3D printed stir bar sorptive extraction coupled with high performance liquid chromatography for trace estrogens analysis in environmental water samples.

BACKGROUND: Any imaginary shape with good preparation reproducibility can be made by 3D printing technology, and it has been applied in various fields. Comparatively, its applications in sample pre-treatment are relatively less, most of which involves making extraction sorbents and producing non-functionalized devices for support assistance. 3D printing has not been applied to fabricate stir bars in stir bar sorptive extraction, mainly due to the lacking of suitable printing feedstocks. This work aimed to fabricate stir bars by 3D printing, reducing the manufacturing cost and steps and improving preparation reproducibility. (90) RESULTS: By using fused deposition modeling technique and porous filament printing feedstock, stir bars were fabricated without any modifications. Adsorption performance of 3D printed stir bars were investigated for substances with different structures and polarities. Five estrogens with adsorption efficiencies of over 80 % were selected as the representatives. The 3D printed stir bars exhibited good preparation reproducibility (2.9-4.4 %) and higher extraction recoveries (73-81 %) for five estrogens than commercial polydimethylsiloxane coated stir bars (13-69 %) in a shorter time (90 vs 120 min). They showed long lifespan (160 times) with good mechanical properties and merited reduced manufacturing cost (0.064 $ per bar) and manual operation. A method of stir bar sorptive extraction coupled with high performance liquid chromatography was proposed for trace analysis of estrogens in environmental water. Under the optimized conditions, the linear ranges for estrogens were 0.5-200 µg/L with LODs of 0.13-0.17 µg/L. (136) SIGNIFICANCE: The feasibility of fused deposition modeling in stir bar fabrication was demonstrated, along with the potential of porous filament printing feedstock as the sorbent for substances with medium polarity. 3D printed stir bars were featured with excellent preparation reproducibility, long lifespan, and good mechanical properties. The stir bar fabrication method can be used for mass production with minimal differences in products performance. (62).

A new α-pyrone from Arthrinium pseudosinense culture medium and its estrogenic activity in MCF-7 cells.

A new α-pyrone analog, arthrifuranone A (1) was isolated from an EtOAc-extract of Arthrinium pseudosinense culture medium. The isolation workflow was guided by a Molecular Networking-based dereplication strategy. The chemical structure of the new compound was elucidated using MS and NMR spectroscopic techniques, and the absolute configuration was established by the Mosher's method and gauge-including atomic orbital NMR chemical shift calculations, followed by DP4 + analysis. The isolated compound was evaluated for its estrogenic activity using the MCF-7 estrogen responsive human breast cancer cells. Compound 1 showed estrogenic activity by increasing the proliferation of MCF-7 cells at the concentration of 3.125 µM via phosphorylation of estrogen receptor-α.

Determination of estrogens and estrogen mimics by solid-phase extraction with liquid chromatography-tandem mass spectrometry.

An analytical method has been developed and validated for the determination of six estrogens and estrogen mimics, namely estriol (E3), bisphenol A (BPA), 17ß-estradiol (E2), estrone (E1), ethynyl estradiol (EE2) and dienestrol (DIE), with frequent occurrence in the natural environment. Solid phase extraction coupled with liquid chromatography tandem mass spectrometry (SPE-LC-MS/MS) using electrospray ionization (ESI) in a negative mode was applied to concentration, identification, and quantification of estrogens and estrogen mimics. The SPE conditions were optimized as the selection of C18 as cartridges and MeOH as an eluent, and the control of solution pH at 9.0. The method was validated by satisfactory recoveries (80-130%) and intra-day and inter-day precision (<18.4%, as relative standard deviation), and excellent linearity for calibration curves (R2 > 0.996). The limits of detection (LODs) for six target estrogenic compounds ranged between 2.5 and 19.2 ng/L. The effects of matrix background on the determination were evaluated in terms of LODs, LOQs, analyte recovery, and slopes of calibration curves in five different water matrices. Matrix effects by tap water were negligible. However, both matrix suppression and enhancement (i.e., E3, E1, DIE) were observed in surface water and wastewater. The positive correlation between LODs and TOC in various water matrices indicated the negative effect of organic pollutants on the method sensitivity. The sum of target estrogenic compounds in environmental samples were within 17-9462 ng/L.

Two new norsesquiterpenoids with estrogenic activity from the stems and leaves of Dioscorea oppositifolia L.

