High-Performance Data Processing Workflow Incorporating Effect-Directed Analysis for Feature Prioritization in Suspect and Nontarget Screening.

Roel C H Vermeulen,Jelle J Vlaanderen,Jeroen Meijer,Marja H Lamoree,Timo Hamers,Tim J H Jonkers,Corine J Houtman

doi:10.1021/acs.est.1c04168

Roel C H Vermeulen, Jelle J Vlaanderen + Show 5 more

Open Access

https://doi.org/10.1021/acs.est.1c04168

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Abstract

Effect-directed analysis (EDA) aims at the detection of bioactive chemicals of emerging concern (CECs) by combining toxicity testing and high-resolution mass spectrometry (HRMS). However, consolidation of toxicological and chemical analysis techniques to identify bioactive CECs remains challenging and laborious. In this study, we incorporate state-of-the-art identification approaches in EDA and propose a robust workflow for the high-throughput screening of CECs in environmental and human samples. Three different sample types were extracted and chemically analyzed using a single high-performance liquid chromatography HRMS method. Chemical features were annotated by suspect screening with several reference databases. Annotation quality was assessed using an automated scoring system. In parallel, the extracts were fractionated into 80 micro-fractions each covering a couple of seconds from the chromatogram run and tested for bioactivity in two bioassays. The EDA workflow prioritized and identified chemical features related to bioactive fractions with varying levels of confidence. Confidence levels were improved with the in silico software tools MetFrag and the retention time indices platform. The toxicological and chemical data quality was comparable between the use of single and multiple technical replicates. The proposed workflow incorporating EDA for feature prioritization in suspect and nontarget screening paves the way for the routine identification of CECs in a high-throughput manner.

Highlights

The focus of chemical screening has shifted over the last decade from targeted analysis of a limited group of known compounds to new screening techniques to detect a broader spectrum of chemicals that are of emerging concern
Chromatography coupled to high-resolution mass spectrometry (HRMS) allows for the detection of several thousands of accurate masses in a sample in a single measurement regardless of their origin and whether they are of concern for environmental or human health.[1]
An experiment-driven prioritization approach is effect-directed analysis (EDA), which guides screening efforts to those chemical features that exhibit a toxicological mechanism of action related to an adverse outcome

Summary

Introduction

The focus of chemical screening has shifted over the last decade from targeted analysis of a limited group of known compounds to new screening techniques to detect a broader spectrum of chemicals that are of emerging concern. Chromatography coupled to high-resolution mass spectrometry (HRMS) allows for the detection of several thousands of accurate masses (features) in a sample in a single measurement regardless of their origin and whether they are of concern for environmental or human health.[1] Nontarget and suspect screening allows for the annotation of features; chemical identification remains challenging and laborious and requires confirmation with analytical standards. Prioritization steps are required to determine which chemical features warrant further identification.[1] An experiment-driven prioritization approach is effect-directed analysis (EDA), which guides screening efforts to those chemical features that exhibit a toxicological mechanism of action related to an adverse outcome. EDA reduces the chemical complexity of the sample and facilitates the high-throughput screening of bioactive chemicals of emerging concern (CECs) in abiotic and biotic samples

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