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Estrogens in aquatic environments pose significant ecological and health risks due to their cumulative effects rather than individual impacts. This study investigates the voltammetric behavior of estrone (E1), 17β-estradiol (E2), estriol (E3), and 17α-ethinylestradiol (EE2), presenting a cost-effective and straightforward method for their simultaneous determination. Using differential pulse voltammetry (DPV) with a boron-doped diamond electrode, the method demonstrates high precision (deviations under 4%) and a linear dynamic range of 15.35-134.55 µmol·L -1 . Integration of a vacuum evaporation step reduced detection limits to 10 -8 mol·L -1 , enabling effective analysis of real water samples. This optimized approach ensures practical applicability for monitoring total estrogen content in aquatic systems, providing an accessible and reliable alternative to conventional methods.

Steroid hormones (SHs) are among the important classes of Contaminants of Emerging Concern (CECs) whose detection in aquatic environments is vital due to their potential adverse health impacts. Their detection is challenging because of their lower stability in natural conditions and low concentrations. This study reports the presence of steroid hormones in a major river system, the Periyar River, in Kerala (India). Water samples were collected from thirty different river locations in the case of SHs and five locations within these in the case of other CECs. These were subjected to LC-MS/MS and LC-Q-ToF/MS analyses. Five SHs, estriol, estrone, 17 β estradiol, progesterone, and hydroxy progesterone, were separated and targeted using MS techniques. The studies of the water samples confirmed the presence of the first three estrogens in different sampling sites, with estrone present in all the sampling sites. The concentration of estrone was detected in the range from 2 to 15 ng/L. Estriol and estradiol concentrations ranged from 1.0 to 5 ng/L and 1-6 ng/L, respectively. The hormones at some selected sites were continuously monitored for seven months. The chosen areas include the feed water sites for the drinking water treatment plants across the river. The monthly data revealed that estrone is the only SHs detected in all the samples in the selected months. The highest concentration of SH was found in August. Twelve CECs belonging to pharmaceuticals and personal care products were identified and quantified. In addition, 31 other CECs were also identified using non-target analysis. A detailed study of the hormone mapping reported here is the first from any South Indian River., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Over the past two decades, steroidal estrogens (SEs) such as 17α-ethylestradiol (EE2), 17β-estradiol (E2),17α-estradiol (17α-E2), estriol (E3) and estrone (E1) have elicited worldwide attention due to their potentially harmful effects on human health and aquatic organisms even at low concentration ng/L. Natural steroidal estrogens exhibit greater endocrine disruption potency due to their high binding effect on nuclear estrogen receptors (ER). However, less has been explored regarding their associated environmental risks and fate. A comprehensive bibliometric study of the current research status of SEs was conducted using the Web of Science to assess the development trends and current knowledge of SEs in the last two decades, from 2001 to 2021 October. The number of publications has tremendously increased from 2003 to 2021. We summarized the contamination status and the associated ecological risks of SEs in different environmental compartments. The results revealed that SEs are ubiquitous in surface waters and natural SEs are most studied. We further carried out an in-depth evaluation and synthesis of major research hotspots and the dominant SEs in the matrices were E1, 17β-E2, 17α-E2, E3 and EE2. Nonetheless, investigations of SEs in soils, groundwater, and sediments remain scarce. This study elucidates SEs distribution, toxicological risks, ecological fate and mitigation measures, which will be beneficial for future monitoring, management, and risk assessment. Further studies are recommended to assess the toxicological risks of different SEs in complex environmental matrices to pursue a more precise and holistic quantitative estimation of estrogenic risk., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The widespread occurrence and ubiquitous distribution of estrogens, i.e., estrone (E1), estradiol (E2), and estriol (E3) in our water matrices, is an issue of global concern. Public and regulatory authorities are concerned and placing joint efforts to eliminate estrogens and related environmentally hazardous compounds, due to their toxic influences on the environmental matrices, ecology, and human health, even at low concentrations. However, most of the available literature is focused on the occurrence of estrogens in different water environments with limited treatment options. Thus, a detailed review to fully cover the several treatment processes is needed. This review comprehensively and comparatively discusses many physical, chemical, and biological-based treatments to eliminate natural estrogens, i.e., estrone (E1), estradiol (E2), and estriol (E3) and related synthetic estrogens, e.g., 17α-ethinylestradiol (EE2) and other related hazardous compounds. The covered techniques include adsorption, nanofiltration, ultrafiltration, ultrasonication, photocatalysis of estrogenic compounds, Fenton, Fenton-like and photo-Fenton degradation of estrogenic compounds, electro-Fenton degradation of estrogenic compounds, ozonation, and biological methods for the removal of estrogenic compounds are thoroughly discussed with suitable examples. The studies revealed that treatment plants based on chemical and biological approaches are cost-friendly for removing estrogenic pollutants. Further, there is a need to properly monitor and disposal of the usage of estrogenic drugs in humans and animals. Additional studies are required to explore a robust and more advanced oxidation treatment strategy that can contribute effectively to industrial-scale applications. This review may assist future investigations, monitoring, and removing estrogenic compounds from various environmental matrices. In concluding remarks, a way forward and future perspectives focusing on bridging knowledge gaps in estrogenic compounds removal are also proposed., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Endocrine-disrupting compounds as pesticides affect the hormonal balance, and this can result in several diseases. Therefore, the analysis of representative hormones with acetamiprid (AC) and azoxystrobin (AZ) was a good strategy for the investigation of the endocrine-disrupting activity of pesticides. Hence, a sensitive and rapid analytical method using liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed. The method was validated for the analysis of AC, AZ, estriol, estrone, progesterone, and testosterone in the serum, testis, and liver of rats. The correlation between the residues of pesticides and the disturbance of the endocrine system was evaluated. The different mass parameters, mobile phase types, analytical columns, injection volumes, and extraction solvents were compared to get the lowest limit of detection of the studied compounds. The detection limits of AC, AZ, estriol, estrone, progesterone, and testosterone were 0.05, 0.05, 1.0, 10, and 1.0 ng/ml, respectively. The method developed was applied to evaluate the changes in these hormones induced by the duration of exposure to AC and AZ in rat testis and serum. The hormones level in rat serum and testis had a significant decrease as they were oral gavage treated with different high concentrations of studied pesticides. Both pesticides were distributed in the body of rats by the multi-compartment model (liver, testis, and serum)., Competing Interests: The authors have declared that no competing interests exist.

