Development of an optimized additive solution containing ascorbate‐2‐ phosphate for the preservation of red cells with retention of 2,3 diphosphoglycerate

Abstract

An additive solution containing adenine, ascorbate-2-phosphate, sodium phosphate, dextrose and saline was developed for packed red cell preservation. The combination of all components was simultaneously optimized so that the resulting solution produced the maximum retention of both red cell ATP and 2,3-diphosphoglycerate (2,3-DPG) concentrations. Fourteen nutrient combinations were tested; each combination was evaluated for 42 days of storage using cells from 3 donors. The nutrient combinations were chosen with the aid of a computerized experimental design process. Results of the experiments were modeled by regression analysis, and the model was optimized to produce the best formulation for simultaneous maintenance of ATP and 2,3-DPG. The resulting mathematically optimal formulation was tested in the laboratory using 10 units of red cells. With this solution, it was possible to store red cells for 42 days with retention of 45-55% of the initial ATP and 85-150% of the initial 2,3-DPG. Red cell lysis was low (0.8%), and most of the cells were bioconcave discs (by scanning electron microscopy) at the end of storage. The studies were carried out in an efficient manner by using computer-optimized experimental design techniques coupled with multiple regression modeling and subsequent computer optimization of the models. This experimental approach has potential application to may current blood banking procedures. This additive solution should maintain viable red cells for 42 days. In addition, the solution will maintain red cell 2,3-DPG throughout storage.