Analysis of Endocrine Disrupting Steroids: Investigation of Their Release into the Environment and Their Behavior During Bank Filtration

A reliable and sensitive analytical method for the determination of estrone (E1), 17β-estradiol (E2), and the synthetic hormone 17α-ethinylestradiol (EE2) has been established. Samples are concentrated using automated solid-phase extraction and analysis is performed by liquid chromatography with detection by tandem mass spectrometry. Recoveries of all analytes were between 93% and 107%, and limits of quantification (LOQs) between 0.1 and 0.4 ng/L for purified sewage, and surface, ground, and drinking water, and between 1 and 2 ng/L in case of raw sewage. For the investigation of estrogen release into the environment and its behavior during sewage and surface water treatment, and during ground water recharge, samples from municipal waste water treatment plants (WWTPs), a surface water treatment plant, a bank filtration site, and a ground water enrichment (GWE) pond were analyzed. El was found in the raw waste water with an average concentration of 157 ng/L, whereas E2 and EE2 were found with mean concentrations of only 18 and 9 ng/L, respectively. Sewage treatment by municipal WWTPs affected a removal of EE2 (76%), El (92%), and E2 (94%). In the investigated surface water of Berlin, only E1 could be detected at concentrations around or below 1 ng/L. E2 and EE2 were not present in the Berlin surface water above the LOQ of 0.2 ng/L, respectively. Surface water treatment also leads to a significant removal of E1 (> 80%). In the ground water samples from the GWE site near Lake Tegel, only a few samples contained detectable concentrations of E1. These samples were collected from a shallow monitoring well located very close to the bank of the pond. Even a short distance between the bank and observation wells led to concentrations of El below the LOQ showing the potential of ground water recharge systems for the retention of estrogenic steroids depending on the environmental setting.     

Behavior of Steroid Hormones and Conjugates During Wastewater Treatment – A Comparison of Three Sewage Treatment Plants

The fate of the steroid hormones 17 β‐estradiol, estrone, estriol, 16 α‐hydroxyestrone, and β‐estradiol 17‐acetate, the hormone‐conjugates β‐estradiol 3‐sulfate and estrone 3‐sulfate, and the oral contraceptives 17 α‐ethinylestradiol and mestranol were studied during wastewater treatment as wastewater treatment plants are the major source contamination of urban surface waters with steroid hormones. The elimination efficiencies of three different concepts of WWTPs, i. e., activated sludge versus trickling filter, were compared over four weeks at different weather conditions. While larger WWTPs operating on activated sludge eliminated hormones more constantly than smaller WWTPs, heavy rainfall events led to a collapse of the elimination efficiency. By using trickling filter techniques for the treatment of wastewater an elimination of the steroid hormones could not be observed. Additionally, mass flows on a per person basis are compared. In the three experiments, which ran continuously for four weeks each, it turned out that the concentrations of ethinylestradiol and mestranol were below 6 ng/L in all samples. The inflow concentrations were 70 to 82 ng/L (estrone), 17 to 44 ng/L (estradiol), 61 to 130 ng/L (hydroxyestrone), 189 to 255 ng/L (estriol), 10 to 17 ng/L (estrone‐3‐sulfate) and about 28 ng/L (estradiol‐3‐ sulfate). While in the activated sludge treatment plants the elimination of estrone was 90 and 50%, respectively, estrone was formed from precursors in the trickling filter plant. A similar situation occurred for 17β‐estradiol, estrone 3‐sulfate, and estradiol 3‐sulfate. Hydroxyestrone was eliminated with similar efficiencies in all wastewater treatment plants (64 to 82%), as well as estriol (34 to 69%). Accordingly, the emissions of the wastewater treatment plants differed largely and were not attributed to the size of the respective plant, only.     