Background: Polycystic ovary syndrome (PCOS) is an endocrine and metabolic disorder with various manifestations and complex etiology. Follicular fluid (FF) serves as the complex microenvironment for follicular development. However, the correlation between the concentration of steroid in FF and the pathogenesis of PCOS is still unclear., Methods: Twenty steroid levels in FF from ten patients with PCOS and ten women with male-factor infertility undergoing in vitro fertilization were tested by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in order to explore their possibly correlation with PCOS. Meanwhile, the mRNA levels of core enzymes in steroid synthesis pathway from exosomes of FF were also detected by qPCR., Results: The estriol (p < 0.01), estradiol (p < 0.05) and prenenolone (p < 0.01) levels in FF of PCOS group were significantly increased, compared to the normal group, and the progesterone levels (p < 0.05) were decreased in PCOS group. Increased mRNA levels of CYP11A, CYP19A and HSD17B2 of exosomes were accompanied by the hormonal changes in FF. Correlation analysis showed that mRNA levels of CYP11A and HSD17B2 were negatively correlated with percent of top-quality embryos and rate of embryos develop to blastocyst., Conclusion: Our results suggest that increased levels of estrogen and pregnenolone in follicular fluid may affect follicle development in PCOS patients, and the mechanism is partially related to HSD17B1, CYP19A1 and CYP11A1 expression change in FF exosomes.

Hormones as a group of emerging contaminants have been increasingly used worldwide, which has increased their concern at the environmental level in various matrices, as they reach the water bodies through effluents due to the ineffectiveness of conventional treatments. Here we review the environmental scenario of hormones estriol (E3), 17β-estradiol (E2), and 17α-ethinylestradiol (EE2), explicitly their origins, their characteristics, interactions, how they reach the environment, and, above all, the severe pathological and toxicological damage to animals and humans they produce. Furthermore, studies for the treatment of these endocrine disruptors (EDCs) are deepened using electrochemical processes as the remediation methods of the respective hormones. In the reported studies, these micropollutants were detected in samples of surface water, underground, soil, and sediment at concentrations that varied from ng L -1 to μg L -1 and are capable of causing changes in the endocrine system of various organisms. However, although there are studies on the ecotoxicological effects concerning E3, E2, and EE2 hormones, little is known about their environmental dispersion and damage in quantitative terms. Moreover, biodegradation becomes the primary mechanism of removal of steroid estrogens removal by sewage treatment plants, but it is still inefficient, which shows the importance of studying electrochemically-driven processes such as the Electrochemical Advanced Oxidation Processes (EAOP) and electrocoagulation for the removal of emerging micropollutants. Thus, this review covers information on the occurrence of these hormones in various environmental matrices, their respective treatment, and effects on exposed organisms for ecotoxicology purposes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Background: Light-initiated chemiluminescent assays (LICA) are homogeneous assays that are sensitive, specific, and free of separation and washing steps and have high throughput and high precision., Methods: In this research, we developed a competitive method by LICA to achieve accurate quantification of estradiol (E2) in human serum. E2 competed with estriol (E3) for binding to anti-human E2 antibodies. E3 was linked to biotin via bovine serum albumin as a linker. As this assay used competition between the labeled tracer and the analyte, an increase in E2 concentration will cause a signal decrease., Results: The expected detection range of E2 was 20-5000 pg/mL. The analytical and functional sensitivities were 7.16 and 13.7 pg/mL, respectively. The intra- and inter-assay coefficients of variation were both below 15%, and the recovery rate ranged from 97.5% to 106.8%. The interference rates ranged from -3.6% to 5.4% and met detection requirements for E2 in hyperbilirubinemia, hemolysis, and lipemia in clinical samples. In addition, the cross-reactivity rates between E2 and structural analogs and some reproductive hormones varied from 1.9% to 10.6% which showed that LICA is highly specific for E2. Moreover, our results showed high accordance with the IMMULITE 2000 (y = 0.6695x + 47.92, r 2  = .843) and VIDAS systems (y = 1.099x - 821.5, r 2  = .9392)., Conclusion: Our data show that the LICA, which is easy to automate, is a promising technique for quantification of E2 in human serum and could be used for clinical detection., (© 2019 The Authors. Journal of Clinical Laboratory Analysis published by Wiley Periodicals, Inc.)

The mixed random processes of the first order autoregressive process (AR(1)) and white noise have been proved to provide a good approximation of baseline noise in a variety of analytical instruments, and may therefore be useful for estimating precision profiles. This study aims to examine a recently proposed autocorrelation method for estimating three noise parameters involved in the mixed processes (two for AR(1) and one for white noise) of HPLC, which can then be used to calculate the precision profile. This chemometric method was applied to repeatability evaluations of estriol determination using HPLC with UV detection (HPLC-UV). The relative standard deviations (RSDs) of peak area measurements for 5.0 mg/L estriol were observed to be 1.42% for the autocorrelation method and 1.63% for actual repeated measurements of real samples (n = 6). The theoretical RSDs of the autocorrelation method fell within the 95% confidence intervals of the repeated measurements. It is found that the noise parameters are obtained from real chromatographic baseline via the autocorrelation method. Moreover, the instrumental detection limit of estriol based on ISO 11843 was obtained from the precision profile (plot of RSD of measurements against concentration). This is the first paper to describe the autocorrelation method is a practically useful technique for evaluating the precision profile of HPLC-UV analyses without recourse to the repeated measurements of real samples.

This study investigated selected steroidal and phenolic endocrine disrupting compounds (EDCs) in the surface water of the Bahe River (China) using gas chromatography mass spectrometry (GC-MS). Their effect on the wild sharpbelly Hemiculter leucisculus was investigated. The concentrations of 4-t-octylphenol, nonylphenol, bisphenol-A, estrone, 17 β-estradiol, 17 α-Ethinylestradiol, and estriol were up to 126.0, 634.8, 1573.1, 55.9, 23.9, 31.5, and 5.2 ng L -1 in the surface water, and up to 26.4, 103.5, 146.9, 14.2, 9.3, 13.8, and 1.3 ng g -1 in the fish muscle tissue, respectively. High estrogen equivalent levels and hazard quotients were found in the middle and lower reaches of the river, and the pollution in these regions caused enhanced growth conditions, inhibition of gonad growth, and suppression of spermatogenesis in H. leucisculus. The up-regulation of Vitellogenin mRNA expression in male fish, collected from relatively heavily EDCs contaminated areas, indicates a potential estrogenic effect. The differential expression profiles of genes related to steroidogenesis at all sampling sites suggests that these endpoints may play an important role for the pollution monitoring of estrogenic EDCs in the Bahe River., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

