Paper-based isotachophoretic preconcentration technique for low-cost determination of glyphosate.

Electrophoretic microfluidic paper-based analytical devices (e-µPADs) are promising for low-cost and portable technologies, but quantitative detection remains challenging. In this study, we develop a paper-based isotachophoretic preconcentration and separation method for the herbicide glyphosate as a model analyte. The device, consisting of two electrode chambers filled with leading and terminating electrolytes and a nitrocellulose strip as the separation carrier, was illuminated by a flat light source and operated with a voltage supply of 400 V. Detection was accomplished using a simple camera. Colorimetric detection was optimized through competitive complexation between glyphosate, copper ions, and pyrocatechol violet as a dye. The buffer system was optimized using simulations, (i) ensuring the pH was optimal for the demetallation of the blue pyrocatechol violet-copper complex [PV] to the yellow free dye and (ii) ensuring the electrophoretic migration of glyphosate into the slower [PV] for the colorimetric reaction. A new data evaluation method is presented, analyzing the RGB channel intensities. The linear range was between 0.8 and 25 µM, with a LOD of approximately 0.8 µM. The ITP separation preconcentrated glyphosate by a factor of 820 in numerical simulations. The method may be applied to control glyphosate formulations, especially in developing countries where herbicide sales and applications are poorly regulated.

Effects of MCPA and difenoconazole on glyphosate degradation and soil microorganisms.

Modern agriculture relies heavily on pesticide use to meet the demands of food quality and quantity. Therefore, pesticides are often applied in mixtures, leading to a diverse cocktail of chemicals and their metabolites in soils, which can affect non-target organisms such as soil microorganisms. Pesticides are tested for their single effects, but studies on their interactive effects are scarce. This study aimed to determine the effects of up to three simultaneously applied pesticides on the soil microbial community and on their special function in pesticide degradation. Agricultural soil without previous pesticide application was exposed to different mixtures of the herbicide glyphosate (GLP), the phenoxy herbicide MCPA (2-methyl-4-chlorophenoxyacetic acid) and the fungicide difenoconazole (DFC) for up to 56 days. Isotopic and molecular methods were used to investigate effects of the mixtures on the microbial community and to follow the mineralization and utilization of GLP. An initial increase in the metabolic quotient by up to 35 % in the presence of MCPA indicated a stress reaction of the microbial community. The presence of multiple pesticides reduced both gram positive bacterial fatty acid methyl esters (FAMEs) by 13 % and the abundance of microorganisms with the genetic potential for GLP degradation via the AMPA (aminomethylphosphonic acid) pathway. Both the number of pesticides and the identities of individual pesticides played major roles. Surprisingly, an increase in 13C-labelled GLP mineralization of up to 40 % was observed while carbon use efficiency (CUE) decreased. Interactions between multiple pesticides might alter the behavior of individual pesticides and be reflected in the microbial community. Our results highlight the importance of investigating not only single pesticides, but also pesticide mixtures and their interactions.

SWIEET-a salt-free alternative to QuEChERS.

The efficient extraction of various analytes from a wide spectrum of matrices with organic solvents is still a great challenge in analytical chemistry. Especially polar and charged compounds are hard to extract in combination with neutral analytes of intermediate to low polarity. The QuEChERS method is often chosen and has been adapted not only to the analysis of food samples, but also to environmental matrices (soil, wastewater) or biota. In this study, we overcome major drawbacks of QuEChERS such as low recoveries of charged analytes and impairment of downstream analysis by high salt loads. The new extraction method, applicable to liquid and solid samples, is called SWIEET (sugar water isopropanol ethyl nitrile extraction technique). Phase separation of the otherwise miscible extraction solvents water and acetonitrile is achieved by sugaring-out instead of salting-out. Extraction efficiencies were greatly improved by adding isopropanol to the acetonitrile phase. The concentrations of the additives glucose and isopropanol, as well as temperature, were optimized by a design of experiment. Further improvement was achieved through electro- or double-extractions. For all sample types tested (surface water, wastewater treatment plant effluent, tomato, soil, and oats), recoveries and precision were higher with SWIEET than with the established QuEChERS method. From wastewater treatment plant effluent, 75% recovery on average were achieved with our SWIEET method compared to 37% with QuEChERS for a model analyte mixture with polarities of logDpH7 = - 5.7 - 3.5. Higher recoveries and lower standard deviations compared to QuEChERS were achieved especially for polar and charged analytes such as metformin. Handling proved to be easy, since there was no additional solid phase and no tedious weighing of salts.

