Deep Characterisation of the sn-Isomer Lipidome Using High-Throughput Data-Independent Acquisition and Ozone-Induced Dissociation

We report a workflow that enables untargeted characterization of hundreds of sn-resolved glycerophospholipid isomers from biological extracts in less than 20 min. It reveals that sn-isomer populations are tightly regulated and significantly different between cell lines and enables identification of rare lipids containing ultra long chain monounsaturated fatty acyl chains at the sn-1 position. It provides an order of magnitude increases over the number of sn-isomer pairs previously reported and is coupled with an automated data analysis pipeline.