Online solid phase extraction liquid chromatography tandem mass spectrometry (SPE-LC-MS/MS) method for the determination of sucralose in reclaimed and drinking waters and its photo degradation in natural waters from South Florida

Background Sucralose has gained popularity as a low calorie artificial sweetener worldwide. Due to its high stability and persistence, sucralose has shown widespread occurrence in environmental waters, at concentrations that could reach up to several μg/L. Previous studies have used time consuming sample preparation methods (offline solid phase extraction/derivatization) or methods with rather high detection limits (direct injection) for sucralose analysis. This study described a faster and sensitive analytical method for the determination of sucralose in environmental samples. Results An online SPE-LC–MS/MS method was developed, being capable to quantify sucralose in 12 minutes using only 10 mL of sample, with method detection limits (MDLs) of 4.5 ng/L, 8.5 ng/L and 45 ng/L for deionized water, drinking and reclaimed waters (1:10 diluted with deionized water), respectively. Sucralose was detected in 82% of the reclaimed water samples at concentrations reaching up to 18 μg/L. The monthly average for a period of one year was 9.1 ± 2.9 μg/L. The calculated mass loads per capita of sucralose discharged through WWTP effluents based on the concentrations detected in wastewaters in the U. S. is 5.0 mg/day/person. As expected, the concentrations observed in drinking water were much lower but still relevant reaching as high as 465 ng/L. In order to evaluate the stability of sucralose, photodegradation experiments were performed in natural waters. Significant photodegradation of sucralose was observed only in freshwater at 254 nm. Minimal degradation (<20%) was observed for all matrices under more natural conditions (350 nm or solar simulator). The only photolysis product of sucralose identified by high resolution mass spectrometry was a de-chlorinated molecule at m/z 362.0535, with molecular formula C12H20Cl2O8. Conclusions Online SPE LC-APCI/MS/MS developed in the study was applied to more than 100 environmental samples. Sucralose was frequently detected (>80%) indicating that the conventional treatment process employed in the sewage treatment plants is not efficient for its removal. Detection of sucralose in drinking waters suggests potential contamination of surface and ground waters sources with anthropogenic wastewater streams. Its high resistance to photodegradation, minimal sorption and high solubility indicate that sucralose could be a good tracer of anthropogenic wastewater intrusion into the environment.

Assessment of the occurrence and potential risks of antibiotics and their metabolites in South Florida waters using liquid chromatography tandem mass spectrometry

