Unprecedented Ionization Processes in Mass Spectrometry Provide Missing Link between ESI and MALDI

In the field of mass spectrometry, producing intact, highly‐charged protein ions from surfaces is a conundrum with significant potential payoff in application areas ranging from biomedical to clinical research. Here, we report on the ability to form intact, highly‐charged protein ions on high vacuum time‐of‐flight mass spectrometers in the linear and reflectron modes achievable using experimental conditions that allow effective matrix removal from both the sample surfaces and from the charged clusters formed by the laser ablation event. The charge states are the highest reported on high vacuum mass spectrometers, yet they remain at only around a third of the highest charge obtained using laser ablation with a suitable matrix at atmospheric pressure. Other than physical instrument modifications, the key to forming abundant and stable highly‐charged ions appears to be the volatility of the matrix used. Cumulative results suggest mechanistic links between the ionization process reported here and traditional ionization methods of electrospray ionization and matrix‐assisted laser desorption/ionization. Missing link: The ability to form intact, highly‐charged protein ions on high vacuum time‐of‐flight mass spectrometers in the linear and reflective detection modes is reported. Cumulative results suggest mechanistic links between the ionization process reported here and traditional ionization methods of electrospray ionization and matrix‐assisted laser desorption/ionization.