The distribution of lactate between red blood cells (RBC) and plasma was examined at rest, during exercise and 30 min after exercise in six Standardbred horses. Lactate and water concentrations were measured in blood and plasma samples collected prior to exercise, during the last 15 s of each step of an incremental exercise test and at 5 min intervals during the first 30 min after exercise. The mean ratio of RBC lactate concentration (RBCLa) to plasma lactate concentration (PLa) prior to exercise was 1.02 +/- 0.34. Haemoconcentration during exercise was associated with more rapid accumulation of lactate in plasma than in RBCs. Mean whole blood lactate concentration (BLa) was only 59% of PLa in samples collected during exercise. BLa was highly correlated with PLa during exercise (r = 0.98; P < 0.001), but individual PLa values differed from predicted BLa values by up to 2.1 mmol l-1 when PLa exceeded 8 mmol l-1. At each exercise speed and time after exercise there was a large variation between horses in the ratio of BLa/PLa. During exercise at 11 m s-1 the ratio ranged from 0.46-0.73. The BLa/PLa ratio was significantly correlated with increasing exercise intensity, (r = -0.68, P < 0.001) and with haematocrit, (r = -0.69, P < 0.001). The effects of PLa and haematocrit on the BLa/PLa ratio during exercise and the post-exercise period varied greatly between horses. The ratio of lactate concentrations in water of RBCs and plasma varied greatly during and after exercise (0.61-1.22). The ratio also varied considerably between horses, with coefficients of variation ranging from 14-34%. Lactate concentrations in RBC and plasma water vary greatly between horses during and after exercise. We concluded that lactate is unevenly distributed between red blood cells and plasma in blood samples centrifuged immediately after collection, and the variability depends on haematocrit.