Ecological and ecotoxicological responses in the assessment of the ecological status of freshwater systems: A case-study of the temporary stream Brejo of Cagarrão (South of Portugal).

The objective of the study was to assess the integrated use of macroinvertebrate indexes and ecotoxicological parameters in the evaluation of the ecological status of a temporary stream with a strong agricultural influence. Water quality was analysed at two sampling sites along the stream, considering: chemical supporting parameters; hazardous substances (pesticides); benthic macroinvertebrate communities, through quality (Iberian Biological Monitoring Working Party and Iberian Average Score Per Taxon) and multi-metric indices (Southern Portuguese Index of Invertebrates and Ecological Quality Ratio); and ecotoxicological responses using lethal and sub-lethal bioassays. The water chemical characterization showed high levels of organic matter and nutrients, mainly in the dry period ((biochemical oxygen demand (BOD5): 18.5-25.5mgL-1, chemical oxygen demand (COD): 60.8-193.7mgL-1; total phosphorus (TP): 0.17-0.33mgL-1)), which may compromise the support of biological life. In accordance with the physicochemical results, the stream had an ecological status less than good. Of the 25 pesticides analysed, only five, namely terbuthylazine, 2-methyl-chlorophenoxyacetic acid, bentazone, mecoprop and metolachlor were quantified. In general, the concentrations of pesticides detected were low, except at the source of the stream in January 2012 (sum of pesticides 2.29µgL-1), mainly due to the concentration of bentazone (1.77µgL-1), both values surpassing the European Commission threshold values. The analysis of benthic macroinvertebrates showed low levels of abundance and family diversity, with communities dominated by resistant groups to organic pollution and pesticides, such as the Chironomidae family. In general, the reproduction ecotoxicological results showed a very marked decrease in the number of juveniles per female. The Spearman correlation identified pesticides, namely MCPA (R=-0.89; p<0.05), as the main responsible for the observed effect. The results showed the linearity and complementarity of the two groups of biological responses, allowing to cover the interactions between the ecosystem's species and the different types of pollutants.

Cocaine and other illicit drugs in airborne particulates in urban environments: a reflection of social conduct and population size.

Levels of cocaine and other psychoactive substances in atmospheric particulate matter (PM) were determined in urban environments representing distinct social behaviours with regard to drug abuse: night-life, university and residential areas. Three cities (with population>1 million and <0.3 million inhabitants) were selected. Mean daily levels of drugs in PM were 11-336 pg/m3 for cocaine, 23-34 pg/m3 for cannabinoids, and 5-90 pg/m3 for heroin. The highest levels were recorded on weekends, with factors with respect to weekdays of 1-3 for cocaine, 1-2 for cannabinoids and 1.1-1.7 for heroin. Higher levels were detected in the night-life areas, pointing towards consumption and trafficking as major emission sources, and possibly ruling out drug manufacture. The similarities in temporal trends at all sites suggested a city-scale transport of psychoactive substances. Correlations were detected between cocaine and amphetamine consumption (r2=0.98), and between heroin and cannabinoids (r2>0.82).

Evaluation of two aquatic passive sampling configurations for their suitability in the analysis of estrogens in water.

The presence of estrogens in the aquatic environment has been the target of several studies in the last decade. Newly developed passive sampling techniques for polar organic chemicals show great promise for the assessment of long-time exposure of aquatic organisms to emerging contaminants. In the present work, two configurations of the Chemcatcher(®) passive sampler have been tested for their applicability to the analysis of seven estrogens in water. Accumulation experiments in the laboratory, to calculate the uptake rates, and a field trial show that the polar configuration of this device may be used for the efficient sampling and determination of estrogens in water. Time weighted average concentrations were determined in the field trial and compared with spot sampling concentrations. The detection of estriol using passive sampling, although not found with spot sampling, clearly demonstrates the value of this technique in assessing relevant concentrations of estrogens in the aquatic media.

Analysis of phytoestrogens, progestogens and estrogens in environmental waters from Rio de Janeiro (Brazil).

The environment is currently exposed to a large variety of man-made chemicals (e.g. for industrial, medicinal use) which have potential adverse effects to its ecological status. In addition, the densely populated areas represent local high emissions of those chemicals leading to more aggravating consequences. Estrogenic compounds that end-up in environmental water directly affect living organisms by interfering with their endocrine metabolism. The assessment of their presence in the environment requires sensitive and selective analytical methods. Nineteen estrogenic compounds belonging to different classes (5 free estrogens, 6 conjugated estrogens, 3 progestogens and 5 phytoestrogens) have been studied. The analytical methodology developed is based on solid phase extraction followed by liquid chromatography tandem mass spectrometry and has been applied to study the occurrence of the above mentioned analytes in environmental waters from the state of Rio de Janeiro (Brazil). Due to insufficient infra-structure in this region, waste waters are released onto the environment without or with incomplete previous treatment. The results show that high levels of the phytoestrogens daidzein, coumestrol and genistein of up to 366 ng/L and progesterone of up to 47 ng/L could be found in river water. Estrogens and their conjugated derivatives were detected in the lower ng/L range up to 7 ng/L. The main estrogens estrone, estradiol and the synthetic ethinyl estradiol could not be detected. The developed method showed overall good performance with recoveries above 80% (with one exception), limits of detection < or =2 ng/L, good linearity and reproducibility.