Two new norsesquiterpenoids, dioscopposin A (1) and dioscopposin B (2), as well as 21 known compounds (3-23) were isolated from the stems and leaves of Dioscorea oppositifolia L. Their structures were elucidated by detailed analysis of comprehensive spectroscopic data. The absolute configurations were deduced by the comparison of experimental and calculated electronic circular dichroism spectra. Estrogenic activity of all the isolated compounds were evaluated using MCF-7 cells proliferation assay and compounds 2, 3, 7, 13, 15, 16, 18 and 21 exhibited proliferation activity.

Estrogenic Effect of Salvia officinalis Extract on Reproductive Function of Female Mice and Identification of Its Phenolic Content.

BACKGROUND: Natural Phytoestrogens present in plants are effective hormonal replacement therapy. They are converted to estrogenic substances in the gastrointestinal tract, which is considered as the natural alternative to estrogen substitute treatment for postmenopausal women. AIMS AND OBJECTIVE: Salvia officinalis, a herb traditionally used to ameliorate postmenopausal complications, can provide a safe alternative to synthetic pharmaceuticals for the treatment of menopause. Therefore, it is conceivable to detect the possible estrogenic effect of Salvia Officinalis extract as an estrogen replacement therapy in female mice. METHODS: Phytochemical, pharmacological, and immune histopathological techniques are adopted in this study. HPLC is used for the identification of extracted constituents of sage herb. The uterotrophic activity of the extract was determined in immature female mice. Moreover, the mean thickness and luminal epithelium and the photomicrographs of the luminal epithelium of the uterus were also studied. RESULTS: HPLC revealed that quercetin is the major extracted constituent (28.6%) of the total components. Saliva officinalis extract produced a significant increase in the uterine dry weight of equal potency to estrogen. The uterus exhibited a significant increase in luminal epithelial cell height (43.3 ± 6.1µm and 36.5 ± 2.5µm) for estradiol and sage extract, respectively, compared with the control group (18.2 ± 3.5µm). Furthermore, the endometrium showed the lining epithelium formed of a single layer of columnar epithelium. The stroma seemed more voluminous with dilated vasculature. Conversely, the myometrium within the uterus was not affected in any of the experimental groups. CONCLUSION: The sage herbs induced proliferative changes in the uteri of treated mice, which suggest possible estrogenic properties. Saliva officinalis extract can be used as a hormonal replacement for women during menopause and could be further explored for contraceptive use.

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.

Isoflavonoid Profiling and Estrogen-Like Activity of Four Genista Species from the Greek Flora.

As part of our ongoing research on phytoestrogens, we investigated the phytochemical profile and estrogen-like activities of eight extracts from the aerial parts of four Genista species of Greek flora using estrogen-responsive cell lines. Ethyl acetate and methanolic extracts of G. acanthoclada, G. depressa,G. hassertiana, and G. millii were obtained with accelerated solvent extraction and their phytochemical profiles were compared using ultra-high-performance liquid chromatography-high-resolution mass spectrometry (uHPLC-HRMS). Fourteen isoflavonoids, previously isolated from G. halacsyi, were used as reference standards for their identification in the extracts. Thirteen isoflavonoids were detected in both extracts of G. acanthoclada and G. hassertiana, while fewer and far fewer were detected in G. millii and G. depressa, respectively. The ethyl acetate extracts of G. hassertiana and G. acanthoclada displayed 2.45- and 1.79-fold higher, respectively, estrogen-like agonist activity in Ishikawa cells compared to MCF-7 cells at pharmacologically relevant concentrations. Both these extracts, but not that of G. depressa, contained mono- and di-O-ß-d-glucosides of genistein as well as the aglycone, all three of which are known to display full estrogen-like activity at lower-than-micromolar concentrations. The possibility of using preparations rich in G. hassertiana and/or G. acanthoclada extracts as a potentially safer substitute for low-dose vaginal estrogen for menopausal symptoms is discussed.

An emerging powerful technique for distinguishing isomers: Trapped ion mobility spectrometry time-of-flight mass spectrometry for rapid characterization of estrogen isomers.

