Preparation of deep eutectic solvent modified magnetic graphene oxide/metal organic framework nanocomposites for the extraction of three estrogens in cosmetics.

A novel magnetic dispersive solid phase extraction (MDSPE) procedure based on the deep eutectic solvent (DES) modified magnetic graphene oxide/metal organic frameworks nanocomposites (MGO@ZIF-8@DES) was established and used for the efficient enrichment of estradiol, estrone, and diethylstilbestrol in cosmetics (toner, lotion, and cream) for the first time. Then, the three estrogens were separated and determined by UHPLC-UV analysis method. In order to study the features and morphology of the synthesized adsorbents, various techniques such as FT-IR, SEM, and VSM measurements were executed. The MGO@ZIF-8@DES nanocomposites combine the advantages of high adsorption capacity, adequate stability in aqueous solution, and convenient separation from the sample solution. To achieve high extraction recoveries, the Box-Behnken design and single factor experiment were applied in the experimental design. Under the optimum conditions, the method detection limits for three estrogens were 20-30 ng g-1. This approach showed a good correlation coefficient (r more than 0.9998) and reasonable linearity in the range 70-10000 ng g-1. The relative standard deviations for intra-day and inter-day were beneath 7.5% and 8.9%, respectively. The developed MDSPE-UHPLC-UV method was successfully used to determine  three estrogens in cosmetics, and acceptable recoveries in the intervals of 83.5-95.9% were obtained. Finally, three estrogens were not detected in some cosmetic samples. In addition, the Complex GAPI tool was used to evaluate the greenness of the developed pretreatment method. The developed MDSPE-UHPLC-UV method is sensitive, accurate, rapid, and eco-friendly, which provides a promising strategy for determining hormones in different complex samples.

Highly sensitive determination of endocrine disrupting chemicals in foodstuffs through magnetic solid-phase extraction followed by high-performance liquid chromatography-tandem mass spectrometry.

BACKGROUND: Endocrine disrupting chemicals (EDCs), proved to be potential carcinogenic threats to human health, have received great concerns in food field. It was essential to develop effective methods to detect EDCs in food samples. The present study proposed an efficient method to determine trace EDCs including estrone (E1), 17ß-estradiol (E2), estriol (E3) and bisphenol A (BPA) based on magnetic solid-phase extraction (MSPE) coupled high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) in meat samples. RESULTS: Fe3 O4 @COF(TpBD)/TiO2 nanocomposites were synthesized via functionalization of magnetic covalent organic frameworks (COFs) with titanium dioxide (TiO2 ) nanoparticles, and used as absorbents of MSPE to enrich EDCs. The efficient EDCs enrichment relies on π-π stacking interaction, hydrogen bonding, and the interaction between titanium ions (IV, Ti4+ ) and hydroxyl groups in EDCs, which improves the selectivity and sensitivity. Under the optimized conditions, target EDCs were rapidly extracted through MSPE with 5 min. Combining Fe3 O4 @COF(TpBD)/TiO2 based MSPE and HPLC-MS/MS to determine EDCs, good linearities were observed with correlation coefficient (R2 ) ≥ 0.9989. The limits of detection (LODs) and limits of quantification (LOQs) were 0.13-0.41 µg kg-1 and 0.66-1.49 µg kg-1 , respectively. Moreover, the proposed method was successfully applied to real samples analysis. CONCLUSIONS: The established MSPE-HPLC-MS/MS method was successfully applied to determine EDCs in meat samples with rapidness, improved selectivity and sensitivity. It shows great prospects for EDCs detection in other complicated matrices. © 2020 Society of Chemical Industry.

Estrogenic Activity of Mycoestrogen (3ß,5α,22E)-Ergost-22-en-3-ol via Estrogen Receptor α-Dependent Signaling Pathways in MCF-7 Cells.

