Electrospray ionization with higher-energy collision dissociation tandem mass spectrometry toward characterization of ceramides as [M + Li]+ ions: Mechanisms of fragmentation and structural identification

Ceramide is a huge lipid family consisting of diversified structures in which various modifications are seen in the fatty acyl chain and the long chain base (LCB). In this contribution, a higher collision energy (HCD) linear ion-trap mass spectrometric method (LIT MSn) was applied to study the mechanisms underlying the fragmentation processes of ceramide molecules in 12 subclasses, which were desorbed by ESI as the [M thorn Li]thorn ions. Multiple sets of fragment ions reflecting the fatty acyl chain and LCB were observed in the HCD MS2 spectra for all the ceramide classes, resulting in unambiguous definition of the ceramide structures, including the chain length and the modification (alpha-hydroxy-, beta-hydroxy-, omega-hydroxy-FA) of the fatty acyl moiety, and the types of LCB (sphingosine, phytosphigosine, 6-hydroxysphingosine). Thereby, this approach permits differentiation of isomeric structures and ceramide species in the biological specimen can be unveiled in detail. By application of sequential MS3, the double bond position along the fatty acyl chain of the molecule can be located, and a complete structural characterization of ceramides can be achieved. (C) 2020 Elsevier B.V. All rights reserved.

Automated summary

This study utilized HCD MS2 spectra to investigate the fragmentation processes of ceramide molecules, allowing for accurate definition of ceramide structures based on observed sets of fragment ions reflecting the fatty acyl chain and LCB. The approach permits differentiation of isomeric structures and detailed characterization of ceramide species in biological samples.