Assessment of estrogenic activity of leachate from automobile tires with two in vitro bioassays

In this study, a combination of enzyme-linked receptor assay (ELRA) and yeast estrogen screen (YES) assay was firstly applied to determine whether automobile tires immersed in fresh water can leach chemicals, which display estrogenic activity. We optimized ELRA substituting the chromogene substrate by a luminescent one, and found that luminescent ELBA was more sensitive to 17 beta-estradiol (17 beta-E2) with a detection limit of 0.016 mu g/l, compared to 0.088 mu g/l in the chromogene version. In ELRA, all tire leachates obviously showed estrogenic activity, which was increased with duration of immersion. Moreover, the leachate from hackled tires showed more potent estrogenicity than that from the whole ones. In comparison to ELRA, no detectable estrogenic activity was found in all tire leachates with YES assay. The results from YES assay further evidenced that antiestrogenic compounds can be leached from tires. As tire leachates contain estrogenic compounds, they could be important pollution sources, potentially harmful to wildlife and human health. Thus, use of shredded tires as road fill or in landfill sites should arouse our attention.

Comparative study of estrogenic potencies of estradiol, tamoxifen, bisphenol-A and resveratrol with two in vitro bioassays

This study was undertaken to compare the sensitivity of two in vitro screening test methods and to determine the accuracy of predicted response to spiked laboratory water samples. A newly developed enzyme-linked receptor assay (ELRA) and a widely used yeast estrogen screen (YES) assay were selected to evaluate the estrogenic responses. Four natural, pharmaceutical, xenobiotic or phytobiotic chemicals: 17beta-estradiol (E2), tamoxifen, bisphenol-A and resveratrol were examined, and 17beta-E2 was used as a positive control. 17beta-E2 can strongly induce estrogenic response in both test systems, however, ELRA was found to be more sensitive to 17beta-E2 with a detection limit of 0.07 mug/l compared to 0.88 mug/l in YES assay. Similar results were obtained for bisphenol-A and resveratrol, and their estrogen potencies relative to E2 (100%) determined by ELRA were at least 5.6 times greater than produced by YES assay. ELRA was unable to distinguish the anti-estrogen tamoxifen and YES assay is also poor at distinguishing. Comparison of response to spiked laboratory water samples show that ELRA can give accurate determination to all four chemicals with recoveries among 70-120%, while YES can only give accurate determination to 17beta-E2 and bisphenol-A with recoveries among 69-112%. The comparative results provide evidence that ELRA is more suitable for rapid screening estrogenic potency of the environmental samples. Combination of ELRA and mammalian cellular assay will constitute an advantageous test to specify agonistic or antagonistic effects. (C) 2003 Elsevier Ltd. All rights reserved.