Walter F. Kocholaty

Hemoglobin function during blood storage. XV. Effects of metabolic additives inosine and methylene blue on p50 and 2,3-DPG.

The major purpose of transfusing whole blood or red cells is to provide the recipient with an improved ability to transport oxygen to the tissues. Hemoglobin has this capability, but in most mammalian red cells-including the human-2,3-diphosphoglycerate (2,3-DPG) is required for normal oxygen release to the tissues (Chanutin, 1966; Chanutin and Curnish, 1967; Benesch & Benesch, 1967). After the discovery of the mediator of-or cofactor for-normal hemoglobin function; that is, 2,3-DPG, the first studies which looked at this compound in the red cell and the oxygen affinity of hemoglobin during liquid blood storage showed that 2,3-DPG decreased very rapidly during storage in acid citrate dextrose (ACD). The oxygen affinity of hemoglobin increased at the same rate as shown by the decrease in p50 (an inverse though direct measure of oxygen affinity) which was similar in slope of decrease to the decrease in 2,3-DPG during the several weeks which were studied (Akerblom, et al, 1968; Bunn, H. F., et al, 1969). The early work of Chanutin and his coworkers which goes back to 1958, and other studies which relate to the importance of 2,3-DPG for the normal position of the oxyhemoglobin dissociation curve have been reviewed more thoroughly in the preceedings from the first International Symposium on Red Cell Metabolism and Function (Dawson, 1970).

Hemoglobin Function in Stored Blood: VIII. Further Effects of Phosphate on Red Cell ATP and 2,3-DPG.

SummaryInorganic phosphate which is known to stimulate red cell glycolysis is present in one of the preservatives for storing whole blood, citrate-phosphate-dextrose (CPD), but not the other, acid-citrate-dextrose (ACD). Both of these preservatives for liquid storage were developed before 2,3-diphosphoglycerate (2,3-DPG) was found to be necessary for normal hemoglobin function. In a recent study we have shown that very high concentrations of phosphate (10, 15, and 20 mM) were deleterious for maintaining 2,3-DPG. In the present study a lower range of phosphate concentrations (2, 4, 6, and 8 mM) was studied for maintenance of 2,3-DPG and ATP during storage under blood banking conditions. The lowest concentration, 2 mM, which corresponds to CPD was found to be the best concentration for maintaining 2,3-DPG and thus hemoglobin function. Four mM phosphate was not quite as good but better than no phosphate. Six and 8 mM phosphate were considerably worse.ZusammenfassungIm Anschluß an vorangegangene Arbeiten wurde die Anwendung von geringen Phosphatmengen (2–8 mM) zur Stabilisierung von 2,3-DPG und ATP während der Lagerung von Blut unter Blutbankbedingungen geprüft, um die Hämoglobinfunktion aufrecht zu erhalten. Am besten bewährte sich die 2mM-Konzentration des anorganischen Phosphats, während die 4mM-Konzentration weniger wirkungsvoll war. 6–8 mM-Konzentrationen wurden als ungeeignet befunden, die Stabilisierung von 2,3-DPG und ATP zu gewährleisten.

THE CONTROL OF HEMOGLOBIN FUNCTION IN BLOOD STORED FOR TRANSFUSION PURPOSES.

Hemoglobin in the intact red cell has a lower oxygen affinity than hemoglobin in free solution. The main reason for this difference is 2, 3-diphosphoglycerate (2, 3-DPG), but this was not obvious for many years after 2, 3-DPG was found in unusually high concentrations in mammalian red cells (Greenwald, 1925) and in human red cells (Rapoport, 1941). In other cells 2, 3-DPG condentrations are very low. Because the association of hemoglobin with 2, 3-DPG has been established only recently, a brief review of important studies is given in the following paragraph.