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Diagnostic Tests: 24-Hour Urine Cortisol Level

Diagnoses for Which This Is the Most Sensitive Test


The 24-hour urinary free cortisol level is the most sensitive screening test for endogenous hypercortisolism and is the best initial test for diagnosing Cushing’s syndrome. Because cortisol secretion is variable and follows a diurnal rhythm, a 24-hour urine collection provides an integrated measurement of cortisol production over an entire day, making it more reliable than a single serum cortisol level.


What This Substance Is

Cortisol is a glucocorticoid hormone produced by the adrenal cortex. It plays a key role in metabolism, immune regulation, blood pressure control, and the physiologic response to stress. Excess cortisol leads to the characteristic physical and metabolic abnormalities seen in Cushing’s syndrome.


When to Answer Urinary Free Cortisol

You should choose 24-hour urinary free cortisol when the clinical presentation suggests Cushing’s syndrome. Classic features include:


  • Truncal (central) obesity
  • “Buffalo hump”
  • Purple striae
  • Easy bruising
  • Proximal muscle weakness
  • Hypertension or glucose intolerance

In exam questions, the correct best initial test for suspected Cushing’s syndrome is either 24-hour urine cortisol or the overnight dexamethasone suppression test.



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