Optimization of a Method for Measuring 2,3-Diphosphoglycerate in Erythrocytes Using 31P NMR Analysis

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Abstract 2,3-Diphosphoglycerate is present in large quantities in mammalian red blood cells and controls the affinity of hemoglobin to oxygen. The level of 2,3-diphosphoglycerate changes with the development of certain pathologies, such as chronic lung diseases, anemia, and hyperthyroidism. 31P NMR spectroscopy has shown that reducing the number of washings of rat erythrocytes significantly increases the stability of 2,3-diphosphoglycerate in these samples. The level of 2,3-diphosphoglycerate does not change for 3.5 h with a single wash and for 4 h without washing. At the same time, with standard double washing, the content of 2,3-diphosphoglycerate decreases after 2.5 h. Analysis of its distribution between red blood cells and removed plasma, as well as analysis of the serum of one animal, showed that all the 2,3-diphosphoglycerate remains in the suspension of red blood cells and is absent in the plasma and serum. The results we obtained should be taken into account when working with intact erythrocytes.

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