The Thermo Orbitrap Ascend Tribrid is a high-resolution LC-MS system paired with a Vanquish Neo UHPLC or Vanquish Flex HPLC, offering an extended mass range (m/z 40–16,000), ultra-high resolving power (up to 480,000 at m/z 200), sub-ppm mass accuracy, and femtogram-level sensitivity. It supports fast scanning, polarity switching, and a wide dynamic range (>5,000). Advanced capabilities include FAIMS for improved ion separation, multiple fragmentation techniques (CID, HCD, Easy-ETD and UVPD), and both heated electrospray and nanospray ion sources, enabling flexible, high-performance analysis of small molecule, metabolites, lipids, peptides, and proteins. Designed to excel in TMTpro isobaric mass tagging multiplex quantitation (up to 35-plex) using synchronous precursor selection (SPS) with MS3, it can also perform label-free quantitation using Data-Independent Acquisition (DIA) analysis detecting 1000s of proteins from low input samples. Post-translational modification analysis can be performed on a wide range of modifications.