Assessment of thermal and solvent stable SPME fibers for metabolomics studies performed in living systems

Date

2025-05-15

Author

Kahremanoğlu, Kübra
Jaroch, Karol
Szeliska, Paulina
Filipiak, Wojciech
Charemski, Bartłomiej
Żuchowska, Karolina
Çetin, Enes
Eroğlu, Ahmet E.
Bojko, Barbara
Boyacı, Ezel

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Solid phase microextraction (SPME), as a sampling/sample preparation technique, offers unique solutions for the most challenging applications, including metabolomics studies of living systems. However, for global metabolomics it is critical to use an SPME sampler facilitating the extraction of both volatiles and nonvolatiles, which at the same time is compatible with thermal and solvent-assisted desorption. As a promising universal coating, recently hydrophilic-lipophilic balanced (HLB) particles immobilized in PTFE have been introduced as a new SPME sampler to provide a wide-range of analyte coverage and compatibility with solvent and thermal desorption. Thus, making it suitable for both gas and liquid chromatography (GC/LC) based applications. However, its potential in metabolomics has not been investigated to date. In this study, HLB/PTFE SPME fibers were prepared, evaluated with selected polar and non-polar metabolites relevant to biological systems, and validated for cell-line studies. The validation proved that these fibers can extract a wide-range of molecules (LogP: 4.2 to 15.6) with acceptable accuracy (≤19% RE%) and repeatability (intra-day ≤17% and inter-day 12% RSD%). The LOQ was determined to vary between 150.0 and 500.0 ng/mL. Upon validation, the fibers were used in a proof-of-concept study for extraction of endometabolome and exometabolome of melanoma B16F10 and lung cancer LL2 cell lines. The metabolome studies showed that HLB/PTFE fibers provide lower coverage, but for some compounds higher extraction efficiency compared to HLB/PAN fibers used in LC-based metabolomics. Fibers also proved suitable for GC-MS analysis, allowing for the detection of 36 volatile organic compounds in the headspace of the cell lines and RPMI medium.

Subject Keywords

Biological analysis, Cancer cell metabolome, Cell line studies, Metabolomics, Sample preparation, Solid phase microextraction

URI

https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85217042953&origin=inward
https://hdl.handle.net/11511/113693

Collections

Department of Chemistry, Article