An automated on-line SPE-LC-MS/MS method was developed for the quantitation of multiple classes of antibiotics in environmental waters. High sensitivity in the low ng/L range was accomplished by using large volume injections with 10-mL of sample. Positive confirmation of analytes was achieved using two selected reaction monitoring (SRM) transitions per antibiotic and quantitation was performed using an internal standard approach. Samples were extracted using online solid phase extraction, then using column switching technique; extracted samples were immediately passed through liquid chromatography and analyzed by tandem mass spectrometry. The total run time per each sample was 20 min. The statistically calculated method detection limits for various environmental samples were between 1.2 and 63 ng/L. Furthermore, the method was validated in terms of precision, accuracy and linearity. ^ The developed analytical methodology was used to measure the occurrence of antibiotics in reclaimed waters (n=56), surface waters (n=53), ground waters (n=8) and drinking waters (n=54) collected from different parts of South Florida. In reclaimed waters, the most frequently detected antibiotics were nalidixic acid, erythromycin, clarithromycin, azithromycin trimethoprim, sulfamethoxazole and ofloxacin (19.3-604.9 ng/L). Detection of antibiotics in reclaimed waters indicates that they can't be completely removed by conventional wastewater treatment process. Furthermore, the average mass loads of antibiotics released into the local environment through reclaimed water were estimated as 0.248 Kg/day. Among the surface waters samples, Miami River (reaching up to 580 ng/L) and Black Creek canal (up to 124 ng/L) showed highest concentrations of antibiotics. No traces of antibiotics were found in ground waters. On the other hand, erythromycin (monitored as anhydro erythromycin) was detected in 82% of the drinking water samples (n.d-66 ng/L). The developed approach is suitable for both research and monitoring applications.^ Major metabolites of antibiotics in reclaimed wates were identified and quantified using high resolution benchtop Q-Exactive orbitrap mass spectrometer. A phase I metabolite of erythromycin was tentatively identified in full scan based on accurate mass measurement. Using extracted ion chromatogram (XIC), high resolution data-dependent MS/MS spectra and metabolic profiling software the metabolite was identified as desmethyl anhydro erythromycin with molecular formula C36H63NO12 and m/z 702.4423. The molar concentration of the metabolite to erythromycin was in the order of 13 %. To my knowledge, this is the first known report on this metabolite in reclaimed water. Another compound acetyl-sulfamethoxazole, a phase II metabolite of sulfamethoxazole was also identified in reclaimed water and mole fraction of the metabolite represent 36 %, of the cumulative sulfamethoxazole concentration. The results were illustrating the importance to include metabolites also in the routine analysis to obtain a mass balance for better understanding of the occurrence, fate and distribution of antibiotics in the environment. ^ Finally, all the antibiotics detected in reclaimed and surface waters were investigated to assess the potential risk to the aquatic organisms. The surface water antibiotic concentrations that represented the real time exposure conditions revealed that the macrolide antibiotics, erythromycin, clarithromycin and tylosin along with quinolone antibiotic, ciprofloxacin were suspected to induce high toxicity to aquatic biota. Preliminary results showing that, among the antibiotic groups tested, macrolides posed the highest ecological threat, and therefore, they may need to be further evaluated with, long-term exposure studies considering bioaccumulation factors and more number of species selected. Overall, the occurrence of antibiotics in aquatic environment is posing an ecological health concern.^

Assessment of the Occurrence and Potential Risks of Antibiotics and their Metabolites in South Florida Waters Using Liquid Chromatography Tandem Mass Spectrometry

An automated on-line SPE-LC-MS/MS method was developed for the quantitation of multiple classes of antibiotics in environmental waters. High sensitivity in the low ng/L range was accomplished by using large volume injections with 10-mL of sample. Positive confirmation of analytes was achieved using two selected reaction monitoring (SRM) transitions per antibiotic and quantitation was performed using an internal standard approach. Samples were extracted using online solid phase extraction, then using column switching technique; extracted samples were immediately passed through liquid chromatography and analyzed by tandem mass spectrometry. The total run time per each sample was 20 min. The statistically calculated method detection limits for various environmental samples were between 1.2 and 63 ng/L. Furthermore, the method was validated in terms of precision, accuracy and linearity. The developed analytical methodology was used to measure the occurrence of antibiotics in reclaimed waters (n=56), surface waters (n=53), ground waters (n=8) and drinking waters (n=54) collected from different parts of South Florida. In reclaimed waters, the most frequently detected antibiotics were nalidixic acid, erythromycin, clarithromycin, azithromycin trimethoprim, sulfamethoxazole and ofloxacin (19.3-604.9 ng/L). Detection of antibiotics in reclaimed waters indicates that they can’t be completely removed by conventional wastewater treatment process. Furthermore, the average mass loads of antibiotics released into the local environment through reclaimed water were estimated as 0.248 Kg/day. Among the surface waters samples, Miami River (reaching up to 580 ng/L) and Black Creek canal (up to 124 ng/L) showed highest concentrations of antibiotics. No traces of antibiotics were found in ground waters. On the other hand, erythromycin (monitored as anhydro erythromycin) was detected in 82% of the drinking water samples (n.d-66 ng/L). The developed approach is suitable for both research and monitoring applications. Major metabolites of antibiotics in reclaimed wates were identified and quantified using high resolution benchtop Q-Exactive orbitrap mass spectrometer. A phase I metabolite of erythromycin was tentatively identified in full scan based on accurate mass measurement. Using extracted ion chromatogram (XIC), high resolution data-dependent MS/MS spectra and metabolic profiling software the metabolite was identified as desmethyl anhydro erythromycin with molecular formula C36H63NO12 and m/z 702.4423. The molar concentration of the metabolite to erythromycin was in the order of 13 %. To my knowledge, this is the first known report on this metabolite in reclaimed water. Another compound acetyl-sulfamethoxazole, a phase II metabolite of sulfamethoxazole was also identified in reclaimed water and mole fraction of the metabolite represent 36 %, of the cumulative sulfamethoxazole concentration. The results were illustrating the importance to include metabolites also in the routine analysis to obtain a mass balance for better understanding of the occurrence, fate and distribution of antibiotics in the environment. Finally, all the antibiotics detected in reclaimed and surface waters were investigated to assess the potential risk to the aquatic organisms. The surface water antibiotic concentrations that represented the real time exposure conditions revealed that the macrolide antibiotics, erythromycin, clarithromycin and tylosin along with quinolone antibiotic, ciprofloxacin were suspected to induce high toxicity to aquatic biota. Preliminary results showing that, among the antibiotic groups tested, macrolides posed the highest ecological threat, and therefore, they may need to be further evaluated with, long-term exposure studies considering bioaccumulation factors and more number of species selected. Overall, the occurrence of antibiotics in aquatic environment is posing an ecological health concern.

