Whole-body alanine kinetics were studied using continuous infusions of [15N]-, [3,3,3-2H3]-, [1-13C]-, and [3-13C]alanine tracers in healthy male subjects in the postabsorptive state. Alanine kinetics were highly dependent on the choice of isotopically labeled alanine. Highest rates of alanine flux (mean +/- SE) were obtained with the [3,3,3-2H3]alanine (474 +/- 41 mumol X kg-1 X h-1). [1-13C]- and [3-13C]alanine tracers gave intermediate values (297 +/- 12 and 317 +/- 22 mumol X kg-1 X h-1, respectively). The slowest rates of alanine turnover were measured with [15N]alanine (226 +/- 7 mumol X kg-1 X h-1). These results emphasize the heterogeneous metabolism of different portions of the alanine molecule and the importance of choosing an appropriate alanine tracer for studying different aspects of alanine metabolism.