Determination of phenolic endocrine disrupting chemicals and acidic pharmaceuticals in surface water of the Pearl Rivers in South China by gas chromatography-negative chemical ionization-mass spectrometry

An analytical method for phenolic endocrine disrupting chemicals and acidic pharmaceuticals in river water was developed using gas chromatography mass spectrometry (GC-MS) coupled with negative chemical ionization (NCI) technique, and used for the determination of these compounds in the Pearl Rivers (Liuxi, Zhujiang and Shijing Rivers). Derivatization using pentafluorobenzoyl chloride (PFBOCl) and pentafluorobenzyl bromide (PFBBr) before GC-MS analysis were applied and optimized for phenolic compounds and acidic compounds, respectively. The target compounds were analyzed for river waters from the upstream to downstream of the Pearl Rivers. Phenolic compounds 4-tert-octylphenol (4-t-OP), 4-nonylphenol (4-NP), bisphenol-A (BPA), estrone (E1), estradiol (E2) and triclosan (TCS) were detected at trace or low levels in the water samples from Liuxi River and Zhujiang River. Diethylstilbestrol (DES) was not detected in the Pearl Rivers. The highest concentrations of the phenolic compounds were found in Shijing River, and they were 3150 ng/L for 4-t-OP, 11,300 ng/L for 4-NP, 1040 ng/L for BPA, 79 ng/L for E1, 7.7 ng/L for E2 and 355 ng/L for TCS, respectively. Only a few acidic pharmaceuticals were detected at low concentrations in water from Liuxi River and Zhujiang River, but the highest concentrations for the acidic pharmaceuticals were also found in Shijing River. The highest concentrations detected for clofibric acid, ibuprofen, gemfibrozil, naproxen, mefenamic acid and diclofenac were 17 ng/L, 685 ng/L, 19.8 ng/L, 125 ng/L, 24.6 ng/l and 150 ng/L, respectively. The results suggest Liuxi and Zhujiang Rivers are only slightly contaminated and can be used as drinking water sources, but Shijing River is heavily polluted by the wastewater from nearby towns.     

Analysis of environmental endocrine disrupting chemicals using the E-screen method and stir bar sorptive extraction in wastewater treatment plant effluents

Endocrine disrupting chemicals (EDCs) have become a major issue in the field of environmental science due to their ability to interfere with the endocrine system. Recent studies show that surface water is contaminated with EDCs, many released from wastewater treatment plants (WWTP). This pilot study used biological (E-screen assay) and chemical (stir bar sorptive extraction-GC-MS) analyses to quantify estrogenic activity in effluent water samples from a municipal WWTP and in water samples of the recipient river, upstream and downstream of the plant.The E-screen assay was performed on samples after solid phase extraction (SPE) to determine total estrogenic activity; the presence of estrogenic substances can be evaluated by measuring the 17-β-estradiol equivalency quantity (EEQ). Untreated samples were also assayed with an acute toxicity test (Vibrio fischeri) to study the correlation between toxicity and estrogenic disruption activity.Mean EEQs were 4.7 ng/L (± 2.7 ng/L) upstream and 4.4 ng/L (± 3.7 ng/L) downstream of the plant, and 11.1 ng/L (± 11.7 ng/L) in the effluent. In general the WWTP effluent had little impact on estrogenicity nor on the concentration of EDCs in the river water. The samples upstream and downstream of the plant were non-toxic or weakly toxic (0 < TU < 0.9) while the effluent was weakly toxic or toxic (0.4 < TU < 7.6). Toxicity and estrogenic activity were not correlated.At most sites, industrial mimics, such as the alkylphenols and phthalates, were present in higher concentrations than natural hormones. Although the concentrations of the detected xenoestrogens were generally higher than those of the steroids, they accounted for only a small fraction of the EEQ because of their low estrogenic potency. The EEQs resulting from the E-screen assay and those calculated from the results of chemical analyses using estradiol equivalency factors were comparable for all samples and closely correlated.     