We investigated the occurrence, removal and mass balance of 8 endocrine disrupting compounds (EDCs), including estrone (E1), estradiol (E2), estriol (E3), ethinylestradiol (EE2), triclosan (TCS), triclocarbon (TCC), 4-n-nonyl phenol (NP) and 4-n-octyl phenol (OP), along with 5 of their transformation products (TPs), including 4-hydroxy estrone (4-OH E1), 4-hydroxy estradiol (4-OH E2), methyl triclosan (MeTCS), carbanilide (NCC), dichlorocarbanilide (DCC) in a wastewater treatment plant. Generally, E3 showed the highest concentrations in wastewater with median value of 514 ng/L in influent, while TCS and TCC showed highest level in sludge and suspended solids (SS) with median value of 960 and 724 μg/kg, respectively. Spatial variations were observed along each unit of the wastewater treatment processes for dissolved analytes in wastewater and adsorbed analytes in suspended solids and sludge. Special emphasis was placed to understand the mass load of EDCs and their TPs to the wastewater treatment unit and mass loss during the wastewater treatment processes. Mass loss based on both aqueous and suspended phase concentration revealed that majority of these chemicals were significantly removed during the treatment process except for TCS, TCC, and three of their TPs (MeTCS, NCC, DCC), which were released or generated during the treatment process. Mass load results showed that 42.4 g of these EDCs and their TPs entered this wastewater treatment system daily via influent, whereas 6.15 g and 7.60 g were discharged through effluent and sludge., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Drinking water safety is continuously threatened by the emergence of numerous toxic organic pollutants (TOPs) in environmental waters. In this study, an approach integrating in vitro bioassays and chemical analyses was performed to explore toxicological profiles of representative source and drinking water from waterworks of the Yangtze River (Yz), Taihu Lake (Th), and the Huaihe River (Hh) basins in eastern China. Overall, 34 of 96 TOPs were detected in all water samples, with higher concentrations in both source and drinking water samples of Hh, and pollutant profiles also differed across different river basins. Non-specific bioassays indicated that source water samples of Hh waterworks showed higher genotoxicity and mutagenicity than samples of Yz and Th. An EROD assay demonstrated dioxin-like toxicity which was detected in 5 of 7 source water samples, with toxin concentration levels ranging from 62.40 to 115.51 picograms TCDD equivalents per liter of water (eq./L). PAHs and PCBs were not the main contributors to observed dioxin-like toxicity in detected samples. All source water samples induced estrogenic activities of 8.00-129.00 nanograms 17β-estradiol eq./L, and estrogens, including 17α-ethinylestradiol and estriol, contributed 40.38-84.15% of the observed activities in examined samples. While drinking water treatments efficiently removed TOPs and their toxic effects, and estrogenic activity was still observed in drinking water samples of Hh. Altogether, this study indicated that the representative source water in eastern China, especially that found in Hh, may negatively affect human health, a finding that demonstrates an urgent requirement for advanced drinking water treatments., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

In this study, a method for the simultaneous determination of two steroid hormones, 17β-estradiol (E2) and estriol (E3), and a hormone mimicking polycarbonate, bisphenol-A (BPA), was developed and validated. This was thereafter used for the determination of the levels of the hormones in surface water collected around some livestock farms. The sensitivity of the method allowed the LODs and LOQs of the hormones and mimic hormone in the range 1.14-2.510 and 3.42-7.53 μg/L, respectively. The results revealed wide variability in the concentrations of E2 and E3, while BPA was not detected at any of the sampling stations. The concentration of E3 ranged between <1.14 and 45.5 μg/L (N = 120) in station 2 water. The highest concentration of E2 (15.7 μg/L, N = 80) was observed in water from station 1. The varied concentrations may be connected with the nature and sources of release, inconsistencies in analyte distribution due to dynamics of water flow pattern and the physical/chemical properties of the receiving water bodies.

The occurrence and estrogenic activities of steroid estrogens, such as the natural estrone (E1), 17β estradiol (E2), and estriol (E3), as well as the synthetic 17α-ethynylestradiol (EE2), were investigated in eight sampling points along the Langat River (Malaysia). Surface water samples were collected at 0.5 m and surface sediment 0-5 cm from the river surface. Instrument analysis of steroid estrogens was determined by UPLC-ESI-MS with an ultra-performance liquid chromatograph (Perkin Elmer FX15) coupled to a Q Trap function mass spectrophotometer (model 3200: AB Sciex). Steroid estrogen concentrations were higher in the Langat River sediments than those in its surface water. In surface water, E1 was not detected in any sampling point, E2 was only detected in two midstream sampling points (range 0-0.004 ng/L), E3 in three sampling points (range 0-0.002 ng/L), and EE2 in four sampling points (range 0-0.02 ng/L). E1 and E2 were detected in sediments from all sampling points, E3 in five sampling points, while EE2 only in one midstream sample (3.29E-4 ng/g). Sewage treatment plants, farming waste, and agricultural activities particularly present midstream and downstream were identified as potential sources of estrogens. Estrogenic activity expressed as estradiol equivalents (EEQs) was below 1 ng/L in all samples for both surface water and sediment, indicating therefore a low potential estrogenic risk to the aquatic environment. Although the health risks are still uncertain for drinking water consumers exposed to low levels of steroid estrogen concentrations, Langat River water is unacceptable for direct drinking purposes without treatment. Further studies of endocrine disruptors in Malaysian waters are highly recommended.

The intake of contaminated foods is an important exposure pathway for endocrine disrupting chemicals (EDCs). However, data on the occurrence of EDCs in foodstuffs are sporadic and the resultant risk of co-exposure is rarely concerned. In this study, 450 food samples representing 7 food categories (mainly raw and fresh food), collected from three geographic cities in China, were analyzed for eight EDCs using high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). Besides estrone (E1), other EDCs including diethylstilbestrol (DES), nonylphenol (NP), bisphenol A (BPA), octylphenol (OP), 17β-estradiol (E2), 17α-ethinylestradiol (EE2), and estriol (E3) were ubiquitous in food. Dose-dependent relationships were found between NP and EE2 (r=0.196, p<0.05), BPA (r=0.391, p<0.05). Moreover, there existed a correspondencebetween EDCs congener and food category. Based on the obtained database of EDCs concentration combined with local food consumption, dietary EDCs exposure was estimated using the Monte Carlo Risk Assessment (MCRA) system. The 50th and 95th percentile exposure of any EDCs isomer were far below the tolerable daily intake (TDI) value identically. However, the sum of 17β-estradiol equivalents (∑EEQs) exposure in population was considerably larger than the value of exposure to E2, which implied the underlying resultant risk of multiple EDCs in food should be concern. In conclusion, co-exposure via food consumption should be considered rather than individual EDCs during health risk evaluation., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Novel chemiluminescent (CL) imaging microtiter plates with high-throughput, low-cost, and simple operation for detection of four biomarkers related to Down's syndrome screening were developed and evaluated. To enhance the sensitivity of CL immunosensing, soybean peroxidase (SBP) was used instead of horseradish peroxide (HRP) as a label enzyme. The microtiter plates were fabricated by simultaneously immobilizing four capture monoclonal antibodies, anti-inhibin-A, anti-unconjugated oestriol (anti-uE3), anti-alpha-fetoprotein (anti-AFP), and beta anti-HCG (anti-β-HCG), on nitrocellulose (NC) membrane to form immunosensing microtiter wells. Under a sandwiched immunoassay, the CL signals on each sensing site of the microtiter plates were collected by a charge-coupled device (CCD), presenting an array-based chemiluminescence imaging method for detection of four target antigens in a well at the same time. The linear response to the analyte concentration ranged from 0.1 to 40 ng/mL for inhibin-A, 0.075 to 40 ng/mL for uE3, 0.2 to 400 ng/mL for AFP, and 0.4 to 220 ng/mL for β-HCG. The proposed microtiter plates possessed high-throughput, good stability, and acceptable accuracy for detection of four antigens in clinical serum samples and demonstrated potential for practical applicability of the proposed method to Down's syndrome screening. Graphical Abstract Schematic evaluation of the microtiter plater for simultaneous detection of the four biomarkers.