Armillariella tabescens (Scop.) Sing., a mushroom of the family Tricholomataceae, has been used in traditional oriental medicine to treat cholecystitis, improve bile secretion, and regulate bile-duct pressure. The present study evaluated the estrogen-like effects of A. tabescens using a cell-proliferation assay in an estrogen-receptor-positive breast cancer cell line (MCF-7). We found that the methanol extract of A. tabescens fruiting bodies promoted cell proliferation in MCF-7 cells. Using bioassay-guided fractionation of the methanol extract and chemical investigation, we isolated and identified four steroids and four fatty acids from the active fraction. All eight compounds were evaluated by E-screen assay for their estrogen-like effects in MCF-7 cells. Among the tested isolates, only (3ß,5α,22E)-ergost-22-en-3-ol promoted cell proliferation in MCF-7 cells; this effect was mitigated by the ER antagonist, ICI 182,780. The mechanism underlying the estrogen-like effect of (3ß,5α,22E)-ergost-22-en-3-ol was evaluated using Western blot analysis to detect the expression of extracellular signal-regulated kinase (ERK), phosphatidylinositol 3-kinase (PI3K), Akt, and estrogen receptor α (ERα). We found that (3ß,5α,22E)-ergost-22-en-3-ol induced an increase in phosphorylation of ERK, PI3K, Akt, and ERα. Together, these experimental results suggest that (3ß,5α,22E)-ergost-22-en-3-ol is responsible for the estrogen-like effects of A. tabescens and may potentially aid control of estrogenic activity in menopause.

Enhanced adsorption of steroid estrogens by one-pot synthesized phenyl-modified mesoporous silica: Dependence on phenyl-organosilane precursors and pH condition.

In this study, the phenyl-modified mesoporous materials were successfully synthesized using phenyl-organosilanes (trimethoxyphenylsilane and triethoxyphenylsilanea) by one-pot co-condensation method for the removal of estrone (E1), 17ß-estradiol (E2), and 17α-ethinyl estradiol (EE2). Both the triethoxyphenylsilane-modified material (20%EtPh-MCM-41) and trimethoxyphenylsilane-modified material (20%MePh-MCM-41) could rapidly achieve equilibrium in 30 min at low adsorbent dosage of 0.025 g L-1. But the different hydrolysable groups of trimethoxyphenylsilane and triethoxyphenylsilane led to the discrepancies in physicochemical properties of the 20%EtPh-MCM-41 and 20%MePh-MCM-41, and thus affected adsorption performance. The 20%EtPh-MCM-41 exhibited the faster estrogen adsorption rates expressed in pseudo-second-order kinetic constant than the 20%MePh-MCM-41 due to the more hydrophobicity. Conversely, the 20%MePh-MCM-41 had much more estrogen adsorption capacities than the 20%EtPh-MCM-41 because of the more available adsorption sites. The addition of the phenyl-organosilane improved estrogen adsorption by π-π and hydrophobic interactions, and the Langmuir-model-based maximum adsorption amounts could reach 99.02, 83.47, and 53.60 mg g-1 for EE2, E2, and E1, respectively. But excessive concentration of phenyl-organosilane decreased adsorption capacities due to poor pore structure. Alkaline solution, which induced estrogen deprotonation and negative surface charge of absorbents, inhibited estrogen adsorption by electrostatic repulsion and the decreased hydrophobic interaction, but acidic and neutral solutions, ionic strength, and humic acid did not significantly affect estrogen removal. This work not only showed the high potential of trimethoxyphenylsilane-modified MCM-41 used in water purification for steroid estrogens, but also demonstrated the suitable selection of organosilane precursors was key in producing favorable materials with designed functionality.

Study of the adsorption of endocrine disruptor compounds on typical filter materials using a quartz crystal microbalance.