北京污水厂进、出水中内分泌干扰物的分布

以三种酚类化合物(4-OP、4-n-NP、BPA)、五种雌激素(E1、E2、E3、17α-E2、EE2)为目标物,对其在北京市三个污水处理厂进、出水中的浓度及工艺流程中的分布、迁移进行了研究.结果表明,污水处理厂出水中浓度最高的物质是BPA、EE2,分别为(56～140)、(78～115)ng/L.BPA和天然雌激素(17α-E2除外)主要被生物降解去除,而对EE2的去除主要发生在初级处理过程,去除率约为63%.两种烷基酚在污水处理厂并不能被有效去除.与欧美国家、日本等相比,北京市污水处理厂进、出水中的内分泌干扰物浓度偏高,尤其是出水中的雌激素浓度较高,具有一定的环境风险.     

An assessment of estrogenic organic contaminants in Canadian wastewaters

A suite of 30 primarily estrogenic organic wastewater contaminants was measured in several influent/effluent wastewater samples from four municipal wastewater treatment plants and effluents from one bleached kraft pulp mill (BKME) using an ultra-trace analytical method based on gas chromatography-high resolution mass spectroscopy (GC-HRMS). In vitro recombinant yeast assay detection of the estrogenic equivalent (EEq) on whole and solid phase extracted (SPE) and fractionated wastewater was also performed. 19-norethindrone was the most frequently detected and abundant (26-224 ng/L) of all the synthetic estrogens/progesterones in the influent samples. 17alpha-ethinylestradiol was the more frequently detected synthetic estrogen/progesterone in the effluents occurring at or below 5 ng/L with some sporadic occurrences of up to 178 ng/L. The greatest levels of steroidal estrogens in municipal effluents were E1>E2>E3 which were all <20 ng/L. Nonylphenol and di(2-ethylhexyl) phthalate were found to be the highest non-steroidal synthetic compounds surveyed in both municipal influent and effluent samples, both occurring at 6-7 microg/L in municipal effluents. BKME contained relatively large amounts of the plant sterol stigmasterol (4 microg/L) but low amounts of fecal sterols, and steroidal estrogens (E2 only at 6 ng/L) when compared to the municipal effluents. In vitro EEq in the wastewater surveyed ranged from 9-106 ng E2/L and ranked from municipal influent>municipal effluent approximately BKME, with most of the estrogenicity fractionating in the 100% methanol SPE fraction followed by a secondary amount in the diethyl ether (for municipal) or methyl-tert butyl ether (for BKME) SPE fractions. Most correlations between chemical and in vitro estrogenic equivalency were weak (p>0.05 in most cases). Unexpected inverse correlations between in vitro estrogenic activity and concentrations of the estrogenic contaminant bisphenol A were found which likely contributed to the weakness of these correlations. A modified toxicity identification and evaluation procedure was continued with the SPE extracts from the more potent 100% methanol SPE fractions of municipal effluent. High performance liquid chromatography band elution retention times, based on in vitro estrogen detection, indicated that steroidal estrogens such as E2 were responsible for most of the estrogenicity of the samples. Subsequent collection and GC-MS analysis of active bands did not confirm the presence of steroidal estrogens, but expanded the possibility of phthalate esters (i.e. dibutyl phthalate) and natural sterols (i.e. beta-sitosterol) contributing to the overall estrogenic load.     

Sulfamethoxazole contamination of a deep phreatic aquifer

Groundwater samples were obtained from the water table region of a phreatic aquifer (unsaturated zone depth up to 28 m) under land irrigated with wastewater effluents for about 5 decades and a relatively deep pumping well (109 m), used as a drinking water source till 2007, located downstream (1300 m) of wastewater effluent and sludge infiltration facilities. Sulfamethoxazole (SMX) concentrations in secondary effluents varied between 90 and 150 ng/L. SMX was extracted using SPE and was analyzed by HPLC-MS/MS. SMX (maximum concentration of 37 ng/L) was detected in the water table region, in two monitoring wells, after an unsaturated zone transport period of about 16 years. The maximum SMX concentration detected in the pumping well was of 20 ng/L. These results question wastewater effluent disposal strategies including the suitability of irrigation with effluents on the replenishment area of an aquifer supplying drinking water.     

