There is some evidence for a nutritional interaction between vitamin D and vitamin K status. We have recently reported that serum percentage undercarboxylated osteocalcin (%ucOC; a marker of vitamin K status) was inversely correlated with serum 25-hydroxyvitamin D (25(OH)D) concentration (reflective of vitamin D status) in healthy Danish girls (aged 11-12 years), in line with a similar relationship reported in elderly women. While the causal nature of the relationship between vitamin D status and serum %ucOC has been tested in studies of elderly women, it has not been investigated in children. The objective of the present study was to test the hypothesis that improving vitamin D status significantly lowers serum %ucOC. Serum samples from sixty-seven healthy Danish girls (aged 11-12 years), who participated in a 12-month double-blind, placebo-controlled, vitamin D-3 intervention trial were used for the present study. These girls were a subset of subjects which began and finished the intervention during wintertime, thus avoiding the influence of seasonality on vitamin D status. A total of thirty-three and thirty-four of the girls had been randomised to treatment with 10 mu g vitamin D-3 per d and placebo, respectively, for 12 months. Total osteocalcin and the fraction of ucOC in serum (via enzyme-immunoassay) as well as serum 25(OH)D (via HPLC) were assessed at baseline and end-point. Vitamin D3 supplementation significantly increased serum 25(OH)D (21.6 %; P<0.002) but had no effect on serum %ucOC (P>0.8). In conclusion, the findings of the present intervention study in young girls suggest that vitamin D supplementation does not affect serum %ucOC, a marker of vitamin K status.