Trichloroacetic acid (TCA) precipitability of 125I in the blood of mice fed 125I.

A recent study showed that after feeding 125I-labelled proteins or free 125I to mice, as much as 40% of total radioactivity in the circulation precipitated upon mixing whole blood with 10% trichloroacetic acid. We examined this potential limitation to the use of radiolabelled tracers for studies on intestinal digestion of proteins and protein uptake, and identified its mechanism. BALB/c mice were gavage-fed or injected intravenously (i.v.) with Na125I. Blood obtained at 15 min was added directly to 10% trichloroacetic acid (TCA) or was processed to obtain serum or plasma. On mixing with 10% TCA, 25-33% of the radioactivity in whole blood was precipitated; less than 2% of the radioactivity in plasma or serum was precipitated. In vitro studies identified hemoglobin as the primary carrier protein participating in this reaction. If hemoglobin was replaced by methemoglobin or cyanomethemoglobin, then the reaction with 125I did not occur, suggesting that iron in the heme group may be the site for 125I binding and that iron must be in its reduced or ferrous form (Fe2+). The administration of non-radioactive NaI in vivo or its addition to reaction mixtures in vitro completely inhibited the precipitation of 125I by hemoglobin in the presence of TCA. Thus the addition of non-radioactive iodide to TCA stock solutions may effectively prevent non-specific binding of 125I to free hemoglobin released unintentionally during venipuncture or at other stages of blood processing.