A convenient method has been developed for the separation of alkaline phosphates (AP) isoenzymes from canine plasma. The various forms of AP activity were extracted by ethanol and separated on an anion exchanger by fast protein liquid chromatography. In this way a complete discrimination was achieved between the increase in plasma AP activity due to liver disease and that due to corticosteroid induction. The corticosteroid-induced form of AP could be separated from the other isoenzymes because of its relative heat stability at 65 degrees C. A quantitation of the contribution of liver and corticosteroid-induced AP isoenzymes to the total plasma AP activity could be made from the respective heat inactivation plots. The separation of the isoenzymes may be valuable in the purification of the different isoenzymes for further characterization.