dia-PASEF combines the advantages of data-independent acquisition (DIA), such as reproducible and accurate protein identification and quantification, with the benefits of ion mobility as additional separation dimension for the high-throughput analysis of complex proteomics samples. The ion mobility information can be used for alignment of precursor and fragment spectra, leading to higher confidence in protein identification. Herein, we apply the dia-PASEF workflow in combination with an Evosep One (Evosep) chromatography system for the high-throughput analysis of eHAP cell line digests, while quantifying spiked-in, oxidized target peptides in Spectronaut TM (Biognosys) on different gradient lengths.