Occurrence and toxicological relevance of pesticides and trace metals in agricultural soils, sediments, and water of the Sogamoso River basin, Colombia.

Historical pesticide use in agriculture and trace metal accumulation have long term impact on soil, sediment, and water quality. This research quantifies legacy and current-use pesticides and trace metals, assessing their occurrence and toxicological implications on a watershed scale in the Sogamoso River basin, tributary of the Magdalena River in Colombia. Organochlorine pesticides (22), organophosphates (7), and azole fungicides (5), as well as trace metals cadmium (Cd), chromium (Cr), copper (Cu), mercury (Hg), nickel (Ni), lead (Pb), and zinc (Zn) were analyzed in croplands and along the river. Toxic units (TU) and hazard quotients (HQ) were calculated to assess the mixture toxicity. Organochlorines were detected in 84% of soils, 100% of sediments, and 80% of water samples. Organophosphates were found in 100% of soil and sediment samples, as well as in 70% of water samples. Azole fungicides were present in 79% of soils, 60% of sediments, and in 10% of water samples. Total pesticide concentrations ranged from 214.2 to 8497.7 µg/kg in soils, 569.6-12768.2 µg/kg in sediments, and 0.2-4.1 µg/L in water. In addition, the use of partition coefficient (Kd) and organic carbon fraction (foc) allowed the distribution analysis for most of the pesticides in sediments, suspended particulate matter (SPM), and water systems, but not for soils. Concentrations of trace metals Cu, Zn, Pb, and Zn exceeded international quality guidelines for agricultural soils in 16% of the samples. Furthermore, Cu and Zn concentrations exceeded sediment quality guidelines in 50 and 90% of the samples, respectively. These findings demonstrate the broad distribution of complex mixtures of trace metals, legacy organochlorines, and current-use pesticides across the basin, indicating that conventional agriculture is a significant source of diffuse pollution. Sustainable agricultural practices are needed to mitigate adverse impacts on ecosystems and human health.

Microbial Contamination in the Coffee Industry: An Occupational Menace besides a Food Safety Concern?

Respiratory abnormalities among workers at coffee roasting and packaging facilities have already been reported; however, little is known about microbiological contamination inside coffee production facilities. This study intends to assess the microbial contamination (fungi and bacteria) in two coffee industries from Brazil with a multi-approach protocol for sampling and for subsequent analyses using four main sources of samples: filtering respiratory protection devices (FRPD) used by workers, settled dust, electrostatic dust cloths (EDC) and coffee beans. The fungal contamination in the assessed industries was also characterized through the molecular detection of toxigenic species and antifungal resistance. Total bacteria contamination presented the highest values in FRPD collected from both industries (7.45 × 104 CFU·m-2; 1.09 × 104 CFU·m-2). Aspergillus genera was widespread in all the environmental samples collected and sections with clinical relevance (Fumigati) and with toxigenic potential (Nigri and Circumdati) were recovered from FRPD. Circumdati section was observed in 4 mg/mL itraconazole. Sections Circumdati (EDC, coffee beans and settled dust) and Nidulantes (EDC, coffee beans and FRPD) were detected by qPCR. Some of the targeted Aspergillus sections that have been identified microscopically were not detected by qPCR and vice-versa. Overall, this study revealed that microbial contamination is a potential occupational risk in the milling stage and should be tackled when assessing exposure and performing risk assessment. In addition, a multi-sampling campaign should be the approach to follow when assessing microbial contamination and FRPD should be included in this campaign. Occupational exposure to mycotoxins should be considered due to high fungal diversity and contamination. A One Health approach should address these issues in order to prevent consumption of coffee crops and beans infected by fungi and, more specifically, to avoid widespread azole resistance.

Adsorption behavior of azole fungicides on polystyrene and polyethylene microplastics.

