The phospholipid profiles of newborn, young adult and aged rat heart and spleen and their in vitro response to endogenous phospholipases at pH 7.4 and 38°C for 60 min were analysed by thin layer chromatography (TLC) technology and densitometry measurement. The noticeable high level of cardiolipin (CL) and its preferential deacylation, as detected by the formation of monolysocardiolipin (MLCL) and concurrent reduction of CL level were the most prevalent lipolytic events of rat cardiac muscle (newborn, young adult and aged) but the least prevalent in rat spleen. The level of ethanolamine plasmalogen (PE) was high in both the rat spleen and cardiac muscle (newborn, young adult and aged). Following in vitro incubation, the reduction in the level of PE and the high level of lyso alkenyl PE produced were most conspicuous in rat spleen (newborn, young adult and aged) and noticeably less in rat cardiac muscle. These data clearly illustrate the differential response of the endogenous substrates (phospholipids) to the endogenous phospholipases of these two tissues, and probably are related to their physiological activities in vivo.