Inhibitory effect of transferrin on serum zinc measurement

Bovine apo-transferrin (Tf) dose-dependently inhibited zinc (Zn) measurement if apo-Tf was added to a Zn standard solution followed by Zn measurement using a commercial Zn assay kit. Incubation of apo-Tf with zinc sulfate results in loss of Tf to inhibit Zn measurement, probably due to the binding of Tf with Zn. After treatment of Zn-binding Tf with ethylenediaminetetraacetic acid (EDTA) to generate apo-Tf, Zn measurement was even more strongly inhibited. However, when Zn standard solution was added to individual serum samples obtained from four dairy cows, the added Zn was almost recovered quantitatively. Apo-Tf had no effect on serum Zn measurement following its addition to serum samples. Apo-Tf and Zn standard solution was added to serum sample added Zn standard solution and apo-Tf, respectively, beforehand. The last added apo-Tf to the mixed solution showed higher Zn recovery (76-96%) as compared with the last added Zn standard solution (33-61%). Bovine serum albumin (BSA) did not affect the Zn recovery test, but apo-Tf inhibited Zn recovery even in the presence of BSA. These results suggested that, although Tf does not always inhibit serum Zn measurement, the Zn content of Zn-bound Tf could not be measured using the present Zn assay. Bovine serum contains Zn-binding protein with higher affinity to Zn than that of Apo-Tf. In addition, BSA does not inhibit the binding of apo-Tf with Zn, suggesting that BSA has lower affinity to Zn than that of apo-Tf.