Using dialysis, gel filtration, isoelectrofocusing and radioaffinity assay, we studied the unsaturated and saturated binders of bile and the biliary concentration of cobalamin (Cbl) and Cbl analogues compared to the corresponding serum concentrations in 7 choledochodomized patients. Bile contained a single saturated or unsaturated R binder with a molecular mass of about 120,000. Differences in the isoelectrofocusing pattern were observed between unsaturated and saturated R binders and could correspond to two secretion origins, mucosal secretion and hepatocyte clearance, respectively. The concentration of total corrinoids is about 4 times higher in bile than in serum, and this could be explained by a hepatic clearance of serum Cbl analogue-R binder complexes, as previously described in the rabbit. Moreover, the enterohepatic circulation of Cbl seems likely in healthy individuals since the saturated biliary R binder is degraded by pancreatic juice.