RATIONALE: Isomer metabolites are involved in metabolic pathways, and their characterization is essential but remains challenging even using high-performance analytical platforms. The addition of ion mobility prior to mass analysis can help to separate isomers. Here, the ability of a recently developed trapped ion mobility spectrometry system to separate metabolite isomers was examined. METHODS: Three pairs of estrogen isomers were studied as a model of isomeric metabolites under both negative and positive electrospray ionization (ESI) modes using a commercial trapped ion mobility spectrometry-TOF mass spectrometer. The standard metabolites were also spiked into human urine to evaluate the efficiency of trapped ion mobility spectrometry to separate isomers in complex mixtures. RESULTS: The estradiol glucuronide isomers (E2 ß-3G and E2 ß-17G) could be distinguished as deprotonated species, while the estradiol epimers (E2 ß and E2 α) and the methoxyestradiol isomers (2-MeO-E2 ß and 4-MeO-E2 ß) were separated as lithiated adducts in positive ionization mode. When performing analyses in the urine matrix, no alteration in the ion mobility resolving power was observed and the measured collision cross section (CCS) values varied by less than 1.0%. CONCLUSIONS: The trapped ion mobility spectrometry-TOF mass spectrometer enabled the separation of the metabolite isomers with very small differences in CCS values (ΔCCS% = 2%). It is shown to be an effective tool for the rapid characterization of isomers in complex matrices.

Molecularly imprinted polymers prepared from a single cross-linking functional monomer for solid-phase microextraction of estrogens from milk.

In this work, a single cross-linking functional monomer, 2,5-divinylterephthalaldehyde, was designed and synthesized to simplify the preparation of molecularly imprinted polymers (MIPs). In the presence of estradiol as a template, MIPs were successfully prepared using 2,5-divinylterephthalaldehyde along with a solvent and initiator. This method reduced most of the complex variables encountered in the traditional synthesis. Characterization of the morphology and structure of the MIPs was performed by scanning electron microscopy, Fourier transform infrared spectroscopy, and Brunauer-Emmett-Teller analysis. Compared with non-imprinted polymers, the MIPs had higher adsorption capacities for five estrogens with imprinting factors above 2.9. The MIPs had high extraction efficiencies, good functional properties, long lifetimes, and good reproducibility, which made them suitable for solid-phase microextraction (SPME). Coupled with ultra-high performance liquid chromatography tandem mass spectrometry, the MIP-based fibers were applied to SPME for the analysis of five estrogens in milk samples. Under the best conditions, the established method had a wide linear range (0.5-10000 ng kg-1), low limits of detection (0.08-0.26 ng kg-1) and quantification (0.26-0.87 ng kg-1), good precision (3.2-8.1%, n = 6), and fiber-to-fiber reproducibility (4.3%-8.8%, n = 3). The MIPs-based fibers can be reused at least 60 times without apparent loss of extraction efficiency. Finally, this method was applied to the determination of target estrogens in milk samples with satisfactory relative recoveries (84.3%-105%, relative standard deviation ≤ 7.8%).

Combination of Counter Current Salting-Out Homogenous Liquid-Liquid Extraction with Dispersive Liquid-Liquid Microextraction for the High-Performance Liquid Chromatographic Determination of Environmental Estrogens in Water Samples.

In this paper, counter current salting-out homogenous liquid-liquid extraction was combined with dispersive liquid-liquid microextraction for the determination of environmental estrogens in water samples by high-performance liquid chromatography. In this method, initially, sodium chloride was filled into a syringe and a mixture of water sample and acetonitrile was driven to pass through the syringe. Due to salting-out effect, fine droplets of acetonitrile went up through the remaining mixture and aggregated as a separated layer on the top. Then, the collected organic phase (acetonitrile) was removed with a syringe and mixed with carbon tetrachloride (extraction solvent). In the second step, the mixed organic phase was rapidly injected into 5 mL of distilled water to further enrich the analytes. Good linearity was obtained in the concentration range of 2.0~200 ng/mL for diethylstilbestrol (DES) and 8.0~200 ng/mL for octylphenol (OP), respectively. Limits of detection were 0.09 ng/mL for DES and 0.20 ng/mL for OP, respectively. Relative standard deviations for intra- and inter-day precisions were less than 2.1 and 3.1%, respectively. Finally, the established method was successfully applied to determine DES and OP in river water, well water, bottled water and campus drinking water samples with recoveries in the range from 81.0 to 105.9%.

Estrogenic phytochemical from Labisia pumila (Myrsinaceae) with selectivity towards estrogen receptor alpha and beta subtypes.