Agricultural plastic films and triazole fungicides are widely used in agricultural production process. Exposure to natural environment, agricultural plastic films will degrade into micron plastic particles, which will adsorb pesticide molecules and may affect their toxicity, biological activity and persistence. The long-term coexistence of microplastics (MPs) and triazole fungicides will bring potential harms to the agricultural ecological environment. Therefore, two kinds of triazole fungicides flusilazole (FLU) and epoxiconazole (EPO) were selected as cases and the adsorption behaviors of them on polystyrene and polyethylene were investigated. A series of factors which could affect the adsorption behavior were evaluated. Specifically, the particle size of MPs could affect its adsorption capacity, and the smaller the particle size, the stronger the adsorption capacity. Moreover, with the increase of pH value from 6.0 to 9.0, the adsorption capacity of MPs to target compounds gradually increased. The effect of ionic strength was evaluated by NaCl, and 0.05% of NaCl was beneficial to the adsorption process, while the continuous increase of NaCl concentration inhibited the adsorption. Oxalic acid and humic acid decreased the adsorption capacity of flusilazole on PE by 15.99-32.00% and PS by 35.02-48.67%, respectively. In addition, compared with the single pesticide system, the adsorption capacity of MPs for flusilazole and epoxiconazole in the binary pesticides system decreased by 36.13-37.93% and 44.36-51.35%, respectively, indicating that competitive adsorption occurred between the two pesticides. Meanwhile, the adsorption process was evaluated by adsorption kinetics and adsorption isotherms and were consistent with pseudo-second-order kinetic model and Freundlich isotherm model, respectively. Finally, several characterization analyses were conducted to investigated the adsorption mechanism, and hydrogen, halogen bonding and hydrophobic interaction proved to play an important role. The study on the adsorption behavior and mechanism of pesticide on MPs was the basis of assessing the risk of joint exposure.

Construction of a magnetic solid-phase extraction method for the analysis of azole pesticides residue in medicinal plants.

In this work, a sensitive and cost-effective method for the quantitative analysis of azole pesticides residues in six medicinal plants was established based on magnetic cyclodextrin crosslinked with tetrafluoroterephthalonitrile (Fe3O4@TFN-CDPs) coupled with high-performance liquid chromatography (HPLC). Through characterization analysis, the outer shell of Fe3O4@TFN-CDPs has observed coating with a network of the polymer and forming a core-shell structure. Under the optimum conditions, the limits of detection (LODs) and limits of qualification (LOQs) of target pesticides were ranged from 0.011 to 0.106 µg Kg-1 and from 0.036 to 0.354 µg Kg-1, respectively. Finally, the achieved recoveries of pesticides in six medicinal samples fluctuated from 60.1% to 102.3%. Altogether, this method based on Fe3O4@TFN-CDPs composites provided a new idea for the analysis of trace pesticides in complicated matrices.

Biodegradation of typical azole fungicides in activated sludge under aerobic conditions.

Widespread use of azole fungicides and low removal efficiency in wastewater treatment plants (WWTPs) have led to the elevated concentration of azole fungicides in receiving environment. However, there was limited research about the removal mechanism of azole fungicides in the biological treatment of WWTPs. Imidazole fungicide climbazole and triazole fungicide fluconazole were selected to investigate the biodegradation mechanism of azole fungicides in activated sludge under aerobic conditions. Climbazole was found to be adsorbed to solid sludge and resulted in quick biodegradation. The degradation of climbazole in the aerobic activated sludge system was fitted well by the first-order kinetic model with a half-life of 5.3 days, while fluconazole tended to stay in liquid and had only about 30% of loss within 77 days incubation. Ten biotransformation products of climbazole were identified by high resolution mass spectrometry using suspect and non-target screening method. But no biodegradation products of fluconazole were identified due to its limited removal. The possible biodegradation pathways for climbazole were proposed based on the products identification and pathway prediction system, and involves oxidative dehalogenation, side chain oxidation and azole ring loss. The findings from this study suggest that it should be a concern for the persistence of fluconazole in the environment.

A comprehensive review on environmental toxicity of azole compounds to fish.

BACKGROUND: Azoles are considered as one of the most efficient fungicides for the treatment of humans, animals, and plant fungal pathogens. They are of significant clinical importance as antifungal drugs and are widely used in personal care products, ultraviolet stabilizers, and in aircraft for its anti-corrosive properties. The prevalence of azole compounds in the natural environment and its accumulation in fish raises questions about its impact on aquatic organisms. OBJECTIVES: The objective of this paper is to review the scientific studies on the effects of azole compounds in fish and to discuss future opportunities for the risk evaluation. METHODS: A systematic literature search was conducted on Web of Science, PubMed, and ScienceDirect to locate peer-reviewed scientific articles on occurrence, environmental fate, and toxicological impact of azole fungicides on fish. RESULTS: Studies included in this review provide ample evidence that azole compounds are not only commonly detected in the natural environment but also cause several detrimental effects on fish. Future studies with environmentally relevant concentrations of azole alone or in combination with other commonly occurring contaminants in a multigenerational study could provide a better understanding. CONCLUSION: Based on current knowledge and studies reporting adverse biological effects of azole on fish, considerable attention is required for better management and effective ecological risk assessment of these emerging contaminants.

