Single‐Injection Multi‐Omics Analysis by Direct Infusion Mass Spectrometry

Combined proteomics, metabolomics, and lipidomics analyses require long liquid chromatography-mass spectrometry (LC-MS) run times, limiting throughput and increasing costs for large-scale studies. Here, we present single-injection multi-omics analysis by direct infusion (SMAD), an integrated platform leveraging ion mobility mass spectrometry and self-developed software tools to enable single injection multi-omics analysis without liquid chromatography. SMAD allows quantification of over 9000 metabolite m/z features and over 1300 proteins from the same sample in less than 5 min. We validated the efficiency and reliability of SMAD with three case studies: (1) mouse macrophages after M1/M2 polarization and senescence, (2) a pilot drug screen in human cells, and (3) large-scale high-throughput drug screening of mammalian cells in 96-well plates. Finally, relationships between proteomic and metabolomic data are discovered by machine learning and validated.