Removal of dissolved estrogen in sewage effluents by β-cyclodextrin polymer

Substances with estrogenic activity are found in effluents of municipal sewage plants and dairy farms. These effluents have the potential to induce feminization in male fish. In this study, cyclodextrin polymers (CDPs) that are insoluble in both polar and non-polar solvents were selected for the removal of dissolved estrogens in the effluent of a municipal sewage plant. The removal capacity of CDPs was high in the order of β-CDP ≥ γ-CDP ? α-CDP. The mechanism for adsorption of estrogens to β-CDP was not only due to a host-guest interaction as molecular recognition by β-cyclodextrin (β-CD), but also due to adsorption by the polymer matrix. β-CDP of 0.2% (w/v) removed 17β-estradiol (E2) of about 70% from 10⁻¹¹ mol/L, and more than 90% from ≥ 10⁻¹⁰ mol/L. The removal ratios of E2 in the presence of cholesterols, which are contained at higher concentrations than estrogens in sewage effluents and are adsorptive competitor for β-CDP, were about 85% at a cholesterol/E2 molar ratio of 100 and > 90% at molar ratios of 0.1, 1, and 10. The effluent from a municipal sewage plant had estrogenic activity corresponding to 5.5 × 10⁻¹¹ molE2/L by yeast two-hybrid assay. The estrogens in the effluent were also removed > 90% by the β-CDP treatment. Therefore, β-CDP is able to remove dissolved estrogens over a wide range of concentrations in the presence of various contaminants such as wastewaters.     

A theoretical estimation of the concentration of steroid estrogens in effluents released from municipal sewage treatment plants into aquatic ecosystems of central-southern Chile

Endocrine disorders associated with sewage effluents have been documented in aquatic species from various regions of the world and sewage treatment works (STWs) are now widely recognized as one of the major discharge source of endocrine disrupting compounds. Steroid estrogens usually emerge as the main contributors to the endocrine disrupting capacity of municipal sewage effluents. Because human wastes are believed to be the primary source of release of steroid estrogens in watercourses, the presence of these compounds in aquatic systems is likely to constitute a pervasive ecological problem. In spite of that, the endocrine disrupting impact of sewage effluents has rarely been investigated in South America. In this paper, we used Johnson and Williams' predictive model to estimate the concentration of steroid estrogens in effluents released from 38 municipal STWs of central-southern Chile and to assess steroid estrogen concentrations in rivers. In STW effluents, we estimated the estrogen concentrations to range from 9.35 to 739.92 ng/L for estrone, 1.03 to 81.74 ng/L for estradiol and 0.38 to 30.56 ng/L for ethynylestradiol. Overall, the predicted estrogen concentrations are significantly higher than those reported for STW effluents in the literature. This can be explained by demographic and sewage flow differences between Chile and industrialized western countries. Predicted steroid estrogen concentrations at river sites indicate that endocrine disruption in fish is likely to occur in the Itata catchment. However, future research is needed to attest this and to evaluate the real impact of the STW discharges into central-southern Chile's marine and freshwater environments.     

Temporal variability of combined sewer overflow contaminants: Evaluation of wastewater micropollutants as tracers of fecal contamination

A monitoring program was initiated for two sewage outfalls (OA and OB) with different land uses (mainly residential versus institutional) over the course of a year. Eleven CSO events resulting from fall and summer precipitations and a mixture of snowmelt and precipitation in late winter and early spring were monitored. Median concentrations measured in CSOs were 1.5 × 10⁶ Escherichia coli/100 mL, 136.0 mg/L of Total Suspended Solids (TSS), 4599.0 ng/L of caffeine (CAF), 158.9 ng/L of carbamazepine (CBZ), in outfall OA and 5.1 × 10⁴ E. coli/100 mL, 167.0 mg TSS/L, 300.8 ng CAF/L, 4.1 ng CBZ/L, in outfall OB. Concentration dynamics in CSOs were mostly related to the dilution by stormwater and the time of day of the onset of overflows. Snowmelt was identified as a critical period with regards to the protection of drinking water sources given the high contaminant concentrations and long duration of events in addition to a lack of restrictions on overflows during this period. Correlations among measured parameters reflected the origins and transport pathways of the contaminants, with E. coli being correlated with CBZ. TSS were not correlated with E. coli because E. coli was found to be mostly associated with raw sewage whereas TSS were additionally from the resuspension of in-sewer deposits and surface runoff. In receiving waters, E. coli remained the best indicator of fecal contamination in strongly diluted water samples as compared to WWMPs because WWMPs can be diluted to below their detection limits.     

