Screening for estrogen active nonylphenols in surface waters by planar solid phase extraction-planar yeast estrogen screen.

Nonylphenols (NP) are ubiquitous in the environment and show toxic effects and estrogenic activity. According to the corresponding framework directive in the field of water quality, isomers of NP (including 4-n-NP and branched 4-NP) are classified as priority hazardous substances and are regulated as a group of chain and ring isomers with a maximum acceptable concentration of 2 μg/L in inland and other surface waters. This study presents a new sensitive and innovative screening approach for estrogen active NP based on high-performance thin-layer chromatography.

Detection of estrogen active compounds in hops by planar yeast estrogen screen.

Hops used in the brewing process of beer for flavoring are known to contain estrogen active compounds (EAC) and to be the source of EAC in beer. The recently developed planar yeast estrogen screen (pYES) with the substrate resorufin-β-d-galactopyranoside (RGP) successfully was applied for the detection of EAC in ethanolic extracts of hops pellet samples. The only pYES positive compound was identified as the hop flavanone prenylnaringenin (PN) by thin-layer chromatography-mass spectrometry.

Logit-log evaluation of planar yeast estrogen screens.

Receptor assays like the yeast estrogen screen (YES) performed in microtiter plates normally provide dose-response curves with a sigmoidal shape in semi-log plots. Such sigmoidal plots can be linearized by the logit function resulting in logit-log plots, as mainly known for the evaluation of enzyme-linked immunosorbent assays and radioimmunoassays. Since the planar yeast estrogen screen (pYES) represents the transfer of the receptor assay YES to high-performance thin-layer chromatography (HPTLC), it was assumed to obtain sigmoidal shaped dose-response curves from the measured signals, which subsequently could be used to generate logit-log plots.

Planar yeast estrogen screen with resorufin-β-d-galactopyranoside as substrate.

For the planar yeast estrogen screen (pYES), 4-methylumbelliferyl-β-d-galactopyranoside was generally employed as substrate, delivering blue fluorescing 4-methylumbelliferone after enzymatic cleavage by the YES reporter β-d-galactosidase as the positive signal for the presence of estrogen active compounds (EAC). As environmental samples like waste water also contain blue fluorescent components, it is difficult to differentiate them from pYES signals. Therefore, resorufin-β-d-galactopyranoside (RGP), providing the orange fluorescing resorufin after enzymatic cleavage, was introduced as pYES substrate to determine EAC.