[Studies on methods for the determination of amino acid requirements for metabolic balance based on the oxidation rate of 14C labeled lysine to 14CO2].

Male experimental rats (100 gm liveweight) were distributed into 10 groups of 8 animals each and received balanced diets, with the exception of lysine which was added to the diets in graded amounts in such a way that the lysine content of the diets ranged from 2.44 to 5.92 gm/16 gm N. After a feeding period of 7 days the animals received 3H- and 14C lysine injected intraperitoneally, 4 animals of each group were investigated for the total CO2 excretion and 14CO2 excretion during the first 2 hrs after the injection and for the urinary excretion of radioactivity (48 hours). The remaining animals in each group were used for determining the plasma amino acids and for establishing the specific radioactivity of free lysine in the liver and muscles after an 1-hour incorporation period. Total CO2 excretion was not found to be influenced by the lysine contents while the level of excretion of 14C activity through CO2 and that of specific 14C activity of CO2 increased with increasing lysine concentrations. This produced a broken curve pattern, showing an increased release of 14CO2 (under maintenance conditions) if the diet contained 4 gm lysine/16 gm N and more. Investigations for the specific 14C activity of free lysine in the liver, the main site of lysine oxidation, showed that the increase in 14CO2 release was due to an enhanced rate of lysine catabolism and was not brought about by changes in the pool volume or in specific radioactivity. The levels of urinary 14C excretion were not found to be related to the lysine content of the diets, whereas the curve pattern of 3H excretion observed 5 to 8 hrs after injection was similar to that of 14CO2 excretion. The lysine content of blood plasma and the content of free lysine in the liver increased continuously with increasing levels of dietary lysine. The methodological studies made in the present paper showed that in scientific research a determination of amino acid requirements on the basis of CO2 oxidation data may be a very exact and sensitive method. It will also yield values for maintenance requirements.