A B S T R A C T The influence of the serum binding protein (DBP) for vitamin D and its metabolites on the concentration of its main ligands, 25-hydroxyvitamin D3 (25-OHD3) and 1,25-dihydroxyvitamin D3 (1,25-[OHh2D3) was studied. The concentration of both 1,25-(OH)2D3 and DBP in normal female subjects (45±14 ng/liter and 333±58 mg/liter, mean+SD, respectively; n = 58) increased during the intake of estro-progestogens (69±27 ng/liter and 488±90 mg/liter, respectively; n = 29), whereas the 25-OHD3 concentration remained unchanged. A positive correlation was found between the concentrations of 1,25-(OH)2D3 and DBP in these women.At the end of pregnancy, the total concentrations of 1,25-(OH)2D3 (97±26 ng/liter, n = 40) and DBP (616 ±84 mg/liter) are both significantly higher than in nonpregnant females and paired cord serum samples (48±11 ng/liter and 266±41 mg/liter, respectively). A marked seasonal variation of 25-OHD3 was observed in pregnant females and their infants, whereas in the same samples the concentrations of both DBP and 1,25-(OH)2D3 remained constant throughout the year.The free 1,25-(OH)2D3 index, calculated as the molar ratio ofthis steroid and DBP, remains normal in women taking estro-progestogens, however, and this might explain their normal intestinal calcium absorption despite a high total 1,25-(OH)2D3 concentration. In pregnancy the free 1,25-(OH)2D3 index remains normal up to 35 wk of gestation, but during the last weeks of Part of this work was presented at