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Identification of putative steroid receptor antagonists in bottled water: combining bioassays and high-resolution mass spectrometry.

Wagner M, Schlüsener MP, Ternes TA, Oehlmann J - PLoS ONE (2013)

Bottom Line: We detected significant antiestrogenicity in 13 of 18 products. 16 samples were antiandrogenic inhibiting the androgen receptor by up to 90%.Since DEHF is antiestrogenic but not antiandrogenic we conclude that additional, yet unidentified EDCs must contribute to the antagonistic effect of bottled water.This illustrates the need to identify novel toxicologically relevant compounds to establish a more holistic picture of the human exposome.

View Article: PubMed Central - PubMed

Affiliation: Department Aquatic Ecotoxicology, Faculty of Biological Sciences, Goethe University Frankfurt, Frankfurt am Main, Germany.

ABSTRACT
Endocrine disrupting chemicals (EDCs) are man-made compounds interfering with hormone signaling and thereby adversely affecting human health. Recent reports provide evidence for the presence of EDCs in commercially available bottled water, including steroid receptor agonists and antagonists. However, since these findings are based on biological data the causative chemicals remain unidentified and, therefore, inaccessible for toxicological evaluation. Thus, the aim of this study is to assess the antiestrogenic and antiandrogenic activity of bottled water and to identify the causative steroid receptor antagonists. We evaluated the antiestrogenic and antiandrogenic activity of 18 bottled water products in reporter gene assays for human estrogen receptor alpha and androgen receptor. Using nontarget high-resolution mass spectrometry (LTQ-Orbitrap Velos), we acquired corresponding analytical data. We combined the biological and chemical information to determine the exact mass of the tentative steroid receptor antagonist. Further MS(n) experiments elucidated the molecule's structure and enabled its identification. We detected significant antiestrogenicity in 13 of 18 products. 16 samples were antiandrogenic inhibiting the androgen receptor by up to 90%. Nontarget chemical analysis revealed that out of 24520 candidates present in bottled water one was consistently correlated with the antagonistic activity. By combining experimental and in silico MS(n) data we identified this compound as di(2-ethylhexyl) fumarate (DEHF). We confirmed the identity and biological activity of DEHF and additional isomers of dioctyl fumarate and maleate using authentic standards. Since DEHF is antiestrogenic but not antiandrogenic we conclude that additional, yet unidentified EDCs must contribute to the antagonistic effect of bottled water. Applying a novel approach to combine biological and chemical analysis this is the first study to identify so far unknown EDCs in bottled water. Notably, dioctyl fumarates and maleates have been overlooked by science and regulation to date. This illustrates the need to identify novel toxicologically relevant compounds to establish a more holistic picture of the human exposome.

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Antiestrogenic (A) and antiandrogenic activity (B) of 18 bottled waters.13 waters significantly inhibit estrogen receptor alpha, 16 samples antagonize androgen receptor (★★★p<0.0001, compared to controls with endogenous ligand). The activity was normalized to controls containing 17β-estradiol or testosterone (0% inhibition) and such without (100% inhibition). The results represent three extracts per sample tested in three experiments with eight replicates each.
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pone-0072472-g001: Antiestrogenic (A) and antiandrogenic activity (B) of 18 bottled waters.13 waters significantly inhibit estrogen receptor alpha, 16 samples antagonize androgen receptor (★★★p<0.0001, compared to controls with endogenous ligand). The activity was normalized to controls containing 17β-estradiol or testosterone (0% inhibition) and such without (100% inhibition). The results represent three extracts per sample tested in three experiments with eight replicates each.

Mentions: Extracted with the optimized method, the majority of bottled water products significantly inhibited human estrogen as well as androgen receptor. In the YAES 13 products were antiestrogenic (Figure 1 A) with an inhibition of 19.2 (±1.97) to 61.1 (±2.09)%. We detected significant antiandrogenic activity in 16 samples in the YAAS (Figure 1 B). Here, antagonistic activity ranged from 19.0 (±1.66) to 92.3 (±0.88)%. The samples’ potential to antagonize estrogen and androgen receptor is significantly correlated (p<0.001, r = 0.937, Figure S3). Tap water extracted as blank did not induce any significant activity documenting that the procedure does not lead to a contamination with antiestrogens or antiandrogens.


