In order to clarify the interaction between C02 diffusion and HC03-shift in the red blood cell (RBC), HC03-shift was measured by using a stopped flow method combined with fluorometry. When HC03-entered the RBC, the intracellular Pco2 increased, causing a secondary outflow of C02. Conversely, when HC03-ions flowed out of the RBC, the resulting decrease of PC02 caused an inward C02 diffusion. The Pco2 change caused by the inward HC03-shift was about 3-to 4-fold that of the outward shift. During the respective in-and outward-shifts, the mean half times of the extracellular pH changes were 0.15 and 0.13 sec. These were approximately twice as long as those of the primary C02 diffusion. The permeability of HC03-across the RBC membrane was obtained by comparing the experimental extracellular pH curve with a numerical solution for C02 and HC03-diffusions accompanied by the hydration and dehydration reactions. Thus the HC03-permeability was determined to be 5>< 10-4 and 7>< 10-4 cm/sec, in the in-and outward-HC03-shifts, respectively. The influence of Cl-concentration on HC03-permeability was tested by reducing the initial Cl-gradient across the RBC membrane. In a physiological Cl-concentration range the HC03-permeability was not affected by the Cl-gradient.Key Words:HC03-shift, C02 diffusion, HC03-permeability, stopped flow method, Cl-shift.In studies to elucidate the rate of gas exchange in vivo, the HC03-permeability across the red blood cell (RBC) membrane is an important parameters. KLOCKE (1976) measured the HC03-shift in RBC suspension by inhibiting the activity of carbonic anhydrase by using acetazolamide. However, because this enzyme is very active inside the RBC (KERNOHAN et a!., 1963), the rate of HC03-shift in the normal RBC seemed not to obey the rate measured in the suspension containing acetazolamide. Thus, we attempted to measure the rate of HC03-shift through extracellular pH by adding carbonic anhydrase into the normal RBC suspension. During the course of the pH measurement using a stopped flow apparatus, it was found that the intracellular HC03-change resulted in the Pco 2 change, causing a secondary C02 diffusion. Thus, from a recorded extracellular pH change which resulted from both the changes in Pco 2 and HC03 we attempted to determine HC03 -permeability. First, by varying the permeability a numerical solution for extracellular pH change was derived from the C02 diffusion equations accompanying the HC03-shift. Then, by comparing the computed pattern with the measured one, HC03-permeability was determined in terms of the transfer coefficient, rj-(HC03-). To obtain a numerical solution of the diffusion equation, estimated the diffusion coefficients for C02 and HC03r inside the RBC, and NnzEKI et al. (1983) measured the transfer coefficient for C02, 'j(C02) across the RBC membrane. SHIM0ucHI et al. (1984) clarified that the Pco2 change caused by the HC03 shift could be computed by a modified Henderson-Hasselbalch equation. Using these parameter values, KAGAWA and MocHlzuKi (1984) obtained...