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

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.

High-Efficiency Effect-Directed Analysis Leveraging Five High Level Advancements: A Critical Review

“…53 The improvement of resolving power (RP) of mass spectrometry can also promote the identification of causal toxicants, the high RP and precise mass accuracy of HRMS facilitate the identification of unknown substances by differentiating ions with similar m/z ratios and determining their molecular structure by the fragmentation patterns. 158 Highresolution mass spectrometers have different mass accuracy in terms of mass deviation. Fourier-transform ion cyclotron resonance (FT-ICR) mass spectrometry (RP ≥ 1,000,000, mass accuracy ≤1 ppm, and scan rate ≤1 Hz) provides the most precise mass accuracy, followed by Orbitrap mass spectrometry (RP ≥ 100,000, mass accuracy ≤2 ppm, and scan rate <45 Hz) and time-of-flight (TOF) mass spectrometry (RP ≥ 20,000, mass accuracy ≤5 ppm, and scan rate <500 Hz).…”

Effect‐Directed Analysis Based on Transthyretin Binding Activity of Per‐ and Polyfluoroalkyl Substances in a Contaminated Sediment Extract

“…Thus, competitive binding of other substances to TTR results in displacement of bound FITC-T 4 from the TTR and consequently more free FITC-T 4 in the assay, which can be quantified as a decrease in fluorescence. The method, which was originally developed by Ren & Guo (2012), was performed as described by Jonkers et al (2022), with one modification, that is, incubations were performed in the dark for 5 min on a plate shaker (600 rpm, Titramax 1000; Heidolph), followed by a 15-min incubation without shaking. The final volume of the assay was 200 µl.…”

Mixture Risk Assessment of Complex Real-Life Mixtures—The PANORAMIX Project

“…Furthermore, to improve HRMS data annotation quality and enhance the identification success rate, different approaches for the identification of chemicals were reported, such as HaloSeeker to aid the identification of halogenated chemicals [ 21 ] and the CECscreen database, to support the annotation of chemicals of emerging concern and their (human) metabolites [ 45 ]. A workflow incorporating EDA for feature prioritisation in SS and NTS was developed and validated [ 46 , 47 ].…”