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The '''single-breath diffusing capacity test''' is the most common way to determine <math>D_L</math>.<ref name="multiple" /> The test is performed by having the subject blow out all of the air that they can, leaving only the [[Lung volumes|residual lung volume]] of gas. The person then inhales a test gas mixture rapidly and completely, reaching the [[Lung volumes|total lung capacity]] as nearly as possible. This test gas mixture contains a small amount of carbon monoxide (usually 0.3%) and a ''tracer gas'' that is freely distributed throughout the alveolar space but which doesn't cross the alveolar-capillary membrane. [[Helium]] and [[methane]] are two such gasses. The test gas is held in the lung for about 10 seconds during which time the CO (but ''not'' the tracer gas) continuously moves from the alveoli into the blood. Then the subject exhales.
The anatomy of the airways means inspired air must pass through the mouth, trachea, bronchi and bronchioles ([[anatomical dead space]]) before it gets to the alveoli where gas exchange will occur; on exhalation, alveolar gas must return along the same path, and so the exhaled sample will be purely alveolar only after a 500 to 1,000 ml of gas has been breathed out.{{uncited|date=April 2018}} While it is algebraically possible to approximate the effects of anatomy (the ''three-equation method''<ref>{{cite journal |vauthors=Graham BL, Mink JT, Cotton DJ | year = 1981 | title = Improved accuracy and precision of single-breath CO diffusing capacity measurements | url = | journal = J Appl Physiol | volume = 51 | issue = 5| pages = 1306–13 | pmid = 7298468 }}</ref>), disease states introduce considerable uncertainty to this approach. Instead, the first 500 to 1,000 ml of the expired gas is disregarded and the next portion which contain gas that has been in the alveoli is analyzed.<ref name="multiple" /> By analyzing the concentrations of carbon monoxide and inert gas in the inspired gas and in the exhaled gas, it is possible to calculate <math>(D_{L_{CO}})</math> according to Equation {{EquationRef|2}}. First, the ''rate'' at which CO is taken up by the lung is calculated according to:
{{NumBlk|::|<math>\dot{V}_{CO} =\frac {\Delta{[CO]} * V_A} {\Delta{t}} </math> . | {{EquationRef|4}} }}
::::The pulmonary function equipment monitors the change in the concentration of CO that occurred during the breath hold, <math>\Delta{[CO]}</math>, and also records the time <math>\Delta{t}</math>.
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