Quantitation of Serum Hemoglobin -binding Capacity Using Cellulose Acetate Membrane Electrophoresis.

I F NO INTRAVASCULAR liberation of intracellular hemoglobin occurs with the transfusion of thawed, recOllstituted frozen erythrocytes, the amount of free supernatant hemoglobin in units of frozen blood that may be infused without producing henioglobinuria is determined principally by the recipient’s plasma llenloglobin-billdillg capacity (1, 2). Oweii et al. (3) reported a method for quantitation of serum haptoglobin which was dependeiit upon the peroxidase activity of methemoglobiii-haptoglobin complex in which the color of the tetra guaiacoquinone generated is unstable. ,Javid and Horowitz’s modification (4) of Lathem and Worley ‘s method (5) for seruni haptoglobin uses paper electrophoresis and requires 14 hr. for the separation of fi-ee (unbound) hemoglobin, haptoglohin (alpha-0 globulin) bound hemoglobm, and methemalbumin (albumin-bound acid hematin) prior to staining with the stable peroxidase stain o-dianisidine (3,31 diniethoxy beiizidine, Eastman Products, Rochester, N. Y.). We have developed a method for the quantitation of serum hemoglobin-binding capacity by the use of cellulose acetate membrane electrophoresis which requires approximately 90 miii. for separation and an additional 45 mm. for drying, staining with o-dianisidine, and scanning of 8 membranes.