An oxindole efflux inhibitor potentiates azoles and impairs virulence in the fungal pathogen Candida auris.

Candida auris is an emerging fungal pathogen that exhibits resistance to multiple drugs, including the most commonly prescribed antifungal, fluconazole. Here, we use a combinatorial screening approach to identify a bis-benzodioxolylindolinone (azoffluxin) that synergizes with fluconazole against C. auris. Azoffluxin enhances fluconazole activity through the inhibition of efflux pump Cdr1, thus increasing intracellular fluconazole levels. This activity is conserved across most C. auris clades, with the exception of clade III. Azoffluxin also inhibits efflux in highly azole-resistant strains of Candida albicans, another human fungal pathogen, increasing their susceptibility to fluconazole. Furthermore, azoffluxin enhances fluconazole activity in mice infected with C. auris, reducing fungal burden. Our findings suggest that pharmacologically targeting Cdr1 in combination with azoles may be an effective strategy to control infection caused by azole-resistant isolates of C. auris.

Application of capillary electrophoresis technique for the enantioseparation of bioactive ferrocene-based compounds versus DFT calculated data.

Herein, a series of bioactive ferrocene-modified N-heterocycles with alkyl linkers was prepared in good to quantitative yields starting from easy accessible ferrocene alcohols and heterocycles under acidic or neutral (for imidazole) conditions in racemic forms. The analytical resolution of a number of bioactive racemic ferrocene azoles 1-6 (where azole = imidazole, pyrazole, and benzotriazole derivatives) into enantiomers was first carried out by CE using sulfobuthylether-ß-CD (captisol) as a chiral selector. The analytical approaches to highly enantiomeric-enriched ferrocene derivatives are based on the formation of their inclusion complexes. The best chiral separation was achieved using zone CE in a quartz capillary. The ACE was used to evaluate the stability constants of captisol complexes with enantiomeric forms of two ferrocene derivatives 1, FcCHMe-imidazole, and 6, FcCHMe-benzotriazole. The optimal conditions for the resolution of the studied (R, S)-ferrocene compounds 1, 2, and 6 were predicted on the basis of the performed quantum chemical calculations and then implemented by the electrophoretic method. A high correlation between density functional theory calculation results and experimental electrophoresis data were obtained. Successful enantioseparation of racemic mixtures is of great importance for the characterization and further applications of drug candidates in enantiopure forms and in the development of clinical treatment. The advantages of the CE procedure make it possible to have important practical value and significance for determining the purity and enantiomeric excess of other ferrocene-containing compounds.

Occurrence and risk assessment of azole antifungal drugs in water and wastewater.

The occurrence of azole antifungals in the environment presents one of the emerging concerns due to their ecotoxicological threat as well as their potential contribution to the evolution of drug resistant fungi in the environment. In this study, the occurrence of eight commonly prescribed azole antifungal drugs was seasonally determined in influent and effluent water samples from three wastewater treatment plants and a drinking water treatment plant in South Africa. In addition, the risk quotient (RQ) method was employed to investigate the potential ecological and human health risks associated with their presence in the wastewater and/or drinking water. Clotrimazole, econazole, fluconazole, itraconazole, ketoconazole and miconazole were detected at least once in the water samples, while posaconazole and voriconazole were not detected in any of the samples for all seasons at which the samples were collected. Fluconazole was detected at higher frequency (about 96%) with a concentration up to 9959.0 ng L-1. Clotrimazole had the second highest frequency of detection (about 33%) with a concentration up to 143.3 ng L-1. Statistically significant temporal variation in clotrimazole (p < 0.05) and spatial variation in fluconazole (p < 0.05) were observed. In general, the preliminary ecological risk assessment based on risk quotient (RQ) calculation indicated that there is currently no high risk against aquatic organisms (Algae, Daphnia and Fish) related to the azole antifungals. Meanwhile, human health risk assessment demonstrated that fluconazole represented high risk in drinking water. Furthermore, risk estimates showed a potential for the detected concentrations of fluconazole and itraconazole in water samples to pose moderate to high risk for development of antifungal drug resistance. Some of the azole antifungal drugs are ubiquitous in the wastewater and future monitoring and validation studies should be conducted for those drugs that seem to pose human health and ecological risks.

