Evaluation of a new simple collection device for sampling of microparticles in exhaled breath.

The microparticle fraction of exhaled breath is of interest for developing clinical biomarkers. Exhaled particles may contain non-volatile components from all parts of the airway system, formed during normal breathing. This study aimed to evaluate a new, simple sampling device, based on impaction, for collecting microparticles from exhaled breath. Performance of the new device was compared with that of the existing SensAbues membrane filter device. The analytical work used liquid chromatography-tandem mass spectrometry methods. The new device collected three subsamples and these were separately analysed from eight individuals. No difference was observed between the centre position (0.91 ng/sample) and the side positions (1.01 ng/sample) using major phosphatidylcholine (PC) 16:0/16:0 as the analyte. Exhaled breath was collected from eight patients on methadone maintenance treatment. The intra-individual variability in measured methadone concentration between the three collectors was 8.7%. In another experiment using patients on methadone maintenance treatment, the sampling efficiency was compared with an established filter device. Compared to the existing device, the efficiency of the new device was 121% greater for methadone and 1450% greater for DPPC. The data from lipid analysis also indicated that a larger fraction of the collected material was from the distal parts. Finally, a study using an optical particle counter indicated that the device preferentially collects the larger particle fraction. In conclusion, this study demonstrates the usefulness of the new device for collecting non-volatile components from exhaled breath. The performance of the device was superior to the filter device in several aspects.

Two techniques to sample non-volatiles in breath-exemplified by methadone.

The particles in exhaled breath provide a promising matrix for the monitoring of pathological processes in the airways, and also allow exposure to exogenous compounds to be to assessed. The collection is easy to perform and is non-invasive. The aim of the present study is to assess if an exogenous compound-methadone-is distributed in the lining fluid of small airways, and to compare two methods for collecting methadone in particles in exhaled breath. Exhaled particles were collected from 13 subjects receiving methadone maintenance treatment. Two different sampling methods were applied: one based on electret filtration, potentially collecting exhaled particles of all sizes, and one based on impaction, collecting particles in the size range of 0.5-7 µm, known to reflect the respiratory tract lining fluid from the small airways. The collected samples were analyzed by liquid chromatography mass spectrometry, and the impact of different breathing patterns was also investigated. The potential contribution from the oral cavity was investigated by rinsing the mouth with a codeine solution, followed by codeine analysis of the collected exhaled particles by both sampling methods. The results showed that methadone was present in all samples using both methods, but when using the method based on impaction, the concentration of methadone in exhaled breath was less than 1% of the concentration collected by the method based on filtration. Optimizing the breathing pattern to retrieve particles from small airways did not increase the amount of exhaled methadone collected by the filtration method. The contamination from codeine present in the oral cavity was only detected in samples collected by the impaction method. We conclude that methadone is distributed in the respiratory tract lining fluid of small airways. The samples collected by the filtration method most likely contained a contribution from the upper airways/oral fluid in contrast to the impaction method.

Characterization of exhaled breath particles collected by an electret filter technique.

Aerosol particles that are present in exhaled breath carry nonvolatile components and have gained interest as a specimen for potential biomarkers. Nonvolatile compounds detected in exhaled breath include both endogenous and exogenous compounds. The aim of this study was to study particles collected with a new, simple and convenient filter technique. Samples of breath were collected from healthy volunteers from approximately 30 l of exhaled air. Particles were counted with an optical particle counter and two phosphatidylcholines were measured by liquid chromatography-tandem mass spectrometry. In addition, phosphatidylcholines and methadone was analysed in breath from patients in treatment with methadone and oral fluid was collected with the Quantisal device. The results demonstrated that the majority of particles are <1 µm in size and that the fraction of larger particle contributes most to the total mass. The phosphatidylcholine PC(16 : 0/16 : 0) dominated over PC(16 : 0/18 : 1) and represented a major constituent of the particles. The concentration of the PC(16 : 0/16 : 0) homolog was significantly correlated (p < 0.001) with total mass. From the low concentration of the two phosphatidylcholines and their relative abundance in oral fluid a major contribution from the oral cavity could be ruled out. The concentration of PC(16 : 0/16 : 0) in breath was positively correlated with age (p < 0.01). An attempt to use PC(16 : 0/16 : 0) as a sample size indicator for methadone was not successful, as the large intra-individual variability between samplings even increased after normalization. In conclusion, it was demonstrated that exhaled breath sampled with the filter device represents a specimen corresponding to surfactant. The possible use of PC(16 : 0/16 : 0) as a sample size indicator was supported and deserves further investigations. We propose that the direct and selective collection of the breath aerosol particles is a promising strategy for measurement of nonvolatiles in breath.