Efficient Low-Cost Procedure for Microextraction of Estrogen from Environmental Water Using Magnetic Ionic Liquids.

In this study, three magnetic ionic liquids (MILs) were investigated for extraction of four estrogens, i.e., estrone (E1), estradiol (E2), estriol (E3), and ethinylestradiol (EE2), from environmental water. The cation trihexyl(tetradecyl)phosphonium ([P66614]+), selected to confer hydrophobicity to the resulting MIL, was combined with tetrachloroferrate(III), ferricyanide, and dysprosium thiocyanate to yield ([P66614][FeCl4]), ([P66614]3[Fe(CN)6]), and ([P66614]5[Dy(SCN)8]), respectively. After evaluation of various strategies to develop a liquid-liquid microextraction technique based on synthesized MILs, we placed the MILs onto a magnetic stir bar and used them as extracting solvents. After extraction, the MIL-enriched phase was dissolved in methanol and injected into an HPLC-UV for qualitative and quantitative analysis. An experimental design was used to simultaneously evaluate the effect of select variables and optimization of extraction conditions to maximize the recovery of the analytes. Under optimum conditions, limits of detection were in the range of 0.2 (for E3 and E2) and 0.5 µg L-1 (for E1), and calibration curves exhibited linearity in the range of 1-1000 µg L-1 with correlation coefficients higher than 0.998. The percent relative standard deviation (RSD) was below 5.0%. Finally, this method was used to determine concentration of estrogens in real lake and sewage water samples.

Remediation efficiency of three treatments on water polluted with endocrine disruptors: Assessment by means of in vitro techniques.

Chemical substances with potential to disrupt endocrine systems have been detected in aquatic environments worldwide, making necessary the investigation about water treatments able to inhibit such potential. The present work aimed to assess the efficiency for removing endocrine disruptors (with estrogenic and androgenic activity) of three simple and inexpensive substrates that could be potentially used in sectors or regions with limited resources: powdered activated carbon (PAC), powdered natural zeolite (ZEO) (both at a concentration of 500 mg L-1) and natural aquatic humic substances (AHS) (at 30 mg L-1). MilliQ-water and mature water from fish facilities (aquarium water, AW), were artificially spiked with 17ß-estradiol (E2), 17α-ethinylestradiol and dihydrotestosterone. Moreover, effluent samples from waste water treatment plants (WWTP) were also submitted to the remediation treatments. Estrogenic and androgenic activities were assessed with two cell lines permanently transfected with luciferase as reporter gene under the control of hormone receptors: AR-EcoScreen containing the human androgen receptor and HER-LUC transfected with the sea bass estrogen receptor. PAC was efficiently removing the estrogenic and androgenic compounds added to milliQ and AW. However, androgenic activity detected in WWTP effluents was only reduced after treatment with ZEO. The higher surface area of PAC could have facilitated the removal of spiked hormones in clean waters. However, it is possible that the substances responsible of the hormonal activity in WWTP have adsorbed to micro and nanoparticles present in suspension that would have been retained with higher efficiency by ZEO that show pores of several microns in size.

Use of a Battery of Chemical and Ecotoxicological Methods for the Assessment of the Efficacy of Wastewater Treatment Processes to Remove Estrogenic Potency.

Endocrine Disrupting Compounds pose a substantial risk to the aquatic environment. Ethinylestradiol (EE2) and estrone (E1) have recently been included in a watch list of environmental pollutants under the European Water Framework Directive. Municipal wastewater treatment plants are major contributors to the estrogenic potency of surface waters. Much of the estrogenic potency of wastewater treatment plant (WWTP) effluents can be attributed to the discharge of steroid estrogens including estradiol (E2), EE2 and E1 due to incomplete removal of these substances at the treatment plant. An evaluation of the efficacy of wastewater treatment processes requires the quantitative determination of individual substances most often undertaken using chemical analysis methods. Most frequently used methods include Gas Chromatography-Mass Spectrometry (GCMS/MS) or Liquid Chromatography-Mass Spectrometry (LCMS/MS) using multiple reaction monitoring (MRM). Although very useful for regulatory purposes, targeted chemical analysis can only provide data on the compounds (and specific metabolites) monitored. Ecotoxicology methods additionally ensure that any by-products produced or unknown estrogenic compounds present are also assessed via measurement of their biological activity. A number of in vitro bioassays including the Yeast Estrogen Screen (YES) are available to measure the estrogenic activity of wastewater samples. Chemical analysis in conjunction with in vivo and in vitro bioassays provides a useful toolbox for assessment of the efficacy and suitability of wastewater treatment processes with respect to estrogenic endocrine disrupting compounds. This paper utilizes a battery of chemical and ecotoxicology tests to assess conventional, advanced and emerging wastewater treatment processes in laboratory and field studies.

