[en] The high potential of exhaled breath for disease diagnosis has been highlighted in numerous studies. However, exhaled breath analysis is suffering from a lack of standardized sampling and analysis procedures, impacting the robustness of inter-laboratory results, and thus hampering proper external validation. The aim of this work was to verify compliance and validate the performance of two different comprehensive two-dimensional gas chromatography coupled to mass spectrometry platforms in different laboratories by monitoring probe metabolites in exhaled breath following the Peppermint Initiative guidelines. An initial assessment of the exhaled breath sampling conditions was performed, selecting the most suitable sampling bag material and volume. Then, a single sampling was performed using Tedlar bags, followed by the trapping of the volatile organic compounds into thermal desorption tubes for the subsequent analysis using two different analytical platforms. The thermal desorption tubes were first analyzed by a (cryogenically modulated) comprehensive two-dimensional gas chromatography system coupled to high-resolution time-of-flight mass spectrometry. The desorption was performed in split mode and the split part was recollected in the same tube and further analyzed by a different (flow modulated) comprehensive two-dimensional gas chromatography system with a parallel detection, specifically using a quadrupole mass spectrometer and a vacuum ultraviolet detector. Both the comprehensive two-dimensional gas chromatography platforms enabled the longitudinal tracking of the peppermint oil metabolites in exhaled breath. The increased sensitivity of comprehensive two-dimensional gas chromatography enabled to successfully monitor over a 6.5 h period a total of 10 target compounds, namely α-pinene, camphene, β-pinene, limonene, cymene, eucalyptol, menthofuran, menthone, isomenthone, and neomenthol.
Disciplines :
Chemistry Cardiovascular & respiratory systems
Author, co-author :
Zanella, Delphine ; Université de Liège - ULiège > Université de Liège - ULiège
Henin, Adèle; Molecular System, Organic & Biological Analytical Chemistry Group, University of Liège, Liège, Belgium
Mascrez, Steven ; Université de Liège - ULiège > TERRA Research Centre
Stefanuto, Pierre-Hugues ; Université de Liège - ULiège > Département de chimie (sciences) > Chimie analytique, organique et biologique
Franchina, Flavio ; Université de Liège - ULiège > Département de chimie (sciences) > Chimie analytique, organique et biologique ; Department of Chemistry, Pharmaceutical, and Agricultural Sciences, University of Ferrara, Ferrara, Italy
Focant, Jean-François ; Université de Liège - ULiège > Département de chimie (sciences) > Chimie analytique, organique et biologique
Purcaro, Giorgia ; Université de Liège - ULiège > TERRA Research Centre > Chimie des agro-biosystèmes
Language :
English
Title :
Comprehensive two-dimensional gas chromatographic platforms comparison for exhaled breath metabolites analysis.
The authors are grateful to Merck Millipore, SepSolve/Markes, VUV Analytics, LECO, and Shimadzu for the support. The study has been conducted in the framework of the “Peppermint Consortium” and the authors gratefully acknowledge this initiative.
Adams F. Hippocratic Writings: Aphorism 4,5. Web Atomics, New York 1994, p. Aphorism 4,5.
Drabińska N, Flynn C, Ratcliffe N, Belluomo I, Myridakis A, Gould O, Fois M, Smart A, Devine T, Costello BDL. A literature survey of all volatiles from healthy human breath and bodily fluids: The human volatilome. J. Breath Res. 2021; 15: 34001.
Breathborne Biomarkers and the Human Volatilome. Elsevier 2020.
Wallace MAG, Pleil JD. Evolution of clinical and environmental health applications of exhaled breath research: Review of methods and instrumentation for gas-phase, condensate, and aerosols. Anal. Chim. Acta. 2018; 1024: 18–38.
Higgins Keppler EA, Jenkins CL, Davis TJ, Bean HD. Advances in the application of comprehensive two-dimensional gas chromatography in metabolomics. TrAC - Trends Anal. Chem. 2018; 109: 275–86.
Franchina FA, Zanella D, Dubois LM, Focant JF. The role of sample preparation in multidimensional gas chromatographic separations for non-targeted analysis with the focus on recent biomedical, food, and plant applications. J. Sep. Sci. 2020; jssc.202000855.
Di Giovanni N, Meuwis MA, Louis E, Focant JF. Untargeted Serum Metabolic Profiling by Comprehensive Two-Dimensional Gas Chromatography-High-Resolution Time-of-Flight Mass Spectrometry. J. Proteome Res. 2020; 19: 1013–28.