Responses of Hexaplex (Murex) trunculus to selected pollutants

We report the results of a recent survey of the concentration of natural estrogens (17beta-estradiol, 17alpha-estradiol, estrone, estriol) and the synthetic estrogen, 17alpha-ethynylestradiol in representative animal wastes and sewage treatment plant (STP) effluents in the Waikato region of New Zealand. Dairy farm effluent samples showed high levels of estradiol (19-1360 ng/L) and its breakdown product estrone (41-3123 ng/L) compared with piggery or goat farm effluents. The combined load for these estrogens (excluding beta epimer) varied from 60 to >4000 ng/L. The piggery effluent provided the lowest total estrogen load (46 ng/L), with estrone accounting for nearly 60% of the measured estrogens in this sample. The synthetic analogue, 17alpha-ethynylestradiol was detected only in one wastewater treatment plant sample, albeit at trace level. An estrogen receptor competitive binding assay was used to test the biological activity of the samples and confirmed that most agricultural waste samples contain high levels of estrogenic compounds. The potential of these wastes to cause endocrine disruption in the receiving ecosystem is unknown at present.     

Perfluorinated compounds in surface waters and WWTPs in Shenyang, China: mass flows and source analysis

Concentrations of 10 perfluorinated chemicals (PFCs) were investigated in the Hun River (HR), four canals, ten lakes, and influents and effluents from four main municipal wastewater treatment plants (WWTPs) in Shenyang, China. Mass flows of four main PFCs were calculated to elucidate the contribution from different sections of the HR. Overall, perfluorooctanoic acid (PFOA) and perfluorohexanoic acid (PFHxA) were the major PFCs in the HR, with ranges of 2.68-9.13 ng/L, and 2.12-11.3 ng/L, respectively, while perfluorooctane sulfonate (PFOS) was detected at lower levels, ranging from 0.40 to 3.32 ng/L. The PFC concentrations in the HR increased after the river passes through two cities (Shenyang and Fushun), indicating cities are an important contributor for PFCs. Mass flow analysis in the HR revealed that PFC mass flows from Fushun are 1.65-5.50 kg/year for C6-C8 perfluorinated acids (PFCAs) and 1.29 kg/year for PFOS, while Shenyang contributed 2.83-5.18 kg C6-C8 PFCAs/year, and 3.65 kg PFOS/year. The concentrations of PFCs in four urban canals were higher than those in the HR, with the maximum total PFCs of 240 ng/L. PFOA and PFOS showed different trends along these canals, suggesting different sources for the two PFCs. Total PFCs in ten lakes from Shenyang were at low levels, with the greatest concentration (56.2 ng/L) detected in a heavily industrialized area. The PFC levels in WWTP effluents were higher than those in surface waters with concentrations ranging from 18.4 to 41.1 ng/L for PFOA, and 1.69-3.85 ng/L for PFOS. Similar PFC profiles between effluents from WWTPs and urban surface waters were found. These results indicate that WWTPs are an important PFC source in surface water. Finally, we found that the composition profiles of PFCs in surface waters were similar to those in tap water, but not consistent with those in adult blood from Shenyang. The calculation on total daily intake of PFOS by adults from Shenyang showed that the contribution of drinking water to human exposure was minor.     

Identification of estrogen-like effects and biologically active compounds in river water using bioassays and chemical analysis

