The objective of the study was to determine the sensitivity with which an adenoma of the pars intermedia of the pituitary gland can be predicted in horses by measuring the basal concentrations of glucose, cortisol, adrenocorticotropin (ACTH), and insulin, the urinary concentration of corticoids, the urinary corticoid:creatinine ratio, and the plasma cortisol concentration after the administration of 25 IU of ACTH intravenously. The records of 24 cases of histologically confirmed equine pituitary pars intermedia adenomas were used. An adenoma of the pars intermedia of the pituitary gland in horses was predicted with a sensitivity of 100, 100, and 92% for basal plasma ACTH (n = 16), the urinary corticoid:creatinine ratio (n = 7), and basal plasma insulin (n = 12), respectively. The sensitivity of the ACTH-stimulation test was 79% (n = 14).