Entwicklung von Enzymimmunoassays zum Nachweis von Phenoxycarbonsäure-Herbiziden in Trink- und Grundwässern Development of Enzyme Immunoassays for the Detection of Phenoxycarboxylic Acid Herbicides in Drinking Water and Groundwater

Competitive solid phase enzyme immunoassays using polyclonal antibodies were developed for the detection of the phenoxycaboxylic acids MCPB [4-(4-chloro-2-methylphenoxy)butyric acid], Mecoprop [2-(4-chloro-2-methylphenoxy)propionic acid], and 2,4-D [(2,4-dichloro-phenoxy)acetic acid] in drinking water and ground water. The carrier protein for the immunization was bovine serum albumin, horseradish peroxidase conjugates were employed as enzyme tracer. For the three antisera, the optimization of detection limits and test sensitivities was our first consideration. For the mecoprop and 2,4-D antisera, the strongest influences were the pH value and the ionic strength, as much as the use of enzyme tracers with lower affinities. The MCPB antiserum reacted with 2,4-DB[4-(2,4-dichlorophenoxy)butyric acid] with equal specificity, either could be detected at 0.02 μg/L (80% B/B₀), middle of the test (50 B/B₀) lying at 0.1 μg/L. The detection limit with mecoprop antiserum was optimized to 0.02 μg/L, the prescribed limit for drinking water of 0.1 μg/L ling at 60 % B/B₀. The strongest cross-reactivity was found for mecoprop methyl ester. MCPB. 2,4-DB, and dichloroprop have crossreactivities of 50%, 6.7%, and 6.3%, respectively. The 2,4-D antiserum reacts less sensitively with 2,4-D, the detection limit being 0.4 μg/L. The 2,4-D isooctyl ester and 2,4-D methyl ester demonstrate as cross-reacting compounds high cross-reactivites of 3630% and 2230%. The cross-reactions of the compounds 2,4-DB, MCPB, and MCPA [(4-chloro-2-methylphenoxy)acetic acid] lie at 52%, 69%, and 41%. 100 ground water tests were spiked within laboratory. All positive samples were correctly identified. Falsely negative results did not appear.     

Levels of estrogenic compounds in Xiamen Bay sediment, China

Concentrations of seven estrogenic compounds, i.e., estrone (E1), 17β-estradiol (E2), 17α-ethynylestradiol(EE2), diethylstilbestrol (DES), nonylphenol (NP), octylphenol (OP) and bisphenol A (BPA), were determined in sediments and pore water from Xiamen Bay in China, and their distributions and potential risks in the Bay were assessed. Total estrogenic compounds concentrations varied from 49.20 to 1230.69 ng/g dw in sediments and from 102.33 to 4376.60 ng/L in pore water. The highest levels of these compounds were found at Yundang Lagoon. The results showed that estrogenic compounds in Xiamen Bay originated mainly from municipal wastewaters. Compared with other areas, Xiamen Bay was contaminated with high levels of estrogen hormones. This contamination poses a potential threat to benthic organisms. Although a good relationship (r = 0.94) was observed between the estrogenic compounds concentrations and total organic carbon (TOC) contents in sediments, which did not indicate that the sediment organic matter favors the accumulation of the detected estrogenic compounds.     

Simultaneous Analysis of Natural Free Estrogens and Their Sulfate Conjugates in Wastewater

Natural estrogens from humans increasingly attract attention because of their strong endocrine disrupting potency. The discharge of sewage water is considered as the most important source of these endocrine disrupting chemicals (EDCs) in the environment. Therefore, a GC‐MS method was developed for the simultaneous analysis of six natural free estrogens and their sulfate conjugates in municipal wastewater, in which natural free estrogens and sulfate conjugates were successfully separated from an Oasis HLB solid phase extraction (SPE) cartridge with two different eluents, and the sulfate conjugates were then transformed to their corresponding free estrogens by acid solvolysis. Before the analysis with GC‐MS, samples were derivatized by N,O‐bis (trimethylsilyl) trifluoroacetamide (BSTFA) plus 1% trimethylchlorosilane (TMCS) at 80°C for 40 min. Satisfactory recoveries ranging from 64 to 112.3% were obtained by spiking ultra‐purified water, raw, and treated municipal wastewater with the six estrogens at 50, 100, and 50 ng/L, respectively. The method was successfully applied to wastewater samples from one WWTP, which suggested that E1 was the dominant natural estrogens in effluent and E3‐3S was one of the conjugates possibly occurring in the effluent.     