Contaminants of emerging concern in drinking water: assessment of the effectiveness of removal by drinking water treatments and possible risks to human health

Contaminants of emerging concern are increasingly detected in the water cycle, with endocrine-disrupting chemicals (EDCs) receiving attention due to their potential to cause adverse health effects even at low concentrations. Although the EU has recently introduced some EDCs into drinking water legislation, most drinking water treatment plants (DWTPs) are not designed to remove EDCs, making their detection and removal in DWTPs an important challenge. The aim of this doctoral project was to investigate hormones and phenolic compounds as suspected EDCs in drinking waters across the Romagna area (Italy). The main objectives were to assess the occurrence of considered contaminants in source and drinking water from three DWTPs, characterize the effectiveness of removal by different water treatment processes, and evaluate the potential biological impact on drinking water and human health. Specifically, a complementary approach of target chemical analysis and effect-based methods was adopted to explore drinking water quality, treatment efficacy, and biological potential. This study found that nonylphenol (NP) was prevalent in all samples, followed by BPA. Sporadic contamination of hormones was found only in source waters. Although the measured EDC concentrations in drinking water did not exceed threshold guideline values, the potential role of DWTPs as an additional source of EDC contamination should be considered. Significant increases in BPA and NP levels were observed during water treatment steps, which were also reflected in estrogenic and mutagenic responses in water samples after the ultrafiltration. This highlights the need to monitor water quality during various treatment processes to improve the efficiency of DWTPs. Biological assessments on finished water did not reveal any bioactivity, except for few treated water samples that exhibited estrogenic responses. Overall, the data emphasize the high quality of produced drinking water and the value of applying integrated chemical analysis and in vitro bioassays for water quality assessment.

Toxic cyanobacteria in water intended for drinking purpose: improvement of cells and toxins’ analyses and of treatments for their removal

Massive proliferations of cyanobacteria in freshwaters have recently increased, causing ecological and economic losses. Their ever-increasing presence in water sources destined to potabilization has become a major threat for public health, since several species can produce harmful toxins (cyanotoxin). Therefore, additional specific measures to improve management and treatment of drinking water(s) are required. The PhD thesis investigates toxic cyanobacteria in drinking waters with a special focus on Emilia-Romagna (Italy), throughout three separated chapters, each with different specific objectives. The first chapter aims at improving the fast monitoring of cyanobacteria in drinking water, which was investigated by testing different models of multi-wavelength spectrofluorometers. Inter-laboratories calibrations were conducted using mono-specific cultures and field samples, and both the feasibility and the technical limitations of such tools were illustrated. The second chapter evaluates the effectiveness of drinking water treatments in removing cyanobacterial cells and toxins. Two chlorinated oxidants (sodium hypochlorite and chlorine dioxide) already in use for pre-oxidation during water potabilization, were tested on cultures of the toxic cyanobacterium Microcystis aeruginosa posing a specific focus on toxin removal and revealing that pre-oxidation can cause the release of toxins and unknown metabolites. Innovative treatments based on non-thermal plasma were also tested, observing an effective and rapid inactivation of cyanobacterial cells. The third chapter presents a study on a cyanobacterium isolated from a drinking water reservoir of Emilia-Romagna and investigated by combining biological, chemical, and genomic methods. Although the strain did not produce any known cyanotoxin, high toxicity of water-extract was observed in bioassays and potential implications for drinking water were discussed. Overall, the PhD thesis offers new insights into toxic cyanobacteria management in drinking water, highlighting best practices for drinking water managers regarding their detection and removal. Additionally, the thesis provides new contributions to the understanding of the freshwater cyanobacteria community in the Emilia-Romagna region.

