Urban Treatment Plants Research Articles

Wastewater as a nutrient source for hydroponic production of lettuce: Summer and winter growth

Solutions combining soilless cropping systems with wastewater reuse can offer benefits in the agriculture sector, reducing pressure on water resources, promoting sustainable production, and reclaiming wastewater. However, assessing the sanitary risks associated with wastewater reuse is of utmost importance. This study aimed to investigate the hydroponic growth of lettuce (Lactuca sativa var. crispa L.) in wastewater from an urban treatment plant with different levels of treatment and evaluate potential sanitary risks. Crop growth took place in a greenhouse, during summer and winter periods, using wastewater after primary (PTW) or secondary (STW) treatment, and a nutrient solution (NS), as control. Physical and chemical water quality parameters, morphological crop growth parameters, and environmental conditions inside the greenhouse were monitored. Toxicity analyses were carried out through cell viability assays with the Caco-2 cell line and total coliforms and Escherichia coli (E. coli) were determined. Wastewater-grown plants achieved acceptable growth, even though presenting lower fresh weight than NS-plants. STW-plants’ growth was limited essentially by nutrient deficiency, and PTW-plants were affected by nutrient deficiency, pH values, solid load, and N-NO2 concentration. Higher temperatures in summer led to faster crop growth, and lower temperatures in winter allowed better nutrient uptake by the crop. Wastewater-grown plants did not evidence toxicity in leaf extracts up to 1 % w/v. Coliform enumeration data indicated an accumulation in plant roots, with high removal from the wastewater. E. coli was not detected on plants’ leaves and total coliforms were within acceptable limits. Furthermore, the results point to an improvement in the wastewater quality, with minimum removal values of 75.2 % BOD5, 83.1 % COD, 43.4 % P, 44.9 % N, and 90.4 % K. The results demonstrated the viability of wastewater reuse for hydroponic production allowing a better understanding of its processes and contributing to mitigating water scarcity for food production, and the impacts of treated wastewater discharge in freshwater courses, particularly those associated with nutrient delivery to aquatic systems.

Exposure of juvenile Chinook salmon to effluent from a large urban wastewater treatment plant. Part 2. Metabolomic profiling

Targeted metabolomic analysis was conducted on juvenile Chinook salmon exposed for 10 days to wastewater effluent (WWE) from a large urban treatment plant. Exposures included five dilutions of WWE (20%, 5.3%, 1.4%, 0.4%, and 0.1%) and a control with 7 replicates per treatment. Liver was extracted from fish and analyzed via liquid chromatography – mass spectrometry (LC-MS) for 361 endogenous metabolites, of which 185 were detected. Control-versus-treatment comparisons identified several metabolites that were associated with altered biochemical pathways observed for all treatments, including several that are important for energy generation and utilization. These altered pathways are crucial for fish health and may be an early indicator of potential adverse effects on growth, reproduction, behavior, and immune function. Juvenile ocean-type Chinook salmon spend several days to weeks in the nearshore estuary where they may encounter high concentrations of WWE contaminants. They are exposed to a wide range of potent pharmaceuticals, personal care products, and industrial compounds from WWE that have the potential to affect physiological homeostasis and disrupt their normal life cycle.

Characteristics and risk assessment of sewage sludge from urban wastewater treatment plants in Shaanxi Province, China.

In this study, an investigation of important properties, including moisture content, pH, plant nutrients, organic matter, mineral oil, and the contents of heavy metals (HMs) in sewage sludge (SS) collected from 32 urban treatment plants in Shaanxi Province, China, was carried out. The test results showed that the pH and the moisture, organic matter, total nitrogen (TN), total phosphorus (TP), and mineral oil contents of the SS varied over different rainfall periods, and most of the indicators met the standard criteria for SS agricultural reuse in China. Principal component analysis (PCA) and correlation analysis indicated that the pollutant characteristics of the SS depended on time span and geographical distribution. The mean contents of Pb, Cd, Cu, Zn, Ni, Cr, Hg, and As in SS were 3.95, 16.38, 5.43, 7.70, 1.31, 1.53, 32.77, and 1.40 times higher than the soil background values, respectively. Speciation analysis showed that the forms of HMs in the SS were significantly different. Assessments based on the geoaccumulation index (Igeo), Nemerow integrated pollution index (NIPI), and potential ecological risk index (RI) suggested that HM pollution risk levels were either uncontaminated or moderately contaminated in some regions and that SS recycled for agricultural applications carried a low risk. In conclusion, certain potential ecological risks exist for SS agricultural utilization in Shaanxi Province, and it is necessary to reduce the HM content before SS resource utilization for land application.