Risk assessment of representative and priority pesticides, in surface water of the Alqueva reservoir (South of Portugal) using on-line solid phase extraction-liquid chromatography-tandem mass spectrometry.

Surface waters located in intensive agricultural areas are more vulnerable to the pesticides contamination, which is a major concern if the water is intended to be used for human consumption. The aim of this study was to evaluate the presence and the distribution of pesticides in the Alqueva reservoir, an important source of water supply (South of Portugal), considering their representativeness in the agricultural practice of the area. For the analysis of pesticides risk impact we used the environmental quality standards in the field of water policy proposed recently by the European Commission. The pesticides belonging to the classes of phenylureas, triazines, chloroacetanilides, organophosphorous and thiocarbamates were analysed by on-line solid phase extraction-liquid chromatography-tandem mass spectrometry. The pesticides more frequently detected were atrazine, simazine, diuron and terbuthylazine. The highest levels of these pesticides were registered in spring, after pesticides treatment, namely in olive-tree and vine crops. The priority pesticides atrazine and diuron reached values above the annual average proposed in the European Union Legislation. The herbicide atrazine reached values that surpassed the proposed maximum allowable concentration (2,000 ng L(-1)). The sampling stations most affected by these pesticides were Sra. Ajuda, Lucefecit and Alcarrache, located in the northern part of the reservoir, closer to Spain where the agricultural activity is more intensive.

Simultaneous determination of selected endocrine disrupters (pesticides, phenols and phthalates) in water by in-field solid-phase extraction (SPE) using the prototype PROFEXS followed by on-line SPE (PROSPEKT) and analysis by liquid chromatography-atmospheric pressure chemical ionisation-mass spectrometry.

In this study, a new procedure, based on on-line solid-phase extraction (SPE) and analysis by liquid-chromatography-atmospheric pressure chemical ionization-mass spectrometry (LC-APCI-MS), has been developed for the simultaneous, multianalyte determination of 21 selected pesticides, phenols and phthalates in water. SPE was carried out on polymeric PLRP-s cartridges by percolating 20 mL-samples. For sample preconcentration, the performance of a prototype programmable field extraction system (PROFEXS) was evaluated against the commercial laboratory bench Prospekt system used for method development. The Profexs is designed for the automated on-site sampling, SPE preconcentration, and storage of up to 16 samples in SPE cartridges. These cartridges are further eluted and on-line analyzed with the Prospekt coupled to the chromatographic system. In the optimized method, where completely on-line SPE-LC-MS analysis of the samples is carried out with the Prospekt in the laboratory, detection limits lower than 100 ng/L, and satisfactory precision (relative standard deviations <25%) and accuracies (recovery percentages >75%) were obtained for most investigated compounds from the analysis of spiked Milli-Q water. The extraction efficiency achieved with the Profexs was comparable to that of the Prospekt for most compounds and somewhat lower for the most apolar analytes, probably due to adsorption on the pump filters. The completely on-line optimized method was applied to the analysis of surface water, ground water and drinking water from a waterworks in Barcelona. Some pesticides and phenols were found in both surface water and groundwater at ng/L or microg/L levels, but not in the final drinking water. Di(2-ethylhexyl)phthalate (DEHP) was present in all samples investigated, including blanks. To the author's knowledge, this is the first work describing the application of a fully automated on-line SPE-LC-MS method for the simultaneous analysis of pesticides, phenols, and phthalates in water, and the second one that examines the possibilities of the prototype Profexs for automated on-site SPE preconcentration of organic pollutants from water samples.

Endocrine disrupting compounds and other emerging contaminants in the environment: a survey on new monitoring strategies and occurrence data.

An overview of Toxicity Identification and Evaluation (TIE) procedures, used for the effect-based analysis of endocrine disrupting compounds (EDCs) in environmental samples, is presented. Future trends in advanced chemical analysis of EDCs and some emerging contaminants are outlined. The review also gives an overview of concentration levels found in environmental samples and discusses the correlation of calculated estrogenicity (based on chemical analysis of target EDCs) with that measured by various bioassays.

Monitoring of endocrine disruptors in surface waters by the yeast recombinant assay.

