Compounds with estrogenic potencies and their adverse effects in surface waters have received much attention. Both anthropogenic and natural compounds contribute to overall estrogenic activity in freshwaters. Recently, estrogenic potencies were also found to be associated with cyanobacteria and their blooms in surface waters. The present study developed and compared the solid phase extraction and LC-MS/MS analytical approaches for determination of phytoestrogens (8 flavonoids - biochanin A, coumestrol, daidzein, equol, formononetin, genistein, naringenin, apigenin - and 5 sterols - ergosterol, β-sitosterol, stigmasterol, campesterol, brassicasterol) and cholesterol in water. The method was used for analyses of samples collected in stagnant water bodies dominated by different cyanobacterial species. Concentrations of individual flavonoids ranged from below the limit of detection to 3.58 ng/L. Sterols were present in higher amounts up to 2.25 μg/L. Biological potencies of these phytoestrogens in vitro were characterized using the hERα-HeLa-9903 cell line. The relative estrogenic potencies (compared to model estrogen - 17β-estradiol) of flavonoids ranged from 2.25E-05 to 1.26E-03 with coumestrol being the most potent. None of the sterols elicited estrogenic response in the used bioassay. Estrogenic activity was detected in collected field water samples (maximum effect corresponding to 2.07 ng/L of 17β-estradiol equivalents, transcriptional assay). At maximum phytoestrogens accounted for only 1.56 pg/L of 17β-estradiol equivalents, contributing maximally 8.5% of the total estrogenicity of the water samples. Other compounds therefore, most likely of anthropogenic origin such as steroid estrogens, are probably the major drivers of total estrogenic effects in these surface waters.
- MeSH
- Water Pollutants, Chemical analysis MeSH
- Cholestadienols MeSH
- Cholesterol analogs & derivatives MeSH
- Estradiol analysis MeSH
- Estrogens analysis MeSH
- Estrone analysis MeSH
- Phytoestrogens analysis MeSH
- Phytosterols MeSH
- Genistein analysis MeSH
- HeLa Cells MeSH
- Isoflavones analysis MeSH
- Humans MeSH
- Receptors, Estrogen metabolism MeSH
- Cyanobacteria drug effects metabolism MeSH
- Sitosterols analysis MeSH
- Fresh Water MeSH
- Sterols analysis MeSH
- Tandem Mass Spectrometry MeSH
- Water MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
Bisphenol A (BPA) is a widely known endocrine disruptor with estrogenic, antiestrogenic or antiandrogenic properties. BPA could interfere with estrogen metabolism as well with receptor-mediated estrogen actions. Both environmental BPA and estrogens may be traced in body fluids, of which, besides the blood plasma, the seminal fluid is of particular interest regarding their possible interactions in the testis. The method for simultaneously determining BPA and estrogens is then needed, taking into account that their concentrations in these body fluid may differ. Here the method was developed and validated for measurements of BPA, estrone (E1), estradiol (E2) and estriol (E3) in blood plasma and seminal plasma using liquid chromatography-tandem mass spectrometry. Due to the phenolic moiety of all compounds, dansyl chloride derivatization could be used. The analytical criteria of the method with respect to expected concentration of the analytes were satisfactory. The lower limits of quantifications (LLOQ) amounted to 43.5, 4.0, 12.7, 6.7 pg/mL for plasma BPA, E1, E2 and E3, and 28.9, 4.9, 4.5, 3.4 pg/mL for seminal BPA, E1, E2 and E3, respectively. The concentrations of individual steroids differed between body fluids. To the best of our knowledge, this is the first method that enabled the measurement of estrogens and BPA together in one run. The concentrations of E1, E2 and for the first time also of E3 in seminal plasma in normospermic men are reported.
- MeSH
- Benzhydryl Compounds analysis blood MeSH
- Chromatography, Liquid methods MeSH
- Endocrine Disruptors analysis blood MeSH
- Estrogens analysis blood MeSH
- Estrone analysis blood MeSH
- Phenols analysis blood MeSH
- Humans MeSH
- Limit of Detection MeSH
- Semen chemistry MeSH
- Tandem Mass Spectrometry methods MeSH
- Check Tag
- Humans MeSH
- Male MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Validation Study MeSH
- MeSH
- Estradiol analysis blood metabolism MeSH
- Estrogen Replacement Therapy MeSH
- Estrogens analysis blood metabolism MeSH
- Estrone analysis blood metabolism MeSH
- Humans MeSH
- Menopause physiology blood metabolism MeSH
- Osteocalcin drug effects blood metabolism MeSH
- Osteoporosis, Postmenopausal MeSH
- Check Tag
- Humans MeSH
- Female MeSH
