Pre-validation of choriogenin H transgenic medaka eleutheroembryos as a quantitative estrogenic activity test method.

Journal Article (Journal Article)

The choriogenin H - EGFP transgenic medaka (Oryzias melastigma) has been used to test estrogenic substances and quantify estrogenic activity into 17β-estradiol (E2) equivalency (EEQ). The method uses 8 eleutheroembryos in 2 ml solution per well and 3 wells per treatment in 24-well plates at 26 ± 1 °C for 24 ± 2 h, with subsequent measurements of induced GFP signal intensity. EEQ measurements are calculated using a E2 probit regression model with a coefficient of determination (R2 ) > 0.90. The selectivity was confirmed evaluating 27 known estrogenic and 5 known non-estrogenic compounds. Limit of quantitation (LOQ), recovery rate and bias were calculated to be 1 ng/ml EEQ, 104% and 4% respectively. Robustness analysis revealed exposure temperature is a sensitive parameter that should be kept at 26 ± 1 °C. The repeatability of intra- and inter-laboratories achieved CV < 30% for most tested food and cosmetics samples. The lot-lot stability was confirmed by the stable EEQ qualitative control (QC, 1 ng/mL E2) and calibration curve results. The stability of standard reagents, samples and sample extracts was also investigated. These data demonstrated this method to be an accurate indicator of estrogenic activity for both chemicals and extracts.

Full Text

Duke Authors

Cited Authors

  • Chen, X; Cheng, SH; Kinoshita, M; de Witte, PA; Liu, J; Hinton, D; Braunbeck, T; Cotgreave, I; Schlenk, D; Gong, Z; El-Nezami, H; Ho, KC; Chan, KF; Xu, S; Yiu, PY; Zhang, H; Wu, D; Chan, YS; Ny, A; Maes, J

Published Date

  • September 2021

Published In

Volume / Issue

  • 629 /

Start / End Page

  • 114311 -

PubMed ID

  • 34302800

Electronic International Standard Serial Number (EISSN)

  • 1096-0309

International Standard Serial Number (ISSN)

  • 0003-2697

Digital Object Identifier (DOI)

  • 10.1016/j.ab.2021.114311


  • eng