Analytical and Bioanalytical Chemistry

, Volume 396, Issue 2, pp 739–750 | Cite as

Human breath analysis: methods for sample collection and reduction of localized background effects

  • Audrey N. Martin
  • George R. FarquarEmail author
  • A. Daniel Jones
  • Matthias Frank
Original Paper


Solid-phase microextraction (SPME) was applied, in conjunction with gas chromatography–mass spectrometry, to the analysis of volatile organic compounds (VOCs) in human breath samples without requiring exhaled breath condensate collection. A new procedure, exhaled breath vapor (EBV) collection, involving the active sampling and preconcentration of a breath sample with a SPME fiber fitted inside a modified commercial breath-collection device, the RTube™, is described. Immediately after sample collection, compounds are desorbed from the SPME fiber at 250 °C in the GC-MS injector. Experiments were performed using EBV collected at −80 °C and at room temperature, and the results compared to the traditional method of collecting exhaled breath condensate at −80 °C followed by passive SPME sampling of the collected condensate. Methods are compared in terms of portability, ease-of-use, speed of analysis, and detection limits. The need for a clean air supply for the study subjects is demonstrated using several localized sources of VOC contaminants including nail polish, lemonade, and gasoline. Various simple methods to supply clean inhaled air to a subject are presented. Chemical exposures are used to demonstrate the importance of providing cleaned air (organic vapor respirator) or an external air source (tubing stretched to a separate room). These techniques allow for facile data interpretation by minimizing background contaminants. It is demonstrated herein that this active SPME breath-sampling device provides advantages in the forms of faster sample collection and data analysis, apparatus portability and avoidance of power or cooling requirements, and performance for sample collection in a contaminated environment.


Extracted ion chromatograms (XIC m/z 93) of breath samples from a single subject (EBV-RT). One sample was collected over 125 mL of lemonade with no modification to the inhaled air (red trace, top) and another sample was collected over 125 mL of lemonade using a respirator (blue trace, bottom). Peaks labeled are identified as monoterpenes and monoterpene alcohols, flavor ingredients present in lemonade.


Bioanalytical methods Breath analysis GC-MS VOC Breath condensate 



Financial support from an LLNL Laboratory-Directed Research and Development (LDRD) grant 05-ERD-053 and a Defense Advanced Research Project Agency (DARPA) seedling project is greatly appreciated. This work was performed under the auspices of the U.S. Department of Energy (DOE) by Lawrence Livermore National Laboratory in part under Contract W-7405-Eng-48 and in part under Contract DE-AC52-07NA27344. A.N.M. performed this research in part while on appointment as a U.S. Department of Homeland Security (DHS) Fellow under the DHS Scholarship and Fellowship Program, administered by the Oak Ridge Institute for Science and Education (ORISE) for DHS through an interagency agreement with DOE. ORISE is managed by Oak Ridge Associated Universities under DOE contract number DE-AC05-06OR23100. All opinions expressed in this paper are the authors’ and do not necessarily reflect the policies and views of DHS, DOE, or ORISE.


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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Audrey N. Martin
    • 1
    • 2
  • George R. Farquar
    • 1
    Email author
  • A. Daniel Jones
    • 2
  • Matthias Frank
    • 1
  1. 1.Lawrence Livermore National LaboratoryLivermoreUSA
  2. 2.Michigan State UniversityEast LansingUSA

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