Integrated analysis of marine biotoxins and contaminants of emerging concern in bivalve mollusks from Santa Catarina, Brazil.

Santa Catarina is the main producer state of oysters and mussels in Brazil, reaching 98 % of national production. To assure the safety of bivalve mollusks production, control programs of marine biotoxins (MBs) have been continuously performed. Herein, the co-occurrence of MBs and contaminants of emerging concern (CECs) in oyster and mussels from the main production sites of Santa Catarina was reported, covering 178 compounds. Samples of wild and non-cultivated oysters and mussels were also assessed. Chemometric tools were used to evaluate and optimize several sample preparation techniques such as solid-liquid, ultrasound assisted, and pressurized liquid extraction. The optimized protocol was based on ultrasound assisted extraction followed by liquid chromatography coupled to tandem mass spectrometry. The results showed the incidence of several CECs and MBs. In the case of MBs, all results were below the regulatory limits for both cultivated and non-cultivated samples. Wild mollusks have shown a higher number of compounds. Regarding CECs, the more frequent compounds were caffeine, diclofenac, meloxicam, and sertraline. Domoic acid and okadaic acid were the main toxins detected. The results highlighted the need of monitoring for MBs and the potential of oyster and mussels as sentinel organisms to risk analysis of CECs in coastal regions. To the best of our knowledge, this is the first method to describe a simultaneous sample preparation and analysis of CECs and MBs in bivalve mollusks, as well as the first report of meloxicam and florfenicol in mussels and oysters.

Quantification of pharmaceuticals in hospital effluent: Weighted ranking of environmental risk using a fuzzy hybrid multicriteria method.

An analytical method for quantification of seventeen pharmaceuticals and one metabolite was validated and applied in the analysis of hospital effluent samples. Two different sampling strategies were used: seasonal sampling, with 7 samples collected bimonthly; and hourly sampling, with 12 samples collected during 12 h. Thus, the variability was both seasonal and within the same day. High variability was observed in the measured concentrations of the pharmaceuticals and the metabolite. The quantification method, performed using weighted linear regression model, demonstrated results of average concentrations in seasonal samples ranged between 0.19 µgL-1 (carbamazepine) and higher than 61.56 µgL-1 (acetaminophen), while the hourly samples showed average concentrations between 0.07 µgL-1 (diazepam) and higher than 54.91 µgL-1 (acetaminophen). It is described as higher because the maximum concentration of the calibration curve took into account the dilution factor provided by DLLME. The diurnal results showed a trend towards higher concentrations in the first and last hours of sampling. The risk quotient (RQ) was calculated using organisms from three different trophic levels, for all the analytes quantified in the samples. Additionally, in order to understand the level of importance of each RQ, an expert panel was established, with contributions from 23 specialists in the area. The results were analyzed using a hybrid decision-making approach based on a Fuzzy Analytic Hierarchy Process (FAHP) and the Technique for Order Preference by Similarity to Ideal Solution (TOPSIS) method, in order to rank the compounds by environmental risk priority. The compounds of greatest concern were losartan, acetaminophen, 4-aminoantipyrine, sulfamethoxazole, and metoclopramide. Comparison of the environmental risk priority ranking with the potential human health risk was performed by applying the same multicriteria approach, with the prediction of endpoints using in silico (Q)SAR models. The results obtained suggested that sulfamethoxazole and acetaminophen were the most important analytes to be considered for monitoring.

Qualitative evaluation of pharmaceuticals and metabolites in hospital effluent: Influence of sample preparation technique and outranking by environmental risk using the ELECTRE method.

