Hourly land-use regression modeling for NO2 and PM2.5 in the Netherlands.

Annual average land-use regression (LUR) models have been widely used to assess spatial patterns of air pollution exposures. However, they fail to capture diurnal variability in air pollution and consequently might result in biased dynamic exposure assessments. In this study we aimed to model average hourly concentrations for two major pollutants, NO2 and PM2.5, for the Netherlands using the LUR algorithm. We modelled the spatial variation of average hourly concentrations for the years 2016-2019 combined, for two seasons, and for two weekday types. Two modelling approaches were used, supervised linear regression (SLR) and random forest (RF). The potential predictors included population, road, land use, satellite retrievals, and chemical transport model pollution estimates variables with different buffer sizes. We also temporally adjusted hourly concentrations from a 2019 annual model using the hourly monitoring data, to compare its performance with the hourly modelling approach. The results showed that hourly NO2 models performed overall well (5-fold cross validation R2 = 0.50-0.78), while the PM2.5 performed moderately (5-fold cross validation R2 = 0.24-0.62). Both for NO2 and PM2.5 the warm season models performed worse than the cold season ones, and the weekends' worse than weekdays'. The performance of the RF and SLR models was similar for both pollutants. For both SLR and RF, variables with larger buffer sizes representing variation in background concentrations, were selected more often in the weekend models compared to the weekdays, and in the warm season compared to the cold one. Temporal adjustment of annual average models performed overall worse than both modelling approaches (NO2 hourly R2 = 0.35-0.70; PM2.5 hourly R2 = 0.01-0.15). The difference in model performance and selection of variables across hours, seasons, and weekday types documents the benefit to develop independent hourly models when matching it to hourly time activity data.

Design of ß-Alanine-Derived Novel C18 Stationary Phase Containing Embedded Urea and Amide Groups for the Separation of Versatile Analytes.

Novel stationary phases have been emerging recently. A ß-alanine-derived embedded urea and amide group-containing C18 phase (Sil-Ala-C18) was prepared for the first time. The media were packed into a 150 × 2.1 mm HPLC column, and the newly designed column was evaluated with the Tanaka and Neue test protocols in reversed-phase liquid chromatography (RPLC) separation mode. Moreover, it was characterized by the Tanaka test protocol in hydrophilic interaction chromatography (HILIC) separation mode. The new phase was characterized by elemental analysis, attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), scanning electron microscopy (SEM), thermogravimetric analysis (TGA), and solid-state 13C cross-polarization magic angle spinning (CP/MAS) NMR spectroscopy at variable temperatures. The chromatographic evaluation involved very good separation of nonpolar shape-constrained isomers, polar and basic compounds in RPLC, and highly polar compounds in HILIC compared to the commercial reference columns. The Sil-Ala-C18 phase was able to separate the challenging ß- and γ-isomers of tocopherol. The phase was also successfully applied for the separation of the isomers of tocopherol (vitamin E) and capsaicinoids from real samples of chili peppers (Capsicum spp.) in RPLC and ascorbic acid (vitamin C) in HILIC.

Multi-2D LC × LC as a Novel and Powerful Implement for the Maximum Separation of Complex Samples.

Achieving complete information about the chemical composition of complex samples requires the use of multianalytical platforms able to maximize the acquisition of high-quality data for unequivocal identification. However, this process requires long analysis times and several instruments. Food analysis is one of the analytical fields where the analysis of very complex samples has a huge impact. One of these complex samples is vermouth, a fortified wine based on the maceration of a large number of herbs, fruits, barks, seeds, and leaves. The application of conventional or even advanced analytical techniques like comprehensive two-dimensional (2D) liquid chromatography (LC × LC) does not provide enough separation power to resolve the complete profile of this sample. In this work, a novel 2DLC strategy called multi-2D LC × LC is developed. This new setup consists of the use of two different columns with different separation properties in the second dimension (2D) that can be selected during the LC × LC analysis accordingly to the chemical nature of the compounds eluted from the first dimension (1D). The vermouth sample was analyzed using a 1D-PFP and a combination of HILIC (from 0 to 30 min) and C18 (from 30 to the end) columns in the 2D. This setup increased both the peak capacity and the orthogonality of the analysis in comparison to the use of only one of the columns in the 2D. Multi-2D LC × LC is presented as an integrated 2DLC tool that maximizes the separation capacity for very complex samples.

Comprehensive two-dimensional gas chromatography with flow modulator coupled via tube plasma ionization to an atmospheric pressure high-resolution mass spectrometer for the analysis of vermouth volatile profile.