Evaluating the potential severity of biogas toxic release, fire and explosion: consequence modeling of biogas dispersion in a large urban treatment plant

Objectives. Biogas production in treatment plants for energy generation has increased in recent years. This study aimed to model the consequence of biogas release in a large urban treatment plant. Methods. The study modeled biogas storage tank consequences in a large urban treatment plant in Iran. Due to potential for biogas harmfulness, three consequences of toxic release, fire and explosion were evaluated. Scenarios were evaluated in the worst-case situation. All modeling steps were performed using PHAST version 7.2. Results. In the case of catastrophic reservoir rupture in summer, distances of 3788.94, 128.86 and 91.72 m from the reservoir in the wind direction will be in the range of 100, 500 and 1000 ppm biogas, respectively. Study of pressure values due to explosion in the catastrophic rupture scenario revealed that distances of 57.19, 14.70 and 115.84 m from the biogas reservoir were in the range of 0.02, 0.13 and 0.2 bar pressure increase, respectively. Conclusion. Due to the treatment plant’s location in a dense urban area, biogas dispersion could lead to exposure of many people to high-risk areas. Therefore, taking control measures comparable with the consequence modeling output can be a practical step toward reducing vulnerability against such incidents.

Застосування добрив на основі осадів стічних вод для додаткового фосфорного живлення рослин

Goal. To substantiate the use of fertilizers based on sewage sludge (SS) to increase soil fertility and crop yields through additional phosphorus nutrition. Methods. Field experiment — to determine the effectiveness of new types of bio-fertilizers based on SS, mathematical — to analyze and evaluate the reliability of the results. Results. In recent decades, there has been a decline in the livestock industry, which in turn has affected the decline in the production of organic fertilizers and, as a consequence, the deterioration of soil fertility and reduced agricultural productivity. This situation has led to the search for new types of fertilizer resources and the development of effective measures to increase the composition of phosphorus compounds available to plants. These are sewage sludge. One of the promising areas for solving the problem of soil enrichment with available phosphorus compounds for plants may be the composting of SS in urban treatment plants. The result of composting is an available organic product with the participation of phosphate-mobilizing microorganisms, which can be used in agricultural production. Conclusions. The prospects of using non-traditional organic fertilizers — SS and composts based on them — in increasing crop yields are shown. Thus, the main application of the studied types of non-traditional organic fertilizers based on SS of biological treatment plants (BTP) “Pivdenna” and “Pivnichna” (Odessa) with vegetable fillers (winter wheat straw, sunflower seed husk) under corn for grain yield provided an increase of yield ability of crop by 31.6–50.0%.

Cavitation treatment of water from a wastewater treatment plant

Hydrodynamic cavitation damages the apparatus by intensive cavitational erosion of the elements in a hydro system. Being acquainted with the mechanisms of effect, hydrodynamic cavitation can be used to intensify technological processes in different industrial areas. The aim of the present work is to investigate the possibilities using hydrodynamic cavitation for wastewater treatment from an urban treatment plant. To intensify the purifying process series of trials, at different levels of intensity of the electrical field in the zone of cavitation, were performed. Additionally presented, the electrical field assists in forcing the process of silver ions separation with the purpose of microbiological purification of the flowing water. Here we present the results of experimental studies conducted at various times and electrical fields intensity. These trials substantiate the idea of the microbiological purification of wastewaters via hydrodynamic cavitation and represent a huge range of opportunities for applying this method in various industrial fields. The basic criterion that supports our statement is the enormous drop of the microbe number by increasing the cavitation treatment time.