The presence of pharmaceuticals and metabolites in effluents has become a serious environmental problem, so it is essential to be able to monitor these microcontaminants using qualitative approaches, as well as to assess the potential environmental risks that such compounds may present. Therefore, in this study, suspect screening analysis was performed of 2030 pharmaceuticals and metabolites in hospital effluent samples, applying different sample preparation techniques. Additionally, a pioneering association of (Q)SAR assessment of identified contaminants with the ELECTRE multi-criteria decision analysis technique made it possible to prioritize analytes according to their environmental risk, in order to enable their inclusion in environmental monitoring programs. The results showed that the most advantageous alternative sample preparation technique consisted of cleanup (100 mg of silica/alumina sorbent) + dispersive liquid-liquid microextraction (7.5 of aqueous matrix, 325 µL of chloroform (extracting solvent), and 500 µL of acetonitrile as dispersing solvent). This procedure resulted in the identification of 70 pharmaceuticals and metabolites in the composite sample tested. The suspect screening analysis found a total of 105 microcontaminants, 28 of them being "confirmed compounds" and 77 being "suspect compounds". Of the compounds identified, 87% were pharmaceuticals and 13% were metabolites. The compounds identified were subsequently evaluated using different open access software packages, considering eight endpoints: mobility, persistence, estrogen receptor binding, wastewater treatment plant total removal, biodegradability, PBT (persistent, bioaccumulation and toxic), mutagenicity, and carcinogenicity. The (Q)SAR prediction results were used as input data for the ELECTRE outranking method. Categorization of the identified compounds by ELECTRE resulted in the kernel (priority compounds) and a further 19 groups. ELECTRE sensitivity evaluation indicated that for all the cases, the kernel and the following two groups coincided. The categorization provided by the ELECTRE method constitutes a highly intuitive decision and choice tool, which can assist in the selection of compounds if subsequent quantitative analysis is to be carried out.

Multi-criteria decision-making techniques associated with (Q)SAR risk assessment for ranking surface water microcontaminants identified using LC-QTOF MS.

Contaminants of emerging concern (CECs) have been a focus of study for years, with investigations revealing the contamination of different environmental matrices (surface water, soil, air, and sediment) by diverse classes of microcontaminants. Understanding the contamination profiles requires identification and risk assessment of the microcontaminants. In the present work, analysis was made of the presence of 3250 compounds in 27 samples from the Conceição River (Rio Grande do Sul State, Brazil), using an SPE-LC-QTOF MS method. In total, 150 microcontaminants (confirmed and suspected) of different classes, especially pesticides and pharmaceuticals, were identified by an initial qualitative analysis. Subsequently, in silico predictions of eight endpoints, using quantitative structure-activity relationship ((Q)SAR) models, were employed to determine the risk of each previously screened microcontaminant. This large amount of (Q)SAR data, frequently with conflicting information in relation to the responses of the different endpoints, makes it difficult to define which microcontaminants should be prioritized for analysis. Therefore, in order to rank the identified microcontaminants by risk assessment, two multi-criteria decision-making (MCDM) ranking techniques (ToxPi and TOPSIS), associated with a weighting method, were performed to establish the order of priority for further quantitative analysis of the most hazardous microcontaminants. The two rankings were statistically similar, especially for the 20 highest priority microcontaminants. Nonetheless, sensitivity tests carried out for the ToxPi and TOPSIS outputs showed higher performance robustness of TOPSIS, compared to ToxPi. This is the first time that such an approach (screening/(Q)SAR/MCDM methods) has been performed in the context of microcontaminant environmental risk evaluation and demonstrated to be an available strategy to help rank the most concern microcontaminants identified in aqueous environment samples.

Pharmaceuticals, pesticides and metals/metalloids in Lake Guaíba in Southern Brazil: Spatial and temporal evaluation and a chemometrics approach.

