Breath Biopsy: Robust and Reproducible Breath Analysis

Published on: 29 Nov 2017, under Breath Biopsy

Exhaled breath contains thousands of volatile organic compounds (VOCs), valuable biomarkers that reflect metabolic processes taking place within a person’s cells and tissues. Measuring VOC biomarkers in breath could offer a completely non-invasive way of monitoring health and diagnosing disease. However, robust breath biomarker discovery requires highly reproducible workflows for breath collection and VOC analysis.

In the past, a high variability between breath sampling techniques has resulted in a lack of reproducible results1,2. The ReCIVA® Breath Sampler, part of Owlstone Medical’s Breath Biopsy® platform, was designed in collaboration with experts in the breathomics field to provide a standardized method to collect exhaled breath samples. By sampling onto sorbent tubes, ReCIVA overcomes the challenges associated with collecting breath in containers such as Tedlar® bags, which have been shown to result in the loss of sampled VOCs.3

Figure 1 ReCIVA Breath Sampler 

Repeatable Breath Sampling

Reproducibility between breath collects ensure comparability between samples and patients. It is particularly important when making longitudinal measurements of VOC biomarkers, where it is vital to capture accurately the real changes in biomarker concentration over time.

Data from our Breath Biopsy laboratory demonstrates the robustness of the breath sampling and analysis workflow, including breath collection using the ReCIVA Breath Sampler and VOC analysis using FAIMS (field asymmetric ion mobility spectrometry). Figure 2 shows measurement of limonene in breath collected in duplicate on two sorbent tubes during two separate breath collects (A and B) from the same individual.

Breath Biopsy Reproducibility
Figure 2. Limonene in breath samples collected on independent sorbent tubes (1 and 2) using ReCIVA and analyzed using GC-FAIMS (Lonestar® VOC Analyzer) shows high reproducibility within a single breath collect. Small changes in limonene peak area are observed between different breath collects (A and B, performed on the same individual), which is to be expected because some VOCs in breath do vary over time.

FAIMS – excellent reproducibility for VOC analysis

The Breath Biopsy platform uses FAIMS to analyse breath samples. To demonstrate FAIMS reproducibility, known quantities of methyl salicylate and limonene standards were loaded onto sorbent tubes and analyzed using GC-FAIMS (Lonestar VOC Analyzer). The experiment was repeated regularly to assess the variation over time (Figure 3). Data for the two compounds over a three month period are shown. The relative standard deviation is 6.1 % for methyl salicylate and 3.4 % for limonene.

FAIMS Breath Biopsy Reproducibility
Figure 3. GC-FAIMS technology shows excellent reproducibility – Standard compounds were loaded onto sorbent tubes at a defined amount and analyzed using GC-FAIMS (Lonestar VOC Analyzer). Over a 3 month period, the relative standard deviation is 6.1 % for methyl salicylate and 3.4 % for D-limonene.

Examples of longitudinal measurements of breath VOCs in the scientific literature –  READ MORE 

Breath Biopsy®

If you want to learn more about Owlstone Medical’s Breath Sampling and Analysis technology, and our Biomarker Services, why not download our free ebook: Breath Biopsy: The Complete Guide?

Breath Biopsy Complete Guide


  1. Risby, Critical issues for breath analysis, J Breath Res, 2008, PDF of article
  2. Herbig and Beauchamp, Towards standardization in the analysis of breath gas volatiles, J Breath Res, 2014,
  3. Beauchamp et al., On the use of Tedlar® bags for breath-gas sampling and analysis, J Breath Res, 2008,

ReCIVA Breath Sampler device

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