What is the recommended protocol for the 1 mg overnight dexamethasone suppression test to screen for autonomous cortisol secretion in a patient with an adrenal mass?

Medical Advisory BoardAll articles are reviewed for accuracy by our Medical Advisory Board
Educational purpose only • Exercise caution as content is pending human review
Article Review Status
Submitted
Under Review
Approved

Last updated: February 20, 2026View editorial policy

Personalize

Help us tailor your experience

Which best describes you? Your choice helps us use language that's most understandable for you.

1 mg Overnight Dexamethasone Suppression Test Protocol

The 1 mg overnight dexamethasone suppression test should be performed by administering 1 mg of dexamethasone orally at 11 PM (23:00h), followed by measurement of serum cortisol at 8 AM the next morning. 1, 2, 3

Test Administration Protocol

  • Timing of dexamethasone administration: Give 1 mg dexamethasone orally between 11 PM and midnight (23:00h) 1, 2, 3

  • Timing of cortisol measurement: Draw blood for serum cortisol at 8 AM (08:00h) the following morning 1, 2, 3

  • Simultaneous dexamethasone level measurement: Measure serum dexamethasone concentration at the same time as the 8 AM cortisol draw to validate adequate drug absorption and patient compliance 2, 4

Interpretation Thresholds

  • Cortisol ≤50 nmol/L (≤1.8 μg/dL): Excludes autonomous cortisol secretion 1, 2, 3, 5

  • Cortisol 51-138 nmol/L (1.8-5.0 μg/dL): Suggests possible autonomous cortisol secretion; requires additional confirmatory testing 1, 2, 3, 5

  • Cortisol >138 nmol/L (>5.0 μg/dL): Indicates autonomous cortisol secretion 1, 2, 3, 5

The 1.8 μg/dL cutoff provides 100% sensitivity and 87% specificity for detecting Cushing's syndrome, making it the optimal threshold for screening 2, 6

Validation of Test Results

Measuring serum dexamethasone levels is critical to avoid false-positive results from inadequate drug absorption or patient non-compliance. 2, 4

  • Valid test: Serum dexamethasone ≥140 ng/dL (≥3.6 nmol/L) confirms adequate drug levels 4

  • Invalid test: Serum dexamethasone <140 ng/dL indicates inadequate suppression and requires test repetition 4

  • Undetectable dexamethasone (<19.5 ng/dL): Most commonly caused by failure to take medication (17%), concurrent glucocorticoid use (25%), or CYP3A4-inducing drugs like anticonvulsants (20%) 4

In a large series, 11.2% of dexamethasone suppression tests were invalid due to low serum dexamethasone levels, highlighting the importance of measuring drug levels to avoid unnecessary repetition and false-positive results 4

Confirmatory Testing for Equivocal Results (Cortisol 51-138 nmol/L)

When the 8 AM cortisol falls in the indeterminate range, additional criteria must be met to confirm autonomous cortisol secretion: 7

  • Basal ACTH <10 pg/mL (or poor ACTH response to CRH stimulation) 7

  • Evening cortisol ≥5 μg/dL measured between 21:00-23:00h 7

If both criteria are met, autonomous cortisol secretion is confirmed; if only one criterion is met, consider additional testing including DHEA-S levels, 24-hour urinary free cortisol, and clinical manifestations 7

Common Pitfalls and How to Avoid Them

  • CYP3A4-inducing medications: Anticonvulsants (phenytoin, carbamazepine, phenobarbital), rifampin, and St. John's wort accelerate dexamethasone metabolism, causing falsely elevated cortisol levels 2, 4

  • Gastrointestinal malabsorption: Prior gastric bypass, inflammatory bowel disease, or celiac disease impair dexamethasone absorption (13% of invalid tests) 4

  • Oral estrogen therapy: Increases corticosteroid-binding globulin, elevating total cortisol measurements and causing false-positive results 2, 5

  • Patient non-adherence: Failure to take dexamethasone accounts for 17% of undetectable drug levels 4

  • Concurrent glucocorticoid use: Exogenous steroids suppress cortisol but may be misinterpreted if dexamethasone levels are not measured 4

Patient Preparation Instructions

  • No fasting required: ACTH and cortisol levels are determined by hypothalamic-pituitary-adrenal axis feedback, not nutritional status 2

  • Discontinue interfering medications: Stop CYP3A4 inducers at least 2 weeks before testing when clinically feasible 2, 4

  • Verify medication compliance: Provide written and verbal instructions emphasizing the importance of taking dexamethasone at the specified time 4

  • Avoid exogenous glucocorticoids: Ensure patient is not using topical, inhaled, or systemic corticosteroids 2, 4

Why This Test Is Preferred

The 1 mg overnight dexamethasone suppression test is the preferred screening method because it is simple, cost-effective, and has excellent diagnostic accuracy when properly validated with serum dexamethasone levels 1, 2, 3. The conventional 2-day low-dose dexamethasone suppression test (0.5 mg every 6 hours for 48 hours) does not improve diagnostic specificity compared to the overnight test and is more cumbersome 8. The 8 mg dexamethasone suppression test largely replicates the results of the 1 mg test and adds no additional diagnostic value for screening autonomous cortisol secretion 9.

References

Guideline

Guideline Directed Topic Overview

Dr.Oracle Medical Advisory Board & Editors, 2025

Guideline

Cushing's Syndrome Classification and Diagnosis

Praxis Medical Insights: Practical Summaries of Clinical Guidelines, 2026

Guideline

Diagnostic Testing for Adrenal Adenoma

Praxis Medical Insights: Practical Summaries of Clinical Guidelines, 2026

Research

Overnight 1-mg Dexamethasone Suppression Test for Screening Cushing Syndrome and Mild Autonomous Cortisol Secretion (MACS): What Happens when Serum Dexamethasone Is Below Cutoff? How Frequent Is it?

Endocrine practice : official journal of the American College of Endocrinology and the American Association of Clinical Endocrinologists, 2023

Guideline

Diagnostic du Syndrome de Cushing

Praxis Medical Insights: Practical Summaries of Clinical Guidelines, 2025

Professional Medical Disclaimer

This information is intended for healthcare professionals. Any medical decision-making should rely on clinical judgment and independently verified information. The content provided herein does not replace professional discretion and should be considered supplementary to established clinical guidelines. Healthcare providers should verify all information against primary literature and current practice standards before application in patient care. Dr.Oracle assumes no liability for clinical decisions based on this content.

Have a follow-up question?

Our Medical A.I. is used by practicing medical doctors at top research institutions around the world. Ask any follow up question and get world-class guideline-backed answers instantly.