Drinking water containing environmental endocrine disruptor compounds (EDCs) endangers human health, and researching the purification process of drinking water for the effective removal of EDCs is vitally important. Filtering plays a crucial role in the bio-adsorption of EDCs, but the adsorption mechanism that occurs between the EDCs and filters remains unclear. In this study, a quartz crystal microbalance (QCM) was employed to elucidate the adsorption mechanism because QCM is a label-free method that possesses high selectivity, high stability, and high sensitivity. The results indicated that a pseudo-first-order kinetic model best fits the adsorption process of four different EDCs, which included bisphenol A (BPA), estrone (E1), estradiol (E2), and sulfamethoxazole (SMZ), on silica (quartz sand), a typical filter material surface. The order of the amount of individual EDCs absorbed on the silica surface was qE2 > qE1 > qSMZ > qBPA and related to their molecular structure, polarity, and chargeability. As the initial EDC concentration increased, the adsorbed amount of the four EDCs on the silica surface increased; however, the initial concentration had little effect on removal efficiency. The calculated Freundlich exponent (1/n) demonstrated SMZ and BPA showed a greater tendency for adsorption than E1 and E2. The mass response time on the surface of the silica gradually increased as the pH increased (from 5.5 to 8.5), indicating the adsorption rate was inhibited by the increase in pH. The addition of electrolytes shortened the mass response time of EDCs on the QCM chip. The pH and ionic strength produced no significant effects on adsorption because hydrophobicity was the primary contributor to adsorption. This study facilitated a better understanding of the interaction between EDCs and filters in water treatment.

Determination of estrone and 17α-ethinylestradiol in digested sludge by ultrasonic liquid extraction and high-performance liquid chromatography with fluorescence detection.

Estrone, 17ß-estradiol and 17α-ethinylestradiol are increasingly recognised as important micropollutants to be monitored in wastewater treatment plants. These estrogens are retained onto sludge due to their high adsorption and since they are largely used in land applications, the monitoring of these chemicals in sludge samples is of great importance. This study describes a method for the determination of estrone and 17α-ethinylestradiol in fresh sludge samples. After spiking fresh digested sludge with estrone and 17α-ethinylestradiol and maintaining in contact during 5, 30 and 60 min, the freeze-dried samples were subjected to ultrasonic liquid extraction, with methanol and acetone, and analysed by high-performance liquid chromatography with fluorescence detection. The average recoveries obtained for estrone and 17α-ethinylestradiol using the different contact times were 103 ± 3 and 97 ± 4%, respectively. Fresh sludge samples from one waste water treatment plant located in Portugal were analysed and estrone was detected in primary fresh sludge, anaerobic digested sludge and dehydrated sludge at a concentration in the range of 1-4.8 µg/g. The method here developed does not require any sample clean-up, being fast and simple, reliable and inexpensive, making possible its application for monitoring the contamination of sludge with these estrogens.

N-propyl functionalized spherical mesoporous silica as a rapid and efficient adsorbent for steroid estrogen removal: Adsorption behaviour and effects of water chemistry.

To achieve an enhanced and selective adsorption of steroid estrogens, the n-propyl functionalization was applied to the mesoporous silica material (MCM-41) according to the physico-chemical property analysis of steroid estrogens. Adsorption behaviour and water chemistry effects were evaluated with the most concerned steroid estrogens: estrone (E1), 17ß-estradiol (E2) and 17α-ethinyl estradiol (EE2) based on the materials characterization. The results showed the uptakes of E1, E2, and EE2 onto the modified MCM-41 were enhanced and accelerated by the n-propyl functionalization, which was positively correlated with the hydrophobicity of the synthesized materials. Kinetic data fitted the pseudo-second-order model well. Based on the Langmuir model, the maximum adsorption capacities of the n-propyl modified MCM-41 were up to 119.87, 88.38, and 86.91 mg g-1 for EE2, E1, and E2, respectively. Importantly, both acid and neutral solutions were beneficial to estrogen removal, but ionic strength and humic acid did not affect the estrogen adsorption. The above results suggested that the n-propyl functionalized MCM-41 would be a promising adsorbent for the rapid and efficient removal of estrogens with the selectivity from natural organic matter like humic acid. Mechanism analysis showed the key role of hydrophobic interaction, and it also confirmed the contribution of the carbonylic lone pair electrons of E1, which helped the formation of stronger hydrogen bonds with silicon hydroxyls and enhanced the dipole-dipole interaction between E1 and the synthesized materials.

Adsorption of estrone with few-layered boron nitride nanosheets: Kinetics, thermodynamics and mechanism.

