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EPIC CODE: LAB3978 Nicotine and Metabolites, Serum

Additional Codes

Sunquest:  NICOSM
Mayo:         NICOS

Reporting Name

Nicotine and Metabolites, S

Useful For

Monitoring tobacco use in a clinical setting

Performing Laboratory

Mayo Clinic Laboratories in Rochester

Specimen Type

Serum Red

Specimen Required

Collection Container/Tube: Red top (serum gel/SST are not acceptable)

Submission Container/Tube: Plastic vial

Specimen Minimum Volume

0.5 mL

Specimen Stability Information

Specimen Type Temperature Time Special Container
Serum Red Refrigerated (preferred) 28 days
  Ambient  28 days
  Frozen  28 days

Reference Values


<3.0 ng/mL



<3.0 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. This test has not been cleared or approved by the US Food and Drug Administration.

CPT Code Information


G0480 (if appropriate)

LOINC Code Information

Test ID Test Order Name Order LOINC Value
NICOS Nicotine and Metabolites, S 90226-2


Result ID Test Result Name Result LOINC Value
21313 Nicotine 3853-9
21314 Cotinine 10365-5

Clinical Information

Fatalities related to tobacco use are a leading cause of death in United States. Nicotine, coadministered in tobacco products such as cigarettes, pipe tobacco, cigars, or chew, is an addicting substance that causes individuals to continue use of tobacco despite concerted efforts to quit. Nicotine stimulates dopamine release and increases dopamine concentration in the nucleus accumbens, a mechanism that is thought to be the basis for addiction for drugs of abuse.


Nicotine-dependent patients use tobacco products to achieve a peak serum nicotine value of 30 to 50 ng/mL, the concentration at which the nicotine high is maximized. Nicotine is metabolized in the liver to cotinine. Cotinine accumulates in serum in proportion to dose and hepatic metabolism (which is genetically determined); most tobacco users accumulate cotinine in the range of 200 to 800 ng/mL. Serum concentrations of nicotine and metabolites in these ranges indicate the patient is using tobacco or is receiving high-dose nicotine patch therapy.


Nicotine is rapidly metabolized, exhibiting an elimination half-life of approximately 2 hours. Cotinine exhibits an apparent elimination half-life of approximately 24 hours. Heavy tobacco users who abstain from tobacco for 2 weeks exhibit serum nicotine values less than 3.0 ng/mL and cotinine less than 3.0 ng/mL.


Passive exposure to tobacco smoke can cause accumulation of nicotine metabolites in nontobacco users. Serum cotinine has been observed to accumulate up to 8 ng/mL from passive exposure.


Tobacco users engaged in programs to abstain from tobacco require support in the form of counseling, pharmacotherapy, and continuous encouragement. Occasionally, counselors may elect to monitor abstinence by biochemical measurement of nicotine and metabolites in serum to verify abstinence. If results of biologic testing indicate the patient is actively using a tobacco product during therapy, additional counseling or intervention may be appropriate.


Serum nicotine concentration in the range of 30 to 50 ng/mL with cotinine in the range of 200 to 800 ng/mL indicates the subject is either actively using a tobacco product or on nicotine replacement therapy.


To discriminate if a patient on nicotine replacement therapy is also actively using a tobacco product, see NICOU / Nicotine and Metabolites, Random, Urine analysis; the presence of anabasine in urine, a tobacco alkaloid not present in nicotine replacement products, indicates recent tobacco use.


Typical findings are as follows:


While using a tobacco product:

-Peak nicotine concentration: 30 to 50 ng/mL

-Peak cotinine concentration: 200 to 800 ng/mL*

*Higher values may be seen in subjects with high cytochrome P450 2D6 activity


Tobacco user after 2 weeks complete abstinence:

-Nicotine concentration: <3.0 ng/mL

-Cotinine concentration: <3.0 ng/mL


Nontobacco user with passive exposure:

-Nicotine concentration: <3.0 ng/mL

-Cotinine concentration: <8.0 ng/mL


Nontobacco user with no passive exposure:

-Nicotine concentration: <3.0 ng/mL

-Cotinine concentration: <3.0 ng/mL


Knowledge of time elapsed between last dose and specimen collection is important for interpretation of test results.

Clinical Reference

1. Dale LC, Hurt RD, Hays JT: Drug therapy to aid in smoking cessation. Tips on maximizing patients' chances for success. Postgrad Med. 1998 Dec;104(6):75-78, 83-84

2. 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 Sep;48(9):1460-1471

3. Nicotine and cotinine.; Updated November 9, 2021 Accessed April 11, 2022. Available at

4. McGrath-Morrow SA, Gorzkowski J, Groner JA, et al: The effects of nicotine on development. Pediatrics. 2020 Mar;145(3):e20191346. doi: 10.1542/peds.2019-1346

Method Description

Nicotine and metabolites are extracted from serum. The extract is quantified by high-performance liquid chromatography-tandem mass spectrometry.(Unpublished Mayo method)

Report Available

2 to 4 days

Specimen Retention Time

2 weeks

Reject Due To

Gross hemolysis OK
Gross lipemia OK
Gross icterus OK

NY State Approved


Method Name

Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS)

Secondary ID



If not ordering electronically, complete, print, and send a Therapeutics Test Request (T831) with the specimen.