Optimisation of sampling parameters for standardised exhaled breath sampling

The lack of standardisation of breath sampling is a major contributing factor to the poor repeatability of results and hence represents a barrier to the adoption of breath tests in clinical practice. On-line and bag breath sampling have advantages but do not suit multicentre clinical studies whereas storage and robust transport are essential for the conduct of wide-scale studies. Several devices have been developed to control sampling parameters and to concentrate volatile organic compounds (VOCs) onto thermal desorption (TD) tubes and subsequently transport those tubes for laboratory analysis. We conducted three experiments to investigate (i) the fraction of breath sampled (whole versus lower expiratory exhaled breath); (ii) breath sample volume (125, 250, 500 and 1000 ml); and (iii) breath sample flow rate (400, 200, 100 and 50 ml min^-1). The target VOCs were acetone and potential volatile biomarkers for oesophago-gastric cancer belonging to the aldehyde, fatty acids and phenol chemical classes. We also examined the collection execution time and the impact of environmental contamination. The experiments showed that the use of exhaled breath-sampling devices requires the selection of optimum sampling parameters. The increase in sample volume has improved the levels of VOCs detected. However, the influence of the fraction of exhaled breath and the flow rate depends on the target VOCs measured. The concentration of potential volatile biomarkers for oesophago-gastric cancer was not significantly different between the whole and lower airway exhaled breath. While the recovery of phenols and acetone from TD tubes was lower when breath sampling was performed at a higher flow rate, other VOCs were not affected. A dedicated 'clean air supply' reduces the contamination from ambient air, but the breath collection device itself can be a source of contaminants. In clinical studies using VOCs to elicit potential biomarkers of gastro-oesophageal cancer, the optimum parameters are 500 mls sample volume of whole breath with a flow rate of 200 ml min^-1.