An estimation of buffer values of human whole blood by titration experiment under the open condition for carbon dioxide gas.

We studied the buffer mechanism of human whole blood by means of a titration experiment under the open condition in which blood is exposed to carbon dioxide gas. Van Slyke proposed the theory of a blood buffer mechanism in 1922 (J Biol Chem 52: 525-570). However, his theory is not applicable to our experiments because it did not take into consideration the effects of changes in volume and the concentration of bicarbonate ion in the buffer as a result of titration. We studied the blood buffer mechanism theoretically by using graphical representation and developed a theory, which we then applied to our experiments. Buffer value, as defined by Van Slyke, is the quantity of the base deltan(OH)/V in the sample without volume change (in gram equivalent per liter) required to change one unit in pH. Based on our blood buffer theory, we obtain the experimental buffer value betaprime prime or minute, which is the reciprocal slope of the dpH-bdV/V (deltan(OH) in the sample with volume change) relation during a titration experiment at a constant carbon dioxide pressure (pCO(2)) with volume change dV and an isotonic NaOH concentration of b = 0.15 M. The true buffer value beta under physiological conditions is estimated to be beta = deltan(OH)/VdpH = betaprime prime or minute (0.15 minus sign [HCO(3)(minus sign)])/0.15. Here, [HCO(3)(minus sign)] is the concentration of bicarbonate ion in blood that can be calculated by using the Henderson-Hasselbalch equation. We determined the agreement of our experimental results with the theoretical results and estimated the quantity m(A) of the buffer component, except for the bicarbonate buffer, and the dissociation constant K(A) of the buffer reaction in a neutral pH range. The true buffer value of whole blood can be estimated by using our theory and the results of precise experiments.