Compiling and reporting data related to the presence of pharmaceuticals and pesticides are crucial means of assessing the risk those chemicals pose to human health and environment. Data sets from different sources were combined using a data fusion approach to produce a spatial and temporal variation of contaminants presents in water from Lake Guaíba (29°55'-30°24' S; 51°01'-51°20' W). Lake Guaíba is a 496 km2 water body situated in the geological depression of Rio Grande do Sul State, Brazil; that is fed by several rivers from the metropolitan area, the 5th largest metro area in Brazil, with approximately 5 million inhabitants. Analytical methodology to quantify pharmaceuticals and pesticides by LC-QTOF-MS and GC-MS/MS was validated for 41 pharmaceutical and 62 pesticides. Furthermore, 27 chemical elements were analyzed by ICP-MS, and physical chemical parameters were determined using established methodologies. All validation parameters were in accordance with the National Institute of Metrology, Standardization, and Industrial Quality. Thirty-five water samples were analyzed from January to August 2019, and 15 pharmaceuticals and 25 pesticides were present in concentrations ranging from 6.00 ng L-1 to 580.00 ng L-1. Twenty-seven elements were analyzed during the same period, and 18 were present in concentrations ranging from 0.2 µg L-1 to 7060 µg L-1. Samples were tagged according to the points and months of collection to identify temporal and spatial patterns. The main findings show that the compounds are distributed throughout the studied area without an apparent regular pattern, suggesting that events in a specific point affect the entire ecosystem. Conversely, temporal variations were well defined, as samples were grouped according to the climatic conditions of the months of collection. Considering the calculated quotient risks, atrazine, cyproconazole, diuron, and simazine showed the highest risk levels for algae; acetaminophen, diclofenac, and ibuprofen showed the highest risk levels for aquatics invertebrates.

Economically feasible strategy for confirmation of pharmaceuticals in hospital effluent using screening analysis.

The contamination of aquatic systems by pharmaceuticals has received considerable attention in recent decades, because these substances are increasingly detected in the environment. This is due to the abundant use of pharmaceuticals by the population and, consequently, their constant introduction into aquatic systems through domestic, industrial, and hospital wastewaters. Hospital effluents have highly complex compositions and present potential toxicity towards the environment. In this work, a screening methodology was developed to evaluate the occurrence of pharmaceutical products in hospital wastewater, using a viable, easy, and economical strategy employing commercial pharmaceutical compounds for screening analysis. Six samplings of hospital wastewater were carried out monthly (from winter until summer). The samples were filtered and pre-concentrated/extracted using solid phase extraction (SPE). The pharmaceuticals screening procedure required the construction of two databases, one for each ionization mode (positive and negative), which contained information that allowed the identification of the presence of these pharmaceuticals in the studied samples. Commercial pharmaceutical compounds were used as analytical standards. Based on this strategy and, using liquid chromatography coupled to high resolution mass spectrometry, it was possible to screen 110 pharmaceuticals and, from these, to confirm the presence of 38 pharmaceuticals in analyzed samples. These results indicate the analytes that should be taken into account in the further development of quantitative methods for pharmaceutical analysis.

Photodegradation of Emerging Contaminant 2-(tiocyanomethylthio) Benzothiazole (TCMTB) in Aqueous Solution: Kinetics and Transformation Products.

Direct photolysis of the emerging contaminant 2-(thiocyanomethylthio) benzothiazole (TMCTB) was performed in aqueous solution at different concentrations with high-pressure mercury lamp (5.0, 8.0, 13.0, 16.0, 20.0, 23.0, 27.0, 35.0, 40.0, 45.0, and 50.0 mg L- 1) and with natural sunlight radiation (6.0, 30.0, and 60.0 mg L- 1). TCMTB underwent rapid degradation by direct photolysis with a high-pressure mercury lamp in aqueous solutions, with 99% removal after 30 min at all concentrations studied. For sunlight photolysis, TCMTB degradation was observed with 96%, 81%, and 64% removal for initial concentrations of 6.0, 30.0, and 60.0 mg L- 1, respectively, after 7 h of exposure to sunlight. The degradation of TCMTB in lab-scale wastewater had kinetic constant and t1/2 in the same order when compared to the photodegradation of TCMTB in aqueous solutions. In addition, the results showed that photolysis with a high-pressure mercury lamp and sunlight were governed by the same kinetic order, however the kinetic parameters showed that degradation with sunlight was 40 times slower than photolysis with the mercury lamp. Twelve transformation products (TP) were identified, and eight of the TP have not been described in the literature. Furthermore, prediction of toxicity with ECOSAR software was carried out for fish, daphnids, and green algae species. It showed that photolytic treatment is efficient for reducing the toxicity of the compound, since the degradation formed compounds with lower toxicity than the primary compound. In conclusion, this study suggests that photolysis is an efficient way to remove the studied contaminant, and it highlights the potential of this technique for the degradation of emerging contaminants in industrial wastewater treatment plants.

