Cumulative Neutral Loss Model for Fragment Deconvolution in Electrospray Ionization High-Resolution Mass Spectrometry Data.

Clean high-resolution mass spectra (HRMS) are essential to a successful structural elucidation of an unknown feature during nontarget analysis (NTA) workflows. This is a crucial step, particularly for the spectra generated during data-independent acquisition or during direct infusion experiments. The most commonly available tools only take advantage of the time domain for spectral cleanup. Here, we present an algorithm that combines the time domain and mass domain information to perform spectral deconvolution. The algorithm employs a probability-based cumulative neutral loss (CNL) model for fragment deconvolution. The optimized model, with a mass tolerance of 0.005 Da and a scoreCNL threshold of 0.00, was able to achieve a true positive rate (TPr) of 95.0%, a false discovery rate (FDr) of 20.6%, and a reduction rate of 35.4%. Additionally, the CNL model was extensively tested on real samples containing predominantly pesticides at different concentration levels and with matrix effects. Overall, the model was able to obtain a TPr above 88.8% with FD rates between 33 and 79% and reduction rates between 9 and 45%. Finally, the CNL model was compared with the retention time difference method and peak shape correlation analysis, showing that a combination of correlation analysis and the CNL model was the most effective for fragment deconvolution, obtaining a TPr of 84.7%, an FDr of 54.4%, and a reduction rate of 51.0%.

Reducing the influence of geometry-induced gradient deformation in liquid chromatographic retention modelling.

Rapid optimization of gradient liquid chromatographic (LC) separations often utilizes analyte retention modelling to predict retention times as function of eluent composition. However, due to the dwell volume and technical imperfections, the actual gradient may deviate from the set gradient in a fashion unique to the employed instrument. This makes accurate retention modelling for gradient LC challenging, in particular when very fast separations are pursued. Although gradient deformation has been addressed in method-transfer situations, it is rarely taken into account when reporting analyte retention parameters obtained from gradient LC data, hampering the comparison of data from various sources. In this study, a response-function-based algorithm was developed to determine analyte retention parameters corrected for geometry-induced deformations by specific LC instruments. Out of a number of mathematical distributions investigated as response-functions, the so-called "stable function" was found to describe the formed gradient most accurately. The four parameters describing the model resemble the statistical moments of the distribution and are related to chromatographic parameters, such as dwell volume and flow rate. The instrument-specific response function can then be used to predict the actual shape of any other gradient programmed on that instrument. To incorporate the predicted gradient in the retention modelling of the analytes, the model was extended to facilitate an unlimited number of linear gradient steps to solve the equations numerically. The significance and impact of distinct gradient deformation for fast gradients was demonstrated using three different LC instruments. As a proof of principle, the algorithm and retention parameters obtained on a specific instrument were used to predict the retention times on different instruments. The relative error in the predicted retention times went down from an average of 9.8% and 12.2% on the two other instruments when using only a dwell-volume correction to 2.1% and 6.5%, respectively, when using the proposed algorithm. The corrected retention parameters are less dependent on geometry-induced instrument effects.

Abiotic and biotic transformation of torasemide - Occurrence of degradation products in the aquatic environment.

The pharmaceutical torasemide is an important loop diuretic and was 2017 one of the ten most prescribed drugs in Germany. Despite its detection in different compartments of the urban water cycle including drinking water, no studies were so far performed to elucidate its fate in the environment and the occurrence of transformation products (TPs). Therefore, we investigated the phototransformation, microbial degradation, transformation with human liver microsomes and anodic oxidation of torasemide to obtain good coverage of environmentally relevant degradation products. Overall sixteen products were identified, covering the following reaction mechanisms: aromatic and aliphatic hydroxylation, including further oxidation to carboxylic acids and quinone imines, amide cleavage, N-dealkylation, N-dearylation, and sulfonamide hydrolysis to sulfonic acids. Especially the formation of quinone imines could be of concern as they are highly reactive electrophiles. Torasemide itself was observed in all investigated wastewater treatment plant (WWTP) samples and wastewater-impacted surface waters. The maximum detected concentration was about 350 ng L-1. Only three of the sixteen transformation products were generally observed in at least one of the samples and the most frequently detected TPs were the human metabolites hydroxytorasemide (TP 364a) and carboxytorasemide (TP 378a). The complete removal of TP 364a during wastewater treatment was in agreement with the results of microbial degradation experiments. TP 364a was most likely transformed into TP 378a, which was microbially less degraded in lab experiments. Based on estimated concentrations, TP 378a could reach about 1 µg L-1 in the investigated wastewater matrices.

Identification of transformation products of denatonium - Occurrence in wastewater treatment plants and surface waters.

Denatonium, one of the bitterest substances known to man, was recently identified as wastewater borne micropollutant in surface waters. Therefore, photodegradation experiments and electrochemical degradation were performed to identify abiotic and putative biotic transformation products (TPs). Indirect rather than direct photodegradation proved to be important for denatonium removal by solar irradiation and produced seven TPs. Amide hydrolysis, hydroxylation, N-dealkylation, and N-dearylation were revealed as the main mechanisms. Anodic oxidation of denatonium was related to the formation of overall ten products and despite considerable different yields, all TPs from indirect photodegradation were mimicked electrochemically. Among them, lidocaine was the only TP detected after conventional wastewater treatment and in surface waters. The occurrence of lidocaine was however associated with its application as local anesthetic rather than to a degradation of denatonium. The absence of additional products suggests that denatonium degradation is negligible under environmental conditions, supporting the previously described persistent nature of this compound. Advanced water treatment techniques however have the potential to degrade denatonium. About 74% of the initial denatonium load was removed from wastewater during pilot-scale ozonation. The degradation of denatonium was accompanied here with the formation of at least two polar products, which are passing unchanged through a sand filter after ozonation. Both substances have completely unknown (toxicological) properties and this study seems to be the first report about their structures in general, as none of them was found in any of the large compound libraries (e.g. PubChem).

