In vitro detection of small molecule metabolites excreted from cancer cells using a Tenax TA thin-film microextraction device

• Thin film microextraction for detection of cell culture semi-volatile metabolites.
• Detection of released and consumed HeLa cell culture metabolites is demonstrated.
• Method covers the metabolites that cannot be preconcentrated in the headspace.

We developed a new device for the in vitro extraction of small molecule metabolites excreted from cancer cells. The extraction device, which was biocompatible and incubated with cancer cells, consists of a thin Tenax TA film deposited on the surface of a cylindrical aluminum rod. The Tenax TA solid phase was utilized for the direct extraction and preconcentration of the small molecule metabolites from a cell culture sample. The device fabrication and the metabolite extraction were optimized, tested, and validated using HeLa cell cultures. Comparison of metabolic profiles with the control measurement from the culture medium enabled detection of metabolites that were consumed or produced by the cell culture. Tentative identification and semi-quantitative investigation of the excreted metabolites were performed by GC–MS analysis. The proposed approach can be a valuable tool for the characterization of low-volatile cancer cell metabolites that are not covered by use of conventional methods based on headspace solid phase microextraction.