Degradation and toxicity assessment of the nonionic surfactant Triton™ X-45 by the peroxymonosulfate/UV-C process.

The degradation and mineralization of the nonionic surfactant octylphenol ethoxylate (OPEO), commercially known as Triton™ X-45, by the peroxymonosulfate (PMS)/UV-C process were investigated. Three different toxicity tests (Daphnia magna, Vibrio fischeri and Pseudokirchneriella subcapitata) as well as the Yeast Estrogen Screen (YES) bioassay were undertaken to evaluate the potential toxic and estrogenic effects of OPEO and its oxidation products. OPEO removal was very fast and complete after 7 min via PMS/UV-C treatment under the investigated reaction conditions (OPEO = 20 mg L(-1) (47 µM); TOC = 12 mg L(-1); PMS = 2.5 mM; initial reaction pH = 6.5; applied UV-C dose = 21 Wh L(-1)). TOC removal also proceeded rapidly; a gradual decrease was observed resulting in an overall TOC removal of 84%. The toxic responses of PMS/UV-C treated OPEO solutions varied according to the test organism used in the bioassay. Daphnia magna was found to be most sensitive to aqueous OPEO, whereas Pseudokirchneriella subcapitata appeared to be the least sensitive one. Daphnia magna and Vibrio fischeri tests revealed that the inhibitory effect of OPEO decreased significantly during the course of treatment. On the other hand, PMS/UV-C oxidation products exhibited a high toxic effect towards Pseudokirchneriella subcapitata (around 60%). YES test results underlined the need for improving the PMS/UV-C treatment performance to remove the estrogenic activity of OPEO and its oxidation products.

Assessment of multiple hormonal activities in wastewater at different stages of treatment.

Changes in the endocrine potency of municipal wastewater at 3 wastewater treatment plants (WWTPs) in Australia were investigated using a panel of in vitro receptor-driven transactivation assays. The assays were based on human estrogen receptor α, androgen receptor, progesterone receptor, glucocorticoid receptor, and peroxisome proliferator-activated receptor γ2. Total removal efficiencies for estrogenic activity in the dissolved phase were 79.8% to 99.4%. Chemical analysis of 17ß-estradiol, estrone, and 17α-ethinylestradiol levels showed that they accounted for the majority of the observed in vitro estrogenic activity in the final effluents but only 18% to 70% of estrogenic activity in the influents. Removal efficiency for androgenic activity was 97.5% to 100%. Endocrine activity levels were low in the final effluent of the WWTP with the lowest catchment population, with only estrogenic activity detected. In the final effluent of the WWTP with an intermediate catchment population, estrogenic, glucocorticoid, and peroxisome proliferator activities were detected. Estrogenic, antiandrogenic, progestagenic, glucocorticoid, and peroxisome proliferator activities were detected in the final effluent of the WWTP with the highest catchment population. The present study confirms the efficacy of secondary and tertiary treatment in reducing the concentrations of endocrine-active compounds in municipal wastewater. Further work is required to determine the possible health risks to aquatic biota posed by multiple hormonal activities present at low levels.

Advanced oxidation kinetics and mechanism of preservative propylparaben degradation in aqueous suspension of TiO2 and risk assessment of its degradation products.

The absolute kinetic rate constants of propylparaben (PPB) in water with different free radicals were investigated, and it was found that both hydroxyl radicals (HO(•)) and hydrated electrons could rapidly react with PPB. The advanced oxidation kinetics and mechanisms of PPB were investigated using photocatalytic process as a model technology, and the degradation was found to be a pseudo-first-order model. Oxidative species, particularly HO(•), were the most important reactive oxygen species mediating photocatalytic degradation of PPB, and PPB degradation was found to be significantly affected by pH because it was controlled by the radical reaction mechanism and was postulated to occur primarily via HO(•)-addition or H-abstraction reactions on the basis of pulse radiolysis measurements and observed reaction products. To investigate potential risk of PPB to humans and aqueous organisms, the estrogenic assays and bioassays were performed using 100 µM PPB solution degraded by photocatalysis at specific intervals. The estrogenic activity decreased as PPB was degraded, while the acute toxicity at three trophic levels first increased slowly and then decreased rapidly as the total organic carbon decreased during photocatalytic degradation.

Improving the performance of membrane bioreactors by powdered activated carbon dosing with cost considerations.

