By means of a 5-week vitamin B-complex supplementation, associations between indices of vitamin B1, B2, and B6 status (activation coefficients [AC] for erythrocyte transketolase, glutathione reductase, and aspartate aminotransferase) and exercise-induced blood lactate concentration were studied. Subjects, 42 physically active college students (18-32 yrs), were randomized into vitamin (n = 22) and placebo (n = 20) groups. Before the supplementation there were no differences in ACs or basal enzyme activities between the groups. The ACs were relatively high, suggesting marginal vitamin status. In the vitamin group, all three ACs were lower (p < 0.0001) after supplementation: transketolase decreased from 1.16 (1.14-1.18) (mean and 95% confidence interval) to 1.08 (1.06-1.10); glutathione reductase decreased from 1.33 (1.28-1.39) to 1.14 (1.11-1.17); and aspartate aminotransferase decreased from 2.04 (1.94-2.14) to 1.73 (1.67-1.80). No changes were found after placebo. Despite improved indices of vitamin status, supplementation did not affect exercise-induced blood lactate concentration. Hence no association was found between ACs and blood lactate. It seems that marginally high ACs do not necessarily predict altered lactate metabolism.