posted on 2022-01-20, 20:11authored byTim J.
H. Jonkers, Jeroen Meijer, Jelle J. Vlaanderen, Roel C. H. Vermeulen, Corine J. Houtman, Timo Hamers, Marja H. Lamoree
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.