THE DISTRIBUTION OF EXTRACORPUSCULAR HEMOGLOBIN IN CIRCULATING PLASMA*†

Abstract

A paper electrophoretic method was developed for quantifying the concentrations of free hemoglobin, protein-bound hemoglobin and methemalbumin in plasma. This method was based upon the electrophoretic separation of these constituents, staining with benzidine-hydrogen peroxide and photometric analysis of the stained filter paper strips. Using this method the distribution of extracorpuscular hemoglobin following intravenous administration was determined in healthy human subjects. Hemoglobin was bound to plasma protein in a concentration which averaged 128 + 25 mg %. When this binding capacity was exceeded, free hemoglobin and methemalbumin appeared in plasma. Protein-bound hemoglobin disappeared from plasma with time at a rate of 15 mg/minute. The rate of disappearance of free hemoglobin was faster than that of protein-bound hemoglobin.