Diversity and antibiotic resistance among Escherichia coli populations in hospital and community wastewater compared to wastewater at the receiving urban treatment plant

Bacterial diversity and antimicrobial resistance patterns among the indicator organism Escherichia coli were monitored in wastewater samples collected over one year from a hospital (HW), a community (CW) and the receiving urban (UW) wastewater treatment plant (WWTP). We compared levels of antibiotic resistance in the different types of wastewater, and identified whether resistant strains were endemic in the wastewater system. If so, implementation of local treatment at certain resistance hotspots (e.g. hospital outlets) could be used to decrease the amount of resistant bacteria in the wastewater. E. coli from HW (n = 2644), CW (n = 2525) and UW (n = 2693) were analyzed by biochemical phenotyping (PhenePlate System) and antimicrobial susceptibility testing to nine antibiotics (AREB System). The phenotypic diversities of the total E. coli populations were similar for all three sites (Simpson's Diversity index, Di = 0.973), however for individual samples, HW showed low diversities (Median Di = 0.800) and the E. coli flora was often dominated by strains that may have originated from the fecal flora of single individuals. The diversities in CW samples was higher (Median Di = 0.936), and UW samples showed similar diversities as the whole collection of isolates (Median Di = 0.971). Resistance to at least one of the nine antibiotics was observed in 45% of the HW isolates, 44% of CW isolates, and 33% of UW isolates. Resistance to gentamicin and chloramphenicol was uncommon (3.2 and 5.3%, respectively), whereas resistance to tetracycline and ampicillin was most common (24% and 31%, respectively). Extended-spectrum beta-lactamase-producing E. coli (ESBL-EC) were more common in HW (11.5%) and in CW (6.9%) compared to UW (3.7%). A high diversity (Di = 0.974) was observed among ESBL-EC isolates from UW (n = 99), indicating absence of any clonal structure among these isolates. Common PhP types of ESBL-EC often dominated in each HW sample, but were not identified across different samples, whereas ESBL-EC in CW showed low diversity (Di = 0.857) and were dominated by a specific PhP type that was found across almost all CW samples. The antibiotic resistance rates were highest in hospital wastewater, but surprisingly they were also high in the studied community wastewater, compared to the urban wastewater. The relative contribution of HW seemed low in terms of dissemination of antibiotic resistant bacteria to the WWTP.

Subtype analysis of Giardia duodenalis isolates from municipal and domestic raw wastewaters in Iran.

A total of 54 raw wastewater samples collected from three urban treatment plants and two slaughterhouses in Tehran, Iran, were assessed for the presence of the Giardia cysts using immunofluorescence with monoclonal antibodies. To characterize the cysts at the molecular level, the three genetic loci were amplified and sequenced. The assemblages A (37.5%) and E (58.3%) were detected in livestock wastewater samples. Assemblage A, which is composed of only G. duodenalis genotype, was detected in 100% of urban wastewater samples. The subassemblages A2, A3, A-I, A-II, and E3 were identified with β-giardin, triose phosphate isomerase, and glutamate dehydrogenase genes. This study is the first to report on G. duodenalis genotypes in aquatic environmental samples in Iran.

First results of Technosols constructed from municipal waste in Vitoria-Gasteiz (Spain)

Construction of Technosols offers interesting alternatives to two current problems in the city of Vitoria-Gasteiz (Spain): waste recycling and degraded plot recovery. To evaluate the viability of their use, 6 types of Technosols were created by mixing four different byproducts from municipal waste treatment plants. The less than 40 mm size fraction material from the municipal construction and demolition waste treatment plant was used as the main ingredient, bio-stabilized material from the solid urban waste treatment plant mixed with triturated pruning was used as organic matter input, recycled bentonites and topsoil from the public plots of Vitoria-Gasteiz (Spain) completed the mixture. Mixes were prepared in triplicate and installed in 48 m<sup>2</sup> cells along with another 4 control cells containing only one of the ingredients at the municipal landfill of Gardelegui. A monitoring program for different parameters on soil, eluates and natural leachates was established to test the Technosols’ capacity to sustain vegetation without negative impacts on the environment. The final objective is to test their ability to restore unused municipal plots. Results from the first year show that Technosols are a suitable option for degraded sites restoration and green infrastructure support. All controlled parameters on soil are within the limits set by autonomic legislation for land use as public park. The eluate analysis concludes for all studied parameters that all mixes would be classified by legislation as inert waste, except for the sulphate concentration (which exceeds the inert waste limit of 1 000 ppm), that currently would label the soil as non-hazardous. In the natural leachate analysis strongly basic pH values were present above 9.5, the limit allowed in Royal Decreet 849/1986, but acidified throughout the year moving towards neutrality, with final values between 7.31 and 7.51. Leachate from CDW30, TS15 and RB30 Technosols showed not allowed values with respect to sulfates and Fe during the last sampling, surpassing the limits of 2 mg/l and 2000 mg/l respectively. All studied Technosols presented a low ecological potential risk (RI < 150) for heavy metals in soil and eluates.

