Assessment of thermal and solvent stable SPME fibers for volatilome investigation through in vitro experiments with cancer cell lines

Abstract

Background

solid phase microextraction (SPME) was introduced to bioanalysis years ago, but still, it did not find its place in routine analysis, despite the many advantages it offers. The limited number of commercial devices and protocols ready to be adopted for multi-residue analysis, i.e. untargeted metabolomics, are among the most important reasons. In this study, HLB-PTFE-coated SPME fibers were optimized for in vitro cell line studies targeting the volatilome of cancer cells.

Results

the HLB-PTFE SPME fibers with 10 mm coating length were prepared using dip coating approach with home-made HLB particles. The fiber sampling optimization and evaluation of its performance was performed in cell line media spiked with 18 representative compounds selected from the previous preliminary study on A549 lung cancer cell line. The results showed that headspace extraction for 20 min at 38 °C followed by 2 min desorption at 230 °C to GC-MS provides a reliable method with detection limits in a range of 2.2–43 ng/mL and repeatability better than 10% for determination of the volatiles in headspace of cell-line media. Moreover, the application of the developed method in B16F10 melanoma and LL2 lung cell lines revealed successful differentiation based on volatilome profiling.

Significance

the satisfactory results of optimization studies and accordance of results obtained here for cell line applications and literature indicate that the method developed with HLB-PTFE-coated SPME fibers and GC-MS approach suit future IVIVE study and meeting 3Rs rules requirements.