Boron nitride Nanosheets (BNNSs) was fabricated with a method of heating the mixture of boric acid and urea in N2 atmosphere and used to remove estrone (E1) from water. The obtained BNNSs exhibited a higher surface area of 896 m2/g, a large pore volume of 0.76 cm3/g, and only few layers (0.398 nm) with the boric acid and urea ratio of 1:80. The layer number of BNNSs decreased from 15 to 4 with the mole ratio of boric acid and urea decreasing from 1:20 to 1:80, which was identified by SEM, TEM, AFM and BET measurements. More importantly, BNNSs presented an outstanding adsorption performance for estrone with the adsorption capacity of 249.15 mg E1/g BNNSs. The adsorption process could be best fitted by pseudo second-order kinetic model and the equilibrium data at different temperatures were well fitted by Langmuir isotherm model. The thermodynamics analysis revealed that E1 adsorption on BNNSs was spontaneous (ΔG = -29.33 kJ mol-1), enthalpy-retarded (ΔH = 29.75 kJ mol-1), entropy-driven (ΔS = 198.26 J mol-1 K-1), and mostly chemical adsorption. The adsorption rates of E1 in water were sharply enhanced with thinner BNNSs as absorbents and removal efficiency by BN-60 regenerated after 6 times was above 95%, it was shown that the surface areas, mesopores and remarkable structure played important roles in the adsorption process. The firmness of E1 onto BNNSs and the stability of adsorption efficiency made BNNSs as a potential absorbent for efficient removal of E1 from wastewater.

Facile preparation of polydopamine-coated imprinted polymers on the surface of SiO2 for estrone capture in milk samples.

Estrone molecularly imprinted polymers were synthesized through the self-polymerization of dopamine on the surface of silica gels, which had the characteristics of mild polymerization conditions, simple reaction procedure and good specific recognition ability for estrone. The estrone molecularly imprinted polymers were characterized by scanning electron microscopy, Fourier transform infrared spectroscopy, thermogravimetric analysis, elemental analysis and nitrogen adsorption-desorption tests. The characterization confirmed that the imprinted polymers were successfully grafted on the surface of silica gels. Through investigating the adsorption performance, the prepared estrone molecularly imprinted polymers exhibited high adsorption capacity, fast mass transfer, as well as excellent selectivity toward estrone. The estrone molecularly imprinted polymers as the solid-phase extraction adsorbent coupled with high-performance liquid chromatography was developed to determine estrone from the milk samples. The developed estrone molecularly imprinted polymer solid-phase extraction with high-performance liquid chromatography method exhibited satisfactory specificity, precision, accuracy and good linearity relationship in the range of 0.2-20 µg/mL. The developed method is simple, fast, effective and high specificity method and it provides a new method to detect the residues of estrone in animal foods.

A MALDI-MS-based quantitative analytical method for endogenous estrone in human breast cancer cells.

The level of endogenous estrone, one of the three major naturally occurring estrogens, has a significant correlation with the incidence of post-menopausal breast cancer. However, it is challenging to quantitatively monitor it owing to its low abundance. Here, we develop a robust and highly sensitive mass-assisted laser desorption/ionization mass spectrometry (MALDI-MS)-based quantitative platform to identify the absolute quantities of endogenous estrones in a variety of clinical specimens. The one-step modification of endogenous estrone provided good linearity (R(2) > 0.99) and significantly increased the sensitivity of the platform (limit of quantitation: 11 fmol). In addition, we could identify the absolute amount of endogenous estrones in cells of the breast cancer cell line MCF-7 (34 fmol/10(6) cells) by using a deuterated estrone as an internal standard. Finally, by applying the MALDI-MS-based quantitative method to endogenous estrones, we successfully monitored changes in the metabolic expression level of estrones (17.7 fmol/10(6) letrozole-treated cells) in MCF-7 cells resulting from treatment with an aromatase inhibitor. Taken together, these results suggest that this MALDI-MS-based quantitative approach may be a general method for the targeted metabolomics of ketone-containing metabolites, which can reflect clinical conditions and pathogenic mechanisms.

Comparative study of emerging micropollutants removal by aerobic activated sludge of large laboratory-scale membrane bioreactors and sequencing batch reactors under low-temperature conditions.

