Metabolite)

NNAL measured in urine (cutoff 47.3 pg/mL) offers unique advantages 2, 1:

  • Half-life of 10-18 days, detectable in urine for 6-12 weeks
  • Highly tobacco-specific
  • Can distinguish NRT use from actual tobacco exposure (critical for patients in cessation programs)
  • Detects smokeless tobacco use

For patients using NRT, add NNAL testing to distinguish therapeutic nicotine use from tobacco exposure. 1

Clinical Screening Tools

Beyond laboratory testing, validated screening questionnaires are available for adolescents and adults 2:

  • CRAFFT 2.1+N: Ten-item questionnaire assessing past-year nicotine use with specific examples (JUUL, Puff Bars, vape pens)
  • BSTAD (Brief Screener for Tobacco, Alcohol, and other Drugs): High sensitivity/specificity for identifying substance use
  • S2BI (Screening to Brief Intervention): Queries past-year frequency with built-in intervention recommendations

Important Clinical Caveats

Cotinine cannot distinguish between NRT and tobacco use, as it detects nicotine from any source. 2 This limitation is clinically significant when monitoring patients in smoking cessation programs who are using nicotine patches, gum, or lozenges.

Self-reported smoking status is frequently inaccurate. Objective cotinine testing identified smokers who falsely reported being nonsmokers in validation studies. 3

For comprehensive nicotine dose estimation after low-level exposure, measuring the sum of multiple metabolites (cotinine + cotinine-glucuronide + trans-3'-hydroxycotinine + 3HC-glucuronide) in 24-hour urine collection provides the strongest correlation (r=0.96) with actual nicotine dose. 4

References

Guideline

Nicotine Testing Guidelines

Praxis Medical Insights: Practical Summaries of Clinical Guidelines, 2025

Guideline

Guideline Directed Topic Overview

Dr.Oracle Medical Advisory Board & Editors, 2025

Research

Validation of self-reported smoking status using saliva cotinine: a rapid semiquantitative dipstick method.

Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology, 2007

Research

Estimation of nicotine dose after low-level exposure using plasma and urine nicotine metabolites.

Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology, 2010

Professional Medical Disclaimer

This information is intended for healthcare professionals. Any medical decision-making should rely on clinical judgment and independently verified information. The content provided herein does not replace professional discretion and should be considered supplementary to established clinical guidelines. Healthcare providers should verify all information against primary literature and current practice standards before application in patient care. Dr.Oracle assumes no liability for clinical decisions based on this content.

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