In conclusion, we present the development of a highthroughput SPE protocol based on 96-well format for faster, cheaper, and more robust omic-scale exposure analysis. This workflow was optimized comprehensively considering a large scale of targeted analytes, and chemical coverage was further compared with a validated PPT method to ensure sufficient analyte coverage for targeted LC-MS/MS and non-targeted LC-HRMS applications. Despite reduced method performance for very polar compounds, the comparison of SPE and PPT workflows clearly indicated that the SPE method is a suitable candidate for large-scale NTA or suspect screening applications, especially due to the outstanding enhancement of sample throughput and instrumental robustness. This is underlined by the comparable overall performance of PPT and SPE workflows for a wide range of analytes, especially when RP-LC is used as a frontend separation technique. A pragmatic balance between reduced matrix complexity and chemical coverage was achieved, and due to the high sample throughput, the presented workflow can be regarded as a suitable candidate for exposome-wide association studies (ExWAS). Future work will include the expansion to even more toxicologically relevant chemicals and testing the capacity of the developed sample preparation workflow in large cohort studies to investigate the impact on instrumental robustness and downtimes.