Franchina FA, Purcaro G, Burklund A, Beccaria M, Hill JE. Evaluation of different adsorbent materials for the untargeted and targeted bacterial VOC analysis using GC×GC-MS. Anal. Chim. Acta. 2019; 1066: 146–53.
Rees CA, Nordick KV, Franchina FA, Lewis AE, Hirsch EB, Hill JE. Volatile metabolic diversity of Klebsiella pneumoniae in nutrient-replete conditions. Metabolomics. 2017; 13: https://doi.org/10.1007/s11306-016-1161-z
Phillips M, Cataneo RN, Chaturvedi A, Kaplan PD, Libardoni M, Mundada M, Patel U, Zhang X. Detection of an Extended Human Volatome with Comprehensive Two-Dimensional Gas Chromatography Time-of-Flight Mass Spectrometry. PLoS One. 2013; 8: 1–8.
Beccaria M, Stefanuto P-H, Purcaro G, Franchina FA, in: Cordero, C. (Ed.), Characterization of Odorant Patterns by Comprehensive Two-Dimensional Gas Chromatography. 2022, pp. 367–409.
Gruber B, Groeger T, Harrison D, Zimmermann R. Vacuum ultraviolet absorption spectroscopy in combination with comprehensive two-dimensional gas chromatography for the monitoring of volatile organic compounds in breath gas: A feasibility study. J. Chromatogr. A. 2016, 1464: 141–6.
Purcaro G, Nasir M, Franchina FA, Rees CA, Aliyeva M, Daphtary N, Wargo MJ, Lundblad LKA, Hill JE. Breath metabolome of mice infected with Pseudomonas aeruginosa. Metabolomics. 2019; 15: 10.
Franchina FA, Mellors TR, Aliyeva M, Wagner J, Daphtary N, Lundblad LKA, Fortune SM, Rubin EJ, Hill JE. Towards the use of breath for detecting mycobacterial infection: A case study in a murine model. J. Breath Res. 2018; 12: https://doi.org/10.1088/1752-7163/aaa016
Adiguzel Y, Kulah H. Breath sensors for lung cancer diagnosis. Biosens. Bioelectron. 2015; 65: 121–38.
Mellors TR, Nasir M, Franchina FA, Smolinska A, Blanchet L, Flynn JL, Tomko J, O'Malley M, Scanga CA, Lin PL, Wagner J, Hill JE. Identification of Mycobacterium tuberculosis using volatile biomarkers in culture and exhaled breath. J. Breath Res. 2019; 13: 0–6.
Blanchet L, Smolinska A, Baranska A, Tigchelaar E, Swertz M, Zhernakova A, Dallinga JW, Wijmenga C, Van Schooten FJ. Factors that influence the volatile organic compound content in human breath. J. Breath Res. 2017; 11, https://doi.org/10.1088/1752-7163/aa5cc5
Beauchamp JD, Pleil JD. Simply breath-taking? Developing a strategy for consistent breath sampling. J. Breath Res. 2013; 7: https://doi.org/10.1088/1752-7155/7/4/042001
Henderson B, Ruszkiewicz DM, Wilkinson M, Beauchamp JD, Cristescu SM, Fowler SJ, Salman D, Francesco FDi, Koppen G, Langejürgen J, Holz O, Hadjithekli A, Moreno S, Pedrotti M, Sinues P, Slingers G, Wilde M, Lomonaco T, Zanella D, Zenobi R, Focant J-F, Grassin-Delyle S, Franchina FA, Malásková M, Stefanuto P-H, Pugliese G, Mayhew C, Thomas CLPP. A benchmarking protocol for breath analysis: the peppermint experiment. J. Breath Res. 2020; 14: 046008.