The Nackdong River is the longest river in South Korea and passes through major cities that have several industrial complexes, including chemical, electric, and petrochemical complexes, and municipal characteristics such as apartment complexes. Along the river, the Gumi region has an electric industrial complex and an apartment complex that may be possible point sources of xenoestrogens such as phenolic compounds. To identify the causative chemicals for estrogenic activity in the river water of this region, bioassay-directed chemical analysis was performed. All samples from six sampling sites (an upstream point: S1; hot spot points: S2-1, S2-2, and S2-3; and downstream points: S3, and S4) showed estrogenic activity in the E-screen assay, with bio-EEQs (17β-E₂-equivalent quantities) ranging from 25.35-677.15 pg/L. Samples from S2-2, the sampling point downstream of the junction of stream water, and domestic and industrial wastewater, contained the highest estrogenic activity. Since the bio-EEQ of the organic acid fraction (F2) of the S2-2 sample had the highest activity (823.25 pg-EEQ/L) and F2 may contain phenolic compounds, GC-MS analyses for phenolic xenoestrogens were conducted with the organic acid fractions of the river water samples. Six estrogenic phenolic chemicals, 4-NP, BPA, 4-t-OP, 4-t-BP, 4-n-OP, and 4-n-HTP, were detected, with the highest concentrations (I-EEQ) found in S2-2 (231.80 pg/L). Among these phenolic chemicals, 4-NP was the most potent estrogen (bio-EEF; 8.12 × 10^− 5) and acted as a full agonist. Furthermore, 4-NP was present at levels (2.0 µg/L in S2-2) that can induce VTG induction in fish (>1 µg/L). In addition, we confirmed that river water (S2-2) significantly increased serum VTG levels in crucian carp (Carassius auratus) in a fish exposure experiment under laboratory conditions. Therefore, phenolic xenoestrogens, especially 4-NP, may be the main causative compounds responsible for the estrogenic effect on the Nackdong River.     

Occurrence of seven artificial sweeteners in the aquatic environment and precipitation of Tianjin,China

Seventy water samples, including wastewaters, tap waters, fresh surface waters, coastal waters, groundwaters, and precipitation samples, from Tianjin, China, were analyzed for seven commonly used artificial sweeteners (ASs). The concentrations of the investigated ASs were generally in the order of wastewater treatment plant (WWTP) influent > WWTP effluent > surface water > tap water > groundwater ≈ precipitation, while the composition profiles of ASs varied in different waters. Acesulfame, sucralose, cyclamate, and saccharin were consistently detected in surface waters and ranged from 50 ng/L to 0.12 mg/L, while acesulfame was the dominant AS in surface and tap waters. Aspartame was found in all of the surface waters at a concentration up to 0.21 μg/L, but was not found in groundwaters and tap waters. Neotame and neohesperidin dihydrochalcone were less frequently detected and the concentrations were low. The concentrations of the ASs in some of the surface waters were of the same order with those in the WWTP influents, but not with the effluents, indicating there are probably untreated discharges into the surface waters. The ASs were detected in precipitation samples with high frequency, and acesulfame, saccharin, and cyclamate were the predominant ASs, with concentrations ranging from 3.5 ng/L to 1.3 μg/L. A gross estimation revealed that precipitation may act as a source for saccharin and cyclamate in the surface environment of Tianjin city. Moreover, the presence of ASs in the atmosphere was primarily assessed by taking 4 air samples to evaluate their potential source in precipitation.     

Occurrence of androgens and progestogens in wastewater treatment plants and receiving river waters: Comparison to estrogens

Research has shown that exposure to androgens and progestogens can cause undesirable biological responses in the environment. To date, however, no detailed or direct study of their presence in wastewater treatment plants has been conducted. In this study, nine androgens, nine progestogens, and five estrogens were analyzed in influent and final effluent wastewaters in seven wastewater treatment plants (WWTPs) of Beijing, China. Over a period of three weeks, the average total hormone concentrations in influent wastewaters were 3562 (Wujiacun WWTP)-5400 ng/L (Fangzhuang WWTP). Androgens contributed 96% of the total hormone concentrations in all WWTP influents, with natural androgen (androsterone: 2977 ± 739 ng/L; epiandrosterone: 640 ± 263 ng/L; and androstenedione: 270 ± 132 ng/L) being the predominant compounds. The concentrations of synthetic progestogens (megestrol acetate: 41 ± 25 ng/L; norethindrone: 6.5 ± 3.3 ng/L; and medroxyprogesterone acetate: 6.0 ± 3.2 ng/L) were comparable to natural ones (progesterone: 66 ± 36 ng/L; 17α,20β-dihydroxy-4-progegnen-3-one: 4.9 ± 1.2 ng/L; 21α-hydroxyprogesterone: 8.5 ± 3.0 ng/L; and 17α-hydroxyprogesterone: 1.5 ± 0.95 ng/L), probably due to the wide and relatively large usage of synthetic progestogens in medical therapy. In WWTP effluents, androgens were still the dominant class accounting for 60% of total hormone concentrations, followed by progestogens (24%), and estrogens (16%). Androstenedione and testosterone were the main androgens detected in all effluents. High removal efficiency (91-100%) was found for androgens and progestogens compared with estrogens (67-80%), with biodegradation the major removal route in WWTPs. Different profiles of progestogens in the receiving rivers and WWTP effluents were observed, which could be explained by the discharge of a mixture of treated and untreated wastewater into the receiving rivers.     

