Trapped Ion Mobility Spectrometry (TIMS) separates ions based on their mobility across an electric field and with subsequent release based on gas phase mobility. Parallel accumulation serial fragmentation (PASEF) synchronizes TIMS with MS/MS precursor selection allowing fragmentation of multiple precursors per scan. dia-PASEF-MS is reported to have higher sensitivity and quantitative accuracy. As an applied bioanalytical tool in clinical research, MS based biomarker discovery involves quantifying hundreds of plasma proteins. Here, we present a robust workflow combining dia-PASEF-MS with a standardized liquid chromatography platform on a timsTOF for quantitative and accurate biomarker discovery.After implementation of a robust dia-PASEF workflow, we identified candidate markers of myocardial-demand ischemia for earlier therapeutic intervention in patients with clinical suspicion of a heart attack.