Selectivity, sensitivity, high-throughput as well as rapid peak detection and assignation are key requirements to be fulfilled in metabolomics and bioanalysis. Nowadays, meeting the analytical challenges raised by complex samples and/or biological matrices is less prone to performance compromise thanks to the development of a new generation of hyphenated platforms. Initiated a decade ago with liquid chromatography (LC), the combination of optimized instruments able to operate at ultra-high pressure with columns packed with sub-2 µm particles (UHPLC) have drastically improved the separation efficiency <i>per</i> time unit. Meanwhile direct coupling with modern mass spectrometry (MS) detectors displaying improved acquisition rate has considerably increased both detection speed and sensitivity.<br/>The resurgence of supercritical fluid chromatography (SFC) can be seen as a continuity of the current quest for improved analytical performance. Profiting of a new generation of dedicated systems, this approach allows to achieve equivalent or even higher chromatographic performance than UHPLC thanks to both low viscosity of its mobile phase and full compatibility with modern chromatographic supports (UHPSFC). Moreover, SFC is particularly advantageous in terms of separation selectivity tuning. Indeed, the compatibility of SFC mobile phase with a broad spectrum of column chemistries offers almost infinite possibilities to play with retention while extending the analyzable compounds space already covered by LC.<br/>Despite these benefits, the use of SFC-MS for the analysis of complex sample and/or biological matrices has been scarcely reported. The present contribution will focus on tackling tenacious myths and misconceptions around the technique applicability for complex sample analysis. First, the successful interfacing of UHPSFC with MS detection will be illustrated. Advantages and drawbacks of such hyphenation will be addressed in terms of performance, sensitivity and robustness. Then, the applicability of UHPSFC-MS/MS as a screening approach to highlight doping agents in biological samples will be demonstrated. Method selectivity, sensitivity, linearity and matrix effects obtained for more than 200 forbidden substances will also be discussed in details and critically compared with UHPLC-MS/MS results. Finally, the relevance of the use of combined approach involving both UHPSFC and UHPLC coupled with high-resolution MS to maximize plant metabolites profiling will be demonstrated.

New insights in modern SFC-MS for the analysis of complex samples and biological matrices

253rd National Meeting (2017)

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