Determination of the contamination by azole antimycotics in hospital and house sewage - a pilot project for the city of Dresden.

Medicininal compounds and their metabolites are known to end up in sewerage and may slip through the cleaning process. Azole antimycotics are frequently used in hospitals, in particular for patients with cancer or immunosuppression. The aim of the study was to determine whether measurable azole antimycotic concentrations were introducted in the sewarage drain of an acute care hospital with special interest in oncology and hematology and the extent of removal of antimycotics by the sewerage treatment plant. For this, the concentrations of three commonly used azole antimycotics were measured in the effluent of the sewerage drain at the University Hospital Dresden, as well as in the influent and effluent of the main sewerage treatment plant of the city. To extrapolate the theoretical influent to the sewerage treatment plant, prescription from the regio`s main health insurance the AOK Sachsen and the hospital consumption data were used. Measurable concentrations were obtained for fluconazole and ketoconazole in the influent and effluent of the sewerage treatment plant. Voriconazole's concentrations were under the lower limit of quantification. To determine the azole clearance of the treatment plant a sludge sample was investigated. Sufficient clearance was detected for ketoconazole but not for fluconazole. The consumption and prescription rates were collected and correlated with the measured concentrations. In result, only fluconazole's concentrations provided a good match with the prescription and consumption data.

Construction of a hydrazone-linked chiral covalent organic framework-silica composite as the stationary phase for high performance liquid chromatography.

Covalent organic frameworks (COFs), as an emerging class of crystalline porous organic polymers, have great potential for applications in chromatographic separation owning to their fascinating crystalline structures and outstanding properties. However, development of COF materials as novel stationary phases in high performance liquid chromatography (HPLC) is just in its infancy. Herein, we report the design and construction of a new hydrazone-linked chiral COF, termed BtaMth COF, from a chiral hydrazide building block (Mth) and present a one-pot synthetic method for the fabrication of BtaMth@SiO2 composite for HPLC separation of isomers. The as-synthesized BtaMth chiral COF displays good crystallinity, high porosity, as well as excellent chemical stability. Meanwhile, the fabricated HPLC column by using BtaMth@SiO2 composite as the new stationary phase exhibits high resolution performances for the separation of positional isomers including nitrotoluene and nitrochlorobenzene, as well as cis-trans isomers including beta-cypermethrin and metconazole. Additionally, some effects such as the composition of the mobile phase and column temperature for HPLC separations on the BtaMth@SiO2 packed column also have been studied in detail. The successful applications indicate the great potentials of hydrazone-linked chiral COF-silica composite as novel stationary phase for the efficient HPLC separation.

YcaO-Dependent Posttranslational Amide Activation: Biosynthesis, Structure, and Function.

With advances in sequencing technology, uncharacterized proteins and domains of unknown function (DUFs) are rapidly accumulating in sequence databases and offer an opportunity to discover new protein chemistry and reaction mechanisms. The focus of this review, the formerly enigmatic YcaO superfamily (DUF181), has been found to catalyze a unique phosphorylation of a ribosomal peptide backbone amide upon attack by different nucleophiles. Established nucleophiles are the side chains of Cys, Ser, and Thr which gives rise to azoline/azole biosynthesis in ribosomally synthesized and posttranslationally modified peptide (RiPP) natural products. However, much remains unknown about the potential for YcaO proteins to collaborate with other nucleophiles. Recent work suggests potential in forming thioamides, macroamidines, and possibly additional post-translational modifications. This review covers all knowledge through mid-2016 regarding the biosynthetic gene clusters (BGCs), natural products, functions, mechanisms, and applications of YcaO proteins and outlines likely future research directions for this protein superfamily.

Azole-resistant Aspergillus fumigatus harboring TR34/L98H, TR46/Y121F/T289A and TR53 mutations related to flower fields in Colombia.

