What is the purpose and procedure of the Adrenocorticotropic Hormone (ACTH) stimulation test?

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ACTH Stimulation Test: Purpose and Procedure

The ACTH stimulation test (cosyntropin test) is the recommended diagnostic test for evaluating adrenal insufficiency, with the high-dose (250-μg) protocol being preferred over the low-dose (1-μg) test due to comparable diagnostic accuracy, easier administration, and more standardized reference ranges. 1, 2

Purpose of the Test

  • The ACTH stimulation test is used to diagnose adrenal insufficiency by evaluating the adrenal cortex's ability to respond to stimulation with synthetic ACTH (cosyntropin) 3
  • It helps differentiate between primary adrenal insufficiency (adrenal gland dysfunction) and secondary adrenal insufficiency (pituitary dysfunction) 2
  • The test is indicated for screening adrenocortical insufficiency in adults and pediatric patients, in combination with other diagnostic tests 3
  • It is particularly valuable in evaluating patients with suspected Critical Illness-Related Corticosteroid Insufficiency (CIRCI) 1

Test Procedure

Preparation

  • Discontinue glucocorticoids and spironolactone on the day of testing 3
  • For long-acting glucocorticoids, discontinuation should occur for a longer period before testing 3
  • Estrogen-containing medications should be stopped 4-6 weeks before testing as they may elevate plasma total cortisol levels 3

Administration Protocol

  • High-dose protocol (recommended): 250 μg of cosyntropin administered intravenously or intramuscularly 1, 2
  • For pediatric patients: 0.125 mg for patients under 2 years; 0.25 mg for patients 2-17 years 3
  • Blood samples are collected for serum cortisol measurement at baseline (0 minutes) and at 30 and 60 minutes after cosyntropin administration 3, 4

Interpretation

  • In primary adrenal insufficiency: A peak cortisol level below 18 μg/dL (500 nmol/L) at 30 or 60 minutes indicates adrenal insufficiency 1, 2
  • For newer specific cortisol assays: Updated cutoff values of 14-15 μg/dL are recommended instead of the historical 18 μg/dL threshold 5
  • In equivocal cases: A delta cortisol (change from baseline) of <9 μg/dL after cosyntropin administration may indicate CIRCI 1
  • A baseline cortisol <2 μg/dL is highly predictive of subnormal stimulated cortisol values 5

Clinical Applications

  • Primary Adrenal Insufficiency: The test shows low cortisol and elevated ACTH levels 1
  • Secondary Adrenal Insufficiency: Shows low cortisol with low or normal ACTH levels 1, 2
  • Critical Illness-Related Corticosteroid Insufficiency (CIRCI): The test helps identify patients who may benefit from corticosteroid therapy 1
  • Immune-Related Adverse Events: Used to diagnose adrenal insufficiency in patients receiving immune checkpoint inhibitor therapy 1

Advantages of High-Dose vs. Low-Dose Protocol

  • The Society of Critical Care Medicine and European Society of Intensive Care Medicine recommend the high-dose (250-μg) ACTH test over the low-dose (1-μg) test 1, 2
  • Advantages of high-dose protocol:
    • Easier to administer (no preparation required at bedside) 1
    • More standardized protocol with established reference ranges 2
    • Comparable diagnostic accuracy to low-dose testing 1
    • Commercial availability of 250-μg ampoules 6

Potential Pitfalls and Limitations

  • Cortisol binding globulin levels can affect total cortisol measurements, potentially leading to diagnostic inaccuracies 3, 7
  • Recent pituitary injury cases may show false-negative results with the ACTH stimulation test 6
  • The test should be performed within the first 3 days after initiation of empiric glucocorticoid therapy for reliable results 8
  • Medications affecting cortisol or cortisol binding globulin levels can interfere with test interpretation 3, 7
  • The time of day when the test is performed may affect results, though the high-dose test is less affected by this variable 7

Special Considerations

  • In critically ill patients, a random plasma cortisol <10 μg/dL may be diagnostic of CIRCI without requiring an ACTH stimulation test 1
  • Treatment of suspected acute adrenal insufficiency should never be delayed for diagnostic procedures 1
  • All patients diagnosed with adrenal insufficiency should receive education on stress dosing and carry emergency identification 1

References

Guideline

Guideline Directed Topic Overview

Dr.Oracle Medical Advisory Board & Editors, 2025

Guideline

Diagnosis of Adrenal Insufficiency in Hypopituitarism

Praxis Medical Insights: Practical Summaries of Clinical Guidelines, 2025

Research

Intramuscular ACTH stimulation test for assessment of adrenal function.

The Journal of the Association of Physicians of India, 2013

Research

Adrenocorticotropic hormone stimulation test during high-dose glucocorticoid therapy.

Endocrine practice : official journal of the American College of Endocrinology and the American Association of Clinical Endocrinologists, 2009

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.

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