Equations and nomogram for the relationship of human blood p50 to 2,3-diphosphoglycerate, CO2, and H+

We describe a new method for tonometry of small amounts of blood (up to 0.25 mL) at known pO2, PCO2, and temperature, in small, reusable, closed Pyrex flasks. Equilibrated blood is analyzed for oxygen saturation, pH, and organic phosphate concentration with standard techniques, and its p50 (the pO2 at which hemoglobin is half-saturated with oxygen) is determined with full control of all the variables known to affect it. The SD in the measurement of p50 is 0.044 kPa (0.33 mmHg). We made 63 determinations of p50 on normal human blood under different conditions of pH and pCO2, and with different concentrations of 2,3-diphosphoglycerate and ATP. Empirical equations and a nomogram were derived, which allow the calculation of p50 from known values of pCO2, pH, and [2,3-DPG]/[Hb4] molar ratio with a SD of 97 and 114 Pa (0.73 and 0.86 mmHg), respectively.