Comparison of natural estrogen removal efficiency in the conventional activated sludge process and the oxidation ditch process

The presence of natural estrogens, 17beta-estradiol (E2), estrone (E1) and estriol (E3), as well as estrogenic activity in wastewater influents and secondary effluents were investigated in 20 full-scale wastewater treatment plants in Japan. In all of the influent samples, natural estrogens were detected at concentrations above the minimum limits of detection (0.5ng/L). The concentrations of natural estrogens detected in the effluent of oxidation ditch plants were generally lower than previously reported values. On the other hand, in the conventional activated sludge plants, increments of E1 during biological treatment were frequently observed although E2 and E3 were removed effectively in the process. The removal rates of natural estrogens or estrogenic activity show no observed statistical relationship with the solids retention time (SRT) and the hydraulic retention time (HRT). However, the plants with high SRT or HRT generally showed high and stable removal of both natural estrogens and estrogenic activity.     

Occurrence and removal of N-nitrosamines in wastewater treatment plants

The presence of nitrosamines in wastewater might pose a risk to water resources even in countries where chlorination or chloramination are hardly used for water disinfection. We studied the variation of concentrations and removal efficiencies of eight N-nitrosamines among 21 full-scale sewage treatment plants (STPs) in Switzerland and temporal variations at one of these plants. N-nitrosodimethylamine (NDMA) was the predominant compound in STP primary effluents with median concentrations in the range of 5-20 ng/L, but peak concentrations up to 1 μg/L. N-nitrosomorpholine (NMOR) was abundant in all plants at concentrations of 5-30 ng/L, other nitrosamines occurred at a lower number of plants at similar levels. From concentrations in urine samples and domestic wastewater we estimated that human excretion accounted for levels of <5 ng/L of NDMA and <1 ng/L of the other nitrosamines in municipal wastewater, additional domestic sources for <5 ng/L of NMOR. Levels above this domestic background are probably caused by industrial or commercial discharges, which results in highly variable concentrations in sewage. Aqueous removal efficiencies in activated sludge treatment were in general above 40% for NMOR and above 60% for the other nitrosamines, but could be lower if concentrations were below 8-15 ng/L in primary effluent. We hypothesize that substrate competition in the cometabolic degradation explains the occurrence of such threshold concentrations. An additional sand filtration step resulted in a further removal of nitrosamines from secondary effluents even at low concentrations. Concentrations released to surface waters were largely below 10 ng/L, suggesting a low impact on Swiss water resources and drinking water generation considering the generally high environmental dilution and possible degradation. However, local impacts in case a larger fraction of wastewater is present cannot be ruled out.     

Determining estrogenic steroids in Taipei waters and removal in drinking water treatment using high-flow solid-phase extraction and liquid chromatography/tandem mass spectrometry