Effects of powdered activated carbon (PAC) dosing on the overall performance of membrane bioreactors (MBR) were investigated in two bench-scale submerged MBRs. Positive impacts of PAC dosing on membrane fouling and the removal of 17beta-estradiol (E2) and 17alpha-ethyinylestradiol (EE2) were demonstrated over a six-month stable operational period. PAC dosing in the MBR increased the removal rates of E2 and EE2 by 3.4% and 15.8%, respectively. The average soluble extracellular polymeric substances (EPS) and colloidal total organic carbon (TOC) concentrations in the PAC-MBR sludge was 60.1% and 61.8% lower than the control MBR sludge, respectively. Lower soluble EPS and colloidal TOC concentrations in the PAC-MBR sludge resulted in a slower rate of trans-membrane pressure (TMP) increase during MBRs operation, which could prolong the lifespan of membranes. Cost assessment showed that PAC dosing could reduce the operating cost for membrane cleaning and/or membrane replacement by about 25%. The operating cost for PAC dosing could be offset by the benefit from its reducing the cost for membrane maintenance.

Assessment of in vivo estrogenic response and the identification of environmental estrogens in the Yangtze River (Nanjing section).

Environmental estrogens in the aquatic environment have been shown to be responsible for the feminization of fish. The estrogenic content of the Yangtze River (Nanjing section--referred to as the studied area herein) was assessed using a combination of bioassay and chemical analysis. The in vivo bioassay was conducted by exposing adult male goldfish (Carassius auratus) to different concentrations of river water (25%, 50% and 100%) sampled from three representative sections of the studied area. Chemical analysis of estrogens in water from the three representative sections was conducted using solid phase extraction-gas chromatograph (SPE-GC) detection. The assay showed significant serum vitellogenin (VTG) and 17beta-estradiol (E2) induction and gonad atrophy in the treated fish. The strength of in vivo estrogenic responses in the three representative sections is in the order of Jiangxinzhou section>Daqiao section>Sanchahe section. The result is consistent with the levels of water estrogens determined from the chemical analysis. Steroidal estrogens were the major causal agents responsible for the estrogenic responses in the Jiangxinzhou and Daqiao sections, while phenolic estrogens were the main contributors in the Sanchahe section. The results of these in vivo bioassay and chemical analysis demonstrate that fish in the Yangtze River are exposed to environmental estrogens and are at a risk of feminization.

Assessment of trace organic chemical removal by a membrane bioreactor using gas chromatography/mass spectrometry and a yeast screen bioassay.

A membrane bioreactor (MBR) was assessed for the removal of estrogens, androgens, and a selection of pharmaceuticals and personal care products. The biomass and aqueous components of the MBR were investigated to determine whether removal was by biodegradation or by adsorption to the biomass. Removal was monitored using chemical analysis by gas chromatography/mass spectrometry (GC-MS) as well as biological analysis using estrogenic and androgenic yeast assays. Results showed that the MBR was effective in removing the compounds of concern from raw influent with removal rates between 78 and 99%. Removal efficiencies were comparable or better than those reported for conventional activated sludge systems, which was attributed to the relatively high sludge retention time of the MBR. The biomass component showed significant concentrations of salicylic acid, triclosan, and 4-tert-octylphenol. Estrogenic and androgenic activity was also measured in the biomass. Estrone was identified as the main compound responsible for the estrogenic activity. It was concluded that the main removal pathway was biodegradation, but sorption to biomass may also be important, particularly for triclosan and 4-tert-octylphenol.

[Behaviors and environmental risk assessment of estrogenic endocrine disruptors in anaerobic domestic wastewater treatment].

Solid phase extraction-LC/MS/MS detection method was applied to study the concentrations and sludge adsorptions of nine estrogenic endocrine disruptors (EEDs) in an upflow anaerobic sludge blanket (UASB) and an anaerobic filter (AF). Nonylphenol equivalent quantity (NEQ) was calculated to evaluate the environmental risk of the EEDs. The results show anaerobic sludge adsorption has positive correlation with IgK(ow) and molecular length of EEDs. 17beta-estradiol (E2) and daidzein reached 83.2% and 90.4% removal rates in UASB, respectively. Genistein in AF is removed more than 80%. The sludge-water partition coefficient (K(p)) of nine EEDs ranged from 0.15-23.3 in UASB and 0.05-159.67 in AF. Biodegradation could be the main elimination pathway for daidzein, genistein, bisphenol A and estriol, by combining analysis of removal data and K(p) values. Environmental risk of EEDs was reduced after anaerobic treatment, as effluent NEQ was below the US EPA nonylphenol criteria (28 microg x L(-1), hour average concentration standard).

[Determination of environmental estrogens in pharmacy wastewater using solid-phase extraction-gas chromatography/mass spectrometry with derivatization].