Mechanistic modeling indicates rapid glyphosate dissipation and sorption-driven persistence of its metabolite AMPA in soil.

Residual concentrations of glyphosate and its main transformation product aminomethylphosphonic acid (AMPA) are often observed in soils. The factors controlling their biodegradation are currently not well understood. We analyzed sorption-limited biodegradation of glyphosate and AMPA in soil with a set of microcosm experiments. A mechanistic model that accounts for equilibrium and kinetic sorption facilitated interpretation of the experimental results. Both compounds showed a biphasic dissipation with an initial fast (up to Days 7-10) and subsequent slower transformation rate, pointing to sorption-limited degradation. Glyphosate transformation was well described by considering only equilibrium sorption. Model simulations suggested that only 0.02-0.13% of total glyphosate was present in the soil solution and thus bioavailable. Glyphosate transformation was rapid in solution (time required for 50 % dissipation of the total initially added chemical [DT50 ] = 3.9 min), and, despite strong equilibrium sorption, total glyphosate in soil dissipated quickly (DT50  = 2.4 d). Aminomethylphosphonic acid dissipation kinetics could only be described when considering both equilibrium and kinetic sorption. In comparison to glyphosate, the model simulations showed that a higher proportion of total AMPA was dissolved and directly bioavailable (0.27-3.32%), but biodegradation of dissolved AMPA was slower (DT50  = 1.9 h). The model-based data interpretation suggests that kinetic sorption strongly reduces AMPA bioavailability, leading to increased AMPA persistence in soil (DT50  = 12 d). Thus, strong sorption combined with rapid degradation points to low risks of glyphosate leaching by vertical transport through soil in the absence of preferential flow. Ecotoxicological effects on soil microorganisms might be reduced. In contrast, AMPA persists, rendering these risks more likely.

Structure and Dynamics of Human Chemokine CCL16-Implications for Biological Activity.

Human C-C motif ligand 16 (CCL16) is a chemokine that is distinguished by a large cleavable C-terminal extension of unknown significance. Conflicting data have been reported concerning its tissue distribution and modulation of expression, rendering the biological function of CCL16 enigmatic. Here, we report an integrated approach to the characterisation of this chemokine, including a re-assessment of its expression characteristics as well as a biophysical investigation with respect to its structure and dynamics. Our data indicate that CCL16 is chiefly synthesised by hepatocytes, without an appreciable response to mediators of inflammation, and circulates in the blood as a full-length protein. While the crystal structure of CCL16 confirms the presence of a canonical chemokine domain, molecular dynamics simulations support the view that the C-terminal extension impairs the accessibility of the glycosaminoglycan binding sites and may thus serve as an intrinsic modulator of biological activity.

Picomolar detection limits for glyphosate by two-dimensional column-coupled isotachophoresis/capillary zone electrophoresis-mass spectrometry.

