Quantitative Exposomics Targeting over 200 Toxicants and Key Biomarkers at the Picomolar Level

The exposome encompasses
environmental exposures throughout life
and significantly impacts health and disease. Exposure chemicals,
present at trace levels, are frequently quantified using targeted
LC–MS/MS. Many existing methods are limited to a narrow range
of analyte classes or lack sufficient sensitivity for exposomic analyses,
and applicability to large sample cohorts for exposome-wide association
studies (ExWAS) remains to be demonstrated. Here, we present a scalable,
fit-for-purpose next-generation human biomonitoring (HBM) workflow
for analyzing >230 biomarkers in urine, plasma, and serum using
solid-phase
extraction in 96-well plates and LC–MS/MS. Moreover, a complementary
conceptual framework for validation criteria of assays designed to
analyze large panels of highly diverse compounds at trace levels is
proposed. Method robustness was evaluated, demonstrating extraction
recovery (60–130%), matrix effects (SSE, 60–130%), inter-/intraday
precision (RSD <30%), and high sensitivity (limit of detection
<0.1 ng/mL) for 59–80% of the analytes across the investigated
biological matrices. To showcase the method’s applicability
in epidemiological studies, 200 urine samples from pregnant women
in a longitudinal pregnancy cohort were analyzed. More than 100 analytes
including PFAS, drugs, air pollutants, pesticides, flame retardants,
mycotoxins, industrial products, food processing contaminants, plastics-related
chemicals, and phytotoxins, were detected, several for the first time
in a U.S. urinary biomonitoring study. With its broad analyte coverage,
ultimate sensitivity, robustness, and high sample throughput, this
method meets the performance requirements for future large-scale ExWAS
applications in public and personalized prevention research.