What is the correct method to reconstitute a 0.25 mg Cortrosyn (cosyntropin) lyophilized vial with sterile 0.9% sodium chloride, including the resulting concentration, appropriate dosing aliquots for high‑dose and low‑dose ACTH stimulation tests, and storage recommendations?

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Cortrosyn (Cosyntropin) Reconstitution Protocol

Standard Reconstitution Method

Reconstitute the 0.25 mg Cortrosyn lyophilized vial by adding 1 mL of sterile 0.9% sodium chloride to produce a final concentration of 0.25 mg/mL (250 mcg/mL). 1

Step-by-Step Reconstitution Procedure

  • Sanitize the vial stopper with 70% isopropanol before needle puncture 1
  • Use sterile technique throughout the preparation process, avoiding direct contact contamination of sterile needles and syringes 1
  • Add 1 mL of sterile 0.9% sodium chloride (without phenol preservative preferred) to the lyophilized vial 1
  • Mix gently until the powder is completely dissolved—do not shake vigorously 1
  • Perform visual inspection after mixing to ensure complete dissolution and absence of particulate matter 1
  • Warm to room temperature before administration if the solution was refrigerated 1

Dosing Aliquots for ACTH Stimulation Tests

High-Dose (Standard) Test

Administer the entire 0.25 mg (250 mcg) dose intravenously or intramuscularly for the standard cosyntropin stimulation test. 2, 3 This is the FDA-approved dose and is recommended over low-dose testing due to easier practical administration, comparable diagnostic accuracy, and regulatory approval. 2

Low-Dose Test (When Specifically Indicated)

For a 1 mcg low-dose test, dilute the reconstituted solution further:

  • Draw up 0.1 mL of the 0.25 mg/mL solution (containing 25 mcg)
  • Add to 2.4 mL of sterile 0.9% sodium chloride to create a 2.5 mL solution containing 10 mcg/mL
  • Administer 0.1 mL of this diluted solution (1 mcg dose) 4

Note: The low-dose test requires bedside dilution, making it less practical for routine clinical use, and the high-dose test is preferred in standard practice. 2 However, in patients with obesity (BMI >30), a weight-adapted dose of 0.2 mcg/kg may be more appropriate than the fixed 1 mcg dose. 5

Storage Recommendations

Unreconstituted Vials

  • Store sealed lyophilized vials at room temperature with desiccation if using bulk powder, though FDA-approved prepackaged vials do not require desiccation or freezing 1

Reconstituted Solutions

  • Refrigerate at 4°C immediately after reconstitution 1
  • Use within 24 hours when reconstituted with sterile 0.9% sodium chloride without phenol 1
  • Discard any unused solution remaining in the vial or syringe after administration 1
  • Do not store reconstituted solutions at room temperature for extended periods, as this accelerates degradation 1

Critical Reconstitution Considerations

Avoid adding preservatives such as phenol to the diluent unless specifically required by the manufacturer, as preservative-free sterile saline is preferred and widely used. 1 The pH of cosyntropin in normal saline is weakly to moderately acidic, and buffered solutions should not be used as they are less stable. 1

Label the reconstituted vial clearly with the drug name, concentration, reconstitution date, and expiration time. 1

References

Guideline

Guideline Directed Topic Overview

Dr.Oracle Medical Advisory Board & Editors, 2025

Guideline

Diagnosing Adrenal Insufficiency in Hypo-osmolar Hyponatremia

Praxis Medical Insights: Practical Summaries of Clinical Guidelines, 2026

Guideline

Adrenal Insufficiency Diagnosis

Praxis Medical Insights: Practical Summaries of Clinical Guidelines, 2025

Research

Low-Dose ACTH Stimulation Test in Obesity: A Randomized Dose Assessment.

International journal of endocrinology, 2022

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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