Four emerging micropollutants ibuprofen, diclofenac, estrone (E1) and 17α-ethinylestradiol (EE2) were studied in large laboratory-scale wastewater treatment plants (WWTPs) with high nitrifying activity. Activated sludge (AS) with sludge retention times (SRTs) of 12days and 14days in sequencing batch reactors (SBRs) and 30days, 60days and 90days in membrane bioreactors (MBRs) were examined at 8°C and 12°C. Concentrations of pharmaceuticals and their main metabolites were analysed in liquid phase and solid phase of AS by liquid chromatography-tandem mass spectrometry (LC-MS/MS). A remarkable amount of contaminants were detected in solids of AS, meaning the accumulation of micropollutants in bacterial cells. The biodegradation rate constants (Kbiol) were affected by SRT and temperature. MBR with a 90-day SRT showed the best results of removal. Conventional SBR process was inefficient at 8°C showing Kbiol values lower than 0.5lgSS(-1)d(-1) for studied micropollutants.

Derivatization of estrogens enhances specificity and sensitivity of analysis of human plasma and serum by liquid chromatography tandem mass spectrometry.

Estrogens circulate at concentrations less than 20pg/mL in men and postmenopausal women, presenting analytical challenges. Quantitation by immunoassay is unreliable at these low concentrations. Liquid chromatography tandem mass spectrometry (LC-MS/MS) offers greater specificity and sometimes greater sensitivity, but ionization of estrogens is inefficient. Introduction of charged moieties may enhance ionization, but many such derivatives of estrogens generate non-specific product ions originating from the "reagent" group. Therefore an approach generating derivatives with product ions specific to individual estrogens was sought. Estrogens were extracted from human plasma and serum using solid phase extraction and derivatized using 2-fluoro-1-methylpyridinium-p-toluenesulfonate (FMP-TS). Electrospray in positive mode with multiple reaction monitoring using a QTrap 5500 mass spectrometer was used to quantify "FMP" derivatives of estrogens, following LC separation. Transitions for the FMP derivatives of estrone (E1) and estradiol (E2) were compound specific (m/z 362→238 and m/z 364→128, respectively). The limits of detection and quantitation were 0.2pg on-column and the method was linear from 1-400pg/sample. Measures of intra- and inter-assay variability, precision and accuracy were acceptable (<20%). The derivatives were stable over 24h at 10°C (7-9% degradation). Using this approach, E1 and E2, respectively were detected in human plasma and serum: pre-menopausal female serum (0.5mL) 135-473, 193-722pmol/L; male plasma (1mL) 25-111, 60-180pmol/L and post-menopausal female plasma (2mL), 22-78, 29-50pmol/L. Thus FMP derivatization, in conjunction with LC-MS/MS, is suitable for quantitative analysis of estrogens in low abundance in plasma and serum, offering advantages in specificity over immunoassay and existing MS techniques.

Bioaugmentation Mitigates the Impact of Estrogen on Coliform-Grazing Protozoa in Slow Sand Filters.

Exposure to endocrine-disrupting chemicals (EDCs), such as estrogens, is a growing issue for human and animal health as they have been shown to cause reproductive and developmental abnormalities in wildlife and plants and have been linked to male infertility disorders in humans. Intensive farming and weather events, such as storms, flash flooding, and landslides, contribute estrogen to waterways used to supply drinking water. This paper explores the impact of estrogen exposure on the performance of slow sand filters (SSFs) used for water treatment. The feasibility and efficacy of SSF bioaugmentation with estrogen-degrading bacteria was also investigated, to determine whether removal of natural estrogens (estrone, estradiol, and estriol) and overall SSF performance for drinking water treatment could be improved. Strains for SSF augmentation were isolated from full-scale, municipal SSFs so as to optimize survival in the laboratory-scale SSFs used. Concentrations of the natural estrogens, determined by gas chromatography coupled with mass spectrometry (GC-MS), revealed augmented SSFs reduced the overall estrogenic potency of the supplied water by 25% on average and removed significantly more estrone and estradiol than nonaugmented filters. A negative correlation was found between coliform removal and estrogen concentration in nonaugmented filters. This was due to the toxic inhibition of protozoa, indicating that high estrogen concentrations can have functional implications for SSFs (such as impairing coliform removal). Consequently, we suggest that high estrogen concentrations could impact significantly on water quality production and, in particular, on pathogen removal in biological water filters.

