Test ID CORTU Cortisol, Free, 24 Hour, Urine
Necessary Information
24-Hour volume (in milliliters) is required.
Specimen Required
Supplies: Urine Tubes, 10-mL (T068)
Submission Container/Tube: Plastic, urine tube
Specimen Volume: 5 mL
Collection Instructions:
1. Collect urine for 24 hours.
2. Add 10 g of boric acid as preservative at start of collection.
Additional Information: See Urine Preservatives-Collection and Transportation for 24-Hour Urine Specimens for multiple collections.
Secondary ID
8546Useful For
Preferred screening test for Cushing syndrome
Diagnosis of pseudo-hyperaldosteronism due to excessive licorice consumption
Test may not be useful in the evaluation of adrenal insufficiency
Special Instructions
Method Name
Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS)
Reporting Name
Cortisol, Free, USpecimen Type
UrineSpecimen Minimum Volume
3 mL
Specimen Stability Information
Specimen Type | Temperature | Time | |
---|---|---|---|
Urine | Refrigerated (preferred) | 14 days | |
Frozen | 28 days | ||
Ambient | 7 days |
Clinical Information
Cortisol is a steroid hormone synthesized from cholesterol by a multienzyme cascade in the adrenal glands. It is the main glucocorticoid in humans and acts as a gene transcription factor influencing a multitude of cellular responses in virtually all tissues. Cortisol plays a critical role in glucose metabolism, maintenance of vascular tone, immune response regulation, and in the body's response to stress. Its production is under hypothalamic-pituitary feedback control.
Only a small percentage of circulating cortisol is biologically active (free), with the majority of cortisol inactive (protein bound). As plasma cortisol values increase, free cortisol (ie, unconjugated cortisol or hydrocortisone) increases and is filtered through the glomerulus. Urinary free cortisol (UFC) in the urine correlates well with the concentration of plasma free cortisol. UFC represents excretion of the circulating, biologically active, free cortisol that is responsible for the signs and symptoms of hypercortisolism.
UFC is a sensitive test for the various types of adrenocortical dysfunction, particularly hypercortisolism (Cushing syndrome). A measurement of 24-hour UFC excretion, by liquid chromatography-tandem mass spectrometry (LC-MS/MS), is the preferred screening test for Cushing syndrome. LC-MS/MS methodology eliminates analytical interferences including carbamazepine (Tegretol) and synthetic corticosteroids, which can affect immunoassay-based cortisol results.
Reference Values
0-2 years: not established
3-8 years: 1.4-20 mcg/24 hours
9-12 years: 2.6-37 mcg/24 hours
13-17 years: 4.0-56 mcg/24 hours
≥18 years: 3.5-45 mcg/24 hours
Use the factor below to convert from mcg/24 hours to nmol/24 hours:
Conversion factor
Cortisol: mcg/24 hours x 2.76=nmol/24 hours (molecular weight=362.5)
For SI unit Reference Values, see https://www.mayocliniclabs.com/order-tests/si-unit-conversion.html
Cautions
Acute stress (including hospitalization and surgery), alcoholism, depression, and many drugs (eg, exogenous cortisone, anticonvulsants) can obliterate normal diurnal variation, affect response to suppression/stimulation tests, and increase baseline levels.
This test has limited usefulness in the evaluation of adrenal insufficiency.
This methodology (liquid chromatography-tandem mass spectrometry) eliminates analytical interferences including carbamazepine (Tegretol) and synthetic corticosteroids.
Renal disease (decreased excretion) may cause falsely low 24-hour urinary free cortisol values.
Improper collection may alter results. For example, a missed morning collection may result in false-negative tests; an extra morning collection (ie, >24 hours) may give false-positive results.
Twenty-four hour urinary free cortisol values may be elevated to twice the upper limit of the normal range during pregnancy.
Patients with exogenous Cushing syndrome caused by ingestion of hydrocortisone will not have suppressed cortisol values.
Day(s) Performed
Monday through Friday
Report Available
2 to 5 daysPerforming Laboratory

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
82530