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HelpTest Code NICOU Nicotine and Metabolites, Random, Urine
Additional Codes
Cotinine
Reporting Name
Nicotine and Metabolites, UUseful For
Monitoring tobacco use
Monitoring patients on nicotine-replacement therapy for concurrent use of tobacco products
Performing Laboratory
Mayo Clinic Laboratories in Rochester
Specimen Type
UrineSpecimen Required
Supplies: Sarstedt Aliquot Tube, 5 mL (T914)
Container/Tube: Plastic urine container
Submission Container/Tube: Plastic, 5-mL tube
Specimen Volume: 3 mL
Collection Instructions:
1. Collect a random urine specimen.
2. No preservative.
Specimen Minimum Volume
2 mL
Specimen Stability Information
| Specimen Type | Temperature | Time |
|---|---|---|
| Urine | Refrigerated (preferred) | 28 days |
| Frozen | 365 days | |
| Ambient | 28 days |
Reference Values
Non-tobacco user with no passive exposure:
Nicotine <5.0 ng/mL
Cotinine
<5.0 ng/mL
Anabasine
<2.0 ng/mL
Nornicotine
<2.0 ng/mL
Day(s) Performed
Monday through Sunday
Test Classification
This test was developed and its performance characteristics determined by Mayo Clinic in a manner consistent with CLIA requirements. It has not been cleared or approved by the US Food and Drug Administration.CPT Code Information
80323
G0480 (if appropriate)
LOINC Code Information
| Test ID | Test Order Name | Order LOINC Value |
|---|---|---|
| NICOU | Nicotine and Metabolites, U | 43125-4 |
| Result ID | Test Result Name | Result LOINC Value |
|---|---|---|
| 82510 | Nicotine | 3854-7 |
| 21321 | Cotinine | 10366-3 |
| 21324 | Nornicotine | 33917-6 |
| 21323 | Anabasine | 33915-0 |
Interpretation
Urine nicotine in the range of 1000 to 5000 ng/mL with cotinine in the range of 1000 to 8000 ng/mL indicates the subject is either actively using a tobacco product or on high-dose nicotine patch therapy.
The presence of anabasine and nornicotine indicates a subject on patch therapy who is actively using a tobacco product.
Typical findings are as follows:
While using a tobacco product:
-Peak nicotine concentration: 1000 to 5000 ng/mL
-Peak cotinine concentration: 1000 to 8000 ng/mL
-Anabasine concentration: 10 to 500 ng/mL
-Nornicotine concentration: 30 to 900 ng/mL
Tobacco user after 2 weeks complete abstinence:
-Nicotine concentration: <30 ng/mL
-Cotinine concentration: <50 ng/mL
-Anabasine concentration: <2.0 ng/mL
-Nornicotine concentration: <2.0 ng/mL
Nontobacco user with passive exposure:
-Nicotine concentration: <20 ng/mL
-Cotinine concentration: <20 ng/mL
-Anabasine concentration: <2.0 ng/mL
-Nornicotine concentration: <2.0 ng/mL
Nontobacco user with no passive exposure:
-Nicotine concentration: <5.0 ng/mL
-Cotinine concentration: <5.0 ng/mL
-Anabasine concentration: <2.0 ng/mL
-Nornicotine concentration: <2.0 ng/mL
Clinical Reference
1. Moyer TP, Charlson JR, Enger RJ, et al. Simultaneous analysis of nicotine, nicotine metabolites, and tobacco alkaloids in serum or urine by tandem mass spectrometry, with clinically relevant metabolic profiles. Clin Chem 2002;48(9):1460-1471
2. Dale LC, Hurt RD, Hays JT. Drug therapy to aid in smoking cessation. Tips on maximizing patients' chances for success. Postgrad Med. 1998;104(6):75-78, 83-84
3. Rudasingwa G, Kim Y, Lee C, Lee J, Kim S, Kim S. Comparison of nicotine dependence and biomarker levels among traditional cigarette, heat-not-burn cigarette, and liquid e-cigarette users: Results from the Think Study. Int J Environ Res Public Health. 2021;18(9):4777. doi:10.3390/ijerph18094777
4. Sharma P, Sane N, Anand SD, Marimutthu P, Benegal V. Assessment of cotinine in urine and saliva of smokers, passive smokers, and nonsmokers: Method validation using liquid chromatography and mass spectrometry. Indian J Psychiatry. 2019;61(3):270-276. doi:10.4103/psychiatry.IndianJPsychiatry_61_18
5. AACC: Nicotine and cotinine. Testing.com; Updated February 2, 2023, Accessed October 7, 2025. Available at www.testing.com/tests/nicotine-and-cotinine/
Report Available
2 to 5 daysMethod Name
Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS)
Forms
If not ordering electronically, complete, print, and send a Therapeutics Test Request (T831) with the specimen.