Resolving Challenges in Steroid Analysis with High-Resolution SLIM and Mobility-Aligned Fragmentation

Resolving Challenges in Steroid Analysis with High-Resolution SLIM and Mobility-Aligned Fragmentation
Sabrina M. Cramer, Tim Causon
MOBIE offers a 4- to 10-fold increase in native IM Rp across the full m/z range of interest for steroid analysis, leading to better discrimination of isomeric steroids. Routine CCS calibration on MOBIE in combination with application-specific correction of data allows existing steroid CCSN2 databases to be used for data analysis. Mobility-aligned fragmentation provides cleaner fragment mass spectra in DIA mode, improving identification confidence without compromising the duty cycle.
MOBIE provides a major boost in IM resolving power across the full m/z range of interest encompassed by steroid analysis. Building on the history of consensus calibrants, CCSN2 values from MOBIE are extremely consistent with expectations for non-uniform field IM technology, displaying excellent agreement with CCSN2 database values when using established internal correction strategies. Minor differences of <0.2% between isomers can also be confirmed and partially resolved with MOBIE. The resolution advantage of SLIM can be further exploited for complex samples for filtering EICs to reduce matrix interferences, improve signal-to-noise, and avoid generation of chimeric fragment mass spectra with mobility-aligned fragmentation that negatively influence data processing.