Evaluation of Cryptosporidium oocyst and Giardia cyst removal efficiency from urban and slaughterhouse wastewater treatment plants and assessment of cyst viability in wastewater effluent samples from Tehran, Iran

Five municipal and domestic wastewater treatment plants, most of which had secondary treatment systems formed by activated sludge, were studied during 2013–2014 in Tehran. The study was done in order to evaluate their efficiency in terms of removal of Cryptosporidium and Giardia by (oo)cyst recovery in effluent samples using immunofluorescence with monoclonal antibodies. Results showed that mean concentrations of cysts in the influent samples always outnumbered mean concentrations of oocysts (883.3 ± 4,16.7–3,191.7 ± 1,067.2 versus 4.8 ± 6.2–83.8 ± 77.3 (oo)cysts/L), and that lower concentrations of (oo)cysts were recorded in summer, and higher levels in autumn, and that the difference was statistically significant (t-test, P <0.05) only in wastewater from slaughterhouses. Results for removal percentages of all the plants ranged from 76.7 to 92.1% for cysts and from 48.9 to 90.8% for oocysts. There was more reduction of (oo)cysts at the urban treatment plant by activated sludge-A2O-sand filtration than at plants with conventional activated sludge and activated sludge-trickling filter, however, this difference was not statistically significant for cysts and oocysts (ANOVA, P > 0.05). Infections in mice inoculated with cysts obtained from urban wastewater effluent demonstrated presence of infectious Giardia cysts. Results demonstrate limited efficiency of conventional wastewater treatment processes at physico-chemical removal of (oo)cysts.

Detection of parasitic particles in domestic and urban wastewaters and assessment of removal efficiency of treatment plants in Tehran, Iran.

BackgroundIn recent years, decreasing annual rainfalls in some countries and population growth have led to a shortage of freshwater resources. Thus, recycled wastewaters has been suggested for agricultural activities. Contamination of wastewaters with pathogens is a major concern for the use of these waters. This study aimed to (i) investigate the occurrence of helminth eggs and protozoan (oo)cysts in human and livestock wastewaters, and (ii) evaluate the parasite removal efficiencies of urban and slaughterhouse treatment plants in Tehran province, Iran. One hundred and eight samples were collected from five urban and domestic wastewater treatment plants in Iran. Wastewater samples were concentrated by centrifugal-concentration and filtration methods.ResultsThe quantity of helminths egg and protozoa (oo)cyst per liter of urban raw wastewater ranged from 1.2 × 101 to 2.9 × 101 and from 9.6 × 102 to 1.9 × 103, respectively. The number of eggs and (oo)cysts per liter of animal raw wastewater ranged from 1.6 × 103 to 4.9 × 103 and 3.1 × 104 to 6.0 × 104, respectively. The helminths and protozoa identified in urban treatment plants included hookworms, Hymenolepis and Rhabditis (or probably Strongyloides), Entamoeba, Isospora, Giardia, Chilomastix and Cryptosporidium, while in slaughterhouses Trichuris, Trichostrongylus, Moniezia, Dicrocoelium, Fasciola, Entamoeba, Cryptosporidium, Eimeria and Giardia were isolated. The overall removal efficiency of eggs and (oo)cysts in the treatment plants ranged from 94.8 to 95.7% and from 79.3 to 85.8%, respectively.ConclusionThe study results revealed that the efficacy of removal of nematode eggs, and not protozoan (oo)cysts, in urban wastewater treatment plants, is in compliance with the WHO parasitological guideline (<1 nematode per liter) required for unrestricted irrigation.

Modification of Activated Sludge Cohesion by Enzymatic Catalysis Process and Perspectives

It is possible to transform the structure of organic compounds so as to modify some of their properties to make them more “elastic”, for example. Many solid-liquid separation processes use the cohesion property of the solid in suspension to increase the performance of the separation system. In this work, we examine some beneficial aspects that enzyme catalysis may reveal when acting on the strength of cohesion of a suspension. The topic concerns the activated sludge (AS) produced by wastewater of urban (or industrial) treatment plants. We used urban sludge and industrial liquid enzymes produced by the firm NOVO. To accelerate the research process, we employed enzyme mixtures (we called “enzyme cocktails”) that highlighted the active mixtures, but until now we have not characterized specifically active enzymes. Despite a significant number of experiences, it is clear that given the number of different enzymes available and the almost infinite possible ratios of cocktails, only a small number of conceivable methods have been tested (and even less optimized). Nevertheless, a few cocktails were active and promising enough to improve solids contact processes in various industrial processes. We have found a range of combinations that increases the sludge cohesion coefficient and considered this enzyme treatment to strengthen the separation by solid contact structures mainly to tackle important hydraulic shock.

