| Title: | Capturing Actively Produced Microbial Volatile Organic Compounds from Human-Associated Samples with Vacuum-Assisted Sorbent Extraction |
| Author(s): | Phan J; Kapcia J; Rodriguez CI; Vogel VL; Cardin DB; Dunham SJB; Whiteson K; |
| Address: | "Department of Molecular Biology and Biochemistry, University of California Irvine. Department of Ecology and Evolutionary Biology, University of California Irvine. Entech Instruments Inc. Department of Molecular Biology and Biochemistry, University of California Irvine; katrine@uci.edu" |
| ISSN/ISBN: | 1940-087X (Electronic) 1940-087X (Linking) |
| Abstract: | "Volatile organic compounds (VOCs) from biological samples have unknown origins. VOCs may originate from the host or different organisms from within the host's microbial community. To disentangle the origin of microbial VOCs, volatile headspace analysis of bacterial mono- and co-cultures of Staphylococcus aureus, Pseudomonas aeruginosa, and Acinetobacter baumannii, and stable isotope probing in biological samples of feces, saliva, sewage, and sputum were performed. Mono- and co-cultures were used to identify volatile production from individual bacterial species or in combination with stable isotope probing to identify the active metabolism of microbes from the biological samples. Vacuum-assisted sorbent extraction (VASE) was employed to extract the VOCs. VASE is an easy-to-use, commercialized, solvent-free headspace extraction method for semi-volatile and volatile compounds. The lack of solvents and the near-vacuum conditions used during extraction make developing a method relatively easy and fast when compared to other extraction options such as tert-butylation and solid phase microextraction. The workflow described here was used to identify specific volatile signatures from mono- and co-cultures. Furthermore, analysis of the stable isotope probing of human associated biological samples identified VOCs that were either commonly or uniquely produced. This paper presents the general workflow and experimental considerations of VASE in conjunction with stable isotope probing of live microbial cultures" |
| Keywords: | Bacteria Gas Chromatography-Mass Spectrometry/methods Humans Solid Phase Microextraction/methods Solvents *Staphylococcal Infections Staphylococcus aureus *Volatile Organic Compounds/analysis; |
| Notes: | "MedlinePhan, Joann Kapcia, Joseph 3rd Rodriguez, Cynthia I Vogel, Victoria L Cardin, Daniel B Dunham, Sage J B Whiteson, Katrine eng R01 HL136647/HL/NHLBI NIH HHS/ Research Support, N.I.H., Extramural Video-Audio Media 2022/06/21 J Vis Exp. 2022 Jun 1; (184). doi: 10.3791/62547" |