The impact of solids retention time (SRT) on estrone (E1), 17β-estradiol (E2), estriol (E3) and 17α-ethinylestradiol (EE2) removal in an activated sludge plant (ASP) was examined using a pilot plant to closely control operation. Exsitu analytical methods were simultaneously used to enable discrimination of the dominant mechanisms governing estrogen removal following transitions in SRT from short (3d) to medium (10d) and long (27d) SRTs which broadly represent those encountered at full-scale. Total estrogen (∑EST, i.e., sum of E1, E2, E3 and EE2) removals which account for aqueous and particulate concentrations were 70±8, 95±1 and 93±2% at 3, 10 and 27d SRTs respectively. The improved removal observed following an SRT increase from 3 to 10d was attributable to the augmented biodegradation of the natural estrogens E1 and E2. Interestingly, estrogen biodegradation per bacterial cell increased with SRT. These were 499, 1361 and 1750ng 10(12) viable cells(-1)d(-1). This indicated an improved efficiency of the same group or the development of a more responsive group of bacteria. In this study no improvement in absolute ∑EST removal was observed in the ASP when SRT increased from 10 to 27d. However, batch studies identified an augmented biomass sorption capacity for the more hydrophobic estrogens E2 and EE2 at 27d, equivalent to an order of magnitude. The lack of influence on estrogen removal during pilot plant operation can be ascribed to their distribution within activated sludge being under equilibrium. Consequently, lower wastage of excess sludge inherent of long SRT operation counteracts any improvement in sorption., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The range of exposure rates to the steroidal estrogens estrone (E1), beta-estradiol (E2), estriol (E3), and ethinyl estradiol (EE2) in the aquatic environment was investigated by modeling estrogen introduction via municipal wastewater from sewage plants across the US. Model predictions were compared to published measured concentrations. Predictions were congruent with most of the measurements, but a few measurements of E2 and EE2 exceed those that would be expected from the model, despite very conservative model assumptions of no degradation or in-stream dilution. Although some extreme measurements for EE2 may reflect analytical artifacts, remaining data suggest concentrations of E2 and EE2 may reach twice the 99th percentile predicted from the model. The model and bulk of the measurement data both suggest that cumulative exposure rates to humans are consistently low relative to effect levels, but also suggest that fish exposures to E1, E2, and EE2 sometimes substantially exceed chronic no-effect levels., (Published by Elsevier Ltd.)

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the determination of estrone, 17beta-estradiol, estriol in animal liver and kidney tissues. The sample was extracted by tert-butyl methyl ether, and the extract was evaporated by nitrogen at 45 degrees C. The residue was redissolved in hexane-dichloromethane (6:4, v/v), then purified on a silica solid-phase extraction column. The eluant was evaporated by nitrogen, dissolved in acetonitrile-water (7:3, v/v) and then analyzed by LC-MS/MS. The separation of estrogens was performed on a Poroshell 120 EC-C18 column with the mobile phases of acetonitrile and water with a gradient elution. The identification and quantification of estrogens were carried out by negative electrospray ionization in the multiple reaction monitoring mode using external standard method. The calibration curves showed good linearity in the range of 1.0-20.0 microg/kg with the correlation coefficients above 0.99. The limit of quantification was 1.0 microg/kg for each estrogen. The average recoveries of the estrogens spiked at 1.0, 2.0, 10.0 microg/kg levels in different matrices were between 70.2% and 114%, and the relative standard deviations were between 2.01% and 14.5%. The method is simple, rapid, sensitive, good in purification effect. It is suitable for the confirmation and quantification of estrogens in animal liver and kidney tissues.

Concentrated animal feeding operations have been recognized as one of the most important contributors of natural estrogens which show significant endocrine-disrupting properties in aquatic environments. In this study, the concentrations of 17α-estradiol (17α-E2), 17β-estradiol (17β-E2), estrone (E1), and estriol (E3) in several matrices, including soils (surface and deep), sediments (surface and deep), and groundwaters, around a typical dairy farm were surveyed using gas chromatography/mass spectrometry. Of the two farmlands, surface and subsurface sediments in waste lagoon and along effluent drainage drench, the concentrations of 17α-E2, 17β-E2, and E1 ranged from below detection limit to the highest level of 6.60 μg/kg, except that E3 was not detectable. Three estrogens of 17α-E2, 17β-E2, and E1 with the concentrations of 3.18-31.61 ng/L were observed in two groundwater samples. The results clearly demonstrated the vertical migration and horizontal transport of estrogens in the investigated area. Within 750-m distance, it was observed the attenuation of 17α-E2, 17β-E2, and E1 along the effluent route and the horizontal migration of estrogens was less than 1,350 m in this survey.

Natural estrogens such as estrone (E1), 17β-estradiol (E2), estriol (E3), and the synthetic one, 17α-ethinylestradiol (EE2), are excreted by humans and animals and enter into environment through discharge of domestic sewage effluents and disposal of animal waste. The occurrence of these substances in aquatic ecosystems may affect the endocrine system of humans and wildlife so it has emerged as a major concern for water quality. Extensive research has being carried out during the last decades on the efficiency of the degradation and/or removal of these hormones in sewage treatment plants (STPs). Conventional and advanced treatments have been investigated by different authors for the elimination of estrogens from water. This paper aims to review the different processes and treatments that have been applied for the elimination of E1, E2, E3 and EE2 from water. With this purpose, physical, biological and advanced oxidation processes (AOP) have been addressed., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The occurrence of free and conjugated estrogens was examined in a survey of eleven sewage treatment plants (STPs) and their discharge water in the United Kingdom using grab sampling. The STPs included trickling filter with and without tertiary treatment, and activated sludge with tertiary treatment. For three activated sludge plants both influent and effluent samples were compared. For a further 8STPs only the effluent was examined. The estrone-3-sulphate, estradiol-3-sulphate and estriol-3-sulphate concentrations (up to 20 ng L(-1)) were typically 5-fold that of the respective free estrogen concentration in the effluents. This represents a substantial additional 'potential' estrogen load arriving in the receiving waters. Estrone-3-glucuronide was found at 9 ng L(-1), estradiol-3-glucuronide at 7 ng L(-1), and estriol-3-glucuronide at 32 ng L(-1) in sewage influent. Except on one occasion, no glucuronide conjugates could be found in the effluent. The results suggest in most cases glucuronide conjugates will be completely transformed in sewage treatment whilst sulphate conjugates will only be partially removed., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Estrogens constitute a recognized group of environmental emerging contaminants which have been proven to induce estrogenic effects in aquatic organisms exposed to them. Low removal efficiency in wastewater treatment plants results in the presence of this type of contaminants in surface waters and also even in finished drinking water. This manuscript reviews the environmental occurrence of natural (estrone, estradiol and estriol) and synthetic (ethynyl estradiol) estrogens in different water matrices (waste, surface, ground and drinking water), and their removal mainly via chemical oxidative processes. Oxidative treatments have been observed to be very efficient in eliminating estrogens present in water; however, disinfection by-products (DBPs) are generated during the process. Characterization of these DBPs is essential to assess the risk that drinking water may potentially pose to human health since these DBPs may also have endocrine disrupting properties. This manuscript reviews the DBPs generated during oxidative processes identified so far in the literature and the estrogenicity generated by the characterized DBPs and/or by the applied disinfection technology., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

