A comprehensive risk management on human exposure to cyanotoxins, whose production is actually unpredictable, is limited by reliable analytical tools for monitoring as many toxic algal metabolites as ...possible. Two analytical approaches based on a LC-QTOF system for target analysis and suspect screening of cyanotoxins in freshwater were presented. A database with 369 compounds belonging to cyanobacterial metabolites was developed and used for a retrospective data analysis based on high resolution mass spectrometry (HRMS). HRMS fragmentation of the suspect cyanotoxin precursor ions was subsequently performed for correctly identifying the specific variants. Alternatively, an automatic tandem HRMS analysis tailored for cyanotoxins was performed in a single chromatographic run, using the developed database as a preferred precursor ions list. Twenty-five extracts of surface and drinking waters contaminated by cyanobacteria were processed. The identification of seven uncommon microcystins (M(O)R, MC-FR, MSer7-YR, D-Asp3MSer7-LR, MSer7-LR, dmAdda-LR and dmAdda-YR) and 6 anabaenopeptins (A, B, F, MM850, MM864, oscyllamide Y) was reported. Several isobaric variants, fully separated by chromatography, were pointed out. The developed methods are proposed to be used by environmental and health agencies for strengthening the surveillance monitoring of cyanotoxins in water.
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•Rapid structural screening and quantitation of target cyanotoxins.•First database developed for identifying 369 algal metabolites.•Possibility to perform automatic analysis of cyanotoxins in various matrices.•Uncommon variants of cyanotoxins evidenced in freshwaters by LC tandem HRMS.
•Ecotoxicological impact of bisphenol a analogues.•BPA analogues-treated cells show chloroplast ultrastructural alterations.•Bisphenol a analogues induce ROS production and lipid ...accumulation.•Microalgae bioaccumulate bisphenol a analogues.
As well-known, microalgae have a pivotal role in aquatic environments, being the primary producer. In this study, we investigated the effects of Bisphenol A (BPA) analogues on cell ultrastructure, reactive oxygen species (ROS) production and photosynthetic pigment responses in the diatom Phaeodactylum tricornutum. Microalgae were exposed during both exponential and stationary growth phases to an environmental relevant concentration (300 ng/L) of three differing BPA analogues (BPAF, BPF, and BPS) and their mixture (100 ng/L of each compound). Bioaccumulation of such compounds in microalgae was also analysed. During the stationary growth phase, a significant increase in the percentage of cells with hydrogen peroxide production was recorded after exposure to both BPS and MIX. Conversely, no significant effects on total chlorophylls and carotenoids were observed. During exponential growth phase we observed that control cultures had chloroplasts with well-organized thylakoid membranes and a central pyrenoid. On the contrary, the culture cells treated with BPA analogues and MIX showed chloroplasts characterized by evident dilation of thylakoid membranes. The presence of degeneration areas in the cytoplasm was also recorded. During the stationary growth phase, control and culture cells were characterized by chloroplasts with a regular thylakoid system, whereas BPA analogues-exposed cells were characterized by a deep degradation of the cytoplasm but showed chloroplasts without evident alterations of the thylakoid system. Lipid bodies were visible in treated microalgae. Lastly, microalgae bioaccumulated mainly BPS and BPF, alone or in the MIX. Overall, results obtained revealed that BPA analogues can affect some important biochemical and ultrastructure features of microalgae, promoting ROS production. Lastly, the capability of microalgae to bioaccumulate bisphenols suggest a potential ecotoxicological risk for filter-feeders organisms.
EDTA and soluble Cr(III) are usually both present in wastewaters coming from treatment plants handling tannery effluents. A well-established method to determine EDTA is based on the conversion of ...free and complexed EDTA into its Fe(III) complex. This procedure gives inconsistent data when Cr(III)-EDTA is present. This fact was here demonstrated by studying the kinetics of the exchange reaction between Fe(III) and Cr(III)-EDTA at 90 °C and various pH values, from acidic to neutral. The reaction is very slow (several weeks); the slow kinetics of conversion of Cr(III)-EDTA to Fe(III)-EDTA is even more accentuated at room temperature and the low concentrations of reactants in wastewaters. The presence of EDTA complexes of Fe(III) and Cr(III) was demonstrated in industrial effluents and wastewaters by developing a selective method based on liquid chromatography-tandem mass spectrometry (LC-MS/MS), which was able to detect free and complexed EDTA at concentration levels <1 μM. A systematic underestimation of the EDTA expressed as Fe(III) complex was demonstrated in samples containing Cr(III)-EDTA. Cr(III)-EDTA was identified for the first time as a component of wastewater samples at a concentration level of about 2 μM and turned out to be an inert species that significantly contributes to the final soluble Cr amount. This study gives new insights into the inertness of Cr(III) toward metal exchange equilibria of EDTA complexes, resolves a bias in the analysis of total EDTA in samples containing Cr(III)-EDTA, allowing the direct determination of free and complexed EDTA by LC-MS.