Capillary electrophoresis-mass spectrometry often lacks sufficient limits of detection for trace substances in the environment due to its low loadability. To overcome this problem, we conducted a feasibility study for column-coupling isotachophoresis to capillary electrophoresis-mass spectrometry. The first dimension isotachophoresis preconcentrated the analytes. The column-coupling of both dimensions was achieved by a hybrid capillary microfluidic chip setup. Reliable analyte transfer by voltage switching was enabled by an in-chip capacitively coupled contactless conductivity detector placed around the channel of the common section between two T-shaped crossings in the chip connecting both dimensions. This eliminated the need to calculate the moment of analyte transfer. A commercial capillary electrophoresis-mass spectrometry instrument with easily installable adaptations operated the setup. Prior to coupling isotachophoresis with capillary zone electrophoresis-mass spectrometry, both dimensions were optimized individually by simulations and verified experimentally. Both dimensions were able to stack/separate all degradation products of glyphosate, the most important herbicide applied worldwide. The first dimension isotachophoresis also removed phosphate, which is a critical matrix component in many environmental samples. Enrichment and separation of glyphosate and its main degradation product aminomethylphosphonic acid by the two-dimensional setup provided an excellent limit of detection of 150 pM (25 ng/L) for glyphosate.

The wastewater micropollutant carbamazepine in insectivorous birds-an exposure estimate.

Insects with aquatic life stages can transfer sediment and water pollutants to terrestrial ecosystems, which has been described for metals, polyaromatic hydrocarbons, and polychlorinated chemicals. However, knowledge of the transfer of aquatic micropollutants released by wastewater treatment plants is scarce despite some preliminary studies on their occurrence in riparian spiders. In our study, we address a major analytical gap focusing on the transfer of the micropollutant carbamazepine from the larvae to the adult midges of Chironomus riparius using an optimized QuEChERS extraction method and HPLC-MS/MS applicable to both life stages down to the level of about three individuals. We show that the uptake of carbamazepine by larvae is concentration-dependent and reduces the emergence rate. Importantly, the body burden remained constant in adult midges. Using this information, we estimated the daily exposure of insectivorous tree swallows as terrestrial predators to carbamazepine using the energy demand of the predator and the energy content of the prey. Assuming environmentally relevant water concentrations of about 1 µg/L, the daily dose per kilogram of body weight for tree swallows was estimated to be 0.5 µg/kg/day. At places of high water contamination of 10 µg/L, the exposure may reach 5 µg/kg/day for this micropollutant of medium polarity. Considering body burden changes upon metamorphosis, this study fills the missing link between aquatic contamination and exposure in terrestrial habitats showing that wastewater pollutants can impact birds' life. Clearly, further analytical methods for biota analysis in both habitats are urgently required to improve risk assessment.

13C assimilation as well as functional gene abundance and expression elucidate the biodegradation of glyphosate in a field experiment.

Glyphosate (N-phosphonomethylglycine; GLP) and its main metabolite AMPA (aminomethylphosphonic acid), are frequently detected in relatively high concentrations in European agricultural topsoils. Glyphosate has a high sorption affinity, yet it can be detected occasionally in groundwater. We hypothesized that shrinkage cracks occurring after dry periods could facilitate GLP transport to greater depths where subsoil conditions slow further microbial degradation. To test this hypothesis, we simulated a heavy rainfall event (HRE) on a clay-rich arable soil. We applied 2.1 kg ha-1 of 100% 13C3, 15N-labeled GLP one day before the simulated rainfall event. Microbial degradation of translocated GLP over a 21-day period was assessed by quantifying 13C incorporation into phospholipid fatty acids. Microbial degradation potential and activity were determined by quantifying the abundance and expression of functional genes involved in the two known degradation pathways of GLP; to AMPA (goxA) or sarcosine (sarc). We confirmed that goxA transcripts were elevated in the range of 4.23 x 105 copy numbers g-1 soil only one day after application. The increase in AMPA associated with a rise in goxA transcripts and goxA-harboring microorganisms indicated that the degradation pathway to AMPA dominated. Based on 13C-enrichment 3 h after the HRE, fungi appeared to initiate glyphosate degradation. At later time points, Gram+-bacteria proved to be the main degraders due to their higher 13C-incorporation. Once GLP reached the subsoil, degradation continued but more slowly. By comparing GLP distribution and its microbial degradation in macropores and in the bulk soil, we demonstrated different time- and depth-dependent GLP degradation dynamics in macropores. This indicates the need for field studies in which soil properties relevant to GLP degradation are related to limiting environmental conditions, providing a realistic assessment of GLP fate in soils.