Comparative endocrine disrupting compound removal from real wastewater by UV/Cl and UV/H2O2: Effect of pH, estrogenic activity, transformation products and toxicity.

Extensive use of endocrine disruptor compounds (EDCs) and their release through various pathways into the environment are emerging environmental concerns. In this context, H2O2 and chlorine UV-based treatments were carried out to evaluate their efficiency in the removal of the bisphenol A (BPA), 17ß-estradiol (E2) and 17α-ethinylestradiol (EE2) at 100 µg L-1 from ultrapure water and from wastewater treatment plants (WWTP). Photolysis was performed under different irradiation sources, i.e. UVC and UVA. The effect of H2O2 (3 and 30 mg·L-1), free chlorine concentrations (1 and 2 mg·L-1) and pH (5, 7 and 9) were also investigated. Toxicity (Raphidocelis subcapitata) and estrogenic activity (yeast estrogen screen - YES assay) were assessed during the processes. Compound removal at optimal operating parameters reached 100% after 15 and 2 min for UVC/H2O2 (pH 9 and 3 mg L-1 of H2O2), and UVC/Cl (pH 9 and 2 mg L-1 of chlorine), respectively. Total organic carbon (TOC) removal achieved 37% and 45% for the H2O2 and Cl-UV based process, respectively. The in vitro YES assay indicated that the formed by-products were non-estrogenic compounds, while the toxicity evaluation revealed high cell growth inhibition due to UVC/Cl byproducts. During the UV-based processes, 30 transformation products (TPs) were identified, in which three new chlorinated TPs from E2 and EE2 may be responsible for toxicity effects. EDC degradation by UV/Cl is faster than by UV/H2O2, although chlorinated toxic byproducts were also formed during the UV/Cl process.

Presence of antibiotic resistance genes and its association with antibiotic occurrence in Dilúvio River in southern Brazil.

It is known that antibiotics are widely used in human and veterinary medicine. In some countries the use is controlled, however few restrictions to their use are enforced in many countries. Antibiotics and their metabolites can reach the water bodies through sewage systems, especially in those countries with partial or absent wastewater treatment systems. The overuse and misuse of antibiotics has been linked with the increase of antibiotic resistant bacteria. The relation between the occurrence of antibiotics and resistance genes in surface waters has been widely studied worldwide evincing the great importance of this subject. In this work, a methodology for quantification of 40 antibiotics of 5 different classes, in river water, by SPE-LC-MS/MS was validated. Samples were taken during a two-year period from Dilúvio River, a stream that crosses the city of Porto Alegre (RS - Brazil) and receives in nature domestic effluent. The methodology met the requirements of validation, with Limit of Quantification varying from 20 ng L-1 to 100 ng L-1. A total of 48 samples was analyzed for the presence of antibiotics for two years. From the 40 antibiotics analyzed, 8 of them (Azithromycin, Cephalexin, ciprofloxacin, clindamycin, norfloxacin, sulfadiazine, sulfamethoxazole and trimethoprim) were present in all sampling points in the range of

Degradation of pharmaceuticals in different water matrices by a solar homo/heterogeneous photo-Fenton process over modified alginate spheres.

A solar homo/heterogeneous photo-Fenton process using five materials (Fe(II), Fe(III), mining waste, Fe(II)/mining waste, and Fe(III)/mining waste) supported on sodium alginate was used as a strategy to iron dosage for the degradation of eight pharmaceuticals in three different water matrices (distilled water, simulated wastewater, and hospital wastewater). Experiments were carried out in a photoreactor with a capacity of 1 L, using 3 g of iron-alginate spheres and an initial hydrogen peroxide concentration of 25 mg L-1, at pH 5.0. All the materials prepared were characterized by different techniques. The Fe(III)-alginate spheres presented the best pharmaceutical degradation after a treatment time of 116 min. Nineteen transformation products generated during the solar photo-Fenton process were identified by liquid chromatography coupled to quadrupole time-of-flight mass spectrometry, using a purpose-built database developed for detecting these transformation products. Finally, the transformation products identified were classified according to their toxicity and predicted biodegradability.

