Open Access Research article

Storage conditions for stability of offline measurement of fractional exhaled nitric oxide after collection for epidemiologic research

Yoshiko Yoda1, Naruhito Otani1, Hideki Hasunuma12, Hiroshi Kanegae2 and Masayuki Shima1*

Author Affiliations

1 Department of Public Health, Hyogo College of Medicine, 1-1 Mukogawa-cho, Nishinomiya, Hyogo 663-8501, Japan

2 Center for Environmental Information Science, 8-19 Yonban-cho, Chiyoda-ku, Tokyo, 102-0081, Japan

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BMC Pulmonary Medicine 2012, 12:68  doi:10.1186/1471-2466-12-68

Published: 2 November 2012



The measurement of fractional concentration of nitric oxide in exhaled air (FeNO) is valuable for the assessment of airway inflammation. Offline measurement of FeNO has been used in some epidemiologic studies. However, the time course of the changes in FeNO after collection has not been fully clarified. In this study, the effects of storage conditions on the stability of FeNO measurement in exhaled air after collection for epidemiologic research were examined.


Exhaled air samples were collected from 48 healthy adults (mean age 43.4 ± 12.1 years) in Mylar bags. FeNO levels in the bags were measured immediately after collection. The bags were then stored at 4°C or room temperature to measure FeNO levels repeatedly for up to 168 hours.


In the bags stored at room temperature after collection, FeNO levels were stable for 9 hours, but increased starting at 24 hours. FeNO levels remained stable for a long time at 4°C, and they were 99.7% ± 7.7% and 101.3% ± 15.0% relative to the baseline values at 24 and 96 hours, respectively. When the samples were stored at 4°C, FeNO levels gradually decreased with time among the subjects with FeNO ≥ 51 ppb immediately after collection, although there were almost no changes among the other subjects. FeNO levels among current smokers increased even at 4°C, although the values among ex-smokers decreased gradually, and those among nonsmokers remained stable. The rate of increase was significantly higher among current smokers than among nonsmokers and ex-smokers from 9 hours after collection onwards.


Storage at 4°C could prolong the stability of FeNO levels after collection. This result suggests that valid measurements can be performed within several days if the samples are stored at 4°C. However, the time course of the changes in FeNO levels differed in relation to initial FeNO values and cigarette smoking.

Cigarette smoking; Epidemiologic research; Exhaled nitric oxide; Offline measurement; Refrigeration; Storage conditions; Wheezing