Heavy rainfall following a summer drought stimulates soil redox dynamics and facilitates rapid and deep translocation of glyphosate in floodplain soils.

We present field data on the effects of heavy rainfall after drought on the mobility of glyphosate and redox conditions in a clayey floodplain soil. By applying glyphosate together with deuterated water as conservative tracer in combination with time resolved in situ redox potential measurements, the spatial and temporal patterns of water infiltration and pesticide transport as well as the concomitant changes of the redox conditions were revealed. Our findings demonstrate that shrinkage cracks in dry soils can serve as effective transport paths for atmospheric oxygen, water and glyphosate. The rain intensity of a typical summer storm event (approx. 25 mm within one hour) was sufficient to translocate deuterated water and glyphosate to the subsoil (50 cm) within 2 hours. Soil wetting induced partial closure of the shrinkage cracks and stimulated microbial activity resulting in pronounced dynamics of in situ soil redox conditions. Redox potentials in 40 to 50 cm depth dropped permanently to strongly reducing conditions within hours to days but fluctuated between reducing and oxidizing conditions in 10 to 30 cm depth. Our findings highlight the close link between the presence of macropores (shrinkage cracks), heavy rainfall after drought, redox dynamics and pesticide translocation to the subsoil and thus call for further studies addressing the effects of dynamic redox conditions as a limiting factor for glyphosate degradation.

Phosphate addition enhances alkaline extraction of glyphosate from highly sorptive soils and aquatic sediments.

BACKGROUND: Analytical constraints complicate environmental monitoring campaigns of the herbicide glyphosate and its major degradation product aminomethylphosphonic acid (AMPA): their strong sorption to soil minerals requires harsh extraction conditions. Coextracted matrix compounds impair downstream analysis and must be removed before analysis. RESULTS: A new extraction method combined with subsequent capillary electrophoresis-mass spectrometry for derivatization-free analysis of glyphosate and AMPA in soil and sediment was developed and applied to a suite of environmental samples. It was compared to three extraction methods from literature. We show that no extraction medium reaches 100% recovery. The new phosphate-supported alkaline extraction method revealed (1) high recoveries of 70-90% for soils and aquatic sediments, (2) limits of detections below 20 µg kg-1 , and (3) a high robustness, because impairing matrix components (trivalent cations and humic acids) were precipitated prior to the analysis. Soil and sediment samples collected around Tübingen, Germany, revealed maximum glyphosate and AMPA residues of 80 and 2100 µg kg-1 , respectively, with residues observed along a core of lake sediments. Glyphosate and/or AMPA were found in 40% of arable soils and 57% of aquatic sediment samples. CONCLUSION: In this work, we discuss soil parameters that influence (de)sorption and thus extraction. From our results we conclude that residues of glyphosate in environmental samples are easily underestimated. With its possible high throughput, the method presented here can resolve current limitations in monitoring campaigns of glyphosate by addressing soil and aquatic sediment samples with critical sorption characteristics.

Capillary electrophoresis applied for the determination of acidity constants and limiting electrophoretic mobilities of ionizable herbicides including glyphosate and its metabolites and for their simultaneous separation.

Thermodynamic acidity constants and limiting ionic mobilities were determined for polyprotic non-chromophore analytes using capillary electrophoresis with capacitively coupled contactless conductivity detection. It was not necessary to work with buffers of identical ionic strength as ionic strength effects on effective electrophoretic mobilities were corrected by modeling during data evaluation (software AnglerFish). The mobility data from capillary electrophoresis coupled to conductivity detection were determined in the pH range from 1.25 to 12.02 with a high resolution (36 pH steps). With this strategy, thermodynamic acidity constants and limiting ionic mobilities for various acidic herbicides were determined, sometimes for the first time. The model analytes included glyphosate, its metabolites, and its acetylated derivates (aminomethyl phosphonic acid, glyoxylic acid, sarcosine, glycine, N-acetyl glyphosate, N-acetyl aminomethyl phosphonic acid, hydroxymethyl phosphonic acid). The obtained data were used in simulations to optimize separations by capillary electrophoresis. Simulations correlated very well to experimental results. With the new method, the separation of glyphosate from interfering components like phosphate in beer samples was possible.