Antibiotic resistance, antimicrobial residues and bacterial community composition in urban wastewater

This study was based on the hypothesis that the occurrence of antimicrobial residues and antibiotic resistant bacteria in the sewage could be correlated with the structure and composition of the bacterial community and the antibiotic resistance loads of the final effluent. Raw and treated wastewater composite samples were collected from an urban treatment plant over 14 sampling dates. Samples were characterized for the i) occurrence of tetracyclines, penicillins, sulfonamides, quinolones, triclosan, arsenic, cadmium, lead, chromium and mercury; ii) antibiotic resistance percentages for tetracycline, sulfamethoxazole, ciprofloxacin and amoxicillin and iii) 16S rRNA gene-DGGE patterns. The data of corresponding samples, taking into account the hydraulic residence time, was analyzed using multivariate analysis.Variations on the bacterial community structure of the final effluent were significantly correlated with the occurrence of tetracyclines, penicillins, sulfonamides, quinolones and triclosan in the raw inflow. Members of the class Epsilonproteobacteria presented positive correlations with those antimicrobials, whereas negative correlations were observed with Beta and Gammaproteobacteria and Firmicutes. Antibiotic resistance percentages presented different trends of variation in heterotrophs/enterobacteria and in enterococci, varied over time and after wastewater treatment. Antibiotic resistance was positively correlated with the occurrence of tetracyclines residues and high temperature. A relationship between antibiotic residues, bacterial community structure and composition and antibiotic resistance is demonstrated. Further studies, involving more wastewater treatment plants may help to elucidate this complex relationship.

Anaerobic Removal of Trace Organic Contaminants in Sewage Sludge: 15 Years of Experience

Trace organic contaminants (TOCs) correspond to a broad range of molecules generated either directly or indirectly by human activity. Even though TOCs are found at low concentrations in the environment, they often accumulate by biomagnification and bioaccumulation into biological organisms and cause irreversible damages in biological systems through direct or indirect toxic effects such as endocrine disruption and tumour initiation. This manuscript presents the main findings of over fifteen years of research focusing on biological removal of various TOCs found in sewage sludge from urban treatment plants. A special focus of the research was made on microbial processes in complex anaerobic ecosystems. Four families of compounds mostly retrieved in urban plants were studied: the polycyclic aromatic hydrocarbons (PAHs), the polychlorobiphenyls (PCBs), the phthalic acid esters (PAEs), and the nonylphenol ethoxylates (NPEs). It was observed that the microbial capability for removing low amounts of TOCs required a long adaptation time and was often limited by the bioavailability of these compounds. In fact, the overall biodegradation resulted from the numerous interactions existing between the matrix (organic matter) and the microbial ecosystems according to the physico-chemical sorption properties of these compounds. Mechanistic aspects were also tackled in depth and specific models were developed for better understanding the network of interactions between TOCs, microorganisms, and organic matter. These findings could be extrapolated to other ecosystems such as soils and sediments. Finally, it was shown that microbial cometabolism was essential for TOC removal, and the concept of bioavailability was not only dependent on the nature, the level, and the sorption properties of TOCs but was also strongly dependent on the nature and the concentration of the sludge organic matter. Specific parameters were proposed for better evaluating the fate of TOCs in microbial anaerobic processes and technological solutions for efficient removal of these compounds were also proposed.