Resistance to triazoles in Aspergillus fumigatus has been reported in azole-naive patients in Europe, Asia, Australia and North America. This resistance has been linked to fungicide-driven mutations in the cyp51A gene and its promoter region. We investigated the presence of environmental azole-resistant A. fumigatus strains related to the use of azole fungicides in Colombia. Soil samples were collected from flower beds, flower fields and public gardens from the outskirts, suburbs and city centre of Bogotá. Out of the 86 soil samples taken, 17 (19.8%) grew A. fumigatus of whom eight (9.3%) contained 40 strains able to grow on azole-containing itraconazole and/or voriconazole supplemented media. All but one triazole-resistant strains were isolated from soil samples collected from flower fields and flower beds (39/40). Importantly, the majority had the TR46/Y121F/T289A, TR34/L98H, and TR53 molecular resistance mechanisms and one azole resistant strain had a wild-type cyp51A gene. Soil samples from flower fields and beds contained 4 azole fungicides (penconazole, difenoconazole, tetraconazole and tebuconazole) above the limit of detection. Our findings underline the need for extensive investigations to determine azole-resistant A. fumigatus prevalence in both clinical and environmental samples in other regions of Latin America.

Biomolecular Interaction Assays Identified Dual Inhibitors of Glutaminase and Glutamate Dehydrogenase That Disrupt Mitochondrial Function and Prevent Growth of Cancer Cells.

Glutaminase (KGA/isoenzyme GAC) is an emerging and important drug target for cancer. Traditional methods for assaying glutaminase activity are coupled with several other enzymes. Such coupled assays do not permit the direct and stringent characterization of specific glutaminase inhibitors. Ebselen was identified as a potent 9 nM KGA inhibitor in the KGA/glutamate oxidase (GO)/horse radish peroxidase (HRP) coupled assay but showed very weak activity in inhibiting the growth of glutamine-dependent cancer cells. For rigorous characterization, we developed a direct kinetic binding assay for KGA using bio-layer interferometry (BLI) as the detection method; Ebselen was identified as a GDH inhibitor but not a KGA inhibitor. Furthermore, we designed and synthesized several benzo[d][1,2]selenazol-3(2H)-one dimers which were subjected to SAR analysis by several glutaminolysis specific biochemical and cell based assays. Novel glutamate dehydrogenase (GDH) or dual KGA/GDH inhibitors were discovered from the synthetic compounds; the dual inhibitors completely disrupt mitochondrial function and demonstrate potent anticancer activity with a minimum level of toxicity.

Analysis of azole fungicides in fish muscle tissues: Multi-factor optimization and application to environmental samples.

Azole fungicides have been reported to be accumulated in fish tissue. In this study, a sensitive and robust method using high-performance liquid chromatography-tandem mass spectrometry combined with ultrasonic extraction, solid-liquid clean-up, liquid-liquid extraction and solid-phase extraction (SPE) for enrichment and purification have been proposed for determination of azole fungicides in fish muscle samples. According to the results of non-statistical analysis and statistical analysis, ethyl acetate, primary secondary amine (PSA) and mixed-mode cation exchange cartridge (MCX) were confirmed as the best extraction solvent, clean-up sorbent and SPE cartridge, respectively. The satisfied recoveries (81.7-104%) and matrix effects (-6.34-7.16%), both corrected by internal standards, were performed in various species of fish muscle matrices. Method quantification limits of all azoles were in the range of 0.07-2.83ng/g. This optimized method was successfully applied for determination of the target analytes in muscle samples of field fish from Beijiang River and its tributaries. Three azole fungicides including climbazole, clotrimazole and carbendazim were detected at ppb levels in fish muscle tissues. Therefore, this analytical method is practical and suitable for further clarifying the contamination profiles of azole fungicides in wild fish species.

Estandarización y validación en Colombia de una metodología basada en HPLC para la determinación de la concentración sérica de posaconazol / Standardisation and validation of an HPLC method for determining serum posaconazole levels in Colombia