The analysis of complex samples is a big analytical challenge due to the vast number of compounds present in these samples as well as the influence matrix components could cause in the methodology. In this way, comprehensive two-dimensional gas chromatography coupled to mass spectrometry (GC × GC-MS) is a very powerful tool to achieve the characterization of complex samples. Nevertheless, due to possible coelutions occurring in these matrices, mixed spectra are generally obtained with electron ionization (EI) which could extremely complicate the identification of the analytes. Thereby, new methodology setups are required to improve the confidence on the identification in non-targeted determinations. Here, we present a high-throughput methodology consisting of GC × GC with flow modulation coupled to high-resolution atmospheric pressure mass spectrometry (HRMS) via a novel tube plasma ion source (TPI). The flow modulator allows to easily automate the GC × GC method compared to traditional cryo-modulators, while the soft ionization provided by TPI helps to preserve the [M]+• or [M+H]+ ions, thus increasing the confidence in the identification. Additionally, the combination of a flow modulation with an atmospheric pressure mass spectrometer significantly improves the sensitivity over flow modulated GC × GC-EI-MS methods because no split is required. This methodology was applied to the analysis of a complex sample such as vermouth where the volatile profile is usually considered by consumers as a product quality indicator since it raises the first sensations produced during its consumption. Using this approach, different classes of compounds were tentatively identified in the sample, including monoterpenes, terpenoids, sesquiterpenoids and carboxylic acid, and carboxylate esters among others, showing the great potential of a GC × GC-TPI-qTOF-MS platform for improving the confidence of the identifications in non-targeted applications.

Development of a Tube Plasma Ion Source for Gas Chromatography-Mass Spectrometry Analysis and Comparison with Other Atmospheric Pressure Ionization Techniques.

A tube plasma ionization (TPI) open-air source for gas chromatography-mass spectrometry (GC-MS) was developed. This source is based on an inverse low temperature plasma configuration where the pin inner electrode is applying the high voltage and the grounded electrode is the housing itself. The ionization possibilities were tested by using an EPA mix of priority contaminants, showing that 68% of the analytes could undergo both proton-transfer and charge-exchange reactions. The potential of using different discharge gases (He and Ar) to ionize the analytes and auxiliary gases (He, N2, O2, and synthetic air) to transport the ions toward the MS was carefully investigated. Additionally, the addition of water was also tested to show the different ionization trends in the TPI source. Finally, the ionization by TPI under both dry and wet conditions was compared with other gas-phase atmospheric pressure ionization sources showing TPI could ionize a wider range of compounds (97%) than atmospheric pressure chemical ionization (APCI, 95%) and atmospheric pressure photoionization (APPI, 87%). Besides, the detection capability of TPI was better than APCI and APPI, achieving instrumental limits of detection down to 3 fg on column, which demonstrates the great potential of this ionization source for GC-MS determinations.

Downregulation of eNOS and preserved endothelial function in endothelial-specific arginase 1-deficient mice.

Arginase 1 (Arg1) is a ubiquitous enzyme belonging to the urea cycle that catalyzes the conversion of l-arginine into l-ornithine and urea. In endothelial cells (ECs), Arg1 was proposed to limit the availability of l-arginine for the endothelial nitric oxide synthase (eNOS) and thereby reduce nitric oxide (NO) production, thus promoting endothelial dysfunction and vascular disease. The role of EC Arg1 under homeostatic conditions is in vivo less understood. The aim of this study was to investigate the role of EC Arg1 on the regulation of eNOS, vascular tone, and endothelial function under normal homeostatic conditions in vivo and ex vivo. By using a tamoxifen-inducible EC-specific gene-targeting approach, we generated EC Arg1 KO mice. Efficiency and specificity of the gene targeting strategy was demonstrated by DNA recombination and loss of Arg1 expression measured after tamoxifen treatment in EC only. In EC Arg1 KO mice we found a significant decrease in Arg1 expression in heart and lung ECs and in the aorta, however, vascular enzymatic activity was preserved likely due to the presence of high levels of Arg1 in smooth muscle cells. Moreover, we found a downregulation of eNOS expression in the aorta, and a fully preserved systemic l-arginine and NO bioavailability, as demonstrated by the levels of l-arginine, l-ornithine, and l-citrulline as well as nitrite, nitrate, and nitroso-species. Lung and liver tissues from EC Arg1 KO mice showed respectively increase or decrease in nitrosyl-heme species, indicating that the lack of endothelial Arg1 affects NO bioavailability in these organs. In addition, EC Arg1 KO mice showed fully preserved acetylcholine-mediated vascular relaxation in both conductance and resistant vessels but increased phenylephrine-induced vasoconstriction. Systolic, diastolic, and mean arterial pressure and cardiac performance in EC Arg1 KO mice were not different from the wild-type littermate controls. In conclusion, under normal homeostatic conditions, lack of EC Arg1 expression is associated with a down-regulation of eNOS expression but a preserved NO bioavailability and vascular endothelial function. These results suggest that a cross-talk exists between Arg1 and eNOS to control NO production in ECs, which depends on both L-Arg availability and EC Arg1-dependent eNOS expression.

