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

ABSTRACT 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 s ingle‐injection m ulti‐omics a nalysis by d irect 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.