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

Publication ,  Journal Article
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 ...
Published in: Analytical biochemistry
September 2021

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.

Duke Scholars

Published In

Analytical biochemistry

DOI

EISSN

1096-0309

ISSN

0003-2697

Publication Date

September 2021

Volume

629

Start / End Page

114311

Related Subject Headings

  • Regression Analysis
  • Protein Precursors
  • Oryzias
  • Limit of Detection
  • Estradiol
  • Egg Proteins
  • Cell Extracts
  • Biosensing Techniques
  • Biochemistry & Molecular Biology
  • Animals, Genetically Modified
 

Citation

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Chen, X., Cheng, S. H., Kinoshita, M., de Witte, P. A., Liu, J., Hinton, D., … Maes, J. (2021). Pre-validation of choriogenin H transgenic medaka eleutheroembryos as a quantitative estrogenic activity test method. Analytical Biochemistry, 629, 114311. https://doi.org/10.1016/j.ab.2021.114311
Chen, Xueping, Shuk Han Cheng, Masato Kinoshita, Peter A. de Witte, Jianjun Liu, David Hinton, Thomas Braunbeck, et al. “Pre-validation of choriogenin H transgenic medaka eleutheroembryos as a quantitative estrogenic activity test method.Analytical Biochemistry 629 (September 2021): 114311. https://doi.org/10.1016/j.ab.2021.114311.
Chen X, Cheng SH, Kinoshita M, de Witte PA, Liu J, Hinton D, et al. Pre-validation of choriogenin H transgenic medaka eleutheroembryos as a quantitative estrogenic activity test method. Analytical biochemistry. 2021 Sep;629:114311.
Chen, Xueping, et al. “Pre-validation of choriogenin H transgenic medaka eleutheroembryos as a quantitative estrogenic activity test method.Analytical Biochemistry, vol. 629, Sept. 2021, p. 114311. Epmc, doi:10.1016/j.ab.2021.114311.
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. Pre-validation of choriogenin H transgenic medaka eleutheroembryos as a quantitative estrogenic activity test method. Analytical biochemistry. 2021 Sep;629:114311.
Journal cover image

Published In

Analytical biochemistry

DOI

EISSN

1096-0309

ISSN

0003-2697

Publication Date

September 2021

Volume

629

Start / End Page

114311

Related Subject Headings

  • Regression Analysis
  • Protein Precursors
  • Oryzias
  • Limit of Detection
  • Estradiol
  • Egg Proteins
  • Cell Extracts
  • Biosensing Techniques
  • Biochemistry & Molecular Biology
  • Animals, Genetically Modified