Study and comparison of polydopamine and its derived carbon decorated nanoparticles in the magnetic solid-phase extraction of estrogens.

Surface functionalization enabled by bioinspired polydopamine (PDA) is recognized as a convenient route for fabrication of multifunctional nanoparticles. In the present work, magnetic nanoparticles with polymer (Fe3O4@PDA) and carbon shell (Fe3O4@C) were prepared by self-oxidation of dopamine, and carbonization of the PDA coating. The performance of the two magnetic sorbents in the extraction and determination of four estrogens, estrone (E1), estradiol (E2), estriol (E3) and diethylstilbestrol (DES) from water samples in the form of magnetic solid-phase extraction was investigated. Orthogonal array design was utilized to facilitate the optimization of the proposed sample preparation approach. The highest extraction capabilities of the two sorbents were achieved under different experimental conditions. Fe3O4@PDA was shown to be superior to Fe3O4@C in the enrichment of estrogens, suggesting stronger interactions were established between the PDA coating and the target compounds. The extraction and desorption operations were enabled more conveniently by magnetic separation and the extracts were analyzed by high-performance liquid chromatography coupled with ultraviolet and fluorescence detection. The limits of detection achieved in the proposed method were in the range of 0.072-0.15ng/mL for E1 and DES, and 0.0017-0.0062ng/mL for E2 and E3. Good precision (>0.9995) was obtained with the linearity ranging from 0.2 to 100ng/mL, and from 0.01 to 5ng/mL. The method developed was assessed by analysis of the estrogens in tap water, drain water and bottled mineral water samples.

Vortex-assisted hollow fibre liquid-phase microextraction technique combined with high performance liquid chromatography-diode array detection for the determination of oestrogens in milk samples.

A rapid, simple, sensitive and environmentally friendly method has been developed for the determination of three oestrogens (17ß-estradiol (17ß-E2), estrone (E1), and diethylstilbestrol (DES)) in milk samples by using vortex-assisted hollow fibre liquid-phase microextraction (VA-HF-LPME) and high performance liquid chromatography. Method is based on the microextraction of oestrogens from sample solution into 15 µL of nonanoic acid as extracting agent, which is placed inside the hollow fibre followed by vortex-mixing. Vortex provided effective and mild mixing of sample solution and increased the contact between analytes and boundary layers of the hollow fibre, thereby enhancing mass transfer rate and leading to high recovery of target analytes. The extraction equilibrium is achieved within 2 min. Parameters influencing the recovery were investigated and optimized. The proposed technique provided good linearity (>0.9984), repeatability (RSD = 2.56-4.38), low limits of detection (0.06-0.17 ng mL(-1)), and high enrichment factor (330).

Development of a novel magnetic molecularly imprinted polymer coating using porous zeolite imidazolate framework-8 coated magnetic iron oxide as carrier for automated solid phase microextraction of estrogens in fish and pork samples.

A high-performance magnetic molecularly imprinted polymer (MIP) coating using zeolite imidazolate framework-8 coated magnetic iron oxide (Fe3O4@ZIF-8) as a carrier was developed for simultaneous automated solid phase microextraction of four estrogens in 24 food samples. The coating material, abbreviated as MZMIP, was synthesized through time-efficient layer-by-layer assembling of ZIF-8 and MIP film on Fe3O4 particles. It was characterized and automatically coated on the surface of SPME fibers by electromagnetic bonding. The extraction performance, reusability, repeatability, and validity of the MZMIP-SPME system was evaluated for high-throughput analysis of estrone (E1), estradiol (E2), estriol (E3), and ethinylestradiol (EE2). Various factors affecting the quality of MZMIP coating were optimized. Compared with traditional magnetic MIP coating based on Fe3O4@SiO2 carrier, the MZMIP coating exhibited high extraction capacity and quick adsorption and desorption kinetics to E1, E2, E3, and EE2 owing to the larger amount of imprinting sites in MZMIP. Under optimum conditions, the proposed system requires only 25min for pretreatment of all 24 samples (62.5s per sample). The limits of detection and quantitation of the proposed automated system for analysis were found to range from 0.4 to 1.7 and 1.1 to 6.2ngg(-1), respectively. During analysis of spiked fish and pork, the new coating showed better recovery and selectivity compared with Fe3O4@SiO2@MIP (MMIP) and commercially available SPME. The results indicated that the MZMIP coating could be effectively employed for pretreatment of ultra-trace level of estrogens in food.