Occurrence of selected estrogens in mangrove sediments

This paper presents results related to the occurrence and distribution of estrogens along the Brazilian coast. Three mangrove areas were chosen to evaluate the presence of estrogens in surface sediments of mangrove forests. The presence of estrogens was observed in all studied sites. 17-α-Ethinylestradiol (EE2), a synthetic estrogen, was the most common and has been found in higher concentration (0.45-129.78 ng/g) compared to 17-β-estradiol (E1) and estrone (E2) (both being natural estrogens). The concentrations of E1 and E2 ranged from 0.02 to 49.27 ng/g and 0.03 to 39.77 ng/g, respectively. Theoretically, under anaerobic conditions EE2 can be reduced to E1 even in environments such as sediments of mangrove forests, which are essentially anaerobic. Even if the concentrations of estrogens seem to be insignificant in some samples, the effects remain uncertain.     

Nachweis von Industriechemikalien (HPS,BPS und SPS) im Oberflächenwasser

Occurrence of Industrial Chemicals (HPS, BPS, and SPS) in Surface Water The paper gives the results of water examinations for different phenylsulfonamides. Random samples taken every month between May 1999 and August 2000 from surface water out of the river Rhine (kilometer 838), the river Ruhr (Mülheim Styrum) and the river Emscher (Oberhausen center) were tested for the corrosion inhibiting agent 6‐[methyl(phenylsulfonyl)amino]‐hexanoic acid (HPS) as well as its metabolites 4‐[methyl‐(phenylsulfonyl)amino]‐butanoic acid (BPS) and sarkosin‐N‐(phenylsulfonyl) (SPS). Furthermore, the sewage plant effluents of two municipal wastewater treatment plants from the rural area were also included in the monitoring program. The analytical method includes solid‐phase extraction (SPE), a derivatization step as well as gas chromatography mass spectrometry (GC‐MS). SPS is regularly found in all investigated surface waters, but only occasionally in the effluents of the two rural sewage plants. The median values for SPS amount to 0.09 μg/L in the river Rhine, 0.60 μg/L in the river Ruhr, and 0.70 μg/L in the river Emscher. BPS can only be found in the river Ruhr (median value: 0.08 μg/L) and in the river Emscher (median value: 0.41 μg/L). HPS was regularly found in a surface water for the first time. This substance can be detected in the Emscher through the whole measurement period. The median value for HPS amounts to 1.78 μg/L. Aditionally, the validation characteristics of an alternative analytical method including solid‐phase microextraction (SPME) is worked out. The fully automated process includes an on‐fiber methylation step and the GC‐MS. The repeatability standard deviation of the process amounts to RSD < 12%. Detection limits between 0.07 and 0.70 μg/L are achieved.     

Bildung bromhaltiger Desinfektionsnebenprodukte nach Chlorung von Bodenseewasser in Abhängigkeit von Aufbereitungsschritten und der Bromidkonzentration

Formation of Brominated Disinfection By-products after Chlorination of Water from Lake Constance Depending on Treatment Steps and Bromide Concentration The formation of halogenated disinfection by-products (DNP) (trihalomethanes (THM), haloacetic acids (HES), haloacetonitriles (HAN), haloketones (HKE) and chloropicrin (TCNM)) was studied after chlorination (0.5 mg/L) of raw water and drinking water from Lake Constance. The objectives of this study were to investigate the effect of bromide ion on the distribution of DNP-species resulting from chlorination and to draw up a bromide mass-balance. The treatment of water from Lake Constance with ozone decreased the formation of DNP about 35% and of total organic halogens (AOX) about 45%. This decline in DNP-formation is mainly due to the formation of chloroform, dichloro- and trichloroacetic acid. Though there is only a small amount of bromide (6.2 μg/L) in water from Lake Constance, about 32% of THM, 16% of HES, and 30% of HAN contained bromine after chlorination of treated water. The addition of bromide ion (up to 1 mg/L) shifted the DNP-composition to more brominated species. The distribution of the DNP-species depends on the initial molar bromide-to-chlorine ratio. A mathematic model showed the same pattern of DNP-species depending on the initial molar bromide-to-chlorine ratio that was observed experimentally in microstrained raw water and drinking water from Lake Constance. After chlorination of microstrained raw water, 27% of the bromide were incorporated in known DNP, 34% in unknown substances, and about 39% of bromide were not converted to organic bromine. After chlorination of treated water, only 16% of the bromide were incorporated in unknown substances, whereas 34% were found in known DNP. About 50% were found as bromide again. After addition of bromide (up to 25 μg/L) to ozonated and treated water from Lake Constance, between 60 and 72% of the bromide that had reacted after chlorination were found in analyzed low-molecular DNP.     

