LC-MS Meets MS Imaging: Using CCS for Confident Lipid Annotation

This webinar took place on May 26, 2021

Webinar Overview

The involvement of lipids in many biological and physiological processes makes their analysis of paramount importance. High-resolution mass spectrometry (HR-MS) combined with fragmentation experiments can identify lipid species. However, to differentiate isomeric lipids or isobaric interferences, powerful additional separation techniques, often (ultra)high performance liquid chromatography, are required.

Trapped ion mobility-mass spectrometry (TIMS) for the separation of ions after matrix-assisted laser desorption/ionization (MALDI) has become a useful tool in lipid research, increasing the reliability of lipid identification using complementary collision cross section (CCS) data.

In this study, we created a tissue-specific CCS database using hydrophilic interaction liquid chromatography coupled to electrospray TIMS-MS. With the help of this database, we were able to distinguish isomeric lipid classes clearly, such as phosphatidylglycerol (PG) and its regio-isomer bis(monoacylgylcero)phosphate (BMP), as well as isobaric interferences due to adduct formation during the MALDI process. The developed workflow and the application to a mouse spleen sample for confident lipid identification applying MALDI-TIMS-MS imaging will be presented.

In this webinar, you will learn how to:

  • Use trapped ion mobility separation to identify isomeric lipids confidently
  • Separate BMP and PG lipids using TIMS
  • Use CCS to visualize confident lipid annotations spatially on tissue
  • Build tissue-specific CCS databases


Heiko Hayen
Professor for Analytical Chemistry,
University of Münster, Germany


For Research Use Only. Not for use in clinical diagnostic procedures.