A pilot scale comparison of advanced oxidation processes for estrogenic hormone removal from municipal wastewater effluent.

This study investigates the oxidation of selected endocrine disrupting compounds (estrone, 17ß-estradiol, estriol and 17α-ethinylestradiol) during ozonation and advanced oxidation of biologically treated municipal wastewater effluents in a pilot scale. Selected estrogenic substances were spiked in the treated wastewater at levels ranging from 1.65 to 3.59 µg · L(-1). All estrogens were removed by ozonation by more than 99% at ozone doses ≥1.8 mg · L(-1). At a dose of 4.4 · mg L(-1) ozonation reduced concentrations of estrone, 17ß-estradiol, estriol and 17α-ethinylestradiol by 99.8, 99.7, 99.9 and 99.7%, respectively. All tested advanced oxidation processes (AOPs) achieved high removal rates but they were slightly lower compared to ozonation. The lower removal rates for all tested advanced oxidation processes are caused by the presence of naturally occurring hydroxyl radical scavengers - carbonates and bicarbonates.

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.

Evaluation of a molecularly imprinted polymer for determination of steroids in goat milk by matrix solid phase dispersion.

A molecularly imprinted polymer-matrix solid-phase dispersion methodology for simultaneous determination of five steroids in goat milk samples was proposed. Factors affecting the extraction recovery such as sample/dispersant ratio and washing and elution solvents were investigated. The molecularly imprinted polymer used as dispersant in the matrix solid-phase dispersion procedure showed high affinity to steroids, and the obtained extracts were sufficiently cleaned to be directly analyzed. Analytical separation was performed by micellar electrokinetic chromatography using a capillary electrophoresis system equipped with a diode array detector. A background electrolyte composed of borate buffer (25mM, pH 9.3), sodium dodecyl sulfate (10mM) and acetonitrile (20%) was used. The developed MIP-MSPD methodology was applied for direct determination of testosterone (T), estrone (E1), 17ß-estradiol (17ß-E2), 17α-ethinylestradiol (EE2) and progesterone (P) in different goat milk samples. Mean recoveries obtained ranged from 81% to 110%, with relative standard deviations (RSD)≤12%. The molecularly imprinted polymer-matrix solid-phase dispersion method is fast, selective, cost-effective and environment-friendly compared with other pretreatment methods used for extraction of steroids in milk.

Removal of endocrine disrupting compounds in a lab-scale anaerobic/aerobic sequencing batch reactor unit.

The fate and removal of six selected endocrine disrupting compounds in a lab-scale anaerobic/aerobic (A/O) sequencing batch reactor (SBR), operating at 5 days, solids retention time (SRT) were investigated. A carbamazepine (CBZ), acetaminophen (ATP), diltiazem (DTZ), butyl benzyl phthalate (BBP), estrone and progesterone mix was spiked as model endocrine disrupting compounds (EDC) into domestic wastewater obtained from a nearby sewage treatment plant. The influent, effluent and sludge samples from the SBR unit were analysed by using an LC/MS/MS instrument equipped with electrospray ionization. More than 80% removal was observed for all the EDCs tested. It was found that biodegradation is the most important mechanism for BBP, ATP and progesterone. Biodegradation constants were calculated according to the simplified Monod model for these compounds. The DTZ seemed to have lower rate of biodegradation. The CBZ appeared totally resistant to biodegradation. However, it presented a high rate of sorption onto the sludge and was thereby treated. This contradicts with the literature studies.