Denatonium - A so far unrecognized but ubiquitous water contaminant?

Denatonium is one of the bitterest substances known to man and therefore applied in numerous consumer products to prevent an accidental or intentional consumption. So far no information was available on the occurrence of this compound in the environment. A sensitive targeted method was developed and applied to water samples taken in the federal state of Baden-Württemberg, Germany. Denatonium was detected in 100% of the investigated 22 wastewater treatment plant (WWTP) effluents with a maximum concentration of 341 ng L-1. Additionally, water samples were taken from the Ammer river over a period of one week and all wastewater impacted samples showed denatonium at concentrations up to 195 ng L-1. Retrospective analysis of high-resolution mass spectrometric measurements of WWTP effluents from Italy and Switzerland confirmed and therefore point to an international occurrence of denatonium as anthropogenic contaminant.

Assessment of N-Oxide Formation during Wastewater Ozonation.

Worldwide, ozonation of secondary wastewater effluents is increasingly considered in order to decrease the load of organic contaminants before environmental discharge. However, despite the constantly growing knowledge of ozonation over the past few years, the characterization of transformation products (TPs) is still a major concern, particularly because such TPs might remain biologically active. It has been shown for selected tertiary amine pharmaceuticals that they react with ozone and form the corresponding N-oxides. This study therefore applies liquid chromatography-high resolution mass spectrometry (LC-HRMS) to assess the overall N-oxide formation during the pilot-scale ozonation of a secondary wastewater effluent from a major city in Germany. Sample analysis by LC-HRMS revealed the occurrence of 1,229 compounds, among which 853 were precursors attenuated by ozone and 165 were TPs. Further examination of precursors and TPs using Kendrick mass and Kendrick mass defect analysis revealed 34 pairs of precursors and products corresponding to a mono-oxygenation. Among these, 27 pairs (16% of all TPs) were consistent with N-oxides since the TP had a higher retention time than the precursor, a characteristic of these compounds. Using high resolution tandem mass spectrometry, 10 of these N-oxides could be identified and were shown to be stable during a subsequent filtration step.

Application and characterization of electroactive membranes based on carbon nanotubes and zerovalent iron nanoparticles.

Carbon nanotube (CNT) membranes were produced from multi-walled CNTs by a filtration technique and used for the removal of the betablocker metoprolol by adsorptive and reactive processes. The reactivity of CNT membranes was enhanced by nanoparticulate zero-valent iron (NZVI) which was deposited on the CNT membranes by pulsed voltammetry applying defined number of pulses (Fe-CNT (100) and Fe-CNT (400) membranes). Surface analysis with SEM showed iron nanoparticle sizes between 19 and 425 nm. Pore size distribution for the different membranes was determined by capillary flow porometry (Galwick fluid). Pore size distribution for all membranes was similar (40 nm), which resulted in a water permeability typical for microfiltration membranes. Metoprolol was removed by the CNT membrane only by sorption, whereas the Fe-CNT membrane revealed also metoprolol degradation due to Fenton type reactions. Further application of electrochemical potentials on both the CNT and the Fe-CNT membranes improved the removal efficiencies to 74% for CNT membranes at 1 V and to 97% for Fe-CNT (400) membranes at 1 V. Seven transformation products have been identified for metoprolol by high-resolution mass spectrometry when electrochemical degradation was performed with CNT and Fe-CNT membranes. Additionally, two of the identified transformation products (TPs) were also observed for Fe-CNT membranes without the application of electrochemical potential. However, only 10% of the degraded metoprolol could be explained by the formation of TPs.

Comparison of 1,2-dichloroethane, dichloroethene and vinyl chloride carbon stable isotope fractionation during dechlorination by two Dehalococcoides strains.

Carbon stable isotope fractionation during 1,2-dichloroethane (1,2-DCA), dichloroethene (DCE) and vinyl chloride (VC) dechlorination was analysed for two Dehalococcoides strains, Dehalococcoides mccartyi strain 195 (formerly Dehalococcoides ethenogenes strain 195) and D. mccartyi strain BTF08, and used to characterize the reaction. The isotope enrichment factors (εC) determined for 1,2-DCA were -30.8 ± 1.3‰ and -29.0 ± 3.0‰ for D. mccartyi strain BTF08 and D. mccartyi strain 195, respectively. Enrichment factors (εC) determined for chlorinated ethenes with strain BTF08 were -28.8 ± 1.5‰ (VC), -30.5 ± 1.5‰ (cis-DCE) and -12.4 ± 1.1‰ (1,1-DCE). Product, ethene, related enrichment factors (εC1,2-DCA-ethene) calculated for 1,2-DCA (-34.1 and -32.3‰ for strain BTF08 and strain 195, respectively) were similar to substrate based enrichment factors (εC1,2-DCA), supporting the hypothesis that ethene is the direct product of 1,2-DCA dichloroelimination but that VC was a side product as result of branching in the reaction.