Hydrocortisone Acetate

This article is compiled based on the United States Pharmacopeia (USP) – 2025 Edition

Issued and maintained by the United States Pharmacopeial Convention (USP)

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C₂₃H₃₂O₆ 404.50 

Pregn-4-ene-3,20-dione, 21-(acetyloxy)-11,17-dihydroxy-, (11β)-; 

Cortisol 21-acetate CAS RN®: 50-03-3; UNII: 3X7931PO74. 

1 DEFINITION 

Hydrocortisone Acetate contains NLT 97.0% and NMT 102.0% of C₂₃H₃₂O₆, calculated on the dried basis. 

2 IDENTIFICATION 

Change to read: 

Spectroscopic Identification Tests 〈197〉, Infrared Spectroscopy: 197M

Change to read: 

Spectroscopic Identification Tests 〈197〉, Ultraviolet-Visible Spectroscopy: 197U

Analytical wavelength: 242 nm 

Sample solution: 10 µg/mL in methanol 

Acceptance criteria: Absorptivities, calculated on the dried basis, do not differ by more than 2.5%. 

3 ASSAY 

Procedure 

Mobile phase: Butyl chloride, water-saturated butyl chloride, tetrahydrofuran, methanol, and glacial acetic acid (475:475:70:35:30) Standard solution: 0.10 mg/mL of USP Hydrocortisone Acetate RS in Mobile phase 

Sample solution: 0.10 mg/mL of Hydrocortisone Acetate in Mobile phase 

Chromatographic system 

(See Chromatography 〈621〉, System Suitability.) 

Mode: LC 

Detector: UV 254 nm 

Column: 3.9-mm × 30-cm; 10-µm packing L3 

Flow rate: 1 mL/min 

Injection size: 10 µL 

System suitability 

Sample: Standard solution 

Suitability requirements 

Tailing factor: NMT 2.0 

Relative standard deviation: NMT 2.0% 

Analysis 

Samples: Standard solution and Sample solution 

Calculate the percentage of hydrocortisone acetate (C₂₃H₃₂O₆) in the portion of Hydrocortisone Acetate taken:

Result = (r_U/r_S) × (C_S/C_U) × 100 

r_U = peak response of hydrocortisone acetate from the Sample solution

r_S= peak response of hydrocortisone acetate from the Standard solution

C_S = concentration of USP Hydrocortisone Acetate RS in the Standard solution (mg/mL)  

C_U = nominal concentration of hydrocortisone acetate in the Sample solution (mg/mL) 

Acceptance criteria: 97.0%–102.0% on the dried basis 

4 IMPURITIES 

Residue on Ignition 〈281〉 

Sample: 100 mg 

Acceptance criteria: NMT 0.5% 

Organic Impurities 

Solution A: Acetonitrile and water (20:80) 

Solution B: Acetonitrile and water (70:30) 

Mobile phase: See Table 1. 

Table 1 

Diluent: Acetonitrile, glacial acetic acid, and water (700:1:300) 

Standard solution: 5 µg/mL of USP Hydrocortisone Acetate RS in Diluent 

Sample solution: 1 mg/mL of Hydrocortisone Acetate in Diluent 

Chromatographic system 

(See Chromatography 〈621〉, System Suitability.) 

Mode: LC 

Detector: UV 254 nm 

Column: 4.6-mm × 15-cm; 3-µm packing L1 

Flow rate: 1 mL/min 

Injection size: 10 µL 

System suitability 

Sample: Standard solution 

Suitability requirements 

Relative standard deviation: NMT 5.0% 

Analysis 

Samples: Standard solution and Sample solution 

Calculate the percentage of each impurity in the portion of Hydrocortisone Acetate taken: 

Result = (r_U/r_S) × (C_S/C_U) × 100 

r_U = peak response of each impurity from the Sample solution

r_S= peak response from the Standard solution 

C_S = concentration of USP Hydrocortisone Acetate RS in the Standard solution (mg/mL)  

C_U = nominal concentration of hydrocortisone acetate in the Sample solution (mg/mL) 

Acceptance criteria 

Individual impurities: NMT 1.0% 

Total impurities: NMT 2.0% 

5 SPECIFIC TESTS 

Optical Rotation, Specific Rotation〈781S〉 

Sample solution: 10 mg/mL in dioxane 

Acceptance criteria: Between +158° and +167° at 20° 

Loss on Drying 〈731〉: Dry a sample in vacuum at 60° for 3 h: it loses NMT 1.0% of its weight. 

6 ADDITIONAL REQUIREMENTS 

Packaging and Storage: Preserve in well-closed containers. 

USP Reference Standards 〈11〉 

USP Hydrocortisone Acetate RS 

Pregn-4-ene-3,20-dione, 21-(acetyloxy)-11,17-dihydroxy-, (11β)-. 

C₂₃H₃₂O₆ 404.50