Comparative study for analysis of carbohydrates in biological samples.

This work presents a comparative study for the analysis of carbohydrates for four common chromatographic methods, each coupled to mass spectrometry. Supercritical fluid chromatography (SFC), hydrophilic interaction liquid chromatography (HILIC), reversed-phase liquid chromatography (RP-LC) and gas chromatography (GC) with detection by triple quadrupole mass spectrometer (QqQ-MS) are compared. It is shown that gas chromatography and reversed-phase liquid chromatography, each after derivatisation, are superior to the other two methods in terms of separation performance. Furthermore, comparing the different working modes of the mass spectrometer, it can be determined that a targeted analysis, i.e. moving from full scan to single ion monitoring (SIM) and multiple reaction monitoring (MRM), results in an improvement in the sensitivity as well as the repeatability of the method, which has deficiencies especially in the analysis using HILIC. Overall, RP-LC-MS in MRM after derivatisation with 1-phenyl-3-methyl-5-pyrazolone (PMP) proved to be the most suitable method in terms of separation performance, sensitivity and repeatability for the analysis of monosaccharides. Detection limits in the nanomolar range were achieved, which corresponds to a mass concentration in the low µg/L range. The applicability of this method to different biological samples was investigated with various herbal liquors, pectins and a human glycoprotein.

Differentiation of industrial hemp strains by their cannabinoid and phenolic compounds using LC × LC-HRMS.

Cannabis is an ancient plant that has been used for therapeutic and recreational purposes. Nowadays, industrial hemp, a variety with low concentration of the psychoactive cannabinoid Δ9-tetrahydrocannabinol (THC) and high concentration of non-psychoactive cannabinoids, is getting more and more interest in the food, pharmaceutical, and cosmetic industry. However, cannabis not only contains cannabinoids as bioactive components but also other metabolites like terpenes and phenolic compounds, and the content of these interesting secondary metabolites greatly differs with the genetic variety of the plant. Due to the huge complexity of composition of the cannabis matrix, in this work, a comprehensive two-dimensional liquid chromatography (LC × LC) method has been developed as a very power separation technique coupling a pentafluorophenyl (PFP) and a C18 in the first and second dimensions. Two industrial hemp strains (cookie and gelato) were analyzed to determine the difference in their content of cannabinoids and phenolic compounds. To do this, a new demodulation process was applied for the first time to transform 2D raw data into 1D data which allowed carrying out the chemometric analysis needed to determine the statistical differences between the hemp strains. The cookie strain presented a total of 41 cannabinoid markers, while the gelato strain presented more representative phenolic compounds, in total 24 phenolic compounds were detected as potential markers of this sample. These differences in the chemical composition could determine the industrial destiny of the different hemp strains.

Capillary electrophoresis and molecular modeling of the chiral separation of aromatic amino acids using α/ß-cyclodextrin and 18-crown-6.

In this work, chiral separation of enantiomers of three amino acids was achieved using capillary electrophoresis technique with α-cyclodextrin (αCD) as a running buffer additive. Only tryptophan has exhibited baseline separation in the presence of αCD, while the enantiomers of the other two amino acids, phenylalanine and tyrosine, were only partially separated. The addition of 18-crown-6 (18C6) as a second additive imparted only slight improvement to the separation of all enantiomers. On the other hand, all three racemic amino acid mixtures demonstrated no indication of separation when the larger cavity cyclodextrin members, ß- and γCD, are used as running buffer chiral additives. However, remarkable improvements in the separation of the enantiomers of phenylalanine and tyrosine were obtained when 18C6 is used together with ßCD as a running buffer additive. Surprisingly, tryptophan enantiomers were not separated by the dual additive system of cyclodextrin and crown ether. Using electrospray ionization mass spectrometry (ESI-MS), all amino acids were found to form stable binary complexes with individual hosts as well as ternary compounds involving the crown ether and the cyclodextrin. Furthermore, we used molecular dynamics (MD) simulations to build a clear picture about the interaction between the guest and the hosts. Most of these complexes remained stable throughout the simulation times, and the molecular dynamics study allowed better understanding of these supramolecular assemblies.

Development of an inverse low-temperature plasma ionization source for liquid chromatography/mass spectrometry.