Antecedentes. Hasta la fecha, Colombia no cuenta con un servicio especializado de medición de concentraciones séricas de antifúngicos, procedimiento esencial para el adecuado manejo del tratamiento de las infecciones invasivas por hongos. Objetivos. Estandarizar y validar un protocolo simple, sensible y específico, basado en la cromotografía líquida de alta eficiencia, que, cumpliendo con los parámetros recomendados por la Food and Drug Administration, permita detectar y cuantificar concentraciones séricas de posaconazol. Métodos. Se usó un equipo de cromotografía líquida de alta eficiencia Agilent, serie 1200, con detector ultravioleta de matriz de diodos y columna analítica Eclipse-XDB-C18. Como estándar primario se utilizó posaconazol SCH56592 (lote IRQ-PAZ-10-X-103), y como control interno, itraconazol (lote ZR051211PUC921). La validación se hizo teniendo en cuenta los parámetros recomendados por la Food and Drug Administration (selectividad, curvas de calibración, recuperación, exactitud, precisión, sensibilidad, reproducibilidad y estabilidad de la muestra) para este tipo de métodos. Resultados. Los parámetros cromatográficos más adecuados fueron los siguientes: temperatura de la columna, 25°C; detección ultravioleta, 261nm; volumen de inyección, 50μl; flujo, 0,8ml/min; tiempo de migración, 10min; fase móvil, acetonitrilo:agua (70:30). Los tiempos de retención finales fueron de 3,4 y 7,2min para posaconazol e itraconazol, respectivamente, con un rango de cuantificación amplio y confiable, desde 0,125 hasta 16μg/ml. Bajo estas condiciones el método fue selectivo, el R2 de las curvas de calibración fue≥0,99 y el porcentaje de recuperación fue del 98,7%, con un porcentaje del coeficiente de variación inferior al 10%. El porcentaje de error relativo en la exactitud, así como el porcentaje del coeficiente de variación en la precisión, fueron inferiores al 15%, cumpliendo así con los criterios de aceptación recomendados por la Food and Drug Administration. Conclusiones. La selectividad y pureza de la señal cromatográfica obtenida, así como los límites de detección y cuantificación estandarizados, hacen de esta metodología una excelente herramienta para el seguimiento terapéutico de los pacientes bajo tratamiento con posaconazol (AU)

Background. Colombia currently does not have a specialised service for measuring antifungal levels in serum, which is of prime importance for the proper treatment and correct management of invasive fungal infections. Aims. To standardise and validate a simple, sensitive, and specific protocol, based on high performance liquid chromatography, complying with the parameters recommended by the Food and Drug Administration, to detect, identify, and quantify serum concentrations of posaconazole. Methods. A high performance liquid chromatography Agilent series-1 200 equipment was used with ultraviolet diode array detector and analytical column-Eclipse XDB-C18. Posaconazole-SCH56592 (batch IRQ-PAZ-10-X-103) was used as the primary control and itraconazole (batch ZR051211PUC921) was used as an internal control. The validation was performed taking into account all criteria recommended by the Food and Drug Administration (selectivity, calibration curves, recovery, accuracy, precision, sensitivity, reproducibility, and stability of the sample). Results. The most suitable chromatographic conditions were the following: column temperature 25°C, ultraviolet detection at 261nm, 50μl injection volume, flow volume 0.8ml/min, 10min running time, mobile phase of acetonitrile:water (70:30), and final retention times of 3.4 and 7.2min for posaconazole and itraconazole, respectively, with a wide and reliable quantification range (0.125μg/ml to 16μg/ml). Using these parameters, the method was selective, R2 in the calibration curves was≥0.99, and the percentage recovery was 98.7%, with a coefficient of variation less than 10%. The relative error for accuracy and the coefficient of variation for precision were less than 15%, all meeting the acceptance criteria recommended by the Food and Drug Administration. Conclusions. The selectivity and chromatographic purity of the obtained signal, as well as the standardised limits of detection and quantification, make this method an excellent tool for therapeutic monitoring of patients treated with posaconazole (AU)

Identification and determination of chlorinated azoles in sludge using liquid chromatography quadrupole time-of-flight and triple quadrupole mass spectrometry platforms.

Four antimycotic drugs (tioconazole, TCZ; sertaconazole, STZ; fenticonazole, FTZ and itraconazole, ITZ) and the fungicide imazalil (IMZ) are determined in sludge from sewage treatment plants (STPs) following a bottom-up analytical strategy. First, sludge extracts, obtained under different sample preparation conditions, were analyzed by liquid chromatography (LC) quadrupole time-of-flight (QTOF) mass spectrometry (MS). A non-target search strategy, combined with the use of the chlorine mass filter, permitted to detect several chlorinated pollutants including the above referred azoles, which either had not been previously reported (TCZ, STZ, FTZ and ITZ), or scarcely investigated (IMZ), in this environmental compartment. Then, the sample preparation procedure was validated using standards of these compounds and their sensitive and selective determination was performed by LC-MS/MS, based on a QqQ system. Under final working conditions, quantitative extraction yields were attained with negligible changes in ionization efficiencies between sample extracts and standards; therefore, the above compounds were quantified against authentic standard solutions, with absolute recoveries in the range from 75 to 124%, achieving a limit of quantification of 2ngg-1. Analysis of sludge from 10 municipal STPs demonstrated the ubiquity of the identified chlorinated azoles with average concentrations from 31ngg-1, for IMZ, to more than 200ngg-1, for ITZ.