Labisia pumila var. alata (Myrsinaceae) or "Kacip fatimah" is a famous Malay traditional herb used for the maintenance of women's health. The extracts of L.pumila displayed estrogenic activity in rats. Nonetheless, the estrogenic bioactives were not identified. The aim of the study is to identify estrogenic compounds contributing to the established estrogenic activity. Bioactivity-guided-isolation method guided the isolation of pure bioactives. The hexane extract was subjected to a series of silica gel flash and open column chromatography with increasing amount of ethyl acetate in hexane or methanol in chloroform. Each fraction or pure compounds were evaluated on it's estrogen receptor (ER) binding activity with the fluorescence polarization competitive ERα and ERß binding assay kit. Cytotoxic assay using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay method was used to establish the cytotoxic activity of the compounds. Four alkyl resorcinols and a dimeric 1,4-benzoquinone, namely belamcandol B (1), 5-pentadec-10'-(Z)-enyl resorcinol (2), 1,3-dihydroxy-5-pentadecylbenzene (3), 5-(heptadec-12'-(Z)-enyl) resorcinol (4) and demethylbelamcandaquinone B (5) were identified with selective binding affinities towards either ERα or ERß exhibiting selectivity ratio from 0.15-11.9. Alkyl resorcinols (2)-(4) exhibited cytotoxic activity towards HL60 cells with IC50 values from 19.5-22.0 µM. Structural differences between compounds influence the binding affinities to ER subtypes. Further study is needed to establish the agonist or antagonist effect of these compounds on various tissues and to identify if these compounds exert cytotoxic activity through the ERs. When consuming L.pumila as a complementary medicine, careful consideration regarding it's estrogenic compound content should be given due consideration.

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.

In vitro bioanalytical evaluation of removal efficiency for bioactive chemicals in Swedish wastewater treatment plants.

Chemical contamination of wastewater is a problem of great environmental concern, as it poses a hazard to both the ecosystem and to human health. In this study, we have performed a bioanalytical evaluation of the presence and removal efficiency for bioactive chemicals in wastewater treatment plants (WWTPs), using in vitro assays for toxicity endpoints of high relevance for human health. Water samples were collected at the inlet and outlet of five Swedish WWTPs, all adopting a treatment technology including pretreatment, primary treatment (sedimenation), seconday treatment (biological processes), post-sedimentation, and sludge handling. The water samples were analyzed for cytotoxicity, estrogenicity, androgenicity, aryl hydrocarbon receptor (AhR) activity, oxidative stress response (Nrf2) and the ability to activate NFĸB (nuclear factor kappa-light-chain-enhancer of activated B cells) signaling. We observed clear androgenic and estrogenic activities in all inlet samples. Androgenic and estrogenic activities were also observed in all outlet samples, but the activities were lower than the respective inlet sample. AhR activity was observed in all samples, with higher activities in the inlet samples compared to the outlet samples. The removal efficiency was found to be high for androgenic (>99% for two plants and 50-60% for two plants) and estrogenic (>90% for most plants) compounds, while the removal efficiency for AhR-inducing compounds was 50-60% for most plants and 16% for one plant.

Derivatization enhances analysis of estrogens and their bioactive metabolites in human plasma by liquid chromatography tandem mass spectrometry.

Estrogens regulate many diverse biological processes in health and disease. They circulate at a wide range of concentrations in females generating several active metabolites (hydroxy and methoxyestrogens). The metabolites are assumed to be present in much lower levels and are thought to contribute to diseases such as pulmonary arterial hypertension (PAH). Estrogen metabolites are challenging to quantify in plasma and currently available immunoassays are non-specific. Here we have developed and validated a novel assay to simultaneously quantify parent estrogens and their metabolites by mass spectrometry (MS). Estrogens were extracted from human plasma using solid phase extraction and derivatized using 1-(5-fluoro-2, 4-dinitrophenyl)-4-methylpiperazine (PPZ) before quaternization by methylation ("MPPZ"). MPPZ derivatives were separated and quantified by liquid chromatography tandem MS (LC-MS/MS) in positive electrospray ionization mode, using a QTrap 6500 + coupled to a Shimadzu Nexera X2. Separation was achieved using an ACE Excel 2 C18-PFP column (2 µm, 2.1 mm × 150 mm). The limits of quantification (LOQ) were 0.43-2.17 pg on column with a linear range from 2 or 10 - 2000 pg mL-1. Intra and inter-day precision and accuracy were acceptable (<20% at LOQ and <15% above). These derivatives demonstrated minimal degradation upon short-term storage at 15 °C (<20%) and longer term at -20 °C (<20%). Using this approach, estrone (E1) and estradiol (E2) were detected in plasma (0.5 mL) from healthy women and those with PAH but downstream metabolites 16-hydroxy-E1, 16-hydroxy-E2, 2-methoxy-E1 and 4-methoxy-E1 were only detected in plasma from diseased patients. These findings will next be tested robustly in large patient cohorts. This novel LC-MS/MS analysis of estrogens and their bioactive metabolites, using MPPZ derivatization, opens doors for the simultaneous analysis of a panel of estrogens in human plasma, across the endogenous range of concentrations encountered in health and disease.