RATIONALE: An argon inverse low-temperature plasma (iLTP) ionization source for liquid chromatography/tandem mass spectrometry was developed. The iLTP is constructed from simple chromatographic supply materials and is implemented into an atmospheric pressure chemical ionization (APCI) source replacing the APCI discharge needle electrode. The newly developed ion source was coupled to an ultra-high-performance liquid chromatography (UHPLC) system. METHODS: The argon iLTP was characterized by optical emission spectroscopy. The soft ionization of selected standards was also demonstrated by direct infusion experiments. In addition to the use of argon as the discharge gas, helium, synthetic air, and oxygen were used, which were tested for their performance using testosterone and vitamin D3 . RESULTS: Spectroscopic measurements of the argon plasma were conducted, demonstrating the main emission band of argon metastables with corresponding energies of 11.53 eV and 11.72 eV. Infusion experiments indicate a gentle ionization by iLTP, e.g. caffeine, testosterone, reserpine, vitamin D3 , and 25-hydroxyvitamin D3 , which resulted in the corresponding protonated molecules. The splitless coupling with UHPLC (possible flow rates >1000 µL min-1 ) shows promising results in interday repeatability (n = 10) for the substances with a relative standard deviation of less than 5% and limits of detection for caffeine, testosterone, reserpine, vitamin D3 , and 25-hydroxyvitamin D3 of 10 ng L-1 , 50 ng L-1 , 500 ng L-1 , 5 µg L-1 , and 5 µg L-1 , respectively. CONCLUSIONS: The argon iLTP ion source presented in this work shows promising approaches in the field of ionization of small organic molecules. The mechanism related to the discharge gas argon has not been elucidated so far and further investigations are needed. The iLTP ion source shows a very good performance with UHPLC coupling, even at increased flow rates. It could be shown that an argon iLTP can compete with the helium dielectric barrier discharge (DBD) preferred in the literature, making it a more economical choice.

A comparison of associations with childhood lung function between air pollution exposure assessment methods with and without accounting for time-activity patterns.

BACKGROUND: To investigate associations between annual average air pollution exposures and health, most epidemiological studies rely on estimated residential exposures because information on actual time-activity patterns can only be collected for small populations and short periods of time due to costs and logistic constraints. In the current study, we aim to compare exposure assessment methodologies that use data on time-activity patterns of children with residence-based exposure assessment. We compare estimated exposures and associations with lung function for residential exposures and exposures accounting for time activity patterns. METHODS: We compared four annual average air pollution exposure assessment methodologies; two rely on residential exposures only, the other two incorporate estimated time activity patterns. The time-activity patterns were based on assumptions about the activity space and make use of available external data sources for the duration of each activity. Mapping of multiple air pollutants (NO2, NOX, PM2.5, PM2.5absorbance, PM10) at a fine resolution as input to exposure assessment was based on land use regression modelling. First, we assessed the correlations between the exposures from the four exposure methods. Second, we compared estimates of the cross-sectional associations between air pollution exposures and lung function at age 8 within the PIAMA birth cohort study for the four exposure assessment methodologies. RESULTS: The exposures derived from the four exposure assessment methodologies were highly correlated (R > 0.95) for all air pollutants. Similar statistically significant decreases in lung function were found for all four methods. For example, for NO2 the decrease in FEV1 was -1.40% (CI; -2.54, -0.24%) per IQR (9.14 µg/m3) for front door exposure, and -1.50% (CI; -2.68, -0.30%) for the methodology which incorporates time activity pattern and actual school addresses. CONCLUSIONS: Exposure estimates from methods based on the residential location only and methods including time activity patterns were highly correlated and associated with similar decreases in lung function. Our study illustrates that the annual average exposure to air pollution for 8-year-old children in the Netherlands is sufficiently captured by residential exposures.

Progress towards an OECD reporting framework for transcriptomics and metabolomics in regulatory toxicology.

Omics methodologies are widely used in toxicological research to understand modes and mechanisms of toxicity. Increasingly, these methodologies are being applied to questions of regulatory interest such as molecular point-of-departure derivation and chemical grouping/read-across. Despite its value, widespread regulatory acceptance of omics data has not yet occurred. Barriers to the routine application of omics data in regulatory decision making have been: 1) lack of transparency for data processing methods used to convert raw data into an interpretable list of observations; and 2) lack of standardization in reporting to ensure that omics data, associated metadata and the methodologies used to generate results are available for review by stakeholders, including regulators. Thus, in 2017, the Organisation for Economic Co-operation and Development (OECD) Extended Advisory Group on Molecular Screening and Toxicogenomics (EAGMST) launched a project to develop guidance for the reporting of omics data aimed at fostering further regulatory use. Here, we report on the ongoing development of the first formal reporting framework describing the processing and analysis of both transcriptomic and metabolomic data for regulatory toxicology. We introduce the modular structure, content, harmonization and strategy for trialling this reporting framework prior to its publication by the OECD.

