Assessing the feasibility of breath collection from conscious young children to support volatilome analysis: insights into age limitations and breath sampling requirements

Breath volatile organic compounds (VOCs) are increasingly under consideration as biomarkers of respiratory disease. Although numerous studies have identified VOCs that distinguish patient groups, a lack of standardisation among published studies has impeded translation into clinical diagnostics. Standardised breath collection protocols have been proposed for adults and children aged >4 years, but optimal methods for collecting breath from younger children remain to be determined. The aim of this study was to assess the feasibility and acceptability of breath sampling among a young paediatric cohort. A total of 61 children (age 6 months-12 years) were recruited prospectively to observational studies of chronic cough at two study sites. Mixed expiratory breath was collected into 1 l Tedlar Bags using either a drinking straw, mouthpiece, or mask. After concentrating onto thermal desorption tubes, the breath was analysed using two-dimensional gas chromatography coupled with time-of-flight mass spectrometry. Breath collection via a mouthpiece was highly feasible for children aged >2 years. Mask-based collection was required for younger children but was poorly tolerated. Drinking straw-based collections were unsuitable for some children aged <4 years due to challenges maintaining a sufficient seal. At least 700 ml of breath was sampled from 72.6% of children. The number of peaks per sample, total peak area per sample, and composition of breath VOCs were all consistent with successful breath sampling. The high feasibility of breath collection via a mouthpiece in our study suggests established protocols designed for children aged over 4 years can be used with confidence for children from as young as 2 years of age.