The abiotic transformation of estrogens, including estrone (E1), estradiol (E2), estriol (E3) and ethinylestradiol (EE2), in the presence of model vegetable matter was confirmed in this study. Batch experiments were performed to model the catalytic conversion of E1, E2, E3 and EE2 in synthetic wastewater. Greater than 80% reduction in the parent compounds was achieved for each target chemical after 72h with the remaining concentration distributed between aqueous and solid phases as follows: 13% and 7% for E1, 10% and 2% for E2, 6% and 2% for E3, and 8% and 3% for EE2, respectively. Testosterone, androstenedione and progesterone were also monitored in this study, and their concentrations were found to be in agreement with initially spiked amount. Data collected under laboratory conditions provided the basis for implementing new abiotic wastewater treatment technologies that use inexpensive materials., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Estrogens, which contribute greatly to the endocrine-disrupting activity in sewage, are partially sorbed onto particulate matter during sewage treatment. We thus investigated the occurrence of estrogens in different kinds of sludge and throughout a plant-scale anaerobic digestion process. The analytical method was first validated when sorption interaction between spiked estrogens and sludge could occur. Hence, the recovery ratio of estrone (E1), 17beta-estradiol (E2), estriol (E3) and 17alpha-ethinylestradiol (EE2) were determined when added to liquid sludge and mixed under various conditions. We show that minor non-extractable residues were formed (5-10%), suggesting that the sorption interaction established with sludge did not limit estrogen extraction. Estrogen concentrations measured in collected samples varied with sludge type. Secondary sludge showed higher E1 contents than primary sludge: respectively, 43 and 8 ng g(-1) dry weight (dw). Two pathways of E1 production during secondary treatment are proposed to explain such a result. Higher estrogen concentrations were found in secondary sludge from a conventional plant (55 ng g(-1)dw) compared to those from an advanced plant (13 ng g(-1)dw). Based on estimated estrogen concentrations in sewage, we conclude that operating parameters play a role in the sorption of estrogens during secondary treatment. Also, the hydrophobic properties of the estrogens influenced the individual adsorption of each molecule. Thus, E3 showed the highest estimated concentrations in sewage but very low concentrations in sludge. Finally, plant-scale anaerobic digestion showed low efficiency (<40%) for removing estrogens and, regarding the final dewatering process, concentrations increased for E2 and EE2., (Copyright (c) 2010 Elsevier Ltd. All rights reserved.)

Objectives: To provide estimates and confidence intervals for the performance (detection and false-positive rates) of screening for Down's syndrome using repeated measures of biochemical markers from first and second trimester maternal serum samples taken from the same woman., Design: Stored serum on Down's syndrome cases and controls was used to provide independent test data for the assessment of screening performance of published risk algorithms and for the development and testing of new risk assessment algorithms., Setting: 15 screening centres across the USA, and at the North York General Hospital, Toronto, Canada., Participants: 78 women with pregnancy affected by Down's syndrome and 390 matched unaffected controls, with maternal blood samples obtained at 11-13 and 15-18 weeks' gestation, and women who received integrated prenatal screening at North York General Hospital at two time intervals: between 1 December 1999 and 31 October 2003, and between 1 October 2006 and 23 November 2007., Interventions: Repeated measurements (first and second trimester) of maternal serum levels of human chorionic gonadotrophin (hCG), unconjugated estriol (uE3) and pregnancy-associated plasma protein A (PAPP-A) together with alpha-fetoprotein (AFP) in the second trimester., Main Outcome Measures: Detection and false-positive rates for screening with a threshold risk of 1 in 200 at term, and the detection rate achieved for a false-positive rate of 2%., Results: Published distributional models for Down's syndrome were inconsistent with the test data. When these test data were classified using these models, screening performance deteriorated substantially through the addition of repeated measures. This contradicts the very optimistic results obtained from predictive modelling of performance. Simplified distributional assumptions showed some evidence of benefit from the use of repeated measures of PAPP-A but not for repeated measures of uE3 or hCG. Each of the two test data sets was used to create new parameter estimates against which screening test performance was assessed using the other data set. The results were equivocal but there was evidence suggesting improvement in screening performance through the use of repeated measures of PAPP-A when the first trimester sample was collected before 13 weeks' gestation. A Bayesian analysis of the combined data from the two test data sets showed that adding a second trimester repeated measurement of PAPP-A to the base test increased detection rates and reduced false-positive rates. The benefit decreased with increasing gestational age at the time of the first sample. There was no evidence of any benefit from repeated measures of hCG or uE3., Conclusions: If realised, a reduction of 1% in false-positive rate with no loss in detection rate would give important benefits in terms of health service provision and the large number of invasive tests avoided. The Bayesian analysis, which shows evidence of benefit, is based on strong distributional assumptions and should not be regarded as confirmatory. The evidence of potential benefit suggests the need for a prospective study of repeated measurements of PAPP-A with samples from early in the first trimester. A formal clinical effectiveness and cost-effectiveness analysis should be undertaken. This study has shown that the established modelling methodology for assessing screening performance may be optimistically biased and should be interpreted with caution.