Levels of endocrine disrupting compounds in South China Sea

The occurrence of estrogens in the aquatic environment has become a major concern worldwide because of their strong endocrine disrupting potency. In this study, concentrations of four estrogenic compounds, estrone (E1), 17β-estradiol (E2), 17α-ethynylestradiol (EE2), estriol (E3) were determined with liquid chromatography-tandem mass spectrometry analyses in surface water from South China Sea, and distributions and potential risks of their estrogenic activity were assessed. The estrogenic compounds E1, E2 and E3 were detected in most of the samples, with their concentrations up to 11.16, 3.71 and 21.63 ng L⁻¹. However, EE2 was only detected in 3 samples. Causality analysis, EEQ values from chemical analysis identified E2 as the main responsible compounds. Based on the EEQ values in the surface water, high estrogenic risks were in the coastal water, and low estrogenic risks in the open sea.     

Production of Drinking Water from Highly Contaminated Surface Waters: Removal of Organic,Inorganic, and Microbial Contaminants Applying Mobile Membrane Filtration Units

In military out of area missions of the Bundeswehr, it can be necessary to produce drinking water even from highly polluted surface waters containing a variety of organic, inorganic, and microbiological contaminants. Thus, mobile drinking water purification systems must be able to remove such contaminants as far as possible to meet the requirements of the German and European drinking water regulation/directive. Presently, two novel drinking water purification units applying membrane filtration undergo intensive long‐term trials carried out by the Bundeswehr. If these trials positively proof the functionality of these units and their ability to remove all possible contaminants they shall substitute so far available devices which use large amounts of chemicals and charcoal filtration for water purification.In the course of a research project, the functionality of the new devices and their efficacy to remove high amounts of algae, microbes, and organic and inorganic pollutants are additionally tested in “worst‐case” field studies. In September 2000, the first mobile drinking water purification unit was tested at the Teltowkanal in Berlin, Germany.This canal was chosen because it carries high burdens of municipal sewage effluents. The results from the fatigue test confirmed the ability of the water purification unit to reduce the concentrations of all contaminants meeting the maximum tolerance levels set by the German/European drinking water regulation.The pre‐filtration device was very effective in removing algae and solid particles to protect the membranes from clogging and to enable an almost maintenance‐free operation. Residues of pharmaceuticals and some other organic contaminants have almost totally been removed from the surface water where they were detected at individual concentrations up to the μg/L‐level.     

Pestizide in Oberflächenwässern — Befunde in Talsperren und in Uferfiltraten Pesticides in Surface Water — Analytical Results for Drinking Water Reservoirs and Bank Filtrate Waters

The results of a series of investigations are presented, performed by the governmental chemical laboratory Karlsruhe (CLUA) in order to determine pesticides in drinking water for purposes of food monitoring. 231 samples of 8 municipal water supplies (groundwater wells/no bank filtrate) and 34 private water supplies close to the bank of the river Rhine (groundwater wells/partly bank filtrate) were analysed. The sampling sites were located between Mannheim and Greffern, Germany. In 1986 high amounts of atrazine and simazine were determined in the drinking water wells of several private water supplies, especially at the sampling sites in the south. The likewise high chloride content of the samples was taken as a proof of bank filtrate contributing to the water. The decrease of atrazine load in the river water of the Rhine since 1987 has its parallel in the analytical results for the concerned drinking water wells. Obviously the pesticide contamination of water from bank filtrate pumping wells is mainly depending on the pesticide load of the river water. The application of fertilizers and pesticides in the fields seems not to be a significant source of the pesticide contamination of dringing waters derived from bank filtrate water. For reasons of public health a monitoring of pesticide content in bank filtrate pumping wells of private water supplies is recommended.     