Performance of free-flow field-step electrophoresis as cleanup step for the non-target analysis of environmental water samples.

For the analysis of low concentrations of micropollutants in environmental water samples, efficient sample enrichment and cleanup are necessary to reduce matrix effects and to reach low detection limits. For analytes of low and medium polarity, solid-phase extraction is used, but robust methods for the preconcentration of highly polar or ionizable analytes are scarce. In this work, field-step electrophoresis (FSE) was developed as an environmental sample cleanup technique for ionizable micropollutants and ionic transformation products. The FSE electrolyte system preconcentrated 15 acidic model analytes (pKa from -2.2 to 9.1) present in aqueous samples in two fractions by factors of 5-10. Simultaneously, highly mobile matrix compounds were removed including inorganic ions such as sulfate and chloride. The fractions were either directly injected for downstream analysis by reversed-phase liquid chromatography (RPLC) or further processed by evaporative preconcentration with subsequent reconstitution in an organic solvent suitable for separation methods like hydrophilic interaction chromatography. The FSE/RPLC-MS method exhibited high quantitative precision with RSDs of 3-6%. The method was successfully applied to a spiked river water sample and its performance compared with common solid-phase extraction and evaporative concentration, demonstrating a high analyte coverage. FSE combined with non-target screening by RPLC-MS revealed a strong reduction in matrix load especially at low retention times. Seventeen compounds were identified in the FSE fractions sampled at the field step boundary by retention time, accurate mass, and mass fragments. Suspect screening by FSE/RPLC-MS was facilitated by FSE's selectivity for anionic compounds.

Advancements in capacitance-to-digital converter-based C4 D technology for detection in capillary electrophoresis using amplified excitation voltages and comparison to classical and open-source C4 Ds.

This work introduces new hardware configurations for a capacitively coupled contactless conductivity detector (C4 D) based on capacitance-to-digital conversion (CDC) technology for CE. The aim was to improve sensitivity, handling, price, and portability of CDC-based C4 D detectors (CDCD) to reach LODs similar to classic C4 Ds with more sophisticated electric circuits. To achieve this, a systematic study on the CDCDs was carried out including a direct comparison to already established C4 D setups. Instrumental setups differing in electrode lengths, measurement modes, and amplification of excitation voltages were investigated to achieve LODs for alkali metal ions of 4 to 12 µM, similar to LODs obtained by classic C4 D setups. Lowest LODs were achieved for a setup with two 10 mm electrodes at a distance of 0.2 mm and an excitation voltage of 24 V. The detection head was exceptionally lightweight with only 2.6 g and covered only 20 mm of the capillary on total. This allowed the use of multiple detectors along the separation path to enable spatial tracking of analytes during separation. The entirely battery-powered detector assembly weighs less than 200 g, and the data are transmitted wirelessly for possible portable applications. The freely accessible hardware and software were optimized for fully automated measurements with real time data plotting and allowed handling multidetector setups. The new developments were applied to quantify the potassium salt of glyphosate in its herbicide formulation.

Do environmentally relevant concentrations of fluoxetine and citalopram impair stress-related behavior in zebrafish (Danio rerio) embryos?