Simultaneous determination of parabens and synthetic musks in water by stir‐bar sorptive extraction and thermal desorption‐gas chromatography‐mass spectrometry

This study focuses on a method for simultaneously determining personal care products in a wide range of polarities in different water matrices. The method is based on stir-bar sorptive extraction followed by thermal desorption-gas chromatography-mass spectrometry. Prior to extraction, the parabens were acetylated to improve their affinity for the polydimethylsiloxane phase of the stir bar. The method showed good linearity, repeatability and reproducibility between days for all compounds and limits of detection at low ng/L levels (between 0.02 and 0.3 ng/L). The proposed method is also environmentally friendly, because it does not use organic solvents, and reduces the risk of external pollution, due to the minimal manipulation of the sample required. The method developed was successfully applied for the analysis of personal care products in different kinds of water matrices: influents and effluents of urban and industrial wastewater treatment plants, effluents of a reverse osmosis treatment plant and river waters. The influents of urban treatment plants generally showed the highest values for synthetic musks, with concentrations of up to 2219 ng/L of galaxolide, whereas the highest concentrations of parabens were detected in the industrial treatment plants influents.

Subchronic Toxicity Evaluation of a Treated Urban Sewage Sludge

Disposal of tons of sludge produced daily by sewage treatment plants in large cities is a serious problem. Because recycling and application in agriculture have been proposed, the Brazilian National Environmental Council (CONAMA, 2006) issued a legal norm that regulates the use of the sewage sludge (SS) in crops. Due to the complex chemical nature of such products, characterization by analytical methods for health and environmental risk assessment has severe limitations. To overcome such limitations, it is necessary to (1) assess the toxicological potential of SS and (2) identify possible adverse effects in vivo in order to provide critical information for future environmental regulations. The present study was conducted to determine the potential toxicity of SS obtained from a representative urban treatment plant located in the São Paulo State, Brazil. Male and female Wistar rats were fed ad libitum a pelleted diet containing varying amounts of SS. No relevant clinical, hematological, urinary, or gross organ morphological alterations were observed in both genders of rats orally exposed to SS at up to 3.8 g/kg/d for 90 d. Sewage slude produced increased incidence of centrilobular hepatocyte hyperplasia at the high dose and significantly increased aspartate aminotransferease (AST) activities at all doses in both genders. Although the present data indicate some liver involvement, these alterations were considered adaptative and not toxicologically relevant, as the responses were relatively mild, not dose dependent, and no other parameters were markedly affected. The present results may contribute to the establishment of protocols for potential usage in SS agricultural soil application.

Investigation of the acclimatization period: example of the microbial aerobic degradation of volatile organic compounds (VOCs)

The aim of this study is to better evaluate the occurrence of an acclimatization-enrichment period, defined as a selection period of consortia having the capability to biodegrade pollutants. In order to perform this evaluation, two experimental strategies were carried out and the results were studied carefully. Two laboratory-scale reactors were inoculated with activated sludge from an urban treatment plant. During the experiment, these reactors were supplied with a gaseous effluent containing VOCs. For both reactors, the composition is different. Three parameters were monitored to characterize the microflora: bacterial activities, bacterial densities, and the genetic structure of Bacteria and Eukarya domains (Single Strand Conformation Polymorphism fingerprint). The obtained results showed that the resultant biodegradation functions were equivalent. The bacterial community structure differs even if six co-migrated peaks were observed. These data suggest that the microbial communities in both reactors were altered differently in response to the treatment but developed a similar capacity to remove VOCs at the issue of this period. Furthermore, it is suggested that the experimental strategies developed in this work lead to an enrichment in terms of functionality and microbial diversity almost equivalent.

Identification and Measurement of Illicit Drugs and Their Metabolites in Urban Wastewater by Liquid Chromatography−Tandem Mass Spectrometry

Residues of illicit drugs and their metabolites that are excreted by humans may flow into and through wastewater treatment plants. The aim of this study was to develop a method for the determination of cocaine, amphetamines, morphine, cannabinoids, methadone, and some of their metabolites in wastewater. Composite 24-h samples from urban treatment plants were enriched with deuterated internal standards before solid-phase extraction. High-pressure liquid chromatography tandem mass spectrometry with multiple reaction monitoring was used for quantitation. Recoveries were generally higher than 80%, and limits of quantifications were in the low nanograms-per-liter range for untreated and treated wastewater. The overall variability of the method was lower than 10% for untreated and 5% for treated wastewater. The method was applied to wastewater samples coming from two treatment plants in Italy and Switzerland. Quantification ranges were found to be 0.2-1 microg/L for cocaine and its metabolite benzoylecgonine, 80-200 ng/L for morphine, 10 ng/L for 6-acetylmorphine, 60-90 ng/L for 11-nor-9-carboxy-Delta9-tetrahydrocannabinol, 10-90 ng/L for methadone and its main metabolite 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine, and lower than 20 ng/L for amphetamines. As previously reported for cocaine, this method could be useful to estimate and monitor drug consumption in the population in real time, helping social scientists and authorities to combat drug abuse.