The estrogens estrone (E1), 17alpha-estradiol (E2alpha), 17beta-estradiol (E2beta), and estriol (E3) are natural sex hormones produced by humans and animals. In addition, there are some synthetic estrogens, such as 17alpha-ethinylestradiol (EE2), used for contraception purposes. These compounds are able to produce endocrine disruption in living organisms at nanogram-per-liter levels. In both humans and animals, estrogens are excreted in urine and feces, reaching the natural environment through discharge from sewage treatment plants (STP) and manure disposal units. In STPs, hormone removal depends on the type of treatment process and on different parameters such as the hydraulic and sludge retention times. Thus, hormone elimination rates vary from 0% to 90% in different STPs. Animals are also an important source of estrogens in the environment. Indeed, animals produce high concentrations of hormones which will end up in manure which is typically spread on land. Hence, waste-borne animal hormones may transfer these pollutants to the soil. The purpose of this review is to highlight the significance for both health and the environment of pollution by estrogens and critically review the existing knowledge on their fate and removal in different treatment processes. Relevant information on the microbial degradation of hormones and metabolic pathways is also included.

This article describes the development of a short pre-treatment method that allows the simultaneous analysis of free estrogens (estrone, 17beta-estradiol, estriol and 17 alpha-ethynylestradiol) and their sulphate and glucuronide conjugated forms. For a range of matrices, from sewage effluent to river water, the developed methodology based on solid-phase extraction and fractionation technique with ultra-performance liquid chromatography system showed effective separation of the targeted estrogens. The detection limits of this method ranged from 0.2 to 0.8 ng L(-1) for river water. The recoveries for river water and sewage effluent varied from 63% to 127%. The problems of matrix effects and ion suppression or enhancement were allowed quantitatively for in the analysis using standard addition. The developed method was used successfully to detect estrogens and their conjugates in both raw and treated wastewater, and river water at a location in Japan. High concentrations of the free estrogens estrone, 17beta-estradiol and estriol were found in the influent (22.6, 77.2, 64.6 ng L(-1), respectively) but only E1 was still present at a high concentration in the effluent which was reflected in the downstream river concentration. Estrone-3-sulphate was detected up to 18.0 ng L(-1) in influent water sample and 1.1 ng L(-1) in downstream water. For the sulphate conjugates, removal efficiencies varied from 35 to 88%. Glucuronide conjugates were detected only once in the sewage influent.

An analytical method based on ultra performance liquid chromatography has been developed for the simultaneous determination of 15 hormones (triamcinolone, hydrocortisone, prednisone, cortisone, 6a-methylprednisolone, betamethasone, dexamethasone, prednisolone acetate, hydrocortisone acetate, estriol, beta-estradiol, estrone, diethylstilbestrol, testosterone and progesterone) in cosmetics. Various cosmetic samples were extracted with methanol under ultrasonication, and then cleaned up by a Oasis HLB solid phase extraction cartridge. The 15 hormones were separated and detected within 6 min on a Waters ACQUITY UPLC BEH C18 column (1.7 microm, 2.1 mm x 50 mm) by gradient elution with acetonitrile and water system. In the range of 1-25 ng, the calibration curves of 15 hormones showed good linearity with the correlation coefficients larger than 0.9995. The mean recoveries at the three spiked levels (2, 10, 20 mg/kg) were 88.2%-102.4% with the relative standard deviations of 1.6%-7.4%.

In this work four different commercially available enzyme-linked immunosorbent assays (ELISA) (from Japan EnviroChemicals, Ltd., Tokyo, Japan) were evaluated in terms of performance for the rapid screening of estrogens in different water matrices, including natural and spiked samples from urban wastewater, river water and ground water. All four test kits are based on monoclonal antibodies. The compounds detected by these immunoassays are (1) 17-beta-estradiol, (2) estrone, (3) 17-alpha-ethynyl estradiol and (4) estrogens in general, with high recognition properties for 17-beta-estradiol, estrone and estriol. Standards were prepared in water containing 10% (v/v) methanol. The IC50 (corresponding to the 50% of the effective concentration) values, the dynamic ranges, and the limits of detection of the ELISA kits were 0.060-0.304 microg/L, 0.05-5 microg/L and 0.05 microg/L, respectively. All samples were extracted by solid-phase extraction (SPE) beforehand, and the evaluation was carried out by comparing the results obtained by ELISA with those obtained by HPLC-MS/MS using a triple quadrupole (QqQ) instrument. In addition, two different solid-phase extraction procedures were carried out and compared. Except for moderate overestimation in the results observed with the ELISA kits in the analysis of complex wastewater samples, the results obtained using all of the tested techniques were generally in very good agreement.

An efficient method for analyzing seven sexual hormones in cosmetics, namely, estriol, estradiol-17beta, estrone, testosterone, methyl-testosterone, progesterone and diethylstilbestrol by reversed-phase high performance liquid chromatography was developed. Samples were saponified with 20 g/L sodium hydroxide. The sexual hormones were then extracted with dichloromethane-acetic acetate (40:1, v/v) under acidic conditions (pH 3, adjusted with 1 mol/L HCl). An XTerra RP18 column was employed and a mixture of water-methanol-acetonitrile (50:32:18, v/v) was used as mobile phase. The seven sexual hormones were detected at 230 nm. The average spiked recoveries for the seven sexual hormones ranged from 75.6% to 97.8% with relative standard deviations of 1.9% to 7.2%. The linear ranges of determination were from 5 to 50 mg/L with correlation coefficients of 0.9999, and the limits of detection were from 3.7 to 12 ng. The method is suitable to simultaneously assay the seven sexual hormones in cosmetics with simplicity and accuracy in a single injection.

Objective: Method comparison studies were performed in order to move a semi-automated prostate specific antigen (PSA) immunoassay and a manual unconjugated estriol (uE3) immunoassay to an automated chemistry immunoassay analyzer. The results of the two method comparison studies are compared., Design: Serum samples collected on patients with physician orders for PSA or uE3 were assayed by both methods. PSA samples were assayed on a Hybritech Tandem Photon ERA and on two Beckman Coulter Access instruments. UE3 samples were assayed by RIA and on two Beckman Coulter Access instruments. Linear regression analysis was performed on both sets of data and within-run precision and dilution studies were performed on the PSA Access method., Setting: Clinical chemistry laboratory, West Virginia University Hospitals Inc, Morgantown WV., Results: PSA linear regression analysis for the two methods (ERA and Access 1) were y = 1.0008x + 0.0393, r = 0.9976, SE = 0.1319, n = 37 and (ERA and Access 2), y = 1.0019x + 0.0486, r = 0.9964, SE = 0.1632, n = 37. Within-run precision studies for both Access instruments produced acceptable coefficient variations and dilution study results were in PSA reportable range. uE3 linear regression analysis for the two methods (RIA and Access 1) were y = 1.4105x - 0.3741, r = 0.8696, SE = 0.8330, n = 33 and (RIA and Access 2) were y = 1.315x - 0.2292, r = 0.8643, SE = 0.7964, n = 33., Conclusion: The results of the method comparison studies for PSA were acceptable and the automated PSA immunoassay method was adopted. The results of the uE3 comparison studies did not show good correlation; the automated method was not adopted.