Selective serotonin reuptake inhibitors (SSRIs) have been shown to interfere with various physiological functions of aquatic organisms, yet the neuroactive potential of low concentrations of SSRIs in the aquatic environment is unclear. The current study investigated the effects of fluoxetine and citalopram on the visual motor response (VMR) of 107 h old zebrafish (Danio rerio) embryos. Results document a reduction in stress-related swimming activity of zebrafish embryos at environmentally relevant concentration levels, with fluoxetine being more effective than citalopram. Further experiments were designed to elucidate (1) if the lower neuroactive potential of citalopram is due to differences in uptake kinetics, (2) if the metabolite of fluoxetine, norfluoxetine, contributes to the neuroactive potential of fluoxetine, (3) and how SSRIs and their metabolites interact in equimolar mixtures. At the stage of 120 h, zebrafish embryos accumulate citalopram at significantly lower rates (up to 127 times) than fluoxetine. Moreover, it was demonstrated that norfluoxetine reduces the embryonic VMR similarly to fluoxetine resulting in additive effects of these substances on stress-related behavior in zebrafish embryos. In contrast, the interaction of fluoxetine, norfluoxetine and citalopram varied with test concentrations of the equimolar mixtures. Findings provide evidence that environmentally relevant concentrations of fluoxetine reduce stress-related behavior of zebrafish embryos, while these effects may be enhanced by the interaction of multiple SSRIs and their metabolites in environmental exposure scenarios.

Trends in sample preparation and separation methods for the analysis of very polar and ionic compounds in environmental water and biota samples.

Recent years showed a boost in knowledge about the presence and fate of micropollutants in the environment. Instrumental and methodological developments mainly in liquid chromatography coupled to mass spectrometry hold a large share in this success story. These techniques soon complemented gas chromatography and enabled the analysis of more polar compounds including pesticides but also household chemicals, food additives, and pharmaceuticals often present as traces in surface waters. In parallel, sample preparation techniques evolved to extract and enrich these compounds from biota and water samples. This review article looks at very polar and ionic compounds using the criterion log P ≤ 1. Considering about 240 compounds, we show that (simulated) log D values are often even lower than the corresponding log P values due to ionization of the compounds at our reference pH of 7.4. High polarity and charge are still challenging characteristics in the analysis of micropollutants and these compounds are hardly covered in current monitoring strategies of water samples. The situation is even more challenging in biota analysis given the large number of matrix constituents with similar properties. Currently, a large number of sample preparation and separation approaches are developed to meet the challenges of the analysis of very polar and ionic compounds. In addition to reviewing them, we discuss some trends: for sample preparation, preconcentration and purification efforts by SPE will continue, possibly using upcoming mixed-mode stationary phases and mixed beds in order to increase comprehensiveness in monitoring applications. For biota analysis, miniaturization and parallelization are aspects of future research. For ionic or ionizable compounds, we see electromembrane extraction as a method of choice with a high potential to increase throughput by automation. For separation, predominantly coupled to mass spectrometry, hydrophilic interaction liquid chromatography applications will increase as the polarity range ideally complements reversed phase liquid chromatography, and instrumentation and expertise are available in most laboratories. Two-dimensional applications have not yet reached maturity in liquid-phase separations to be applied in higher throughput. Possibly, the development and commercial availability of mixed-mode stationary phases make 2D applications obsolete in semi-targeted applications. An interesting alternative will enter routine analysis soon: supercritical fluid chromatography demonstrated an impressive analyte coverage but also the possibility to tailor selectivity for targeted approaches. For ionic and ionizable micropollutants, ion chromatography and capillary electrophoresis are amenable but may be used only for specialized applications such as the analysis of halogenated acids when aspects like desalting and preconcentration are solved and the key advantages are fully elaborated by further research. Graphical abstract.

Development of a capillary electrophoresis-mass spectrometry method for the analysis of metformin and its transformation product guanylurea in biota.

