Low vitamin B-complex status has been associated with poorer outcome in critically-ill patients. However, these findings have been based on indirect methods. Using direct methods for assessing vitamin status, we examined the effect of B-complex vitamin supplementation by measuring plasma and red blood cell B1, B2 and B6-vitamin concentrations in critically-ill patients.Thiamine diphosphate (TDP), flavin adenine dinucleotide (FAD) and pyridoxal phosphate (PLP) concentrations were measured in plasma and red cells of normal subjects (n=49) and ITU patients (n=41).Compared with the normal subjects, critically-ill patients had higher C-reactive protein and lower albumin concentrations (P<0.001). Also, plasma FAD and PLP were lower (P<0.001) and red cell concentrations of both were higher (P<0.01) in critically-ill patients. Critically-ill patients were grouped according to whether (n=23) or not (n=18) they had been supplemented with B-complex vitamins. Compared with non-supplemented group, the supplemented group had significantly higher red cell TDP and PLP concentrations (P<0.01). Plasma FAD and PLP concentrations did not differ significantly between the groups.The results of the present study suggest that direct measurements of red cell FAD and PLP are more responsive to supplementation than plasma measurements in the critically-ill patient.