Livestock wastes as a source of estrogens and their effects on wildlife of Manko tidal flat,Okinawa

The Manko tidal flat in the southern part of Okinawa Island is an important visiting and wintering area for migratory birds and was added to the Ramsar Convention Register of Wetlands in 1999. This area used to be an inlet extending to the inner part of Naha Port, but recent reclamation projects have restricted its connection to the East China Sea. As is typical in rural regions of subtropical islands, the inhabitants in the Manko basin raise livestock, especially pigs, without employing sufficient waste treatment methods. As sewage treatment works are considered to be one of the main sources of environmental estrogens in urban areas, the significance of livestock farming as a source of estrogens in rural area is examined in this study. In the present study, total estrogenic activities in water and sediment samples from the Manko tidal flat and its basin were measured using a recombinant yeast screen method. Estrogenic activities (equivalent to 17beta-estradiol, E2) were around 10 ng l(-1) in water samples and more than 10 microg kg(-1) in some sediment samples. In addition, the concentrations of estrone (E1) and E2 in water samples measured using LC/MS/MS indicated a high contribution of environmental estrogens from livestock wastes.     

Occurrence and Distribution of Organic Contaminants in the Aquatic System in Berlin. Part III: Determination of Synthetic Musks in Berlin Surface Water Applying Solid-phase Microextraction (SPME) and Gas Chromatography-Mass Spectrometry (GC-MS)

Polycyclic musks and nitro musks were found as environmental pollutants in screening analyses of 30 representative surface water samples collected from rivers, lakes, and canals in Berlin. These synthetic musks, which are used as fragrances in cosmetics, detergents, and other products, are discharged by the municipal sewage treatment plants into Berlinπs surface waters. In particular, the polycyclic musks 1,3,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethylcyclopenta-(g)-2-benzopyrane (HHCB; Galaxolide®, Abbalide®) and 7-acetyl-1,1,3,4,4,6-hexamethyltetraline (AHTN; Tonalide®, Fixolide®) were found in the receiving surface waters at considerable concentrations up to the μg/L-level. In the Wuhle, a small brook almost totally consisting of sewage effluents, maximum concentrations were 12.5 μg/L for HHCB and 6.8 μg/L for AHTN. Additionally, the polycyclic musk 4-acetyl-1,1-dimethyl-6-tert-butylindane (ADBI; Celestolide®, Crysolide®) and 1-tert-butyl-3,5-dimethyl-2,6-dinitro-4-acetylbenzene (musk ketone) were detected at low concentrations in most of the samples. Two other nitro musks, namely 1,1,3,3,5-pentamethyl-4,6-dinitroindene (musk moskene) and 1-tert-butyl-3,5-dimethyl-2,4,6-trinitrobenzene (musk xylene), were only detected in a single surface water sample. Solid-phase microextraction (SPME) with detection by gas chromatography-mass spectrometry (GC-MS) has proven to be a rapid and reliable tool for the screening of synthetic musks in surface and sewage water samples. Internal calibration with a suitable internal standard enables reproducible quantitation of the analytes down to the low ng/L-level.     

Occurrence and Distribution of Organic Contaminants in the Aquatic System in Berlin. Part I: Drug Residues and other Polar Contaminants in Berlin Surface and Groundwater

Several polar contaminants were found in screening analyses of 30 representative surface water samples collected from rivers, lakes, and canals in Berlin. Residues of pharmaceuticals and N-(phenylsulfonyl)-sarcosine originating from various sewage treatment plants effluents were found at concentrations up to the μg/L-level in the surface water, whereas the concentrations of polar pesticides such as dichlorprop and mecoprop were always below 0.1 μg/L. The pharmaceuticals most frequently detected in the surface water samples include clofibric acid, diclofenac, ibuprofen, propiphenazone, and two other drug metabolites. Additional investigations of groundwater wells of a drinking water plant have shown that polar contaminants such as drug residues or N-(phenylsulfonyl)-sarcosine easily leach through the subsoil into the groundwater aquifers when contaminated surface water is used for groundwater recharge in drinking water production.     

