Simultaneous evaluation of benzotriazoles, benzothiazoles and benzenesulfonamides in water samples from the impacted urban Jacarepaguá Lagoon System (Rio de Janeiro, Brazil) by liquid chromatography coupled to electrospray tandem mass spectrometry.

Benzotriazoles, benzothiazoles, and benzenesulfonamides are emerging pollutants stable in aquatic media emitted by anthropogenic sources. Selected compounds of these classes were evaluated in the impacted urban Jacarepaguá Lagoon System (JLS) located in a tropical coastal region of Rio de Janeiro City, Brazil that has experienced a rapid expansion of urban occupation and environmental degradation in recent decades, and it represents a pioneering study of these compounds carried out in Brazilian areas. A method of solid phase extraction followed by ultra-performance liquid chromatography coupled to electrospray tandem mass-spectrometry was implemented to evaluate water samples collected in different water bodies (rivers, lagoons, and channels) of the JLS from March 2017 to May 2018. Limits of quantification (LOQs) ≤ 10.0 ng L-1, method linearity up to 1000 µg L-1, and recoveries between 62 and 121 % at three different levels were obtained. Individual concentrations varied from < LOQ to 5260 ng L-1 (benzotriazole, in May 2018) which also predominated in all river samples. 2-mercaptobenzothiazole predominated in samples taken in lagoons and channels in March 2017, and 2-aminobenzothiazole was never detected. River samples showed total concentrations up to 30 times larger in all sampling campaigns, except March 2017 when the sample taken at Tijuca Lagoon showed the largest total concentration of the compounds studied due to the largest concentration of 2-mercaptobenzothiazole (2505 ng L-1) found in this study. Principal component analysis (PCA) using only composition data was unable to distinguish samples from rivers, and lagoons and channels, but a PCA combining composition data and environmental parameters (pH, Eh, dissolved O2 concentration, temperature, salinity, and conductivity) discriminated the samples according to two groups: rivers and lagoons and channels. The Joá Channel flows directly to the open sea and our data allowed a (preliminary) estimation of the total mass flows of the studied compounds to the open sea, which would vary between 1702 g day-1 (March 2017) to 106 g day-1 (May 2018) and allowed a preliminary estimative based on the geometric mean of input of 87.9 kg year-1, indicating the importance of the drainage area to the contamination of the coastal area, and consequently to ocean pollution.

Effect of pH on the Complex Coacervation and on the Formation of Layers of Sodium Alginate and PDADMAC.

In this study, we investigated the thermodynamic features of a system based on oppositely charged polyelectrolytes, sodium alginate, and poly(diallyldimethylammonium chloride) (PDADMAC) at different pH values. Additionally, a comparison of the effects of the thermodynamic parameters on the growth of the layers based on the same polymers is presented. For this investigation, different techniques were combined to compare results from the association in solution and coassembled layers at the silicon surface. Dynamic light scattering (DLS) and isothermal titration calorimetry (ITC) were used for studies in solution, and the layer-by-layer technique was employed for the preparation of the polymer layers. Ellipsometry and atomic force microscopy (AFM) were used to characterize the layer thickness growth as a function of the solution pH, and interferometric confocal microscopy was employed to analyze the topography and roughness of the films. The titration of both polyelectrolytes in two different sequences of additions confirmed the mechanism; it involved a two-step process that was monitored by varying the enthalpy, as determined by ITC experiments, and the structural evolution of the aggregates into coacervates, according to DLS. The primary process is aggregation to form polyelectrolyte complexes having a smaller hydrodynamic diameter, which abruptly transit toward a secondary process because of the formation of coacervate particles that have a larger hydrodynamic diameter. Independent of pH and the sequence of addition, for the first process, both directions are entropically driven. However, the binding enthalpy (ΔHb) decreased with a decrease in the pH of the solution. The layers grown for the PDADMAC/sodium alginate system demonstrated pH sensitivity with either linear or exponential behavior, depending on the pH values of the polyelectrolyte solutions. The more endothermic process at pH 10 afforded layers with a smaller thickness and with linear growth according to the increase in the number of layers from 5 to 20. Decreases in the pH of the solution resulted in the layers growing exponentially; additionally, a decrease in the ΔHb of the association in the solution was observed. The layer thicknesses measured using ellipsometry and AFM data were in good agreement. Additionally, the influence of pH on the roughness and topography of the films was observed. Films from basic dipping solutions resulted in surfaces that were more homogeneous with less roughness; in contrast, films with more layers and those formed in a low-pH dipping solution were rougher and less homogeneous.

