A comparison has been made between the pH changes in the arterial blood of anaesthetized patients exposed to changes of Pco, above and below the normal range, and those in the same blood equilibrated with similar changes of Pco, in vitro. Changes in the non-respiratory component as represented by standard bicarbonate, were derived from the in vitro equilibrations. There was a linear relationship between log Paooi and pH, and between pH and the plasma bicarbonate concentration, over the whole range studied in vivo. In the presence of wide variations of carbon dioxide tension, nitrous oxide anaesthesia of more than 2 hours duration, supplemented with a muscle relaxant and a potent analgesic, did not produce any significant alteration of the non-respiratory component of acid-base balance. Although changes of standard bicarbonate occurred in response to alterations of Pace these changes are not considered to represent true changes of the non-respiratory component. The application of parameters derived from in vitro equilibrations, to the behaviour of arterial blood in vivo is discussed. The mechanisms of extravascular buffering of induced changes of Pco, are reviewed, and related to the behaviour of arterial blood in equilibrium with extracellular and intracellular tissue fluids.