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Targeted volatolomics of human monocytes: Comparison of 2D-GC/TOF-MS and 1D-GC/Orbitrap-MS methods

K. Zemánková, K. Pavelicová, A. Pompeiano, L. Mravcová, M. Černý, K. Bendíčková, M. Hortová Kohoutková, K. Dryahina, M. Vaculovičová, J. Frič, L. Vaníčková

. 2021 ; 1184 (-) : 122975. [pub] 20211005

Language English Country Netherlands

Document type Journal Article

Persistent link   https://www.medvik.cz/link/bmc22011994

Blood is a complex biological matrix providing valuable information on nutritional, metabolic, and immune status. The detection of blood biomarkers requires sensitive analytical methods because analytes are at very low concentrations. Peripheral blood monocytes play a crucial role in inflammatory processes, and the metabolites released by monocytes during these processes might serve as important signalling molecules and biomarkers of particular physiological states. Headspace solid-phase microextraction (HS-SPME) combined with two different mass spectrometric platforms, two-dimensional (2D) gas chromatography coupled to time-of-flight mass spectrometry (2D-GC/TOF-MS) and one-dimensional gas chromatography coupled to Orbitrap mass spectrometry (GC/Orbitrap-MS), were applied for the investigation of volatile organic compounds (VOCs) produced by human peripheral blood monocytes. An optimized method was subsequently applied for the characterization of changes in VOCs induced by lipopolysaccharides (LPS) and zymosan (ZYM) stimulation. Overall, the 2D-GC/TOF-MS and the 1D-GC/Orbitrap-MS analyses each yielded about 4000 and 400 peaks per sample, respectively. In total, 91 VOCs belonging to eight different chemical classes were identified. The samples were collected in two fractions, conditioned media for monitoring extracellularly secreted molecules and cell pellet samples to determine the intracellular composition of VOCs. Alcohols, ketones, and hydrocarbons were the main chemical classes of the metabolic profile identified in cell fractions. Aldehydes, acids and cyclic compounds were characteristic of the conditioned media fraction. Here we demonstrate that HS-SPME-2D-GC/TOF-MS is more suitable for the identification of specific VOC profiles produced by human monocytes than 1D-GC/Orbitrap-MS. We define the signature of VOCs occurring early after monocyte activation and characterise the signalling compounds released by immune cells into media.

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$a Targeted volatolomics of human monocytes: Comparison of 2D-GC/TOF-MS and 1D-GC/Orbitrap-MS methods / $c K. Zemánková, K. Pavelicová, A. Pompeiano, L. Mravcová, M. Černý, K. Bendíčková, M. Hortová Kohoutková, K. Dryahina, M. Vaculovičová, J. Frič, L. Vaníčková
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$a Blood is a complex biological matrix providing valuable information on nutritional, metabolic, and immune status. The detection of blood biomarkers requires sensitive analytical methods because analytes are at very low concentrations. Peripheral blood monocytes play a crucial role in inflammatory processes, and the metabolites released by monocytes during these processes might serve as important signalling molecules and biomarkers of particular physiological states. Headspace solid-phase microextraction (HS-SPME) combined with two different mass spectrometric platforms, two-dimensional (2D) gas chromatography coupled to time-of-flight mass spectrometry (2D-GC/TOF-MS) and one-dimensional gas chromatography coupled to Orbitrap mass spectrometry (GC/Orbitrap-MS), were applied for the investigation of volatile organic compounds (VOCs) produced by human peripheral blood monocytes. An optimized method was subsequently applied for the characterization of changes in VOCs induced by lipopolysaccharides (LPS) and zymosan (ZYM) stimulation. Overall, the 2D-GC/TOF-MS and the 1D-GC/Orbitrap-MS analyses each yielded about 4000 and 400 peaks per sample, respectively. In total, 91 VOCs belonging to eight different chemical classes were identified. The samples were collected in two fractions, conditioned media for monitoring extracellularly secreted molecules and cell pellet samples to determine the intracellular composition of VOCs. Alcohols, ketones, and hydrocarbons were the main chemical classes of the metabolic profile identified in cell fractions. Aldehydes, acids and cyclic compounds were characteristic of the conditioned media fraction. Here we demonstrate that HS-SPME-2D-GC/TOF-MS is more suitable for the identification of specific VOC profiles produced by human monocytes than 1D-GC/Orbitrap-MS. We define the signature of VOCs occurring early after monocyte activation and characterise the signalling compounds released by immune cells into media.
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$a Pavelicová, Kristýna $u Department of Chemistry and Biochemistry, Faculty of AgriSciences, Mendel University in Brno, Zemědělská 1, CZ-61300 Brno, Czech Republic; Central European Institute of Technology, Brno University of Technology, Purkyňova 123, CZ-61200 Brno, Czech
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$a Pompeiano, Antonio $u Central European Institute of Technology, Brno University of Technology, Purkyňova 123, CZ-61200 Brno, Czech; Departmentof Forest Botany, Dendrology and Geobiocenology, Faculty of Forest and Wood Technology, Mendel University in Brno, Zemědělská 1 CZ-61300, Czech Republic
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$a Mravcová, Ludmila $u Brno University of Technology, Purkyňova 464/118, CZ-61200, Brno, Czech Republic
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$a Černý, Martin $u Department of Molecular Biology and Radiobiology, Phytophthora Research Centre, Faculty of AgriSciences, Mendel University in Brno, Zemedelska 1, CZ-61300 Brno, Czech Republic
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$a Bendíčková, Kamila $u International Clinical Research Centre of St. Anne's University Hospital Brno, Pekařská 53, CZ-656 91 Brno, Czech Republic
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$a Hortová Kohoutková, Marcela $u International Clinical Research Centre of St. Anne's University Hospital Brno, Pekařská 53, CZ-656 91 Brno, Czech Republic
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$a Dryahina, Kseniya $u J. Heyrovsky Institute of Physical Chemistry of the Czech Academy of Sciences, Dolejškova 3, CZ-18223 Prague, Czech Republic
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$a Vaculovičová, Markéta $u Department of Chemistry and Biochemistry, Faculty of AgriSciences, Mendel University in Brno, Zemědělská 1, CZ-61300 Brno, Czech Republic; Central European Institute of Technology, Brno University of Technology, Purkyňova 123, CZ-61200 Brno, Czech
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$a Frič, Jan $u International Clinical Research Centre of St. Anne's University Hospital Brno, Pekařská 53, CZ-656 91 Brno, Czech Republic; Institute of Hematology and Blood Transfusion, U Nemocnice 2094/1, CZ-128 00 Prague, Czech Republic. Electronic address: JAN.FRIC@FNUSA.CZ
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$a Vaníčková, Lucie $u Department of Chemistry and Biochemistry, Faculty of AgriSciences, Mendel University in Brno, Zemědělská 1, CZ-61300 Brno, Czech Republic; Central European Institute of Technology, Brno University of Technology, Purkyňova 123, CZ-61200 Brno, Czech; Departmentof Forest Botany, Dendrology and Geobiocenology, Faculty of Forest and Wood Technology, Mendel University in Brno, Zemědělská 1 CZ-61300, Czech Republic. Electronic address: lucie.vanickova@mendelu.cz
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