| Title: | Headspace sorptive extraction-gas chromatography-mass spectrometry method to measure volatile emissions from human airway cell cultures |
| Author(s): | Yamaguchi MS; McCartney MM; Linderholm AL; Ebeler SE; Schivo M; Davis CE; |
| Address: | "Mechanical and Aerospace Engineering, University of California Davis, Davis, CA 95616, USA. Center for Comparative Respiratory Biology and Medicine, UC Davis Medical School, Davis, CA 95616, USA. Viticulture and Enology, University of California Davis, One Shields Avenue, Davis, CA 95616, USA. Division of Pulmonary and Critical Care Medicine, University of California Davis, Davis, CA 95616, USA. Mechanical and Aerospace Engineering, University of California Davis, Davis, CA 95616, USA. Electronic address: cedavis@ucdavis.edu" |
| Journal Title: | J Chromatogr B Analyt Technol Biomed Life Sci |
| DOI: | 10.1016/j.jchromb.2018.05.009 |
| ISSN/ISBN: | 1873-376X (Electronic) 1570-0232 (Print) 1570-0232 (Linking) |
| Abstract: | "The human respiratory tract releases volatile metabolites into exhaled breath that can be utilized for noninvasive health diagnostics. To understand the origin of this metabolic process, our group has previously analyzed the headspace above human epithelial cell cultures using solid phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS). In the present work, we improve our model by employing sorbent-covered magnetic stir bars for headspace sorptive extraction (HSSE). Sorbent-coated stir bar analyte recovery increased by 52 times and captured 97 more compounds than SPME. Our data show that HSSE is preferred over liquid extraction via stir bar sorptive extraction (SBSE), which failed to distinguish volatiles unique to the cell samples compared against media controls. Two different cellular media were also compared, and we found that Opti-MEM(R) is preferred for volatile analysis. We optimized HSSE analytical parameters such as extraction time (24?ª+h), desorption temperature (300?ª+ degrees C) and desorption time (7?ª+min). Finally, we developed an internal standard for cell culture VOC studies by introducing 842?ª+ng of deuterated decane per 5?ª+mL of cell medium to account for error from extraction, desorption, chromatography and detection. This improved model will serve as a platform for future metabolic cell culture studies to examine changes in epithelial VOCs caused by perturbations such as viral or bacterial infections, opening opportunities for improved, noninvasive pulmonary diagnostics" |
| Keywords: | Cell Line Epithelial Cells/*metabolism Gas Chromatography-Mass Spectrometry/*methods Humans Respiratory Mucosa/*cytology Solid Phase Microextraction Volatile Organic Compounds/*analysis/*isolation & purification/metabolism; |
| Notes: | "MedlineYamaguchi, Mei S McCartney, Mitchell M Linderholm, Angela L Ebeler, Susan E Schivo, Michael Davis, Cristina E eng U01 EB022003/EB/NIBIB NIH HHS/ T32 HL007013/HL/NHLBI NIH HHS/ P30 ES023513/ES/NIEHS NIH HHS/ UG3 OD023365/OD/NIH HHS/ K23 HL127185/HL/NHLBI NIH HHS/ UL1 TR000002/TR/NCATS NIH HHS/ UH3 OD023365/OD/NIH HHS/ Netherlands 2018/05/22 J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Jul 15; 1090:36-42. doi: 10.1016/j.jchromb.2018.05.009. Epub 2018 May 12" |