Combination Therapy with Sulfasalazine and Valproic Acid Promotes Human Glioblastoma Cell Death Through Imbalance of the Intracellular Oxidative Response.

Glioblastoma (GBM) is the most common and aggressive malignant primary brain tumor and still lacks effective therapeutic strategies. It has already been shown that old drugs like sulfasalazine (SAS) and valproic acid (VPA) present antitumoral activities in glioma cell lines. SAS has also been associated with a decrease of intracellular glutathione (GSH) levels through a potent inhibition of xc- glutamate/cystine exchanger leading to an antioxidant deprotection. In the same way, VPA was recently identified as a histone deacetylase (HDAT) inhibitor capable of activating tumor suppression genes. As both drugs are widely used in clinical practice and their profile of adverse effects is well known, the aim of our study was to investigate the effects of the combined treatment with SAS and VPA in GBM cell lines. We observed that both drugs were able to reduce cell viability in a dose-dependent manner and the combined treatment potentiated these effects. Combined treatment also increased cell death and inhibited proliferation of GBM cells, while having no effect on human and rat cultured astrocytes. Also, we observed high protein expression of the catalytic subunit of xc- in all the examined GBM cell lines, and treatment with SAS blocked its activity and decreased intracellular GSH levels. Noteworthy, SAS but not VPA was also able to reduce the [14C]-ascorbate uptake. Together, these data indicate that SAS and VPA exhibit a substantial effect on GBM cell's death related to an intracellular oxidative response imbalance, making this combination of drugs a promising therapeutic strategy.

Decontamination of Mikania glomerata Leaves by Gamma Irradiation: Coumarin Determination by HPLC-DAD, Microbiological Control and Genotoxicological Studies.

Gamma irradiation as a decontaminating physical agent could be an important tool in the production chain of herbal medicines by improving the microbiological quality of raw materials and the safety of final products. This study was undertaken to investigate the genotoxic potential and eventual chemical modifications of a batch of Mikania glomerata raw material decontaminated by different doses of gamma irradiation (2.0, 3.5, and 5.0 kGy), using a cesium-137 source. DNA damage was assessed in vitro by agarose gel electrophoresis in regard to double-chain breaks of plasmid pUC 9.1 DNA and in vivo by micronucleus test in bone marrow cells of Wistar rats. Cytotoxicity in bone marrows was assessed by scoring polychromatic and normochromatic erythrocytes ratio. An HPLC-DAD method was adapted and validated for the enhancement of coumarin selectivity from the other matrix constituents. The microbial load was satisfactorily reduced, leading to sterilization at the highest dose. Genotoxic and cytotoxic effects were not increased in the in vitro and in vivo models. The concentration of coumarin and the chromatographic profiles of the hydroalcoholic plant extracts (ethanol 70% v/v) were not modified after such process. Therefore, this work suggests that gamma irradiation of M. glomerata raw material is suitable and safe for microbiological control purposes at the employed doses.

Street-Like Synthesis of Krokodil Results in the Formation of an Enlarged Cluster of Known and New Morphinans.

"Krokodil" is the street name for a homemade injectable drug that has been used as a cheap substitute for heroin. Codeine is the opioid starting material for krokodil synthesis, and desomorphine is claimed to be the main opioid of krokodil and the main component responsible for its addictive and psychoactive characteristics. However, due to its peculiar manufacture, using cheap raw materials, krokodil is composed of a large and complex mixture of different substances. In order to shed some light upon the chemical complexity of krokodil, its profiling was conducted by reverse phase high performance liquid chromatography coupled to a photodiode array detector (RP-HPLC-DAD) and by liquid chromatography coupled to high resolution tandem mass spectrometry (LC-ESI-IT-Orbitrap-MS). Besides desomorphine, codeine, and morphine, profiting from the high resolution mass spectrometry (HRMS) data, an endeavor to study the morphinans content in krokodil was set for the first time. Considering codeine as the only morphinan precursor and the possible chemical transformations that can occur during krokodil synthesis, the morphinan chemical space was designed, and 95 compounds were defined. By making use of the morphinan chemical space in krokodil, the exact masses featured by HRMS, and the morphinan mass fragmentations patterns, a targeted identification approach was designed and implemented.The proposed 95 morphinans were searched using the full scan chromatogram of krokodil, and findings were validated by mass fragmentation of the correspondent precursor ions (MS2 spectra). Following this effort, a total of 54 morphinans were detected, highlighting the fact that these additional morphinans may contribute to the psychotropic effects of krokodil.

