The purpose of this study was to evaluate a new method to measure closing volume (CV). This new method does not require oxygen or inert gases to be inhaled to obtain the onset of phase 4. Because there are regional differences in the concentrations of the resident alveolar gases (O2, CO2, and N2), there should be an abrupt change in the concentration of these gases at the terminal portion of a prolonged expired vital capacity (VC) that marks the onset of phase 4. Nine normal healthy subjects, 30 to 65 years of age, inspired room air from residual volume (to mimic the maneuver of the standard single breath N2 (SBN2) washout test) to total lung capacity. During the expiration (flow constant at 250 ml.s-1) following a 10-s breath hold at total lung capacity, the exhaled gas was analyzed with a mass spectrometer for fractions of O2, CO2, and N2. Although the onset of phase 4 can be shown as the change in concentration of any of the three alveolar resident gases, oxygen was selected because (1) it demonstrates a greater apex to base concentration gradient than that found with CO2 and N2, and (2) a clear identification of the onset of phase 4 (minimum value of O2 fraction). With this method, the mean +/- SEM of CV was 16.8 +/- 1.52 percent (CV x 100/VC). No significant difference was found among the room air method, SBN2 method, and the helium bolus technique.