Residue of insecticides in foodstuff and dietary exposure assessment of Brazilian citizens.

The goal of this work is to study the presence of 17 different pyrethroids and chlorpyrifos in animal origin food samples, including chicken, beef, fish, eggs and milk. The samples were analyzed by GC/NCI-MS/MS in order to determine their insecticide concentration levels, the relation between the amount of insecticides and the lipid content, as well as their isomeric composition. Bifenthrin, cypermethrin, cyhalothrin, permethrin, deltamethrin and chlorpyrifos have been detected in real samples. The highest levels find were verified for chlorpyrifos (45.7 µg L-1, 17.5 µg kg-1 ww) and cypermethrin (2.75 µg L-1, 14.7 µg kg-1 ww) in milk and beef, respectively. The estimate of daily intake showed that none of these insecticides exceeded the safety limits of the acceptable daily intake values. A strong correlation between the amount of pesticides and lipid content was found for beef, fish and chicken samples. Regarding the isomers, a predominance of cis isomers in samples contaminated with cypermethrin was verified.

Methodology for trace analysis of 17 pyrethroids and chlorpyrifos in foodstuff by gas chromatography-tandem mass spectrometry.

This study aimed to develop an efficient, sensitive, and reliable analytical method for trace analysis of 17 different pyrethroids and chlorpyrifos in the fatty content of animal products, including beef, chicken, eggs, fish, and milk. The method developed is based on an ultrasound extraction using lyophilized samples, a solid phase extraction cleanup with basic alumina and C18 cartridges in tandem, and analysis by gas chromatography coupled to tandem mass spectrometry in negative chemical ionization mode. Recovery values were in the range of 27-128 % with relative standard deviation always below 25 %, and chiral analysis of recovery data showed predominance of isomers of cis form over trans. Limits of detection (LODs) ranged from 0.002 to 6.43 ng g-1 lipid weight (lw), and limits of quantification (LOQs) ranged between 0.006 and 21.4 ng g-1 lw. The developed methodology was used for the analysis of 25 samples of fatty foods. All samples were positive for at least one of the pesticides, chlorpyrifos, bifenthrin, cyhalothrin, permethrin, cypermethrin, or deltamethrin, with mass fraction levels ranging from 0.03 to 270 ng g-1 lw. Graphical Abstract ᅟ.

Effect of 950 MHz UHF electromagnetic radiation on biomarkers of oxidative damage, metabolism of UFA and antioxidants in the livers of young rats of different ages.

PURPOSE: To assess the effect of 950 MHz ultra-high-frequency electromagnetic radiation (UHF EMR) on biomarkers of oxidative damage, as well as to verify the concentration of unsaturated fatty acids (UFA) and the expression of the catalase in the livers of rats of different ages. MATERIALS AND METHODS: Twelve rats were equally divided into two groups as controls (CR) and exposed (ER), for each age (0, 6, 15 and 30 days). Radiation exposure lasted half an hour per day for up to 51 days (21 days of gestation and 6, 15 or 30 days of life outside the womb). The specific absorption rate (SAR) ranged from 1.3-1.0 W/kg. The damage to lipids, proteins and DNA was verified by thiobarbituric acid reactive substances (TBARS), protein carbonyls and comets, respectively. UFA were determined by gas chromatography with a flame ionization detector. The expression of catalase was by Western blotting. RESULTS: The neonates had low levels of TBARS and concentrations of UFA after exposure. There was no age difference in the accumulation of protein carbonyls for any age. The DNA damage of ER 15 or 30 days was different. The exposed neonates exhibited lower expression of catalase. CONCLUSIONS: 950 MHz UHF EMR does not cause oxidative stress (OS), and it is not genotoxic to the livers of neonates or those of 6 and 15 day old rats, but it changes the concentrations of polyunsaturated fatty acid (PUFA) in neonates. For rats of 30 days, no OS, but it is genotoxic to the livers of ER to total body irradiation.