Freshwater arsenic detoxification through selenium-enriched food supplements. A proteomic approach

Dissertação apresentada para a obtenção do grau de doutor em Bioquímica pela Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia

Nitrite biosensing via selective enzymes—a long but promising route

Sensors 2010, 10, 11530-11555

Gestão dos recursos hídricos no processo de desenvolvimento sustentável de Cabo Verde: uma proposta

A água é vital para a sobrevivência, saúde e dignidade do ser humano e uma fonte fundamental para o seu desenvolvimento. As reservas de água potável no mundo estão sob pressão constante embora muitos ainda não tenham acesso a esse precioso líquido para atender ás necessidades básicas. A água é um recurso natural com valor económico, estratégico e social. A percepção da escassez tem levado governos, a reorganizar o ambiente institucional e definir novos direitos de propriedade por meio de um sistema de gestão participativo e descentralizado que estimule a utilização do recurso de forma racional. A investigação está direccionada à gestão e à implementação da Lei n.º 41/II/84 de 18 de Junho, que Aprova o Código de Águas que estabelece as bases gerais do regime jurídico de propriedade, protecção, conservação, desenvolvimento, administração e uso dos recursos. O enfoque da pesquisa é as Instituições Governamentais que lidam com a água e as Associações de Bacias hidrográficas que, com os seus olhares e saberes, contribuem com informações para que se construa uma Proposta de Modelo de Gestão numa perspectiva de sustentabilidade. A proposta de modelo pretende contribuir para uma gestão sustentada. A pesquisa é do tipo qualitativa e usa como metodologia o Estudo de Caso. Foi constatada a complexidade da gestão das águas em Cabo Verde, na qual existem ainda muitos entraves e desafios. A Bacia Hidrográfica é a unidade básica de gestão, onde as Associações, as Câmaras Municipais, os Serviços Autónomos de Água e Saneamento, o Instituto Nacional de Gestão dos recursos Hidricos desempenham o papel de gerí-la e preservá-la. Foram identificados como dificuldades: a posse da água não está bem clara, o que dificulta a gestão da Bacia; os conflitos sociais são frequentes quanto ao uso da água no que tange à poluição; a população rural não está devidamente contemplada na gestão dos recursos hídricos. Por fim, constatou-se que Cabo Verde possui as ferramentas para a evolução da Gestão dos Recursos Hídricos, que é o Código de Água. Water is vital for the survival, health and dignity of the human being and a basic source for development. The drinking waters reserves in the world are under constant pressure. The water can be considered a natural resource with economic, strategically and social values. The perception of the scarcity has taken governments, to reorganize the institutional environment and to define new rights of property by means of participation and decentralized system of management that stimulates the use of the resource in rational form. This investigation addressed the Administration of Water Resources in Cabo Verde and the implementation of Law n.º 41/II/84 of 18 of June, which established the National Policy of Water Resources and the National System of Management. The approach of the research is the Governmental Institutions that deal with the water and the Associations that, at a glance, contribute with information that builds the proposal of water resources management in a sustainability perspective. The proposal has as objective to provide subsidies to advance more and more the research regarding sustainability in the administration of waters. The research was qualitative and it used as methodology the Study Case. We identified as difficulties: the ownership of the water is not well clear; the social conflicts are frequent concerning how to use the water. Finally, it was evidenced that Cape Verde has the tools for the evolution of water resources management, which is the Water Code.