The use of steroid markers to assess sewage contamination of the Black Sea

Analyses of faecal steroids in coastal sediments taken from throughout the Black Sea indicate chronic sewage contamination at some locations. These include Sochi, where concentrations of coprostanol up to 5400 ng g(-1) (dry wt) were recorded, and in the coastal areas adjacent to the Danube delta (2600 ng g(-1) dry wt). Comparatively high values of 5beta/(5beta+5alpha) isomeric ratios of cholestan-3-ol and cholestan-3-one are reported at these locations and are characteristic of sediments contaminated with sewage. Lower concentrations of coprostanol are reported for Bosphorus sediments (12-440 ng g(-1) dry wt) and in the region of Odessa (130-290 ng g(-1) dry wt). Isomeric ratios at these locations also confirm sewage as a significant contributor to steroids in the Bosphorus, but lower values of the ratios indicate only minor sewage inputs in Odessa. In contrast, steroid concentrations and compositions from most Ukrainian sites are characteristic of uncontaminated environments. To place these results into perspective, the range in levels of coprostanol in the Black Sea sediments compares to the lower to mid-range of concentrations reported for coastal sediments on a world-wide basis, with Black Sea sedimentary levels substantially below those previously reported for heavily impacted sites.     

Occurrence of endocrine-disrupting chemicals in riverine sediments from the Pearl River Delta, China

The endocrine-disrupting chemicals (EDCs) was investigated in 28 riverine sediments from the Pearl River system, China and analyzed by an ultrasonication extraction and GC-MS method. The concentrations of 4-tert-octylphenol (OP), 4-nonylphenol (NP), and bisphenol A (BPA) in the sediments were in the ranges of <2.0-210, 107-16198 and <1.7-430 ng/g dw, respectively. The steroid estrogens estrone (E1) and 17β-estradiol (E2) in the sediments ranged from <1.3 to 10.9 ng/gdw and from <0.9 to 2.6 ng/gdw, respectively. The spatial distribution of these chemicals was related to the discharge of domestic and industrial wastewater along the rivers. The positive correlation between EDCs and total organic carbon indicates that sedimentary organic carbon is an important factor in controlling the distributions of EDCs. Compared with other previous studies, the ZR and DR rivers from the PRD were heavily contaminated by APs and BPA.     

Untersuchung der Sorptionseigenschaften des Kationenaustauschers KB-2M mit Makronetzstruktur zur Abtrennung von Zinkionen aus Ab- und Spülwässern

Study of Sorptional Properties of the Cation Exchanger KB-2M with Macroreticular Structure for Recovery of Zinc Ions from Sewage and Rinsing Water Although a number of ion-exchange methods have been employed for the recovery of some transition metals from industrial effluents, knowledge about ion-exchange resins with macroreticular structure is poor. The present paper describes the mechanism of sorption on such exchangers and their application for recovery of zinc from sewage rinsing water. Ion exchanger of macroreticular structure are polymers with long-chained cross-linking agents. We have synthesized carboxylic ion-exchange resins by hydrolysis of copolymerisates of methyl acrylate with different cross-linking agents: divinylbenzene, divinyl sulfide, divinyl ester of ethylene glycol and divinyl ester of di- or triethylene glycol. The sorption process on modifications of the carboxylic resins KB-2 of various structure was studied with different methods: potentiometric titration, infrared spectroscopy, electron microscopy, X-ray structural analysis. The initial zinc concentration in rinsing water was 0.05 mol/L at pH from 3 to 6. For the sorption, 0.2…1.0 g of resin were equilibrated with 100 mL of zinc solution. After equilibrium (12 h), the resin was separated from solution. The zinc ions were determined by atomic absorption spectrometry after stripping with 100 mL of 10% sulfuric acid. The distribution ratio D was calculated (D: mmole of Zn sorbed per gram of resin divided by mmole of Zn per millilitre of solution). By means of infrared spectroscopy, the mechanism of sorption of zinc ions from rinsing water was determined. There may be a possibility of the formation of complexes in the cation-exchange resin phase. It was found out in this paper that the cation-exchanger KB-2M of macroreticular structure is the most effective for the sorption of the Zn²⁺-ions from sewage and rinsing water.     

