The effect of ethyl ether, pentobarbital sodium (Nembutal®) and chloral hydrate anesthesia on the concentrations of glucose, lactate, glycerol, insulin, individual amino acids and urea have been studied in rat plasma, as well as the glycogen content in the animal liver and striated muscle. Control animals were injected with saline solution. Two samples were taken from each animal at 15 and 30 minutes from the beginning of anesthesia. These samples were obtained from the cut tail's tip and from the neck wound produced by beheading the animal. Results show considerable differences between the three sets of animals and their controls as well as in their tail versus neck values. The overall effect of the three anesthetics used were roughly the same in spite of their different actions upon the different parameters studied. This was most marked in the aminograms. Significant changes induced were rather similar in number for all. The metabolite concentration profile in each situation may be used as a reference for possible artifacts induced by any experimental approach using these anesthetics.