Exposomic Biomonitoring of Polyphenols by Non-Targeted Analysis and Suspect Screening

Polyphenols, prevalentin plants and fungi, are investigated intensivelyin nutritional and clinical settings because of their beneficial bioactiveproperties. Due to their complexity, analysis with untargeted approachesis favorable, which typically use high-resolution mass spectrometry(HRMS) rather than low-resolution mass spectrometry (LRMS). Here,the advantages of HRMS were evaluated by thoroughly testing untargetedtechniques and available online resources. By applying data-dependentacquisition on real-life urine samples, 27 features were annotatedwith spectral libraries, 88 with in silico fragmentation,and 113 by MS1 matching with PhytoHub, an online databasecontaining >2000 polyphenols. Moreover, other exogenous and endogenousmolecules were screened to measure chemical exposure and potentialmetabolic effects using the Exposome-Explorer database, further annotating144 features. Additional polyphenol-related features were exploredusing various non-targeted analysis techniques including MassQL forglucuronide and sulfate neutral losses, and MetaboAnalyst for statisticalanalysis. As HRMS typically suffers a sensitivity loss compared tostate-of-the-art LRMS used in targeted workflows, the gap betweenthe two instrumental approaches was quantified in three spiked humanmatrices (urine, serum, plasma) as well as real-life urine samples.Both instruments showed feasible sensitivity, with median limits ofdetection in the spiked samples being 10-18 ng/mL for HRMSand 4.8-5.8 ng/mL for LRMS. The results demonstrate that, despiteits intrinsic limitations, HRMS can readily be used for comprehensivelyinvestigating human polyphenol exposure. In the future, this workis expected to allow for linking human health effects with exposurepatterns and toxicological mixture effects with other xenobiotics.