Wilkinson M, White I, Hamshere K, Holz O, Schuchardt S, Bellagambi FG, Lomonaco T, Biagini D, Di FF, Fowler SJ, Beauchamp JD, Cristescu SM, Focant JF, Franchina FA, Grassin-Delyle S, Hadjithekli A, Henderson B, Koppen GF, Langejürgen J, Malásková M, Mayhew C, Moreno S, Pedrotti M, Pugliese G, Ruszkiewicz DM, Salman D, Sinues P, Slingers GF, Stefanuto PH, Thomas CLP, Wilde M, Zanella D, Zenobi R. The peppermint breath test: A benchmarking protocol for breath sampling and analysis using GC-MS. J. Breath Res. 2021; 15, https://doi.org/10.1088/1752-7163/abd28c
Fiehn O, Robertson D, Griffin J, vab der Werf M, Nikolau B, Morrison N, Sumner LW, Goodacre R, Hardy NW, Taylor C, Fostel J, Kristal B, Kaddurah-Daouk R, Mendes P, van Ommen B, Lindon JC, Sansone SA. The metabolomics standards initiative (MSI). Metabolomics. 2007; 3: 175–8.
Spicer RA, Salek R, Steinbeck C. Compliance with minimum information guidelines in public metabolomics repositories. Sci. Data. 2017; 4: 170137.
Ruszkiewicz DM, Myers R, Henderson B, Yusof H, Meister A, Moreno S, Eddleston M, Darnley K, Nailon WH, McLaren D, Lao YE, Hovda KE, Lam S, Cristescu SM, Thomas CLP. Peppermint protocol: first results for gas chromatography-ion mobility spectrometry. J. Breath Res. 2022; 16: 036004.
Malásková M, Henderson B, Chellayah PD, Ruzsanyi V, Mochalski P, Cristescu SM, Mayhew CA. Proton transfer reaction time-of-flight mass spectrometric measurements of volatile compounds contained in peppermint oil capsules of relevance to real-time pharmacokinetic breath studies. J. Breath Res. 2019; 13: https://doi.org/10.1088/1752-7163/ab26e2
Schmidt E, Bail S, Buchbauer G, Stoilova I, Atanasova T, Stoyanova A, Krastanov A, Jirovetz L. Chemical composition, olfactory evaluation and antioxidant effects of essential oil from Mentha x piperita. Nat. Prod. Commun. 2009; 4: 1107–12.
Lawal O, Ahmed WM, Nijsen TME, Goodacre R, Fowler SJ. Exhaled breath analysis: a review of ‘breath-taking’ methods for off-line analysis. Metabolomics. 2017; 13: 1–16.
Ghimenti S, Lomonaco T, Bellagambi FG, Tabucchi S, Onor M, Trivella MG, Ceccarini A, Fuoco R, Di Francesco F. Comparison of sampling bags for the analysis of volatile organic compounds in breath. J. Breath Res. 2015; 9: https://doi.org/10.1088/1752-7155/9/4/047110
Jia Z, Patra A, Kutty VK, Venkatesan T. Critical review of volatile organic compound analysis in breath and in vitro cell culture for detection of lung cancer. Metabolites. 2019; 9, https://doi.org/10.3390/metabo9030052
Zanella D, Henket M, Schleich F, Dejong T, Louis R, Focant JF, Stefanuto PH. Comparison of the effect of chemically and biologically induced inflammation on the volatile metabolite production of lung epithelial cells by GC×GC-TOFMS. Analyst. 2020; 145: 5148–57.
Franchina FA, Zanella D, Dejong T, Focant J-F. Impact of the adsorbent material on volatile metabolites during in vitro and in vivo bio-sampling. Talanta. 2021; 222: 121569.
Duan Z, Kjeldsen P, Scheutz C. Improving the analytical flexibility of thermal desorption in determining unknown VOC samples by using re-collection. Sci. Total Environ. 2021; 768: 144692.
Cordero C, Rubiolo P, Reichenbach SE, Carretta A, Cobelli L, Giardina M, Bicchi C. Method translation and full metadata transfer from thermal to differential flow modulated comprehensive two dimensional gas chromatography: Profiling of suspected fragrance allergens. J. Chromatogr. A. 2017; 1480: 70–82.
Magagna F, Liberto E, Reichenbach SE, Tao Q, Carretta A, Cobelli L, Giardina M, Bicchi C, Cordero C. Advanced fingerprinting of high-quality cocoa: Challenges in transferring methods from thermal to differential-flow modulated comprehensive two dimensional gas chromatography. J. Chromatogr. A. 2018; 1536: 122–36.
Stilo F, Gabetti E, Bicchi C, Carretta A, Peroni D, Reichenbach SE, Cordero C, McCurry J. A step forward in the equivalence between thermal and differential-flow modulated comprehensive two-dimensional gas chromatography methods. J. Chromatogr. A. 2020; 1627: 461396.