A sensitive method for detecting 17 beta-estradiol (E2) and estriol (E3) in river water has been developed, based on the time-resolved fluoroimmunoassay by using a fluorescent europium chelate label, 4,4'-bis(1",1",1",2",2",3",3"-heptafluoro-4",6"-hexanedion-6"-yl)- chlorosulfo-o-terphenyl (BHHCT)-Eu3+. In the E2 assay, microtiter plates were coated with the E2-bovine serum albumin (BSA) conjugate. The anti-17 beta-estradiol antibody, the biotinylated goat anti-rabbit IgG antibody and the BHHCT-Eu3+ labeled streptavidin (SA)-BSA conjugate were used. In the E3 assay, the goat anti-rabbit IgG antibody was coated on a microtiter plate. The anti-estriol antibody and the BHHCT-Eu3+ labeled E3-BSA conjugate were used. The detection limits for E2 and E3 were 2.3 pg/ml and 4.3 pg/ml, respectively, and the analytical recoveries were 95-120%. Quantitative measurement of estrogens in river water was carried out for Kanda River (Tokyo, Japan) by using the method. The E2 and E3 levels were 32 pg/ml and 5.5 pg/ml, respectively. The detection limits of the present method are in the same orders of magnitude as those of ELISA for E2, and are 1-2 orders of magnitude better for E3.

Aim: To constitute a method to determine the estradiol metabolites in human liver microsome in low concentration of estradiol., Methods: Use high performance liquid chromatography after solvent extraction, evaporation, and reconstitution to separate the metabolites and use a electrochemistry detector to detect the metabolites., Results: With a mobile phase of acetic acid buffer-acetonitrile (50:50, v/v, pH 4.5) at flow rate of 1.0 mL/min and a potential of +0.7 V vs Ag/AgCl, all six composition were well separated and satisfactorily detected. There are E3, 16alpha-OHE1, 2-OHE2, E1, and two unidentified composition. The minimum detectable amount is about 100 p g on column. This method is sensitive enough to detect E1 in a substrate concentration of 1 micromol/L., Conclusion: The method can be used to study the metabolism mechanism of estradiol in liver microsome.

Purpose: To determine whether the results of triple-test analysis differ between spontaneous and IVF pregnancies., Methods: The study population consisted of 140 women with singleton pregnancies, 70 by IVF, and 70 by spontaneous conception. The groups were matched for maternal age, gestational week, and laboratory batch. The levels of all triple-test markers--alpha-fetoprotein (AFP), human chorionic gonadotropin (HCG), and unconjugated estriol (u-E3)--were compared between the groups., Results: Significantly higher HCG levels were detected in the patients with IVF pregnancies than in the control group (1.31 +/- 0.8 vs. 0.95 +/- 0.5 multiple of the medians, respectively, (p < 0.006), but there were no significant differences in AFP or u-E3 levels. Overall, 18.5% of the IVF group were found to be screen-positive as compared with 11.4% of the control group (difference not statistically significant). Only 8 IVF pregnancies (11.4%) reached the lowest calculated risk possible (1:9999) compared with 17 (24.2%) in the control group (p < 0.05)., Conclusions: Our findings support previous data demonstrating elevated maternal serum HCG in IVF patients in comparison with spontaneous ones.

In epidemiologic studies of perinatal exposures, birth weight has been proposed as a proxy variable for intrauterine estrogen exposure. To assess the validity of this assumption, we performed analyses of the association between estriol levels in 188 women in the 17th, 25th, 33rd, and 37th weeks of pregnancy and the birth weights of their infants. We found a general increase in mean cumulative estriol dose with increasing birth weight category throughout pregnancy. In late pregnancy, mean pregnancy estriol level of mothers of infants in the highest birth weight category (>4,500 gm) was twice as high as that of mothers of infants in the lowest category (<2,500 gm), 775 nmol/liter and 392 nmol/liter, respectively. Smoking lowered the maternal estriol levels by 20% or more throughout pregnancy. With smoking and birth weight included in a regression analysis, maternal age, placental weight, and infant ponderal index did not add any explanatory power to the model. Our data suggest that, on an aggregate level, birth weight can be used as a proxy variable of intrauterine estriol exposure.

The ability of breast tumors to synthesize sex steroid hormones is well recognized and their local production is thought to play a role in breast cancer development and growth. The aim of this study was to estimate local intra-tumoral and circulating levels of Estrone (E1), Estrone Sulfate (E1S), Estradiol (E2), Estriol (E3), and Testosterone (T) in 33 pre- and postmenopausal women with primary breast cancer in comparison to 12 pre- and postmenopausal women with benign breast tumors. The mean levels of the studied sex hormones were higher in serum and tumor tissue of breast cancer women than those with benign breast tumors apart from Testosterone which showed a significant decrease in pre- and postmenopausal women with breast cancer (P<0.001for follicular phase, P<0.05 for luteal phase, and P<0.005 for postmenopausal). The levels of the five hormones were significantly higher intra-tumoral than in serum of both benign and malignant breast tumor women with E1S as the predominant estrogen. There was only a positive significant correlation between serum and tumor tissue levels of E1 (rs=0.52, P<0.05 for follicular; rs=0.63, P<0.05 for luteal and rs=0.58, P<0.05 for postmenopausal) and a significant correlation between serum and tumor tissue of T (rs=0.64, P<0.05 for follicular; rs=-0.51, P<0.05 for luteal and rs=-0.81, P<0.04 for postmenopausal).

