Analysis of Adrenocortical Steroids in Pharmaceutical Preparations by High-Pressure Liquid-Liquid Chromatography

The adrenocortical steroids hydrocortisone, cortisone, hydrocortisone acetate, and cortisone acetate were separated by liquid-liquid chromatography. A commercially prepared reverse- phase cyano ethyl silicone column was used in conjunction with a UV precision photometer. Linearity studies were carried out using the internal standard technique and peak height measurements. Responses of each steroid were linear over the working range when equal injection volumes were used. The results of a study of the controlled decomposition of hydrocortisone in basic solution indicated a good separation of drug from the degradation products. Samples of adrenocortical steroids in various dosage forms, primarily creams and ointments, were analyzed by the proposed procedure, yielding a single steroid assay value and an identification and quantitation of each foreign steroid. The sample preparation was a simple dissolution of the steroid in alcohol plus the addition of an internal standard prior to injection.