•Cr(III)-EDTA identified for the first time with a direct measurement in wastewaters.•New HPLC-MS procedure to determine free and complexed EDTA in wastewaters.•Elimination of a systematic bias in the determination of EDTA.•Rationalization of the reaction kinetics of a Cr-EDTA dissociation in water.
Background and Purpose
Gut microbiota is essential for the development of the gastrointestinal system, including the enteric nervous system (ENS). Perturbations of gut microbiota in early life have ...the potential to alter neurodevelopment leading to functional bowel disorders later in life. We examined the hypothesis that gut dysbiosis impairs the structural and functional integrity of the ENS, leading to gut dysmotility in juvenile mice.
Experimental Approach
To induce gut dysbiosis, broad‐spectrum antibiotics were administered by gavage to juvenile (3weeks old) male C57Bl/6 mice for 14 days. Bile acid composition in the intestinal lumen was analysed by liquid chromatography‐mass spectrometry. Changes in intestinal motility were evaluated by stool frequency, transit of a fluorescent‐labelled marker and isometric muscle responses of ileal full‐thickness preparations to receptor and non‐receptor‐mediated stimuli. Alterations in ENS integrity were assessed by immunohistochemistry and Western blot analysis.
Key Results
Antibiotic treatment altered gastrointestinal transit, luminal bile acid metabolism and bowel architecture. Gut dysbiosis resulted in distorted glial network, loss of myenteric plexus neurons, altered cholinergic, tachykininergic and nitrergic neurotransmission associated with reduced number of nNOS neurons and different ileal distribution of the toll‐like receptor TLR2. Functional defects were partly reversed by activation of TLR2 signalling.
Conclusions and Implications
Gut dysbiosis caused complex morpho‐functional neuromuscular rearrangements, characterized by structural defects of the ENS and increased tachykininergic neurotransmission. Altogether, our findings support the beneficial role of enteric microbiota for ENS homeostasis instrumental in ensuring proper gut neuromuscular function during critical stages of development.
The seawater pH measurement is usually quite complicated because that matrix is characterized by a high ionic strength leading to calibration errors if NIST standards are used. For this matrix, ...different pH scales like the “total hydrogen ion concentration scale” (TOT) and the “seawater scale” (SWS), are defined, and suitable synthetic seawater solutions must be prepared according to standard procedures to calibrate the glass electrode. This work provides a new approach to make seawater pH measurements by using the glass electrode calibrated with the NIST standards (pHNIST) converting the pHNIST into the right TOT or SWS scales by using empirical equations derived from theoretical thermodynamic data: pHTOT=pHNIST+0.10383+4.33⋅10−5TS+3.633⋅10−5T2−4.921⋅10−5S2, and pHSWS=pHNIST+0.097733+4.1059⋅10−5TS+3.5437⋅10−5T2−4.941⋅10−5S2, for the TOT and SWS scales, respectively. These equations are functions of two simple experimental parameters, namely, T = temperature (°C) and S = salinity (PSU, (g/L), Practical Salinity Units). These equations were experimentally validated and the uncertainty of pHTOT and pHSWS was demonstrated to have no statistical difference with the corresponding values obtained following the standard operative procedure (SOP) using commercially unavailable seawater-like buffers. The proposed method has therefore the same performances and it is largely preferable as it avoids long and tedious procedures of the synthetic seawater preparations.
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•Easy seawater pH measurement by glass electrode with conventional NIST calibrations.•The use of NIST standards avoids the preparation of the seawater standard solutions.•An easy empirical model for the accurate measurement of the seawater pH.•The computed ΔpH values (between pHTOT and pHNIST) matched the experimental ones.•The overall uncertainty of the pHTOT and the pHSWSvalues is 0.009–0.016 pH units.
New dinuclear di(N-heterocyclic carbene) silver(I), gold(I) and gold(III) complexes have been synthesised and their antiproliferative effects towards various cancer cell lines have been screened. The ...di(N-heterocyclic carbene) ligands have a propylene linker between the carbene moieties and the imidazole backbone has been functionalised with a 1-benzyl- or 1-PEG-1,2,3-triazole ring (PEG=poly(ethylene glycol)) via a CuAAC (copper azido alkyne cycloaddition) reaction. The resulting gold(I) and gold(III) complexes display an antiproliferative activity superior to that of the unfunctionalised pristine complexes together with a higher selectivity towards cancerous cells with respect to healthy cells.