Objective: To make recommendations to physicians providing prenatal care on (1) whether prenatal screening for and diagnosis of Down syndrome (DS) is advisable and (2) alternative screening and diagnosis manoeuvres., Options: "Triple-marker" screening of maternal serum levels of alpha-fetoprotein, human chorionic gonadotropin and unconjugated estriol; fetal ultrasonographic examination; amniocentesis; and chorionic villus sampling (CVS)., Outcomes: Accuracy of detection of DS in fetuses, and risks to the mother, including psychologic distress, and to the fetus from the screening and diagnostic interventions., Evidence: A MEDLINE search for relevant articles published from Jan. 1, 1966, to Mar. 31, 1994, with the use of MeSH terms "Down syndrome," "prenatal diagnosis," "screening," "prevention," "amniocentesis," "chorionic villus sampling," "ultrasonography," "anxiety," "depression" and "psychological stress" and a manual search of bibliographies, recent issues of key journals and Current Contents., Values: The evidence-based methods and values of the Canadian Task Force on the Periodic Health Examination were used. A high value was placed on providing pregnant women with the opportunity to determine whether they are carrying a fetus with DS and to make choices concerning the termination of the pregnancy. The economic issues involved are complex and were not considered., Benefits, Harms and Costs: Triple-marker screening identifies an estimated 58% of fetuses with DS, but it has an estimated rate of true-positive results of 0.1% and of false-positive results of 3.7% (given a risk cut-off of one chance in 190 of DS). These rates vary with maternal age and the risk cut-off chosen. Women with a known risk of having a fetus with DS (e.g., those who have had a previous child with DS) may benefit from a reduction in anxiety after confirmation that their fetus does not have DS. Screening allows women at low risk of having a child with DS to detect fetuses with the syndrome, but may cause psychologic distress if there is a false-positive screening test result. Up to 20% of women with positive results of screening tests may decline to undergo a subsequent amniocentesis. Amniocentesis and CVS are very accurate in diagnosing DS in fetuses and have a very low rate of serious complications for the mother. Amniocentesis is associated with a 1.7% rate of fetal loss when it is performed after 16 weeks' gestation, whereas the rate among controls is 0.7% (for a difference of 1%, 95% confidence interval 0.3% to 1.5%). CVS entails a greater risk of fetal loss than amniocentesis (odds ratio 1.32, 95% confidence interval 1.11 to 1.57). There is little evidence from controlled trials of significant associations between amniocentesis or CVS and neonatal morbidity or malformations; however, samples have been too small to show differences in rare outcomes. Results from some case-control studies suggest that CVS increases the risk of transverse limb deficiency. Costs were not considered because they are beyond the scope of this review., Recommendations: There is fair evidence to offer triple-marker screening through a comprehensive program to pregnant women under 35 years of age (grade B recommendation). Women given detailed information about serum-marker screening show more satisfaction with the screening than those not given this information. There is fair evidence to offer amniocentesis or CVS to pregnant women 35 years of age and older and to women with a history of a fetus with DS or of a chromosome 21 anomaly (grade B recommendation). Information on the limitations and advantages of each procedure should be offered. Triple-marker screening may be offered as an alternative to CVS or amniocentesis to pregnant women over 35., Validation: Recommendations concerning prenatal diagnosis are similar to those of the US Preventive Services Task Force, the Society of Obstetricians and Gynaecologists of Canada, the Canadian College of Medical Geneticists and the Cochrane Pregnancy and Childbirth Group. No previous specific recommendations concerning triple-maker screening exist., Sponsors: These guidelines were developed and endorsed by the Canadian Task Force on the Periodic Health Examination, which is funded by Health Canada and the National Health Research and Development Program.

OBJECTIVES - To develop a standardised approach for analysing Down's syndrome screening performance in clinical practice and to apply it to published intervention trials in order to estimate detection and false positive rates more accurately. METHODS - Peer reviewed intervention trials, grouped by specific combination of analytes, were reanalysed. Revised detection rates were calculated for each study, taking into account both the high spontaneous loss during the last half of pregnancy and the possible under ascertainment of Down's syndrome live births not detected by screening. Collective screening performance was estimated, when possible, using a published methodology based on fitting receiver-operator characteristic curves. RESULTS - Sixteen trials were analysed; 11 using three, and five using two, analytes. Collective screening performance for the triple analyte trials was Down's syndrome detection rates of 57, 64, and 69% at amniocentesis referral rates of 3, 5, and 7% respectively. Four of the five studies involving two analytes performed less well, individually, when compared with the overall performance of the three analyte studies. It was not possible to estimate collective performance for the two analyte studies because there were too few. CONCLUSIONS - Accurate Down's syndrome detection rates are difficult to obtain in intervention trials owing to two potential biases, both of which tend to produce overestimates of the true rates. These sources of bias need to be taken into account when analysing and reporting Down's syndrome intervention trials. The methodology presented here offers the opportunity to achieve a more reliable, standardised estimate of both individual and collective intervention trial screening performance.

An immunological method based on labeling an antigen with an electroactive group detectable by differential pulse polarography has been demonstrated. Estriol labeled with mercuric acetate is electroactive, giving a reduction wave at -300 millivolts versus a standard calomel electrode. Addition of estriol antibody to 4-mercuric acetate estriol diminishes the peak current as a result of the antigen-antibody binding reaction. Separation of free-labeled estriol from antibody-bound-labeled antigen is unnecessary. The method is potentially useful as an analytical immunological technique.

Saliva oestriol, oestradiol, and progesterone concentrations were measured in 23 women who went into spontaneous preterm labour. The patients fell clinically and biochemically into two groups. The 13 who went into preterm labour with intact membranes had a saliva oestriol to progesterone ratio greater than one in every case and greater than the 95th centile for their length of gestation in 12 cases; by contrast, all those who went into spontaneous preterm labour after prolonged rupture of the membranes had an oestriol to progesterone ratio less than one and below the 50th centile for their period of gestation in the one to four days before delivery. Saliva oestradiol to progesterone ratios were randomly distributed throughout the normal range in both groups. It appears that preterm labour without prior prolonged rupture of the membranes is, like term labour, preceded by an increase in the saliva oestriol to progesterone ratio. It may therefore be possible to use this ratio to predict preterm labour.

Free natural estrogens in raw and commercial whole milk were quantitated by radioimmunoassay. The ranges of concentration of estrone, estradiol 17-beta, and estriol were 34 to 55, 4 to 14, and 9 to 31 pg/ml. Proportions of active estrogens (estrone and estradiol) in the fat phases of milk by radioactive tracer on separated milk were 80% and 65%. These findings were supported by radioimmunoassay of skim milk and butter. Equilibrium dialysis of skim milk with hydrogen 3 labeled estrogens showed that 84 to 85% of estrone and estradiol and 61 to 66% of estriol were protein bound. Whey proteins demonstrated a greater binding capacity than casein. This result was confirmed by radioimmunoassay of dry curd cottage cheese and whey. The concentrations in curd were 35, 11, and 6 pg/g. In whey they were 4, 2, and 3 pg/ml. The quantity of active estrogens in dairy products is too low to demonstrate biological activity. Butter was highest with concentrations of 539, 82, and 87 pg/g. Human colostrum demonstrated a maximum concentration of 4 to 5 ng/ml for estrone and estriol and about .5 ng/ml for estradiol. By the 5th day postpartum, they decreased to become similar to cow's milk.

Placental protein 5 (PP5), pregnancy-specific glycoprotein (SP1), pregnancy-associated alpha 2-glycoprotein (SP3) and chorionic gonadotrophin could not be demonstrated in appreciable molar quantities in the soluble fraction from microvillous plasma membrane preparations isolated from the syncytiotrophoblast of full-term human placentae. However, progesterone, total oestriol and placental lactogen may have some association with this membrane.

A pilot study of the endogenous steroid concentrations in human breast tumours was performed. The technique of high-resolution molecular-ion monitoring during combined g.l.c.-mass spectrometry was used to determine oestrone, oestradiol-17beta and oestriol in concentrations above 1ng/g wet wt. of tissue, and dehydroepiandrosterone, testosterone, androsterone (3alpha-hydroxy-5alpha-androstan-17-one) and 3beta-hydroxy-5alpha-androstan-17-one in concentrations exceeding 5ng/g, in extracts of five primary breast tumours.