We utilized the Amicon micropartition system (MPS-1) to prepare ultrafiltrates of serum for evaluating non-protein-bound copper (NPBCu). The method is rapid (a usual turnaround time less than 2 h), practical (only 1 mL of serum is required), and reproducible (average CV = 5.7% within assay and 9.3% between assays). Using this technique, we determined that (a) NPBCu in serum appeared to be fairly stable, showing little variation among subjects in different physiological conditions (normal female, pregnancy, or fetal samples); and (b) of all copper fractions studied, only NPBCu appeared to correlate between maternal concentrations and cord counterparts at delivery. Thus the NPBCu fraction may be appreciably involved in the overall mechanism of copper nutritional supply from mother to fetus.