Measuring internal azole and pyrethroid pesticide concentrations in Daphnia magna using QuEChERS and GC-ECD--method development with a focus on matrix effects.

Pyrethroids are highly toxic towards aquatic macroinvertebrates such as Daphnia magna and can be synergized when co-occurring with azole fungicides. A sensitive analytical method for the measurement of azole-pyrethroid mixtures in aquatic macroinvertebrates is not available at present. We developed and validated an extraction, cleanup, and quantification procedure for four pyrethroid insecticides and four azole fungicides at the picograms per milligram wet weight level in D. magna using a QuEChERS approach and GC-ECD analysis. Short- and long-term matrix effects were analyzed by injection of a series of extracts from D. magna, and the best surrogate standards were identified through correlation analysis of analyte responses. The presence of matrix clearly stabilized the analyte responses (≤6% relative standard deviation of peak area compared to up to 22% when injected without matrix). The sensitivity was high with detection limits and limits of quantification between 58-168 and 119-571 pg mg(wet weight)(-1) for the azoles and 5.8-27 and 12-84 pg mg(wet weight)(-1) for the pyrethroids, respectively. Accuracy (% recovery) was between 95 and 111% and the precision (repeatability) below 10% relative standard deviation for all analytes. In the case of prochloraz, α-cypermethrin, and deltamethrin, normalization to surrogate standards led to a clear improvement of accuracy and precision by up to 8 and 4%, respectively. The method was successfully applied to the measurement of internal α-cypermethrin concentrations in D. magna under environmentally relevant exposure conditions (exposure to a pulse in the micrograms per liter range) with and without co-exposure to propiconazole.

Occurrence, fate and ecological risk of five typical azole fungicides as therapeutic and personal care products in the environment: A review.

Azole fungicides are widely used to treat fungal infection in human. After application, these chemicals may reach to the receiving environment via direct or indirect discharge of wastewaters, thus posing potential risks to non-target organisms. We aimed to review the occurrence, fate and toxicological effects of some representative household azole fungicides in the environment. Azole fungicides were widely detected in surface water and sediment of the aquatic environment due to their incomplete removal in wastewater treatment plants. These chemicals are found resistant to microbial degradation, but can undergo photolysis under UV irradiation. Due to different physiochemical properties, azole fungicides showed different environmental behaviors. The residues of azole fungicides could cause toxic effects on aquatic organisms such as algae and fish. The reported effects include regulation changes in expression of cytochrome P450-related genes and alteration in CYP450-regulated steroidogenesis causing endocrine disruption in fish. Further studies are essential to investigate the removal of azole fungicides by advanced treatment technologies, environmental fate such as natural photolysis, and toxic pathways in aquatic organisms.

Chemometrical tools in the study of the retention behavior of azole antifungals.

Certain chemometrical tools allow an efficient way to provide valuable data to evaluate the retention behavior of analytes in liquid chromatography. In this study of the retention behavior of azole antifungals, the experimental design was applied in combination with artificial neural networks (ANNs). Three potentially significant factors (methanol content, pH of the mobile phase and column temperature) were incorporated in the plan of experiments, defined by central composite design. As the system outputs, the retention factors of all six investigated substances (fluconazole, ketoconazole, bifonazole, clotrimazole, econazole and miconazole) were determined. The pattern for the analyzed behavior of the system was created by employing ANNs. The final, optimized topology of the highly predictive network was 3-8-6. Twelve experiments were used in a training set, whereas a back-propagation algorithm was optimal for network training. The ability of the defined network to predict the retention of the investigated azoles was confirmed by correlations higher than 0.9912 for all analytes. The presented approach allowed the adequate prediction of the retention behavior of azoles, in addition to the extraction of important information for a better understanding of the analyzed system.