A method with capillary electrophoresis coupled to mass spectrometry was optimized to determine the uptake of metformin and its metabolite guanylurea by zebrafish (Danio rerio) embryos and brown trout (Salmo trutta f. fario) exposed under laboratory conditions. Metformin was extracted from fish tissues by sonication in methanol, resulting in an absolute recovery of almost 90%. For the extraction of guanylurea from brown trout, solid-phase extraction was implemented with a recovery of 84%. The use of a mixture of methanol and glacial acetic acid as a non-aqueous background electrolyte was vital to achieve robust analysis using a bare fused-silica capillary with an applied voltage of +30 kV. Problems with adsorption associated with an aqueous background electrolyte were eliminated using a non-aqueous background electrolyte made of methanol/acetic acid (97:3) with 25 mM ammonium acetate (for zebrafish embryos) or 100 mM ammonium acetate (for brown trouts), depending on the sample complexity and matrix influences. High resolution and high separation selectivity from matrix components were achieved by optimization of the ammonium acetate concentration in the background electrolyte. An extensive evaluation of matrix effects was conducted with regard to the complex matrices present in the fish samples. They required adapting the background electrolyte to higher concentrations. Applying this method to extracts of zebrafish embryos and brown trout tissue samples, limits of detection for both metformin and guanylurea in zebrafish embryos (12.2 µg/l and 15 µg/l) and brown trout tissues (15 ng/g and 34 ng/g) were in the low µg/l or ng/g range. Finally, metformin and guanylurea could be both quantified for the first time in biota samples from exposure experiments.

Capillary electrophoresis-mass spectrometry for the direct analysis of glyphosate: method development and application to beer beverages and environmental studies.

In this study, we developed and validated a CE-TOF-MS method for the quantification of glyphosate (N-(phosphonomethyl)glycine) and its major degradation product aminomethylphosphonic acid (AMPA) in different samples including beer, media from toxicological analysis with Daphnia magna, and sorption experiments. Using a background electrolyte (BGE) of very low pH, where glyphosate is still negatively charged but many matrix components become neutral or protonated, a very high separation selectivity was reached. The presence of inorganic salts in the sample was advantageous with regard to preconcentration via transient isotachophoresis. The advantages of our new method are the following: no derivatization is needed, high separation selectivity and thus matrix tolerance, speed of analysis, limits of detection suitable for many applications in food and environmental science, negligible disturbance by metal chelation. LODs for glyphosate were < 5 µg/L for both aqueous and beer samples, the linear range in aqueous samples was 5-3000 µg/L, for beer samples 10-3000 µg/L. For AMPA, LODs were 3.3 and 30.6 µg/L, and the linear range 10-3000 µg/L and 50-3000 µg/L, for aqueous and beer samples, respectively. Recoveries in beer samples for glyphosate were 94.3-110.7% and for AMPA 80.2-100.4%. We analyzed 12 German and 2 Danish beer samples. Quantification of glyphosate and AMPA was possible using isotopically labeled standards without enrichment, purification, or dilution, only degassing and filtration were required for sample preparation. Finally, we demonstrate the applicability of the method for other strong acids, relevant in food and environmental sciences such as N-acetyl glyphosate, N-acetyl AMPA (present in some glyphosate resistant crop), trifluoroacetic acid, 2-methyl-4-chlorophenoxyacetic acid, glufosinate and its degradation product 3-(methylphosphinico)propionic acid, oxamic acid, and others.

Impact of the antidepressant citalopram on the behaviour of two different life stages of brown trout.