Polycyclic aromatic hydrocarbons (PAHs) in street dust of Rio de Janeiro and Niterói, Brazil: Particle size distribution, sources and cancer risk assessment.

Fifteen polycyclic aromatic hydrocarbons (PAHs) were analyzed in two fractions (<63µm and 63-850µm) of street dust samples collected in different urban areas of Rio de Janeiro and Niterói cities, Brazil. Individual PAH concentrations and total PAH concentrations (∑PAH) were evaluated considering geographic and particle size distributions, PAH source and cancer risk. In 53% of the samples, ∑PAH values were higher in the smallest particles than in the largest ones, whereas the inverse was true for 23.5% of the samples. In 23.5% of the samples, both classes of particles showed comparable ∑PAH. The highest ∑PAH concentrations were found in areas of limited air dispersion, resulting in PAH accumulation in street dust. The diagnostic ratios and principal component analysis (PCA) indicated a strong influence of pyrogenic sources on PAH concentration, mainly of vehicular emissions. The cancer risk levels for children and adults, for both particulate size fractions (<63µm and 63µm-850µm), were comparable for dermal contact and by ingestion, and ranged from 10-8 to 10-6, whereas in certain cases the cancer risk level for adults by dermal contact was 2 to 3 times larger than for children. The cancer risk levels via inhalation always ranged from 10-13 to 10-10, so this exposure pathway was almost 104 less severe than through ingestion and dermal contact, and thus negligible.

Development, validation, and application of a method for selected avermectin determination in rural waters using high performance liquid chromatography and fluorescence detection.

Avermectins (AVM) are macrocyclic lactones used in livestock and agriculture. A quantitative method of high performance liquid chromatography with fluorescence detection for the determination of eprinomectin, abamectin, doramectin and ivermectin in rural water samples was developed and validated. The method was employed to study samples collected in the Pito Aceso River microbasin, located in the Bom Jardim municipality, Rio de Janeiro State, Brazil. Samples were extracted by solid phase extraction using a polymeric stationary phase, the eluted fraction was re-concentrated under a gentle N2 flow and derivatized to allow AVM determination using liquid chromatography with fluorescence detection. The excitation and emission wavelengths of the derivatives were 365 and 470nm, respectively, and a total chromatographic run of 12min was achieved. Very low limits of quantification (22-58ngL(-1)) were found after re-concentration using N2. Recovery values varied from 85.7% to 119.2% with standard deviations between 1.2% and 10.2%. The validated method was applied in the determination of AVM in 15 water samples collected in the Pito Aceso River microbasin, but most of them were free of AVM or showed only trace levels of these compounds, except for a sample that contained doramectin (9.11µgL(-1)). The method is suitable for routine analysis with satisfactory recovery, sensitivity, and selectivity.

The harmful chemistry behind "krokodil": Street-like synthesis and product analysis.

"Krokodil" is the street name for a drug, which has been attracting media and researchers attention due to its increasing spread and extreme toxicity. "Krokodil" is a homemade injectable mixture being used as a cheap substitute for heroin. Its use begun in Russia and Ukraine, but it is being spread throughout other countries. The starting materials for "krokodil" synthesis are tablets containing codeine, caustic soda, gasoline, hydrochloric acid, iodine from disinfectants and red phosphorus from matchboxes, all of which are easily available in a retail market or drugstores. The resulting product is a light brown liquid that is injected without previous purification. Herein, we aimed to understand the chemistry behind "krokodil" synthesis by mimicking the steps followed by people who use this drug. The successful synthesis was assessed by the presence of desomorphine and other two morphinans. An analytical gas chromatography-electron impact/mass spectrometry (GC-EI/MS) methodology for quantification of desomorphine and codeine was also developed and validated. The methodologies presented herein provide a representative synthesis of "krokodil" street samples and the application of an effective analytical methodology for desomorphine quantification, which was the major morphinan found. Further studies are required in order to find other hypothetical by-products in "krokodil" since these may help to explain signs and symptoms presented by abusers.

The harmful chemistry behind krokodil (desomorphine) synthesis and mechanisms of toxicity.

