Nitric oxide (NO) is being increasingly used to assess airway inflammation in childhood. The method recommended by the American Thoracic Society workshop is for a prolonged expiration against a resistance. However, this is very difficult to apply in young children. As a result there have been a number of studies in which mixed expired gas has been collected and analyzed for NO content as this requires very little cooperation. This method has, however, yet to be fully validated. The aims of this study were to compare the two sampling techniques of exhaled NO concentrations in asthmatic and healthy children and to assess the correlation between NO levels and spirometry values in asthmatic children We studied 25 control children, mean age 11.5 y, and 20 asthmatics, mean age 12 y. The exhaled NO was sampled using both the single breath technique (SB) and by measuring the NO content in mixed expired air after 1 min tidal breathing (ME). Forced expiratory volume in 1 s (FEV(1)) and expiratory flow rates at 25%, 50%, and 75% of vital capacity (FEF(25), FEF(50), FEF(75), respectively) were measured by compact II spirometer (best of three) in the 20 asthmatic children. The NO level was significantly higher in the asthmatics versus the control children when measured by SB (p = 0.0015) but not when measured by ME (p = 0.1913). The NO results measured by SB were significantly higher than ME results in the asthmatic children (p = 0.008). The NO levels were negatively correlated to FEV(1), FEF(25), FEF(50), and FEV(75) when measured by SB (p < 0.02) but not when measured by ME. The SB but not the ME method for measuring expired NO discriminates between asthmatic and control children and correlates well with the degree of airway obstruction. The use of the ME technique remains unproven.