Cortisol; Cortisone
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Cortisol/Cortisone – Free, 24 hr Urine
Test ID: COCOU U
Test
Method
High Performance Liquid Chromatography/Tandem Mass Spectrometry (LC-MS/MS)
Method Description
The cortisol and cortisone are extracted from the resulting supernatant by an online extraction utilizing high throughput liquid chromatography. This is followed by conventional liquid chromatography and analysis on a tandem mass spectrometer equipped with a heated nebulizer ion source. Deuterated cortisol (d4-cortisol, d7-cortisone) is added to a 0.1 mL sample as an internal standard. Cortisol, Cortisone and d4-cortisol are extracted from the specimens using online turbulent flow high-performance liquid chromatography extraction.
Report Includes
Specimens
Urine
Clinical Utility
Screening test for Cushing syndrome (hypercortisolism).
Assisting in diagnosing acquired or inherited abnormalities of 11-beta-hydroxy steroid dehydrogenase (cortisol to cortisone ratio)
Diagnosis of pseudo-hyperaldosteronism due to excessive licorice consumption.
This test has limited usefulness in the evaluation of adrenal insufficiency.
Test Version
11-Nov-2025
Specimen
Specimens
Urine
Collection Containers
Urine Container (24hr) 4L
Sample Volume
5.0 mL
Minimum Volume
3.0 mL
Collection & Handling
Collection Instructions
Add 10 g of boric acid as preservative at start of collection. Alternatively add 50% Acetic Acid, 25 mL per 24-hour collection or for patients <5 years old, 15 mL per 24-hour collection. Collection without preservative is only acceptable if the specimen is kept cold or frozen during the collection period and then shipped frozen to ICL.
Handling Information
Aliquot into plastic or polypropylene transfer vial. Provide 24h collection volume. Store and send sample frozen.
Stability
| Ambient | 72 hours |
|---|---|
| Refrigerated | 14 days |
| Frozen | 28 days |
Test Version
11-Nov-2025
Performance / Interpretation
Method
High Performance Liquid Chromatography/Tandem Mass Spectrometry (LC-MS/MS)
Method Description
The cortisol and cortisone are extracted from the resulting supernatant by an online extraction utilizing high throughput liquid chromatography. This is followed by conventional liquid chromatography and analysis on a tandem mass spectrometer equipped with a heated nebulizer ion source. Deuterated cortisol (d4-cortisol, d7-cortisone) is added to a 0.1 mL sample as an internal standard. Cortisol, Cortisone and d4-cortisol are extracted from the specimens using online turbulent flow high-performance liquid chromatography extraction.
Turnaround Time
5 days
Results
| Name | Units | Reference Range | Conversion Factor | |
|---|---|---|---|---|
| CORTISOL | mcg/24 h |
|
mcg/24 hours x 2.76=nmol/24 h (molecular weight=362.5) | |
| CORTISONE | mcg/24 h |
|
mcg/24 hours x 2.78=nmol/24 h (molecular weight=360) | |
|
Most patients with Cushing syndrome have increased 24-hour urinary excretion of cortisol and/or cortisone. Further studies, including suppression or stimulation tests, measurement of serum corticotropin (adrenocorticotropic hormone) concentrations, and imaging are usually necessary to confirm the diagnosis and determine the etiology.
Values in the normal range may occur in patients with mild Cushing syndrome or with periodic hormonogenesis. In these cases, continuing follow-up and repeat testing are necessary to confirm the diagnosis. Patients with Cushing syndrome due to intake of synthetic glucocorticoids should have both suppressed cortisol and cortisone. In these circumstances a synthetic glucocorticoid screen might be ordered. Suppressed cortisol and cortisone values may also be observed in primary adrenal insufficiency and hypopituitarism. However, random urine specimens are not useful for evaluation of hypocorticalism. Further, many normal individuals also may exhibit a very low 24-hour urinary cortisol excretion with considerable overlap with the values observed in pathological hypocorticalism. Therefore, without other tests, 24-hour urinary cortisol measurements cannot be relied upon for the diagnosis of hypocorticalism. Patients with 11-beta hydroxy steroid dehydrogenase deficiency may have cortisone to cortisol ratios <1, whereas a ratio of 2:1 to 3:1 is seen in normal patients. Excessive licorice consumption and use of carbenoxolone, a synthetic derivative of glycyrrhizinic acid used to treat gastroesophageal reflux disease, also may suppress the ratio to <1. 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 methodology (liquid chromatography tandem mass spectrometry) eliminates analytical interferences including carbamazepine (Tegretol) and synthetic corticosteroids. This test has limited usefulness in the evaluation of adrenal insufficiency. 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. Renal disease (decreased clearance) may cause falsely low values. Values may be elevated to twice normal in pregnancy. Patients with exogenous Cushing syndrome caused by ingestion of hydrocortisone will not have suppressed cortisol and cortisone values. |
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Test Version
11-Nov-2025
Interface / Setup
HL7 Interface Codes
| Order Code | Result Codes | Units |
|---|---|---|
| COCOU U | 64027Cortisol, U | mcg/24 h |
| 64028Cortisone, U | mcg/24 h |
Test Version
11-Nov-2025