"Krokodil" is the street name for the homemade injectable mixture that has been used as a cheap substitute for heroin. Its use begun in Russia and Ukraine and nowadays is being spread over several other countries. Desomorphine is the semi-synthetic opioid claimed to be the main component of krokodil and considered to be responsible for its psychoactive characteristics. The starting materials for desomorphine synthesis are codeine tablets, alkali solutions, organic solvent, acidified water, iodine and red phosphorus, all of which are easily available in retail outlets, such as supermarkets, drugstores, etc. The resulting product is a light brown liquid that is called krokodil. People who inject krokodil present a great variety of serious signs and symptoms, including thrombophlebitis, ulcerations, gangrene, and necrosis, quickly evolving to limb amputation and death. These effects are thought to result from the toxic components produced as byproducts during the homemade drug synthesis. In this work, we reviewed several aspects of krokodil use, including its epidemiology, pharmacology and the chemical properties of the main active ingredient (desomorphine). To enhance our understanding of the clinical and toxic effects and to support the implementation of harm reduction measures, we also describe the "bathtub chemistry" of krokodil and the content of the final solution.

Characterization of newfound natural luminescent properties of melamine, and development and validation of a method of high performance liquid chromatography with fluorescence detection for its determination in kitchen plastic ware.

Experimental studies of the natural photoluminescence of melamine in aqueous solutions showed that its fluorescence intensity (at 250/365 nm) was appropriated for analytical purposes. The exploitation of such melamine property provided the basis of development of a new, simple, precise and accurate method based on high performance liquid chromatography with fluorescence detection (HPLC-Fluo) to determine melamine in kitchen plastic ware following aqueous extraction using a microwave oven. Optimization of analytical parameters such as solvent composition, pH and extraction conditions led to limits of detection and quantification of melamine of 0.0081 and 0.027 µg mL(-1), respectively, with a linear range up to 10 µg mL(-1). Sample extracts fortified with melamine at three concentration levels produced an average recovery of 98±6%, which was in agreement with the results achieved with a reference HPLC-UV method. Different samples of kitchen plastic ware analyzed by the developed and optimized method showed melamine concentrations in the aqueous extract up to 17 µg mL(-1), which corresponded to 86.0 mg kg(-1) in these utensils. The results obtained indicate that the use of kitchen plastic ware made of melamine can contaminate food with this compound after heating in a microwave oven.

Is it possible to screen for milk or whey protein adulteration with melamine, urea and ammonium sulphate, combining Kjeldahl and classical spectrophotometric methods?

The Kjeldahl method and four classic spectrophotometric methods (Biuret, Lowry, Bradford and Markwell) were applied to evaluate the protein content of samples of UHT whole milk deliberately adulterated with melamine, ammonium sulphate or urea, which can be used to defraud milk protein and whey contents. Compared with the Kjeldahl method, the response of the spectrophotometric methods was unaffected by the addition of the nitrogen compounds to milk or whey. The methods of Bradford and Markwell were most robust and did not exhibit interference subject to composition. However, the simultaneous interpretation of results obtained using these methods with those obtained using the Kjeldahl method indicated the addition of nitrogen-rich compounds to milk and/or whey. Therefore, this work suggests a combination of results of Kjeldahl and spectrophotometric methods should be used to screen for milk adulteration by these compounds.

Antioxidant activity of polyphenols from green and toasted mate tea.

The production and distribution of toasted mate tea in Brazil has increased, which has resulted in its greater consumption. Mate tea is obtained by roasting non-fermented erva-mate in order to produce toasted erva-mate or toasted mate tea. However, although the product is much appreciated, studies of its chemical composition and the concentration of polyphenols, particularly flavonols present in toasted mate tea, are few and often controversial. This paper elucidates some misunderstandings involving the nomenclature of erva-mate and toasted mate, and mainly provides an overview of the composition of polyphenols and antioxidant capacity of toasted mate tea and its raw material, erva-mate, in comparison with other teas, the compositions of which were found in the literature.

Geomicrobiology of cores from Suruí Mangrove--Guanabara Bay--Brazil.

The aim of this work was to quantify the biopolymers associated to esterase enzymes and identify bacterial respiratory activity in four cores collected in Suruí Mangrove, Guanabara Bay - RJ. Biopolymer concentration was 1000 times lower than previously reported in the literature, indicating the need for creating and establishing eutrophication indicative rates and records compatible with tropical coastal systems. The biochemical representative relationships in the cores were equivalent to those from studies on coastal marine environments made in the Northern Hemisphere. The esterase enzymes in the sediment proved efficient in the mineralization of biopolymers, even with preferentially anaerobic metabolic physiology. Despite the lack of incipient geomicrobiological studies, the results highlighted the possible application of microbiology to a better understanding of geological processes.