Determinação de baixas concentrações de cobre em águas naturais por espectrofotometria de absorção atômica em chama usando um sistema SIA com uma coluna de poli(etilenimina)

This paper describes a sequential injection analysis (SIA) set-up coupled to a flame atomic absorption spectrometer (FAAS) to accomplish the determination of low concentrations of copper in drinking waters. Copper is first retained under neutral media in an on-line 29x1.6 mm column filled with poly(ethylenimine) immobilised on silica gel. The retained analyte is then eluted by flowing through the column 250 mL of a nitric acid solution. The selection of 3.85 ml of sample enabled to obtain a detection limit of 0.27 mug/L and a sampling rate of about 24 samples/h. There was a good agrement between the results of 12 samples furnished by the proposed procedure and by electrothermal atomic absorption spectrometry. Repeatability assessment gave a relative standard deviation of 1.3 % after ten replicate analysis of a sample containing about 70 mug/L in copper..

Automated flow-injection method for cadmium determination with pre-concentration and reagent preparation on-line

The spectrophotometric determination of Cd(II) using a flow injection system provided with a solid-phase reactor for cadmium preconcentration and on-line reagent preparation, is described. It is based on the formation of a dithizone-Cd complex in basic medium. The calibration curve is linear between 6 and 300 µg L-1 Cd(II), with a detection limit of 5.4 µg L-1, an RSD of 3.7% (10 replicates in duplicate) and a sample frequency of 11.4 h-1. The proposed method was satisfactorily applied to the determination of Cd(II) in surface, well and drinking waters.

Conflito de usos da água e ocupação do solo da Bacia Hidrográfica do rio Pitimbu no município de Macaíba, RN

Due to the increasing activities and its disordered occupation, the catchment of the Pitimbu river is reason of concern for the population, since the river empties in the lagoon of the Jiqui where 30% of the water of this lagoon are caught for the supplying the city of Natal, playing an important fuction in the supply of water superficial for the capital of the Rio Grande do Norte. The superior stretch of the hydrography catchment of the Pitimbu river - Macaíba/RN, object of this study, is denoted by the eminently agricultural occupation with the use of the irrigated agriculture in the many properties that compose this stretch. Because of this becomes necessary to analyze the availability of the water resources in the catchment of the Pitimbu river, in the superior stretch, collating with the use and occupation of the existing terrain. The stretch in study was divided in eight points which had been performed tests throughout the water and analyzed the parameters physicist-chemistries and heavy metals that are praised by resolution CONAMA nº 357 e, visits in field with photographic survey for characterization of this verifying the situation where if it finds the river. The stretch presented some divergence in the parameters of pH and iron, however the results denote a characteristic of the region. The great amount of slide barrages throughout the river and the wet street was observed, had also verified near the edges the launching of garbage and, the great amount of aquatic vegetation. With this it is concluded that the water of the river can be used for human supplying, therefore inside presents a quality of drinking waters of the standards demanded for resolution CONAMA nº 357, in this way, the stretch in study could be adopted as an Envoiremental Area Protection (APA), to preserve and to guarantee, at least in this stretch, that its condition remains unchanged and continues to supply of permanent form a water of excellent quality

Análise dos fatores responsáveis pela alta salinidade do aqüífero fissural da microbacia da Riacho do Feijão - Pedro Avelino/RN

