Hydrocortisone Ointment

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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1 DEFINITION 

Hydrocortisone Ointment is Hydrocortisone in a suitable ointment base. It contains NLT 90.0% and NMT 110.0% of the labeled amount of hydrocortisone (C₂₁H₃₀O₅). 

2 IDENTIFICATION 

Thin-Layer Chromatographic Identification Test 〈201〉 

Sample solution: Transfer a portion of Ointment, equivalent to 5 mg of hydrocortisone, to a flask. Add 10 mL of methanol, and heat on a steam bath for 5 min with frequent shaking. Cool to solidify the ointment base, and filter. Use the filtrate. 

Acceptance criteria: Meets the requirements 

3 ASSAY 

Procedure 

Diluent: Dilute alcohol (1 in 2) 

Mobile phase: Acetonitrile and water (25:75) 

Standard stock solution: 500 µg/mL of USP Hydrocortisone RS in alcohol 

Standard solution: 50 µg/mL of USP Hydrocortisone RS prepared by mixing Standard stock solution and Diluent (1:9). [Note—If alcohol is used in the nal dilution of the Sample solution, similarly use alcohol instead of aqueous alcohol in the nal dilution of the Standard solution.] Sample solution: Transfer a quantity of Ointment, nominally equivalent to 10 mg of hydrocortisone, to a 150-mL beaker. Add 40 mL of alcohol, and heat on a steam bath while stirring to melt and disperse the Ointment. Cool to room temperature, and filter through glass wool into a 100-mL volumetric flask. Repeat the extraction with two 20-mL portions of alcohol, combining the filtrates in the 100-mL volumetric flask. Add alcohol to volume, and mix. Quantitatively dilute one volume of this solution with an equal volume of water, and pass through a membrane filter of 5-µm pore size. If precipitation occurs on dilution with water, and the solution is still cloudy after filtration, dilute the initial Sample solution with alcohol instead of water. Pass this solution through a membrane filter of 5-µm pore size. Chromatographic system 

(See Chromatography 〈621〉, System Suitability.) 

Mode: LC 

Detector: UV 254 nm 

Column: 3.9-mm × 30-cm; packing L1 

Injection volume: 10–25 µL 

System suitability 

[Note—Adjust the composition of the Mobile phase so that the retention time of hydrocortisone is about 10 min.] Sample: Standard solution 

Suitability requirements 

Relative standard deviation: NMT 3.0% for 5 replicate injections 

Analysis 

Samples: Standard solution and Sample solution 

Calculate the percentage of the labeled amount of hydrocortisone (C₂₁H₃₀O₅) in the portion of Ointment taken:

Result = (r_U/r_S) × (C_S/C_U) × 100 

r_U = peak response from the Sample solution 

r_S = peak response from the Standard solution 

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

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

Acceptance criteria: 90.0%–110.0% 

4 PERFORMANCE TESTS 

Minimum Fill 〈755〉: Meets the requirements 

5 SPECIFIC TESTS 

Microbial Enumeration Tests 〈61〉andTests for Specified Organisms 〈62〉: It meets the requirements of the tests for absence of Staphylococcus aureus and Pseudomonas aeruginosa. 

6 ADDITIONAL REQUIREMENTS 

Packaging and Storage: Preserve in well-closed containers. 

USP Reference Standards 〈11〉 

USP Hydrocortisone RS