Hydrocortisone Acetate Ointment

Corticosteroid.

Hydrocortisone Acetate Ointment contains Hydrocortisone Acetate in a suitable basis.

The ointment complies with the requirements stated under Topical Semi-solid Preparations and with the following requirements.

90.0 to 110.0% of the stated amount.

A. Carry out the method for thin-layer chromatography, Appendix III A using the following solutions.

For ointments containing more than 0.5% w/w of Hydrocortisone Acetate.

(1) Disperse a quantity containing 25 mg of Hydrocortisone Acetate in 5 mL of hot hexane, cool, extract with 10 mL of methanol (90%) and filter. 

(2) Equal volumes of solution (1) and a 0.25% w/v solution of hydrocortisone acetate BPCRS in methanol.

For ointments containing 0.5% w/w or less of Hydrocortisone Acetate.

(1) Disperse a quantity containing 5 mg of Hydrocortisone Acetate in 5 mL of hot hexane, cool, extract with 10 mL of methanol (90%) and filter.

(2) Equal volumes of solution (1) and a 0.05% w/v solution of hydrocortisone acetate BPCRS in methanol.

(a) Use as the coating silica gel G.

(b) Use the mobile phase as described below.

(c) Apply 5 µL of each solution.

(d) After removal of the plate, dry in air and spray with alkaline tetrazolium blue solution.

1.2 volumes of water, 8 volumes of methanol, 15 volumes of ether and 77 volumes of dichloromethane.

The principal spot in the chromatogram obtained with solution (1) corresponds to that in the chromatogram obtained with solution (2) which appears as a single, compact spot.

B. In the Assay, the chromatogram obtained with solution (2) shows a peak with the same retention time as the peak due to hydrocortisone acetate in the chromatogram obtained with solution (1).

Carry out the method for liquid chromatography, Appendix III D using the following solutions.

For ointments containing more than 0.5% w/w of Hydrocortisone Acetate.

(1) Disperse a quantity of the ointment containing 25 mg of Hydrocortisone Acetate in 100 mL of hot hexane, cool and extract with 20 mL of a solution prepared by mixing 3 volumes of methanol and 1 volume of a 15% w/v solution of sodium chloride. Repeat the extraction using two 10-mL quantities of the methanolic sodium chloride solution. Add 5 mL of methanol to the combined extracts and sufficient water to produce 100 mL, mix and filter through a glass microfibre filter (Whatman GF/C is suitable).

(2) Dissolve 25 mg of hydrocortisone acetate BPCRS in 45 mL of methanol, add 5 mL of a 0.5% w/v solution of betamethasone (internal standard) in methanol and sufficient water to produce 100 mL.

(3) Prepare in the same manner as solution (1) but add 5 mL of the internal standard solution in place of the 5 mL of methanol before diluting to volume.

For ointments containing 0.5% w/w or less of Hydrocortisone Acetate.

(1) Disperse a quantity of the ointment containing 5 mg of Hydrocortisone Acetate in 100 mL of hot hexane, cool and extract with 20 mL of a solution prepared by mixing 3 volumes of methanol and 1 volume of a 15% w/v solution of sodium chloride. Repeat the extraction using two 10-mL quantities of the methanolic sodium chloride solution. Add 5 mL of methanol to the combined extracts and sufficient water to produce 100 mL, mix and filter through a glass microfibre filter (Whatman GF/C is suitable).

(2) Dissolve 5 mg of hydrocortisone acetate BPCRS in 45 mL of methanol, add 5 mL of a 0.110% w/v solution of betamethasone (internal standard) in methanol and add sufficient water to produce 100 mL.

(3) Prepare in the same manner as solution (1) but add 5 mL of the internal standard solution in place of the 5 mL of methanol before diluting to volume.

(a) Use a stainless steel column 10 cm × 5 mm) packed with octadecylsilyl silica gel for chromatography (5 µm) (Spherisorb ODS 1 is suitable).

(b) Use isocratic elution and the mobile phase described below.

(c) Use a flow rate of 2 mL per minute.

(d) Use an ambient column temperature.

(e) Use a detection wavelength of 240 nm.

(f) Inject 20 µL of each solution.

50 volumes of methanol and 50 volumes of water.

Calculate the content of C₂₃H₃₂O₆ in the ointment using the declared content of C₂₃H₃₂O₆ in hydrocortisone acetate BPCRS.

Hydrocortisone Acetate Ointment should be protected from light.