Endocrine disruptors exert physiological effects at very low concentrations. Surface waters present often a mixture of high concentrations of low-potency disruptors and low amounts of very powerful ones, making their chemical analysis complicated and expensive. We developed a recombinant yeast assay (RYA) for estrogenic compounds using 96-well microtiter plates. This assay is based on three yeast strains, transformed with self-propagating plasmids. One strain contains an expression plasmid for the human estrogen hormone receptor and an appropriate reporter; it detects estrogenic and antiestrogenic activities. The two other yeast strains, one expressing the human progesterone receptor and a second based on the yeast activator Gal4p, served to analyze the nature of antiestrogenic activities. We applied this technique to water samples from two tributaries on the Llobregat river (NE Spain) as well as from four sewage treatment plants discharging on them. Our results indicate that the efficiency of sewage treatment plants for eliminating estrogenic compounds varied notably, being in at least one case completely inefficient. We also observed a prevalence of an inhibitory activity all through the two rivers; this inhibition was hormone specific. These results were consistent to previously obtained chemical analyses of the same samples, although chemical and in vivo analyses showed rather different levels of sensitivity for some compounds. Our findings demonstrate the utility of the yeast recombinant assay for analyzing complex natural samples; at the same time, they stress the necessity of a panel of different yeast systems to adequately describe endocrine-disruptor activities.

Determination of steroid sex hormones and related synthetic compounds considered as endocrine disrupters in water by fully automated on-line solid-phase extraction-liquid chromatography-diode array detection.

In this study, a procedure for the simultaneous determination in water of six estrogens (estradiol, estriol, estrone, ethynyl estradiol, mestranol, and diethylstilbestrol) and three progestogens (progesterone, norethindrone, and levonorgestrel), selected based on their abundance in the human body, their estrogenic potency, and the extent of their use in contraceptive pills, was developed. The procedure, based on the on-line solid-phase extraction (SPE) of the water sample and subsequent analysis by liquid chromatography/diode array detection (LC/DAD), allows for the monitoring of up to 16 samples in a completely automated, unattended way. The SPE experimental conditions were optimized and the polymeric cartridge PLRP-S selected out of four different cartridges evaluated. The chromatographic separation was carried out on a LiChrospher 100 RP-18 and detection was performed at 200, 225, and 240 nm. The applicability of the method to the analysis of various environmental water samples, including drinking water, groundwater, surface water and sewage treatment plant effluents, was evaluated. Method detection limits were in the range 10-20 ng/l. The method precision and accuracy were satisfactory with recovery percentages ranging from 96 to 111% and relative standard deviations lower than 3%. The technique is also considerably cheap, fast, and easy, and, therefore, very adequate for routing monitoring. To the authors' knowledge it constitutes the first work describing a fully automated, on-line methodology for the continuous monitoring of these compounds in water.

Review of analytical methods for the determination of estrogens and progestogens in waste waters.

Steroid sex hormones and related synthetic compounds, e.g. those used in contraceptive pills, have been shown to be present in the aquatic environment, mainly as a result of inefficient removal in waste-water-treatment plants (WWTP). The concentrations of the compounds, although very low (low ng L(-1)), are sufficient to induce estrogenic responses and alter the normal reproduction and development of wildlife organisms. The compounds have been determined, by a variety of analytical procedures, in the influents and effluents of WWTP, within research programs conducted in different countries to evaluate the efficiency of the waste-water-treatment processes currently in use. Determination of natural and synthetic estrogens and progestogens in waste water is, however, a difficult analytical task, because the very low detection limits required and the complexity of the waste-water matrix necessitate the execution of a considerable number of steps. Thus, in general, complicated, time-consuming extraction and purification processes, usually based on the application of solid-liquid extraction, are performed before final determination by immunoassay, high-performance liquid chromatography, or, most often, gas chromatography coupled with mass spectrometry. This paper reviews the analytical methods so far described for the analysis of these important environmental pollutants in waste water and discusses the key procedural steps, from sampling to analysis, and the techniques most commonly used in the determination.

Determination of steroid sex hormones and related synthetic compounds considered as endocrine disrupters in water by liquid chromatography-diode array detection-mass spectrometry.

In this study, a procedure for the determination of various naturally occurring hormones and of some related synthetic chemicals, commonly used for birth control and treatment of certain hormonal disorders and cancers, in water is described. The procedure includes solid-phase extraction of 0.5 l of water and subsequent analysis of the extract by liquid chromatography with diode array detection and mass spectrometric detection in series (LC-DAD-MS). DAD has been performed at 197, 225, and 242 nm for quantification and confirmatory identification purposes. For MS detection two interfaces--electrospray and atmospheric pressure chemical ionization--in both the positive and the negative ion mode have been tested and the MS parameters influencing the MS signal optimized. DAD and MS have been intercompared for selectivity, sensitivity, precision, and linearity of response. Selected conditions have been applied to the determination of six estrogens (17beta-estradiol, estriol, estrone, ethynylestradiol, mestranol, and diethylstilbestrol) and four progestogens (progesterone, levonorgestrel, norethindrone and ethynodiol diacetate) in several types of water bodies, including sewage influents and effluents, surface water and drinking water. Recoveries greater than 83% and detection limits in the ng/l range have been achieved for most compounds.