The cellular uptake of (14C)-thiamin hydrochloride was studied in the isolated perfused guinea pig heart, using the rapid single circulation, paired-tracer technique, in which D-(3H)-mannitol serves as an extracellular marker. Cellular uptake of this vitamin was estimated by directly comparing venous dilution profiles of (14C) and (3H) radioactivities in the absence and presence of unlabelled thiamin hydrochloride and pyrithiamin hydrobromide. The maximal cellular uptake (Umax) of thiamin was very low (5.31 +/- 1.79%), while in the presence of 10 mM unlabelled thiamin and 1 mM pyrithiamin, Umax was significantly greater (9.71 +/- 1.57% and 12.30 +/- 0.82%, respectively). Our data suggest that there is a saturable mechanism of sarcolemmal thiamin transport out of myocardial cell, while this transport into the cell is unsaturable.