Here we present the development and application of LC-FAIMS-MS/MS based platform for untargeted lipidomics. The combination of normal phase liquid chromatography (NPLC), which can separate different types of phospholipids based on their head groups, and high-resolution MS (HRMS) offers a powerful qualitative and quantitative workflow for serum lipid profiling.We exploited this NPLC-FAIMS-HRMS to analyze the serum lipid profiles in mice infected intranasally with Acinetobacter baumannii, and developed an MS/MS matching tool to identify the detected lipids.