Using a Cyclic Ion Mobility Spectrometer for Tandem Ion Mobility Experiments

Area of Science:

• Analytical Chemistry
• Biochemistry
• Structural Biology

Background:

• Accurate chemical structure characterization is crucial for understanding biological mechanisms.
• Mass spectrometry (MS) alone often insufficient for complex structures like carbohydrates due to isomerism.
• Ion mobility spectrometry (IMS) complements MS by providing conformational sensitivity to isomers.

Purpose of the Study:

• To provide a protocol for generating tandem ion mobility spectrometry (IMS/IMS) data.
• To enable reproducible Collision Cross Section (CCS) calibration for ion mobility data.
• To facilitate the use of IMS/IMS data as structural fingerprints for glycan analysis.

Main Methods:

• Detailed steps for sample preparation for IMS/IMS experiments.
• Building and executing IMS/IMS control sequences on Cyclic IMS instruments.
• Translating IMS arrival times to drift times and extracting mobility information.

Main Results:

• Demonstration of IMS/IMS data generation using a representative glycan.
• Method for accurate CCS calibration of the ion mobility dimension.
• Generation of reproducible mobility profiles acting as glycan fingerprints.

Conclusions:

• The protocol enables straightforward and reproducible IMS/IMS data acquisition.
• IMS/IMS data can be effectively used in molecular networking for glycomics.
• This method enhances structural elucidation of complex carbohydrates.