River water and wastewater treatment plant (WWTP) effluents from metropolitan Taipei, Taiwan were tested for the presence of the pollutants estrone (E1), estriol (E3), 17beta-estradiol (E2), and 17alpha-ethinylestradiol (EE2) using a new methodology that involves high-flow solid-phase extraction and liquid chromatography/tandem mass spectrometry. The method was also used to investigate the removal of the analytes by conventional drinking water treatment processes. Without adjusting the pH, we extracted 1-L samples with PolarPlus C18 Speedisks under a flow rate exceeding 100 mL/min, in which six samples could be done simultaneously using an extraction station. The adsorbent was washed with 40% methanol/60% water and then eluted by 50% methanol/50% dichloromethane. The eluate was concentrated until almost dry and was reconstituted by 20 microL of methanol. Quantitation was done by LC-MS/MS-negative electrospray ionization in the selected reaction monitoring mode with isotope-dilution techniques. The mobile phase was 10 mM N-methylmorpholine aqueous solution/acetonitrile with gradient elution. Mean recoveries of spiked Milli-Q water were 65-79% and precisions were within 2-20% of the tested concentrations (5.0-200 ng/L). The method was validated with spiked upstream river water; precisions were most within 10% of the tested concentrations (10-100 ng/L) with most RSDs<10%. LODs of the environmental matrixes were 0.78-7.65 ng/L. A pre-filtration step before solid-phase extraction may significantly influence the measurement of E1 and EE2 concentrations; disk overloading by water matrix may also impact analyte recoveries along with ion suppression. In the Taipei water study, the four steroid estrogens were detected in river samples (ca. 15 ng/L for E2 and EE2 and 35-45 ng/L for E1 and E3). Average levels of 19-26 ng/L for E1, E2, and EE2 were detected in most wastewater effluents, while only a single effluent sample contained E3. The higher level in the river was likely caused by the discharge of untreated human and farming waste into the water. In the drinking water treatment simulations, coagulation removed 20-50% of the estrogens. An increased dose of aluminum sulfate did not improve the performance. Despite the reactive phenolic moiety in the analytes, the steroids were decreased only 20-44% of the initial concentrations in pre- or post-chlorination. Rapid filtration, with crushed anthracite playing a major role, took out more than 84% of the estrogens. Except for E3, the whole procedure successfully removed most of the estrogens even if the initial concentration reached levels as high as 500 ng/L.     

Comparing steroid estrogen, and nonylphenol content across a range of European sewage plants with different treatment and management practices

The effluent of 17 sewage treatment works (STW) across Norway, Sweden, Finland, The Netherlands, Belgium, Germany, France and Switzerland was studied for the presence of estradiol (E2), estrone (E1), ethinylestradiol (EE2) and nonylphenol (NP). Treatment processes included primary and chemical treatment only, submerged aerated filter, oxidation ditch, activated sludge (AS) and combined trickling filter with activated sludge. The effluent strength ranged between 87 and 846 L/PE (population equivalent), the total hydraulic retention time (HRT) ranged between 4 and 120 h, sludge retention time (SRT) between 3 and 30 d, and water temperature ranged from 12 to 21 °C. The highest estrogen values were detected in the effluent of the STW which only used primary treatment (13 ng/L E2 and 35 ng/L E1) and on one occasion in one of the STW using the AS system (6.5 ng/L E2, 50.5 ng/L E1, but on three other occasions the concentrations in this STW were at least a factor of 6 lower). For the 16 STW employing secondary treatment E2 was only detected in the effluent of six works during the study period (average 0.7-5.7 ng/L). E1 was detected in the effluent of 13 of the same STW. The median value for E1 for the 16 STW with secondary treatment was 3.0 ng/L. EE2 was only detected in two STW (1.1, <0.8-2.8 ng/L). NP could be detected in the effluent of all 14 STW where this measurement was attempted, with a median of 0.31 μg/L and values ranging from 0.05 to 1.31 μg/L. A comparison of removal performance for E1 was carried out following prediction of the probable influent concentration. A weak but significant (α<5%) correlation between E1 removal and HRT or SRT was observed.     

High estrogen concentrations in receiving river discharge from a concentrated livestock feedlot

