Abstract
Characterizing the chemical exposome relies on advanced instrumentation including tandem mass spectrometry coupled to liquid chromatography (LC-MS/MS), and nontargeted analysis (NTA) using high-resolution MS. However, proper sample pretreatment, balancing broad analyte coverage, method robustness, and throughput remain a major bottleneck in exposomics. Here, we developed a robust and scalable solid phase extraction (SPE) protocol for human urine and plasma and optimized it for a panel of 94 highly diverse environmental and food-related contaminants (LogP -0.7 to 6.8). Extraction recoveries (RE) and signal suppression and enhancement (SSE) were determined using targeted LC-MS/MS. Acceptable REs (60-140%) were achieved for >70% of analytes, and acceptable SSE values (60-140%) for 86% and 90% in urine and plasma, respectively. Subsequently, the method was transferred to 96-well plate format, significantly improving throughput to meet the capacity requirements needed for exposome-wide association studies (ExWAS). The established workflow is approximately 10× faster than routinely used metabolomics-based protein precipitation approaches when comparing the estimated total analysis time for 1000 samples. The method's applicability for NTA and suspect screening was tested and compared to a generic protein precipitation protocol using NIST standard reference materials for urine (SRM 3672) and plasma (SRM 1950). Favorable performance was shown for the protein precipitation workflow while the SPE protocol demonstrated promising results. The developed workflow is thus not only superior for future high-throughput targeted exposomics but also offers an option for NTA applications. The presented well-balanced approach is scalable and also applicable to research in the fields of pharmacology, food safety, and systems toxicology.