A method for the determination of estrone (E1), estradiol (E2), ethinylestradiol (EE2) and estriol (E3) in pharmacy wastewater was developed using solid-phase extraction-gas chromatography/mass spectrometry (SPE-GC/MS) with derivatization. The sample was extracted by an SPE column, derivatized by bis (trimethylsilyl) trifluoroacetamide (BSTFA), and analyzed by GC/MS. The detection limits were 1.8-4.7 ng/L, and the relative standard deviations were 2.3%-9.1% (n = 8). The recoveries of above four environmental estrogen compounds were (94.0 +/- 2.9)% to (101 +/- 3.8)%. The method can be applied in the determination of the estrogenic compounds in wastewater samples successfully. The concentrations of EE2 and E1 in wastewater were 396.6 ng/L and 39.9 ng/L, respectively, and the removal rates of EE2 and E1 were 35%-40% after traditional biological treatment. The results demonstrated that the removal efficiency was not satisfactory and the traditional treatment process of wastewater containing estrogen compounds from pharmaceuticals factory should be improved.

An assessment of endocrine disrupting activity changes during wastewater treatment through the use of bioassays and chemical measurements.

The objective of this study was to assess the removal efficiencies of secondary wastewater treatment processes for compounds causing endocrine disrupting activity. The study used bioassays and chemical measurements, such as gas chromatography with mass spectrometry and enzyme immunosorbent assays. A total of seven full-scale water reclamation facilities using different unit operations and two pilot-scale membrane bioreactors were examined. Findings of this study imply that estrogenic disrupting activity in primary effluent is mainly caused by two steroidal hormones (17beta-estradiol and estriol) and, to a lesser extent, by synthetic chemicals, such as bisphenol A, 4-nonylphenol, and 4-tert-octylphenol. During secondary treatment, steroidal hormones were removed to a higher degree than nonylphenol and bisphenol A. The total estrogenic activity was removed by an average of 96%. The remaining concentrations of targeted steroids in secondary effluents, except for estriol, still had the potential to elicit a positive response in the human breast cell cancer assay. For the majority of facilities, the remaining activity was likely attributed to residual concentrations of two steroidal hormones (17beta-estradiol and estriol).

[Equine estrogens vs. esterified estrogens in the climacteric and menopause. The controversy arrives in Mexico]. / Estrógenos equinos vs. estrógenos esterificados en el climaterio y la menopausia. La controversia llega a México.

It exists controversies about if the effects and benefits of the esterified estrogens could be similar to those informed for equines, because its chemical composition and bioavailability are different. Esterified estrogens has not delta 8,9 dehydroestrone, and its absorption and level of maximum plasmatic concentrations are reached very fast. In United States of America and another countries, esterified estrogens has been marketed and using for treatment of climacteric syndrome and prevention of postmenopausal osteoporosis, based on the pharmacopoiea of that country, but the Food and Drug administration (FDA) has not yet authorized up today, a generic version of conjugated estrogens. In Instituto Mexicano del Seguro Social (IMSS) and another institutions of health sector in Mexico, starting in year 2000, it has been used esterified estrogens for medical treatment of climacteric and menopausal conditions. For this reason, in this paper we revised the most recent information about pharmacology, chemical composition, clinical use and costs of the conjugated estrogens with the purpose to guide the decisions to purchase this kind of drugs in Mexican heath institutions.

Assessment of the estrogenic activity of phytoestrogens isolated from bourbon and beer.

Phytoestrogenic substances have previously been isolated and identified in two alcoholic beverages: bourbon and beer. To delineate the relative potencies of the estrogenic substances of plant origin thus far identified in these commonly consumed alcoholic beverages, we evaluated the ability of biochanin A, beta-sitosterol, genistein, and daidzein to bind to cytosolic estrogen receptor binding sites. The in vitro studies demonstrated that each of the contained substances was capable of effectively competing for cytosolic estrogen receptor binding sites of rat liver and uterus. Further, the two phytoestrogenic constituents of bourbon, beta-sitosterol and biochanin A, were less potent than those present in beer. Given the high concentration of beta-sitosterol in bourbon, we chose to evaluate the estrogenicity of beta-sitosterol in vivo using ovariectomized rats. beta-sitosterol was administered either daily or intermittently at 3 doses, based on amounts previously determined to be present in bourbon. The in vivo studies demonstrated that beta-sitosterol is capable of producing a weak estrogenic effect only at the lowest dose (6.2 micrograms/dl) administered intermittently. These responses suggest that beta-sitosterol may be weakly estrogenic at low doses, but is unable to maintain such an effect at higher doses.