Environmental estrogenic chemicals interrupt endocrine systems and generate reproductive abnormalities in wildlife, especially natural and synthetic estrogenic steroid hormones such as 17β-estradiol (E2), estrone (E1), estriol (E3), 17α-ethynylestradiol (EE2), and diethylstilbestrol (DES). Concentrated animal feedlot operations (CAFOs) are of particular concern since large amounts of naturally excreted estrogens are discharged into aquatic environments. This study investigated E2, E1, E3, EE2, and DES with high performance liquid chromatography/tandem mass (HPLC-MS/MS) analyses along Wulo Creek in southern Taiwan, near a concentrated livestock feedlot containing 1,030,000 broiler chickens, 934,000 laying hens, 85,000 pigs, and 1500 cattle. Sampling was performed from December 2008 to May 2009, in which 54 samples were collected. Experimental results indicate that concentrations of EE2 were lower than the limit of detection (LOD), and concentrations of DES were only detected twice. Concentrations ranged from 7.4 to 1267 ng/L for E1, from not detected (ND) to 313.6 ng/L for E2, and from ND to 210 ng/L for E3. E1 had the highest average mass fraction (72.2 ± 3.6%), which was significantly higher than E3 (16.2 ± 1.7%) and E2 (11.5 ± 2.6%). Additionally, the mean E2 equivalent quotient (EEQ) ranged from 17.3 to 137.9 ng-E2/L. Despite having a markedly lower concentration than E1, E2 more significantly contributed (52.4 ± 6.0%) EEQ than E1 (19.7 ± 3.5%). Moreover, the concentrations of E2, E1, and E3 upstream were significantly higher than concentrations downstream, suggesting a high attenuation effect and fast degradation in the study water. Most concentrations in winter season were higher than those of spring season due to the low dilution effect and low microbial activity in the winter season. Based on the results of this study, we recommend further treatment of the wastewater discharge from the feedlot.     

Development of a sensitive E-screen assay for quantitative analysis of estrogenic activity in municipal sewage plant effluents

A simplified proliferation test with human estrogen receptor-positive MCF-7 breast cancer cells (E-screen assay) was optimized and validated for the sensitive quantitative determination of total estrogenic activity in effluent samples from municipal sewage plants. After solid phase extraction of 1 l sewage on either 0.2 g polystyrene copolymer (ENV+) or 1 g RP-C18 material and removal of the solvent, analysis of the extracts in the E-screen assay could be performed without any clean-up step. This was even possible with untreated sewage. Parallel extraction of four sewage samples on both different solid phase materials gave comparable quantitative results in the E-screen. A blank sample did not induce cell proliferation. As additive behaviour of the estrogenic response of single compounds was proven for two different mixtures each containing three xenoestrogens, total estrogenic activity in the sewage samples, expressed as 17 beta-estradiol equivalent concentration (EEQ), could be calculated comparing the EC50 values of the samples with those of the positive control 17 beta-estradiol. The detection limit of the E-screen method was 0.05 pmol EEQ/l (0.014 ng EEQ/l), the limit of quantification 0.25-0.5 pmol EEQ/l (0.07-0.14 ng EEQ/l). In total, extracts of nine effluent and one influent sample from five different municipal sewage plants in South Germany were analyzed in the E-screen. All samples strongly induced cell proliferation in a dose-dependent manner which was completely inhibited by coincubation with 5 nM of the estrogen receptor-antagonist ICI 182,780. The proliferative effect relative to the positive control 17 beta-estradiol (RPE) was between 30 and 101%. 17 beta-Estradiol equivalent concentrations were between 2.5 and 25 ng/l indicating a significant input of estrogenic substances via sewage treatment plants into rivers.     

Occurrence and fate of TMDD in wastewater treatment plants in Germany

The occurrence and fate of 2,4,7,9-tetramethyl-5-decyne-4,7-diol (TMDD) was investigated in four wastewater treatment plants (WWTPs) in Germany. The concentration of TMDD in influents and effluents in the WWTPs ranged from 134 ng/L to 5846 ng/L and from <LOQ to 3539 ng/L correspondingly. Loads determined in influents (10.1 g/d-1142 g/d) and effluents (<LOQ - 425 g/d) indicate that TMDD is partially removed from the wastewater. The elimination rates varied between 33% and 68%. Based on the load analysis, the TMDD effluent discharge of WWTPs investigated varied from 8.29 kg/a to 52.6 kg/a. Day and week profiles were recorded and indicated that TMDD is introduced into the sewage through household and indirect dischargers with high fluctuations. Seasonal variations in the TMDD loads were also analyzed in three of the studied WWTPs. One of the WWTPs demonstrated statistically higher TMDD loads during the warm period (164 g/d) than during the cold period (91.3 g/d), for the others WWTPs any differences could not be established. The input of TMDD during weekends and working days was also studied. The results did not show any significant trend of TMDD discharge into the WWTPs.