Solid-phase extraction of estrogens and herbicides from environmental waters for bioassay analysis-effects of sample volume on recoveries.

Ecotoxicological screening of surface waters can involve multiple analyses using multiple bioassay and chemical analytical methods that require enriched samples to reach low concentrations. Such broad screening of the same sample necessitates sufficient sample volume-typically several liters-to produce a sufficient amount of enriched sample. Often, this is achieved by performing parallel solid-phase extractions (SPE) where extracts are combined into a pool-this is a laborious process. In this study, we first validated our existing SPE method for the chemical recovery of an extended set of compounds. We spiked four estrogenic compounds and 11 herbicides to samples from independent rivers (1 L) and wastewater treatment plant effluents (0.5 L). Then, we investigated the effect of increased sample loading of the SPE cartridges on both chemical and biological recoveries by comparing the validated volumes with four times larger sample volumes (i.e., 4 L river water and 2 L effluent). Samples were analyzed by LC-MS/MS and three bioassays: an estrogen receptor transactivation assay (ERα-CALUX), the combined algae test, and a bacterial bioluminescence inhibition assay. Our existing SPE method was found to be suitable for enriching the extended set of estrogens and herbicides in river water and effluents with near to perfect chemical recoveries (~ 100%), except for the herbicide metribuzin (46 ± 19%). In the large volume river and effluent samples, the biological activities and concentrations of the spiked compounds were between 87 and 104% of those measured with the lower sample loading, which is adequate. In addition, the ratio between the large and original volume SPE method for the non-target endpoint (bacterial bioluminescence inhibition) was acceptable (on average 82 ± 9%). Results indicate that our current water extraction method can be applied to up to four times larger sample volumes, resulting in four times more extract volumes, without significant reductions in recoveries for the tested estrogens and herbicides. Graphical abstract ᅟ.

Trace analysis of estrogens in milk samples by molecularly imprinted solid phase extraction with genistein as a dummy template molecule and high-performance liquid chromatography-tandem mass spectrometry.

Dummy molecularly imprinted polymer microspheres (DMIPMS) towards estrogens were synthesized by Pickering emulsion polymerization employing genistein (GEN) as a dummy template molecule. The FTIR analysis indicated the successful preparation of the imprinted polymers, and the characterization results of scanning electron microscopy and nitrogen adsorption desorption measurement indicated that the obtained DMIPMS are in possess of regular spherical shapes, porous structures and narrow diameter distribution, a BET surface area of 402.74 m2 g-1, a total pore volume of 0.568 cm3 g-1 and a pore diameter of 3.62 nm. The binding capacity and selectivity of DMIPMS were investigated in equilibrium binding experiments and chromatographic evaluation experiments through scatchard analysis and molecular imprinting factor (IF) analysis, respectively. The MIPs showed high binding capacity and excellent selectivity towards seven selected natural and synthetic estrogens, which are Estrone (E1), 17ß-estradiol (ßE2), estriol (E3), ethinylestradiol (EE2), dienestrol(DS), diethylstilbestrol (DES), and hexestrol (HEX). A method for selective determination of seven estrogens in milk samples via dummy molecularly imprinted solid phase extraction coupled with HPLC-MS/MS was developed, which showed good linearity from 2 to 500 µg L-1 with a correlation coefficient (R2) of more than 0.999. The detection limits were within the range of 0.10-0.35 µg L-1 and the recoveries of the seven estrogens at three spiking levels (10,100,250 µg L-1) ranged from 88.9% to 102.3% with relative standard deviation (RSD, n = 5) for intra-day and inter-day assays varied from 0.8% to 4.5%. The developed method is thus proven to be efficient and reliable for regular monitoring of trace estrogens in complex matrices such as milk samples.

