Off-line LCxLC-Mass Spectrometry Workflow using CID and EAD for the Quantification and Comprehensive Structural Double Bond Characterization of Glycerophospholipids in Plasma Samples
Year: 2026
Type: preprint
Abstract: Here we describe an automated workflow using a PAL Robotic Tool Changer (RTC) system for plasma lipid sample preparation and off-line two-dimensional liquid chromatography (LCxLC) separation. The orthogonal chromatographic platform couples hydrophilic interaction liquid chromatography (HILIC) in the first dimension with rapid reversed-phase liquid chromatography (RPLC) in the second dimension, facilitated by automated fraction collection at 10-second intervals into 96-well plates. Lipid profiling and structural identification strategies were evaluated using collision-induced dissociation (CID) and electron-activated dissociation (EAD) across data-dependent acquisition (DDA), dataindependent acquisition (DIA / SWATH), and high-resolution multiple reaction monitoring (HRMRM) modes. Standard Reference Material NIST SRM 1950 (Human Plasma) was employed for lipid quantification and double-bond characterization of phosphatidylcholines (PCs). By combining automated modified Matyas liquid-liquid extraction, and off-line LCxLC, a total of 115 PC’s lipids could be detected in SRM 1950 plasma sample. For 74 PC’s including isomers, double assignments could be proposed based on the EAD spectra.
Source: ChemRxiv
Institution: University of Geneva
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W7220737595