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Updated: Jan 22, 2026

Quantitative Metabolomics of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry
Published on: January 5, 2021
Fast screening and quantitative mass spectral imaging of thin-layer chromatography plates with flowing
Christopher Kuhlmann1, Maximilian Heide1, Carsten Engelhard2,3
1Department of Chemistry and Biology, University of Siegen, Adolf-Reichwein-Str. 2, 57076, Siegen, Germany.
Abstract:
Thin-layer chromatography (TLC) was interfaced to high-resolution mass spectrometry (MS) using a flowing atmospheric-pressure afterglow (FAPA) ambient desorption/ionization source. The influence of different TLC stationary phases on the mass spectral signal response and mass spectral image quality in FAPA-MS was carefully investigated. Specifically, a mixture of selected analgesics (acetaminophen), alkaloids (nicotine and caffeine), and steroids (cortisone) was deposited on different stationary phases (silica plates, RP-modified silica plates, CN-modified silica plates, DIOL-modified silica plates, and NH2-modified silica plates), and TLC plates with different thickness (100, 200, 250, 500, 1000, 2000 μm) of the stationary phase. After analyte separation, mass spectral imaging was performed of the complete TLC plate via FAPA-MS and the detected ion abundance was compared. It was found that TLC plates with larger particle sizes (10-12 μm) and thicker stationary phase layers (e.g., 1000 μm and 2000 μm) led to higher signals (protonated molecules) compared to smaller particles sizes (6-8 μm) and thinner stationary phases (e.g., 100 μm and 200 μm). Instrumental detection limits in the low ng-range/band were determined for TLC-FAPA-MS of caffeine from RP-modified TLC silica plates. Lastly, a quantitative TLC-FAPA-MS method using stable isotope dilution analysis was developed and applied to the quantification of caffeine in energy drinks. Graphical abstract.
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