The diffusing capacity of the lung for carbon monoxide (DL) is affected by changes in alveolar partial pressure of oxygen (PAO2), hemoglobin concentration (Hb), and carboxyhemoglobin concentration (COHb). A number of investigators have derived empiric adjustment equations to account for changes in these variables. We evaluated an adjustment of DL for changes in COHb and PAO2 using a single equation derived from Roughton and Forster's original definitions (J. Appl. Physiol., 1957). Unadjusted DL values declined significantly with rising COHb (-0.938%/percent COHb increase, P less than 0.0001) and rising PAO2 (-0.343%/mm Hg PAO2 increase, P less than 0.0001). Adjusted measured DL using the derived equation showed no significant change with changing COHb and PAO2 levels and provides an acceptable method for adjustment of DL for the effects of varying COHb and PAO2 levels from the standard conditions of COHb% = 0, and PAO2 = 110 mm Hg. Since a similar equation has previously been used to adjust for changes in DL due to anemia, we propose to use a single equation which is theoretically derived and empirically verified to adjust DL measurements for changes in COHb, PAO2 and hemoglobin.