Identification of putative steroid receptor antagonists in bottled water: combining bioassays and high-resolution mass spectrometry.

Wagner M, Schlüsener MP, Ternes TA, Oehlmann J - PLoS ONE (2013)

Antiestrogenic (A) and antiandrogenic activity (B) of 18 bottled waters.13 waters significantly inhibit estrogen receptor alpha, 16 samples antagonize androgen receptor (★★★p<0.0001, compared to controls with endogenous ligand). The activity was normalized to controls containing 17β-estradiol or testosterone (0% inhibition) and such without (100% inhibition). The results represent three extracts per sample tested in three experiments with eight replicates each.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3756062&req=5

pone-0072472-g001: Antiestrogenic (A) and antiandrogenic activity (B) of 18 bottled waters.13 waters significantly inhibit estrogen receptor alpha, 16 samples antagonize androgen receptor (★★★p<0.0001, compared to controls with endogenous ligand). The activity was normalized to controls containing 17β-estradiol or testosterone (0% inhibition) and such without (100% inhibition). The results represent three extracts per sample tested in three experiments with eight replicates each.
Mentions: Extracted with the optimized method, the majority of bottled water products significantly inhibited human estrogen as well as androgen receptor. In the YAES 13 products were antiestrogenic (Figure 1 A) with an inhibition of 19.2 (±1.97) to 61.1 (±2.09)%. We detected significant antiandrogenic activity in 16 samples in the YAAS (Figure 1 B). Here, antagonistic activity ranged from 19.0 (±1.66) to 92.3 (±0.88)%. The samples’ potential to antagonize estrogen and androgen receptor is significantly correlated (p<0.001, r = 0.937, Figure S3). Tap water extracted as blank did not induce any significant activity documenting that the procedure does not lead to a contamination with antiestrogens or antiandrogens.

Bottom Line: We detected significant antiestrogenicity in 13 of 18 products. 16 samples were antiandrogenic inhibiting the androgen receptor by up to 90%.Since DEHF is antiestrogenic but not antiandrogenic we conclude that additional, yet unidentified EDCs must contribute to the antagonistic effect of bottled water.This illustrates the need to identify novel toxicologically relevant compounds to establish a more holistic picture of the human exposome.

View Article: PubMed Central - PubMed

Affiliation: Department Aquatic Ecotoxicology, Faculty of Biological Sciences, Goethe University Frankfurt, Frankfurt am Main, Germany.

ABSTRACT
Endocrine disrupting chemicals (EDCs) are man-made compounds interfering with hormone signaling and thereby adversely affecting human health. Recent reports provide evidence for the presence of EDCs in commercially available bottled water, including steroid receptor agonists and antagonists. However, since these findings are based on biological data the causative chemicals remain unidentified and, therefore, inaccessible for toxicological evaluation. Thus, the aim of this study is to assess the antiestrogenic and antiandrogenic activity of bottled water and to identify the causative steroid receptor antagonists. We evaluated the antiestrogenic and antiandrogenic activity of 18 bottled water products in reporter gene assays for human estrogen receptor alpha and androgen receptor. Using nontarget high-resolution mass spectrometry (LTQ-Orbitrap Velos), we acquired corresponding analytical data. We combined the biological and chemical information to determine the exact mass of the tentative steroid receptor antagonist. Further MS(n) experiments elucidated the molecule's structure and enabled its identification. We detected significant antiestrogenicity in 13 of 18 products. 16 samples were antiandrogenic inhibiting the androgen receptor by up to 90%. Nontarget chemical analysis revealed that out of 24520 candidates present in bottled water one was consistently correlated with the antagonistic activity. By combining experimental and in silico MS(n) data we identified this compound as di(2-ethylhexyl) fumarate (DEHF). We confirmed the identity and biological activity of DEHF and additional isomers of dioctyl fumarate and maleate using authentic standards. Since DEHF is antiestrogenic but not antiandrogenic we conclude that additional, yet unidentified EDCs must contribute to the antagonistic effect of bottled water. Applying a novel approach to combine biological and chemical analysis this is the first study to identify so far unknown EDCs in bottled water. Notably, dioctyl fumarates and maleates have been overlooked by science and regulation to date. This illustrates the need to identify novel toxicologically relevant compounds to establish a more holistic picture of the human exposome.

Show MeSH
Related in: MedlinePlus