Estrogens determination exploiting a SIA-LOV system prior in-port derivatization-large volume injection-programmable temperature vaporization-gas chromatography.

In this work, we present a method for the clean-up, preconcentration and quantification of the four most widely found estrogens (estrone E1, estradiol E2, estriol E3 and ethynyl estradiol EE2) in seawater samples. A sequential injection analysis-lab on valve system (SIA-LOV) has been developed to perform the microsolid phase extraction (µSPE) of the analytes in a fully automated way. After testing different resins and solvents, C18 resin with acetonitrile (ACN) as eluent have been chosen as they provided the best results. Several parameters affecting the extraction have been studied and optimized. Besides, extraction column lifetime has also been checked as it is indicative of the number of consecutive analysis that the column is able to perform before replacing it. Results showed that the same column can be used up to 50 times. Then, the derivatization of the extracts has been performed unattended by exploiting an in-port derivatization of the analytes with N-methyltrimethylsilyltrifluoroacetamide (BSTFA) prior their quantification using large volume injection with programmable temperature vaporization gas chromatography (LVI-PTV-GC-MS).

From micro to macro-contaminants: The impact of low-energy titanium dioxide photocatalysis followed by filtration on the mitigation of drinking water organics.

This study evaluated strategies targeting macro- and micro-organic contaminant mitigation using low-energy titanium dioxide photocatalysis. Energy inputs of 1, 2, and 5 kWh m-3 resulted in incomplete oxidation of macro-organic natural organic matter, signified by greater reductions of UV254 and specific ultraviolet UV absorbance (SUVA) in comparison to dissolved organic carbon (DOC). The rate of UV254 removal was 3 orders of magnitude greater than the rate of DOC degradation. Incomplete oxidation improved operation of downstream filtration processes. Photocatalysis at 2 kWh m-3 increased the bed life of downstream granular activated carbon (GAC) filtration by 340% relative to direct filtration pretreatment. Likewise, photocatalysis operated ahead of microfiltration decreased fouling, resulting in longer filter run times. Using 2 kWh m-3 photocatalysis increased filter run time by 36 times in comparison to direct filtration. Furthermore, levels of DOC and UV254 in the membrane permeate improved (with no change in removal across the membrane) using low-energy photocatalysis pretreatments. While high-energy UV inputs provided high levels of removal of the estrogenic micro-organics estrone (E1), 17ß-estradiol (E2), estriol (E3), and 17α-ethynlestradiol (EE2), low-energy photocatalysis did not enhance removal of estrogens beyond levels achieved by photolysis alone. In the cases of E1 and E3, the addition of TiO2 as a photocatalyst reduced degradation rates of estrogens compared to UV photolysis. Overall, process electrical energy per order magnitude reductions (EEOs) greatly improved using photocatalysis, versus photolysis, for the macro-organics DOC, UV254, and SUVA; however, energy required for removal of estrogens was similar between photolysis and photocatalysis.

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.

History of Estrogen: Its Purification, Structure, Synthesis, Biologic Actions, and Clinical Implications.

This mini-review summarizes key points from the Clark Sawin Memorial Lecture on the History of Estrogen delivered at Endo 2018 and focuses on the rationales and motivation leading to various discoveries and their clinical applications. During the classical period of antiquity, incisive clinical observations uncovered important findings; however, extensive anatomical dissections to solidify proof were generally lacking. Initiation of the experimental approach followed later, influenced by Claude Bernard's treatise "An Introduction to the Study of Experimental Medicine." With this approach, investigators began to explore the function of the ovaries and their "internal secretions" and, after intensive investigations for several years, purified various estrogens. Clinical therapies for hot flashes, osteoporosis, and dysmenorrhea were quickly developed and, later, methods of hormonal contraception. Sophisticated biochemical methods revealed the mechanisms of estrogen synthesis through the enzyme aromatase and, after discovery of the estrogen receptors, their specific biologic actions. Molecular techniques facilitated understanding of the specific transcriptional and translational events requiring estrogen. This body of knowledge led to methods to prevent and treat hormone-dependent neoplasms as well as a variety of other estrogen-related conditions. More recently, the role of estrogen in men was uncovered by prismatic examples of estrogen deficiency in male patients and by knockout of the estrogen receptor and aromatase in animals. As studies became more extensive, the effects of estrogen on nearly every organ were described. We conclude that the history of estrogen illustrates the role of intellectual reasoning, motivation, and serendipity in advancing knowledge about this important sex steroid.