The dinuclear di(N-PEGylated-heterocyclic carbene) gold(I) and gold(III) complexes display a high antiproliferative efficiency, both in terms of activity and selectivity, towards various cancerous cell lines. Display omitted
•New dinuclear diNHC Ag(I), Au(I, III) complexes have been synthesised.•Ligand precursors were functionalised by click chemistry (CuAAC).•Functionalised Au complexes display high antiproliferative activity and selectivity.•The Au(III) complex quickly reacts with GSH to afford Au(I) complex and GSSG.
A highly selective and robust method for simultaneous screening and confirmation of target and non-target phosphodiesterase type 5 (PDE-5) inhibitor analogues within a single chromatographic run in ...counterfeit herbal products was developed. The protocol, based on an easy and rapid extraction with a water/acetonitrile 1 % formic acid solution, followed by sonication and centrifugation, exploits an LC-diode array detector-quadrupole-time-of-flight (DAD-QTOF) system. The extraction method was optimized both at high concentrations and at trace levels. These two situations are typically encountered in pharmaceutical formulations and herbal food supplements. Carryover effects, never reported before and occurring mainly for vardenafil, were overcome using a polymer-based column. An in-house validation was carried out using five blanks of different bulk matrices spiked with seven standard analytes, namely yohimbine, sildenafil, vardenafil, tadalafil, homosildenafil, pseudovardenafil and hydroxyhomovardenafil. Reliable quantitation was possible using a conventional standard solution for all the pharmaceutical and herbal samples considered, as matrix effects were eliminated. Accuracy ranged from 80.9 to 108.1 % with overall relative standard deviation (RSD) <11 % (N = 15), measured at 1.0, 5.0 and 10.0 μg/g. Limits of detection (LODs) obtained ensured the determination of cross contaminations at nanogram per gram levels. A database with 82 PDE-5 inhibitor analogues was implemented for automatic non-target analysis. Among the 26 samples of dietary supplements and herbal remedies bulk marketed for erectile dysfunctions, three samples were found to be contaminated with both registered and unregistered synthetic PDE-5 inhibitors, i.e. yohimbine, sildenafil, dimethylsildenafil and thiodimethylsildenafil or thiomethisosildenafil. The occurrence of such contaminations, both at trace levels and at pharmaceutical dosage, indicates the illicit use of synthetic PDE-5 analogues.
High resolution, high accuracy mass spectrometry is widely used to characterise environmental or biological samples with highly complex composition enabling the identification of chemical composition ...of often unknown compounds. Despite instrumental advancements, the accurate molecular assignment of compounds acquired in high resolution mass spectra remains time consuming and requires automated algorithms, especially for samples covering a wide mass range and large numbers of compounds. A new processing scheme is introduced implementing filtering methods based on element assignment, instrumental error, and blank subtraction. Optional post-processing incorporates common ion selection across replicate measurements and shoulder ion removal. The scheme allows both positive and negative direct infusion electrospray ionisation (ESI) and atmospheric pressure photoionisation (APPI) acquisition with the same programs. An example application to atmospheric organic aerosol samples using an Orbitrap mass spectrometer is reported for both ionisation techniques resulting in final spectra with 0.8% and 8.4% of the peaks retained from the raw spectra for APPI positive and ESI negative acquisition, respectively.
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•Ultra-high resolution mass spectrometry processing scheme from acquisition to analysis.•Method implementable with APPI and ESI ionisation in both polarities.•Example application for environmental samples showing >90% peak filtering.
The effects of the breakdown products of herbicides on aquatic species are largely unknown. In a recent study, we evaluated the effects of glyphosate on the mussel Mytilus galloprovincialis. This ...study was designed to evaluate for the first time the impact of aminomethylphosphonic acid (AMPA) - the main breakdown product of glyphosate - on cellular and biochemical parameters of the mussel Mytilus galloprovincialis. Bivalves were exposed for 7, 14 and 21 days to 1, 10 and 100 μg/L of AMPA and various biomarkers were measured in haemolymph (total haemocyte counts, haemocyte diameter and volume, haemolymph pH, haemocyte proliferation, haemolymph lactate dehydrogenase activity, haemocyte lysate lysozyme and acid phosphatase activities), as well as in gills and digestive gland (superoxide dismutase, catalase, glutathione S-transferase and acetylcholinesterase activities). AMPA concentrations in seawater samples from the experimental tanks were also measured in order to correlate the biomarker responses of mussels with their exposure to the actual concentrations of AMPA. The MANOVA analysis demonstrated that the experimental variables considered (exposure dose, exposure duration, and their interaction) affected significantly biomarker responses. Nevertheless, the two-way ANOVA analysis revealed significant effects of AMPA on most of the biomarkers measured. The overall results of this study demonstrated that AMPA can affect cellular and biochemical parameters in mussels, similarly to glyphosate.
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•Mussels were exposed to various AMPA concentrations.•AMPA affected mostly haemocyte parameters.•AMPA did not affect significantly antioxidant enzymes.•AMPA generally increased AChE activity in gills.
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