The normal human red blood cell, when withdrawn from the circulation, contains a substantial pool of water-soluble organic phosphate compounds, most of which are metabolic intermediates whose concentrations are maintained at constant levels by the glucose metabolism of the cell (1). If blood is stored under blood bank conditions in acid citrate dextrose (ACD) preservative, the organic phosphates of the red cell gradually disappear and inorganic phosphate accumulates. The decline of the organic phosphates coincides with a general deterioration of the cell, for example with a decrease in the glycolytic rate, an inability to concentrate potassium ions and a loss of viability measured by post-transfusion survival (2-4). Several tracted with trichloroacetic acid and the acid was removed with ether. The extract was passed through a 1 X 12 cm. column of Dowex 1-X8 chloride (100 to 200 mesh) which was eluted at 2 ml. per minute with hydrochloric acid and ammonium chloride eluants as indicated in the figures. Fractions of approximately 20 ml. were collected and each was assayed for optical density at 260 m/A and for total phosphorus (9). Spectral measurements at the ultraviolet absorbing peaks indicated that most of the material was adenylate. The anthrone (10), carbazole (11) and orcinol (12) Figure 4 summarizes the changes observed in the phosphate compounds studied as a function of time.The zero time assays revealed several marked differences between the cells collected in ACD in the present study and those red cells collected in heparin anticoagulant (1). Apparently a rapid displacement of the normal equilibrium followed shortly after the blood was mixed with the ACD solution. Actually "zero time" refers to two to three hours of in vitro life for the red cell, mostly at reduced temperature, including the period required for the washing and centrifugation of the 56
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