A Dual-Gated Structures for Lossless Ion Manipulations-Ion Mobility Orbitrap Mass Spectrometry Platform for Combined...

Area of Science:

• Analytical Chemistry
• Biochemistry
• Physical Chemistry

Background:

• High-resolution ion mobility spectrometry-mass spectrometry (HR-IMS-MS) is crucial for analyzing complex biological samples.
• Current HR-IMS and HR-MS analyses are often performed separately due to timescale mismatches.
• A unified approach is needed to enhance the characterization of biological mixtures.

Purpose of the Study:

• To develop and validate a novel platform integrating HR-IMS with HR-MS for simultaneous analysis.
• To overcome the temporal limitations of independent HR-IMS and HR-MS measurements.
• To enable comprehensive characterization of complex biological samples, including proteomics and lipidomics.

Main Methods:

• Coupling an 11 m Structures for Lossless Ion Manipulations (SLIM) module to a Q-Exactive Plus Orbitrap MS using a dual-gated ion injection system.
• Implementing simultaneous SLIM separation, high-resolution Orbitrap mass analysis (up to 140k), and high-energy collision-induced dissociation (HCD).
• Characterizing the platform using phosphazene cations, standard peptides, and complex lipid mixtures.

Main Results:

• The SLIM-Orbitrap platform achieved simultaneous HR-IMS-MS/MS analysis in approximately 25 minutes.
• Demonstrated an average SLIM collision cross-section (CCS) resolving power (Rp^CCS) of ~218 and peak capacity of ~156.
• Successfully separated isobaric lipids and identified peptides, showcasing the system's utility for complex mixture analysis.

Conclusions:

• The developed dual-gated SLIM-Orbitrap platform enables integrated high-resolution ion mobility and mass spectrometry.
• This novel capability significantly advances proteomics and lipidomics by providing high-resolution multimodal data.
• The system lays the groundwork for reference-free identification of unknown ion structures in complex biological samples.