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Metabolites


Title:Application of Thin-Film Microextraction to Analyze Volatile Metabolites in A549 Cancer Cells
Author(s):Filipiak W; Jaroch K; Szeliska P; Zuchowska K; Bojko B;
Address:"Department of Pharmacodynamics and Molecular Pharmacology, Faculty of Pharmacy, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Torun, Dr. Jurasza 2 Str., 85-089 Bydgoszcz, Poland"
Journal Title:Metabolites
Year:2021
Volume:20211014
Issue:10
Page Number: -
DOI: 10.3390/metabo11100704
ISSN/ISBN:2218-1989 (Print) 2218-1989 (Electronic) 2218-1989 (Linking)
Abstract:"Volatile organic compounds (VOCs) have been proposed in the last two decades as biomarkers for disease detection and therapeutic monitoring. Model in vitro experiments with established cell lines are fundamental to clarify whether given VOCs originate from normal human cells or pathogens, including transformed cancer cells. Due to the trace concentrations of target metabolites, adsorptive enrichment is needed before gas chromatography-mass spectrometry (GC-MS) analysis, with solid-phase microextraction (SPME) being perfectly suited for this purpose. Here, a modification of SPME, the thin-film microextraction (TFME) technique, is proposed for analysis of cellular VOCs, which utilizes a planar mesh coated with stationary phase to increase the extraction phase volume and active surface area. In this study, four different adsorbents were compared: carboxen, divinylbenzene, hydrophobic-lipophilic balanced and polydimethylsiloxane. Amongst them, HLB sheets using poly(divinylbenzene-co-N-vinyl-pyrrolidone) skeleton structure proved to be the most versatile, enabling the most sensitive analysis of VOCs with a broad polarity and volatility. For HLB, sampling type (internal static headspace, external bi-directional headspace), extraction temperature and extraction time were also examined. An established method was successfully applied to analyze metabolites produced by A549 cells revealing five volatiles at significantly higher (additionally benzaldehyde at lower) levels in cell culture medium compared to the cell-free reference medium headspace"
Keywords:A549 Gc-ms Spme Tfme VOCs cancer cells cellular metabolites extraction techniques in vitro models volatile metabolites;
Notes:"PubMed-not-MEDLINEFilipiak, Wojciech Jaroch, Karol Szeliska, Paulina Zuchowska, Karolina Bojko, Barbara eng 2017/26/D/NZ6/00136/National Science Center/ Switzerland 2021/10/23 Metabolites. 2021 Oct 14; 11(10):704. doi: 10.3390/metabo11100704"

 
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