The clearance of vitamin D3 from plasma after intravenous injection to rats of intestinal lymph labeled in vivo with radioactive vitamin D3 has been studied. The half-life of the injected vitamin D3 was about 6 min. In double-label experiments the half-life was the same for vitamin D3 as for vitamin A, which is removed from the circulation as chylomicron remnants. Fractionation of the blood plasma according to the density showed that 2 min after the injection of the lymph 30-95% of the radioactive vitamin D3 was recovered in plasma fractions with higher densities than chylomicron remnants (d greater than 1.019 g/ml). The percentage of vitamin D3 in the chylomicron fraction (d less than 1.006 g/ml) of the injected lymph was 66-95%. Taking this into account it could be calculated that the amount of radioactive vitamin D3 transferred from chylomicrons or chylomicron remnants to plasma with d greater than 1.019 g/ml within 2 min varied from 26 to 72%. The transferred radioactivity was recovered in a protein fraction with chromatographic mobilities identical to the binding protein for vitamin D and its metabolites. During the first 13 min after injection the amount of radioactivity recovered in the liver was less than 10%, but from 15 to 55 min the amount increased to about 50% of the injected dose. These findings suggest that the transfer of vitamin D3 from chylomicrons to the alpha-globulin fraction previously observed in vitro is of importance for the in vivo handling of vitamin D3.(ABSTRACT TRUNCATED AT 250 WORDS)