BACKGROUND: Over the last two decades, there has been a constant increase in prescription rates of antidepressants. In parallel, neuroactive pharmaceuticals are making their way into aquatic environments at increasing concentrations. Among the antidepressants detected in the environment citalopram, a selective serotonin reuptake inhibitor, is one of the most commonly found. Given citalopram is specifically designed to alter mood and behaviour in humans, there is growing concern it can adversely affect the behaviour on non-target wildlife. METHODS: In our study, brown trout were exposed to citalopram (nominal concentrations: 1, 10, 100, 1000 µg/L) in two different life stages. Larvae were exposed at 7 and 11 °C from the eyed ova stage until 8 weeks post yolk sac consumption, and juvenile brown trout were exposed for 4 weeks at 7 °C. At both stages we measured mortality, weight, length, tissue citalopram concentration, behaviour during exposure and behaviour in a stressfull environment. For brown trout larvae additionally hatching rate and heart rate, and for juvenile brown trout the tissue cortisol concentration were assessed. RESULTS: During the exposure, both larvae and juvenile fish exposed to the highest test concentration of citalopram (1 mg/L) had higher swimming activity and spent longer in the upper part of the aquaria compared to control fish, which is an indicator for decreased anxiety. Most probably due to the higher swimming activity during the exposure, the juveniles and larvae exposed to 1 mg/L citalopram showed decreased weight and length. Additionally, in a stressful artificial swimming measurement device, brown trout larvae displayed the anxiolytic effect of the antidepressant by reduced swimming activity during this stress situation, already at concentrations of 100 µg/L citalopram. Chemical analysis of the tissue revealed rising citalopram tissue concentrations with rising exposure concentrations. Tissue concentrations were 10 times higher in juvenile fish compared to brown trout larvae. Fish plasma concentrations were calculated, which exceeded human therapeutic levels for the highest exposure concentration, matching the behavioural results. Developmental parameters like hatching rate and heart rate, as well as mortality and tissue cortisol content were unaffected by the antidepressant. Overall, we could trace the pharmacological mode of action of the antidepressant citalopram in the non-target organism brown trout in two different life stages.

Challenges and applications of isotachophoresis coupled to mass spectrometry: A review.

Electrophoretic separations are of growing interest to tackle complex analytical challenges. Nevertheless, capillary electrophoresis, as the most common mode, still suffers from insufficient detection limits due to low capillary loadability. ITP is of growing interest as preconcentration method for capillary electrophoresis and is also interesting to be applied as an independent analytical method. While mass spectrometric detection is common for capillary electrophoresis, the combination of ITP with MS is still a niche technique. In this work, we want to give an overview on isotachophoretic effects in CE-MS and ITP-MS methods, as well as coupling techniques of ITP with CE-MS. The challenges and possibilities associated with mass spectrometric detection in ITP and its coupling to capillary electrophoresis are critically discussed.

Novel approach for the synthesis of a neutral and covalently bound capillary coating for capillary electrophoresis-mass spectrometry made from highly polar and pH-persistent N-acryloylamido ethoxyethanol.

Statically adsorbed or covalently coupled capillary coatings are of crucial importance in capillary electrophoresis-mass spectrometry for the separation of peptides and proteins. So far, published coating strategies and commercially available coated capillaries have a limited pH-stability so that the analysis at strongly acidic pH is limited, or harsh rinsing procedures for biological sample analysis cannot be applied. We here present a capillary coating based on Si-C linkages to N-acryloylamido ethoxyethanol (AAEE) with a new synthetic strategy including LiAlH4 surface reaction. We optimized the coating method with emphasis on stability and reproducibility applying harsh rinsing procedures (strong acid, strong base and organic solvent), using the electroosmotic mobility and separation efficiency of tryptic peptides as performance measure. Complete synthesis is performed in less than 2 days for up to 8 capillaries in parallel of more than 16 m total length. Intra- and inter-batch reproducibility were determined regarding electroosmotic mobility, separation efficiency and migration time precision in CE-MS separations of tryptically digested bovine serum albumin. Coating stability towards rinsing with strong acid (1 mol/L HCl), organic solvent (acetonitrile) and strong base (1 mol/L NaOH) was investigated. Outstanding performance was found for single capillaries. However, inter-capillary reproducibility is discussed critically. The new coating was successfully applied for reproducible CE-MS separation of large proteins in diluted serum, medium-sized peptides and small and highly charged polyamines in fish egg extracts using a very acidic background electrolyte containing 0.75 mol/L acetic acid and 0.25 mol/L formic acid (pH 2.2).