Analytical approach for monitoring endocrine-disrupting compounds in urban waste water treatment plants

The presence of endocrine-disrupting compounds in influent and effluent water samples from four waste water treatment plants located in Italy was studied. The estrogen-like activity of the water samples was measured using a chemiluminescent recombinant yeast assay which is based on genetically engineered yeast cells that express the human estrogen receptor. This receptor, once activated, elicits the expression of the reporter gene lac-Z and, consequently, the production of beta-galactosidase, which is then measured by chemiluminescence. To control and minimize sample matrix effects, an external control based on a modified yeast strain stably expressing beta-galactosidase was developed and also used in the assay. Rapid and sensitive chemiluminescent enzyme immunoassays were also developed and validated for the quantification of 17beta-estradiol, estrone, and estriol in waste water samples. Results from both methods were compared with a reference high-performance liquid chromatography and electrospray ionization tandem mass spectrometry (HPLC ESI-MS-MS) method developed for the quantification of natural estrogens. The recombinant yeast assay revealed a significant estrogenic activity in the influent samples, ranging from 80 to 400 pmol/L 17beta-estradiol equivalents (EEQ), which was reduced by 70-95% in the effluent samples. The yeast assay also showed a systematic 20-30% overestimation of estrogenic activity relative to the HPLC ESI-MS-MS method, suggesting the presence of other compounds in the samples with estrogenic activity. The chemiluminescent enzyme immunoassays showed the presence of estrogens in the influent samples (mean concentrations: 350-450 pmol/L for estrone, 5-100 pmol/L for 17beta-estradiol, 25-300 pmol/L for estriol), with significantly lower concentrations detected in the respective effluent samples. The waste water treatment was able to reduce natural estrogen concentrations by 40-95%, although a high variability was observed. The enzyme immunoassay data correlated well with data obtained by the HPLC ESI-MS-MS method. Although the recombinant yeast assay represents a useful tool for a first-level screening of estrogenic activity due to its simplicity and high analytical throughput, sample matrix effects observed in waste water of industrial origin were found to strongly affect the yeast cells response, even when properly corrected for using the external control, thereby limiting its use to urban waste water. Its integration with chemiluminescent enzyme immunoassays would improve its performance by reducing false negative results, thereby enabling its use in extensive studies monitoring for the presence of endocrine-disrupting compounds in urban treatment plant effluents.

Development of a solid-phase microextraction method for the analysis of phenolic flame retardants in water samples

A solid-phase microextraction (SPME) method for the ultra-trace determination of brominated phenols in aqueous samples has been developed and is reported for the first time to the best of our knowledge. 3,5,3′,5′-tetrabromobisphenol A (TBBPA), the most widely used brominated flame retardant, and other phenolic flame retardants in commercial use, such as 2,4-dibromophenol (2,4-DBP), 2,4,6-tribromophenol (TBP) and pentabromophenol (PBP) have been included as target analytes. The analytical procedure involves the in situ acetylation-SPME and gas chromatography–mass spectrometry (GC–MS) determination of the target analytes. A multi-factor categorical experimental design was created to study the main parameters affecting the extraction efficiency, allowing also the evaluation of interaction effects between factors. The factors studied were type of fiber, extraction mode, exposing the fiber directly into the sample (DSPME) or into the headspace over the sample (HSSPME), and extraction temperature. Carboxen-polydimethylsiloxane (CAR-PDMS) fiber appeared to be the most suitable of the five fibers tested for the extraction of most compounds, excluding PBP and TBBPA for which polydimethylsiloxane (PDMS) was the most efficient coating. The highest response was achieved for both fibers sampling in headspace mode at 100 °C. In order to test the linearity of the method, calibration studies were performed with both CAR-PDMS and PDMS coatings. For both fibers, the method was linear in a range of 2 orders of magnitude, giving relative standard deviation (RSD%) below 10% for most compounds and detection limits at the low pg/mL level. In addition, the feasibility of the method for simultaneous determination of chlorinated and brominated phenols was studied. Finally, the method was applied to several real samples including tap water and effluent and influent waste water samples from an urban treatment plant, in which several phenolic compounds, such as phenol, methylphenols and chlorophenols, could be detected and quantified.