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Thin-layer chromatography previously used to detect synthetic steroids in horse urine has been applied to a study of the metabolism of prednisolone in the horse.

Chromatography of the crude steroid extract was carried out successively in (1) ether, (2) a methylene chloride: dioxane: water system and (3) ethyl acetate. Provisional identification of the metabolites was confirmed by chromatography of the acetates and the bismuthate and chromic acid oxidation products aided by colour reactions with (1) a tetrazolium reagent, (2) vanillin: perchloric acid and (3) a Zimmermann reagent.

Unconjugated steroids detected in the urine after prednisolone administration to the horse were prednisolone, prednisone, cortisol, 20β-dihydroprednisone, 20β-dihydroprednisolone and 11-deoxycortisol. None of these materials was found in normal urine.