TY - JOUR
T1 - High-Throughput Effect-Directed Analysis Using Downscaled in Vitro Reporter Gene Assays to Identify Endocrine Disruptors in Surface Water
AU - Zwart, Nick
AU - Nio, Shan Li
AU - Houtman, Corine J.
AU - De Boer, Jacob
AU - Kool, Jeroen
AU - Hamers, Timo
AU - Lamoree, Marja H.
PY - 2018/4/3
Y1 - 2018/4/3
N2 - Effect-directed analysis (EDA) is a commonly used approach for effect-based identification of endocrine disruptive chemicals in complex (environmental) mixtures. However, for routine toxicity assessment of, for example, water samples, current EDA approaches are considered time-consuming and laborious. We achieved faster EDA and identification by downscaling of sensitive cell-based hormone reporter gene assays and increasing fractionation resolution to allow testing of smaller fractions with reduced complexity. The high-resolution EDA approach is demonstrated by analysis of four environmental passive sampler extracts. Downscaling of the assays to a 384-well format allowed analysis of 64 fractions in triplicate (or 192 fractions without technical replicates) without affecting sensitivity compared to the standard 96-well format. Through a parallel exposure method, agonistic and antagonistic androgen and estrogen receptor activity could be measured in a single experiment following a single fractionation. From 16 selected candidate compounds, identified through nontargeted analysis, 13 could be confirmed chemically and 10 were found to be biologically active, of which the most potent nonsteroidal estrogens were identified as oxybenzone and piperine. The increased fractionation resolution and the higher throughput that downscaling provides allow for future application in routine high-resolution screening of large numbers of samples in order to accelerate identification of (emerging) endocrine disruptors.
AB - Effect-directed analysis (EDA) is a commonly used approach for effect-based identification of endocrine disruptive chemicals in complex (environmental) mixtures. However, for routine toxicity assessment of, for example, water samples, current EDA approaches are considered time-consuming and laborious. We achieved faster EDA and identification by downscaling of sensitive cell-based hormone reporter gene assays and increasing fractionation resolution to allow testing of smaller fractions with reduced complexity. The high-resolution EDA approach is demonstrated by analysis of four environmental passive sampler extracts. Downscaling of the assays to a 384-well format allowed analysis of 64 fractions in triplicate (or 192 fractions without technical replicates) without affecting sensitivity compared to the standard 96-well format. Through a parallel exposure method, agonistic and antagonistic androgen and estrogen receptor activity could be measured in a single experiment following a single fractionation. From 16 selected candidate compounds, identified through nontargeted analysis, 13 could be confirmed chemically and 10 were found to be biologically active, of which the most potent nonsteroidal estrogens were identified as oxybenzone and piperine. The increased fractionation resolution and the higher throughput that downscaling provides allow for future application in routine high-resolution screening of large numbers of samples in order to accelerate identification of (emerging) endocrine disruptors.
UR - http://www.scopus.com/inward/record.url?scp=85045019997&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85045019997&partnerID=8YFLogxK
U2 - 10.1021/acs.est.7b06604
DO - 10.1021/acs.est.7b06604
M3 - Article
AN - SCOPUS:85045019997
VL - 52
SP - 4367
EP - 4377
JO - Environmental Science and Technology
JF - Environmental Science and Technology
SN - 0013-936X
IS - 7
ER -