The fissures aquifer northeast semi-arid Brazilian, present high text frequently of leave, with of low a hídric availability. The research has as objective main to analyze the components that inside influence in the salinity of the waterbearing fissures of an evaluation physicist-chemistry of the water, leading in consideration the physical interventions of the environment. One used techniques of interpretation of image of Landsat satellite -1999 and delimitation of the micro basin through the topographical map SUDENE. One identified waters of the NaCl type with Ca++ and Mg++ in secondary concentrations. The analyzed wells (15), had presented an average salinity of 5.147 mg/L of STD and a well only supplies drinking waters with 319 mg/L of STD. The recharge of the aquifer one if carries through for infiltration in the open fracture of ortognaisse it migmatization. The type and directions of the fracture do not control the STD. Relations between salinity and out let do not exist. The quality of the well of low salinity is identical the superficial waters (aquifer dam and alluvial). The studies of the meteoric erosion processes had evidenced that in the transformations of the rock in ground, the Ca++ and Na+ are taken for superficial waters. The treatment of the data chemical showed that the grade of Na+, Ca++, Mg++ and Cl-are controlled for the evaporation process, from only water that would have the qualities of superficial waters or the well of low salinity. Already the HCO3-grade is controlled for the precipitation of the dolomite. The STD of this aquifer one would be consequence of the high tax of evaporation of dams constructed in regions of plain topography. You leave them precipitated in deep argillaceous ones dry dams are led for the aquifer in first rains. The research suggests some recommendations for the use and exploitation of the water salinity in piscicultura, carcinicultura, culture of the grass-salt (Atriplex sp), among others

Photoelectrocatalytic Removal of Bromate Using Ti/TiO2 Coated as a Photocathode

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of the environment on behavior patterns of laying hens kept in cages

The aim of this paper is to verify the correlation between environmental indicators and behaviors expressed by laying hens kept in cages. The birds react to a severe environment through their behaviors, end the behaviors can be monitored to identify the birds' welfare conditions. The behaviors birds display ere the result of stress caused by the combination of environmental temperature, relative humidity, radiant heat, and air speed (environmental temperature being the most important). In order to check the influence of the environment, an experiment was carried out on a commercial poultry farm, located in the city of Bastos. The study was initiated in March 2007, during four non-consecutive weeks. The birds' behaviors were recorded using video, by cameras installed in the cages. The birds behaviors were identified and noted for the frequency of occurrence for each bird, and the average duration of each behavior (in seconds), using video samples of 15 minutes recorded from 1 PM to 4 PM. The environmental variables collected were: air temperature, concentration of ammonia, relative air humidity, velocity of the air, noise, roof temperature, and light intensity. The observed behaviors were: opening wings, stretching, threatening, ruffling feathers, drinking water, aggressive pecking, eating, running, lying down, stretching head out of the cage, preening, mounting and prostrating. Principal Components Analysis was used to determine associations between the behavior variables and environmental variables described above. In this experiment, there were no significant correlations between behavioral variables and environmental variables.

Effects of the density, aviary type, breed, and age on the behaviors of laying hens kept in cages

An experiment was carried out in order to investigate the behaviors of laying hens due to the environmental factors of: density inside of the cage, aviary type, breed, and age. The experiment was configured as a factorial 4x2x2x2 study, with treatments being four different ages, two different breeds, two different cage densities, and two different aviaries. The birds' behaviors were recorded using video cameras installed in the cages, using samples of 15 minutes recorded from 12 PM to 4 PM. The observed behaviors, frequency and duration of behaviors (measured in seconds) were identified and noted related to each bird. The study was initiated in March 2007, during four non-consecutive weeks. The observed behaviors were: opening wings, stretching, threatening, ruffling feathers, drinking water, aggressive pecking, eating, running, lying down, stretching head out of the cage, preening, mounting, prostrating, and doing nothing (inactivity). Due to the non-normality of the data recorded, the Kruskal-Wallis statistical test of the MINITAB Statistical Software® was used to compare the medians of the variables. For breed factor, only the durations of the eating presented significant differences (p-value< 0.05). For cage density, there was a significant median difference (p-value< 0.05) for almost all behaviors observed. The average length of time of behaviors was higher for the lowest cage density. However, the frequency of behaviors was lmerfor the lowest cage density. The frequency of the behaviors to preen feathers, to lie down, to drink water and to stretch the head were higher in the aviary, where the groups of birds were smaller. The observed behaviors were particularly affected by experimental factors cage density, and aviary type, which directly affects the available space for each bird.