Hydrocortisone Gel

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

2 IDENTIFICATION 

Delete the following: 

Thin-Layer Chromatographic Identification Test 〈201〉 

Sample solution: Transfer a quantity of Gel, equivalent to 5 mg of hydrocortisone, to a flask. Add 5 mL of alcohol, and heat on a steam bath for 5 min, with frequent shaking. Cool, and filter. Use the filtrate. 

Acceptance criteria: Meets the requirements  

Add the following: 

The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.

Add the following: 

The UV spectrum of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.

3 ASSAY 

Change to read: 

Procedure 

Solution A: 0.1% glacial acetic acid in water 

Solution B: 0.1% glacial acetic acid in acetonitrile 

Solution C: Acetonitrile, glacial acetic acid, and water (40:1:60) 

Solution D: Tetrahydrofuran and water (50:50) 

Mobile phase: See Table 1. 

Table 1 

Diluent: Solution C and Solution D (80:20) 

System suitability solution: 0.3 mg/mL of USP Hydrocortisone RS and 6 µg/mL of USP Prednisolone RS in Diluent. Sonicate to dissolve, if necessary. 

Standard solution: 0.3 mg/mL of USP Hydrocortisone RS in Diluent. Sonicate to dissolve, if necessary. 

Sample solution: Nominally 0.3 mg/mL of hydrocortisone prepared as follows. Transfer 3 mg of hydrocortisone from a portion of Gel to a 10- mL volumetric flask. Add 2 mL of Solution D and vortex for about 3 min or until the sample is thoroughly dispersed. Add about 6 mL of Solution C and sonicate for about 10 min with intermittent vortexing and shaking. Dilute with Solution C to volume. Centrifuge a portion of the solution to obtain a clear supernatant. [Note—The use of a centrifuge speed of 3500 rpm for 30 min may be suitable.] Use the supernatant. 

Chromatographic system 

(See Chromatography 〈621〉, System Suitability.) 

Mode: LC 

Detector: UV 246 nm. For Identification B, use a diode array detector in the range of 210–400 nm. 

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

Column temperature: 30° 

Flow rate: 0.8 mL/min 

Injection volume: 10 µL 

System suitability 

Samples: System suitability solution and Standard solution [Note—See Table 2 for relative retention times.] 

Suitability requirements 

Resolution: NLT 1.5 between hydrocortisone and prednisolone, System suitability solution 

Tailing factor: NMT 1.5, Standard solution 

Relative standard deviation: NMT 1.0%, Standard solution 

Analysis 

Samples: Standard solution and Sample solution 

Calculate the percentage of the labeled amount of hydrocortisone (C₂₁H₃₀O₅) in the portion of Gel 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 (µg/mL) 

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

Acceptance criteria: 90.0%–110.0% 

4 IMPURITIES 

Add the following: 

Organic Impurities 

Solution A, Solution B, Solution C, Solution D, Mobile phase, Diluent, System suitability solution, Sample solution, and Chromatographic system: Proceed as directed in the Assay. 

Sensitivity solution: 0.3 µg/mL of USP Hydrocortisone RS in Diluent. Sonicate to dissolve, if necessary. 

Standard solution: 0.0015 mg/mL of USP Hydrocortisone RS in Diluent. Sonicate to dissolve, if necessary. System suitability 

Samples: System suitability solution, Sensitivity solution, and Standard solution 

[Note—See Table 2 for relative retention times.] 

Suitability requirements 

Resolution: NLT 1.5 between hydrocortisone and prednisolone, System suitability solution 

Relative standard deviation: NMT 5.0%, Standard solution 

Signal-to-noise ratio: NLT 10, Sensitivity solution 

Analysis 

Samples: Standard solution and Sample solution 

Calculate the percentage of cortisone or hydrocortisone glyoxal analog in the portion of Gel taken: 

Result = (r_U/r_S) × (C_S/C_U) × (1/F) × 100 

r_U = peak response of cortisone or hydrocortisone glyoxal analog from the Sample solution

r_S = peak response of hydrocortisone from the Standard solution

C_S = concentration of USP hydrocortisone RS in the Standard solution (µg/mL) 

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

F = relative response factor (see Table 2) 

Calculate the percentage of each unspecified impurity in the portion of Gel taken: 

Result = (r_U/r_S) × (C_S/C_U) × 100 

r_U = peak response of each unspecified impurity from the Sample solution

r_S = peak response of hydrocortisone from the Standard solution

C_S = concentration of USP hydrocortisone RS in the Standard solution (µg/mL) 

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

Acceptance criteria: See Table 2. The reporting threshold is 0.1%. 

Table 2 

^a 6β,11β,17,21-Tetrahydroxypregn-4-ene-3,20 dione. 

^b Process impurity included in the table for identication only. Process impurities are controlled in the drug substance and are not to be reported or included in the total impurities of the drug product. 

^c 7α,11β,17,21-Tetrahydroxypregn-4-ene-3,20-dione. 

^d 11β,17-Dihydroxy-3,20-dioxopregn-4-en-21-al. 

^e 11β,17,21-Trihydroxypregna-4,6-diene-3,20-dione. 

^f 17,21-Dihydroxypregna-4-ene-3,11,20-trione. 

^g 17,21-Dihydroxypregn-4-ene-3,20-dione. 

5 PERFORMANCE TESTS 

Minimum Fill 〈755〉: Meets the requirements 

6 SPECIFIC TESTS 

Add the following: 

Microbial Enumeration Tests 〈61〉 and Tests for Specified Microorganisms 〈62〉: The total aerobic microbial count is NMT 102 cfu/g. The total yeasts and molds count is NMT 101 cfu/g. It meets the requirements of the tests for the absence of Staphylococcus aureus and Pseudomonas aeruginosa.  

7 ADDITIONAL REQUIREMENTS 

Packaging and Storage: Preserve in tight containers. 

Change to read: 

USP Reference Standards 〈11〉 

USP Hydrocortisone RS 

USP Prednisolone RS