Simultaneous Analysis of Natural Free Estrogens and Their Conjugates in Wastewater by GC‐MS

A solid‐phase extraction (SPE)‐gas chromatography (GC)‐mass spectrometry (MS) analytical method was developed for the simultaneous analysis of natural free estrogens and their conjugates in wastewater samples. Natural free estrogens and their conjugates in wastewater were successfully separated by the oasis hydrophilic‐lipophilic balance solid phase extraction (Oasis HLB SPE) method, and the conjugates were initially enzyme hydrolyzed by β‐glucuronidase or arylsulfatase from Helix pomatia prior to derivatization. N‐methyl‐N‐(tert‐butyldimethylsilyl)trifluoroacetamide (MTBSTFA) plus 1% tert‐butyldimetheylchlorosilane (TBDMCS) was chosen as the derivatization reagent, and the most appropriate conditions of derivatization were determined to be at 95°C for 90 min. The recovery ratios of nine target chemicals were determined by spiking them in 1 L of ultra‐purified water or the influent of a wastewater treatment plant (WWTP). The recovery ratios of six out of nine for the analytes ranged from 73.3–114.9% with relative standard deviations (RSD) from 1.6–19.9%. The established method was successfully applied to environmental wastewater samples which were collected from one municipal wastewater treatment plant (WWTP) in Osaka, Japan, for the determination of natural free estrogens and their conjugates. In the influent sample, E1, E2, E1‐3S, E3‐3S, and E1‐3G were detected at concentrations of 16.6, 9.6, 8.2, 21.9, and 3.2 ng L^–1, respectively. However, only E1 was detected at a high concentration of 44 ng L^–1 in the effluent sample, suggesting that it is the dominant natural free estrogen in the effluent.     

Bestimmung von Pflanzenbehandlungsmitteln in Wasser durch Online-Fest-Flüssig-Extraktion und HPLC-DAD

Determination of Pesticides in Water by On-line Solid-phase Extraction and HPLC-DAD This paper describes an HPLC on-line procedure for the determination of 29 relevant pesticides in water by diode-array detection using a compact HPLC-system with integrated column-switching. The sample – 5 mL – is injected with a large volume injection system on a precolumn cartridge (RP-C18). Enriched compounds are eluted in frontflush mode by columnswitching to a microbore column and separated with a gradient water/acetonitrile. During desorption the precolumn cartridge is heated at 80°C resulting in a comparable efficiency to the separation with direct injection. The procedure is fully automated for continuous monitoring of drinking-, ground-, and surface water with low contamination levels. Sample preparation is limited to a filtration step with an anion-exchange membrane filter to reduce natural contaminations e.g. humic acids simultaneously. For a concentration of 50 ng/L, the signal to noise ratio varies from 3:1 (Ethidimuron) up to 25:1 (Chloridazon). The standard deviations compare to the standardized HPLC-method DIN EN ISO 11369; the recovery rates are 100% and reproducible. The method was successfully tested on ruggedness with several sequences from a monitoring program. The precolumn cartridge needs replacement after 100 sample injections. Finally, this method was coupled with a benchtop mass-spectrometer (electrospray mode) without changing the chromatographic conditions. With selected ion monitoring (SIM) selectivity and detection sensitivity could be improved considerably compared to DAD-detection.     

Environmentally relevant exposure of 17α-ethinylestradiol impairs spawning and reproductive behavior in the brackish medaka Oryzias melastigma

We assessed the effects of 17α-ethinylestradiol (EE2) on spawning and reproductive behavior of the brackish medaka Oryzias melastigma at environmental concentrations. Breeding pairs were exposed to EE2 at either lower concentrations (0, 1, and 10 ng/L) or higher concentrations (0, 50, and 100 ng/L) for 14 days. Swimming performance and reproductive behavior (following, dancing, and copulation) of the males were analyzed at the end of exposure. Additionally, gonad histology was conducted in the males in the higher EE2 experiment. We found that spawning was significantly inhibited in the 50–100 ng/L EE2 groups. Swimming performance was not affected in all groups. Dancing and copulation were significantly suppressed in the 50–100 ng/L groups. No effect was observed in testis histology in the 50–100 ng/L groups. Our data suggests that reproductive behavior of O. melastigma can be utilized as a behavioral endpoint for ecotoxicology studies of endocrine disruptors in estuarine/marine environments.