Zinc distribution within breast cancer tissue of different intrinsic subtypes.

PURPOSE: To show feasibility of laser ablation inductively coupled mass spectrometry (LA-ICPMS) for analysis of zinc content and concentration in breast cancer tissue and to correlate this with validated prognostic and predictive markers, i.e. histological grading and expression of steroid receptors (estrogen receptor, ER; progesterone receptor, PR) and human epidermal growth-factor receptor 2 (Her2). METHODS: 28 samples of human invasive ductal breast cancer tissue were subclassified into groups of four different intrinsic subtypes according to the expression of ER, PR and Her2 by immunohistological staining and then analyzed for zinc content and distribution by LA-ICPMS applying a calibration technique based on spiked polyacrylamide gels. A correlation of zinc concentration with histological grading and molecular subtypes was analyzed. RESULTS: Consistent with results of a pilot-study LA-ICPMS was feasible to show zinc accumulation in cancerous tissue, even more adjacent healthy stroma was with proportional increase of zinc. Zinc levels were most elevated in triple-positive (TPBC) and in triple-negative (TNB) breast cancers. CONCLUSION: LA-ICPMS was feasible to confirm a connection between zinc and grade of malignancy; furthermore, focusing on a correlation of zinc and intrinsic breast cancer subtypes, LA-ICPMS depicted an upwards trend of zinc for "high-risk-cancers" with highest levels in Her2-positive and in triple-negative (TNBC) disease. The currently uncommon alliance of clinicians and analytical chemists in basic research is most promising to exploit the full potential of diagnostic accuracy in the efforts to solve the enigma of breast cancer initiation and course of disease.

Identification of Chaoborus kairomone chemicals that induce defences in Daphnia.

Infochemicals play important roles in aquatic ecosystems. They even modify food web interactions, such as by inducing defenses in prey. In one classic but still not fully understood example, the planktonic freshwater crustacean Daphnia pulex forms specific morphological defenses (neckteeth) induced by chemical cues (kairomones) released from its predator, the phantom midge larva Chaoborus. On the basis of liquid chromatography, mass spectrometry, and chemical synthesis, we report here the chemical identity of the Chaoborus kairomone. The biologically active cues consist of fatty acids conjugated to the amino group of glutamine via the N terminus. These cues are involved in Chaoborus digestive processes, which explains why they are consistently released despite the disadvantage for its emitter. The identification of the kairomone may allow in-depth studies on multiple aspects of this inducible defense system.

Development of a fast-switching dual (ESI/APCI) ionization source for liquid chromatography/mass spectrometry.

RATIONALE: High-throughput liquid chromatography/mass spectrometry (LC/MS) is an increasing topic in analytical chemistry. Especially the idle time of a mass spectrometer should be reduced for an efficient and cost-saving use. Therefore, a fast-switching dual ion source was developed, which uses the most important ionization techniques at atmospheric pressure, electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI), with one or more LC systems. METHODS: The performance of the developed ion source is shown by infusion experiments and chromatographic analyses of different standard substances. A high-throughput method is demonstrated by coupling two UHPLC systems to the dual ion source with a triple quadrupole mass spectrometer. RESULTS: No decrease in the ion abundance and a stable performance of the mass spectrometer are presented while using the dual ion source. Instrumental limits of detection are 30 ng L-1 for testosterone using ESI and 1 µg L-1 for vitamin D3 using APCI. A fast switching between two UHPLC systems and the dual ion source leads to a high sample throughput of 50 samples in 75 min with relative standard deviations for testosterone and vitamin D3 of 1.5% and 3.8%, respectively. CONCLUSIONS: This work presents the development of a dual ESI and APCI ion source operating simultaneously or in switched mode. The results show sensitive and reliable performance as well as the hyphenation to one or more HPLC systems.

Application of the new at-column dilution (ACD) modulator for the two-dimensional RP×HILIC analysis of Buddleja davidii.

The focus of this study was the analysis of the complex chemical composition from different parts of Buddleja davidii, whose species are commonly known as ornamental plants and herbal medicines in many countries. As an herbal medicine, it has been utilized for stroke treatments, headache, wound healing, neurological disorder, etc. However, the understanding of its chemical matrices is still insufficient. Therefore, an online two-dimensional reversed phase liquid chromatography x hydrophilic interaction liquid chromatography (RPLCxHILIC) system coupled with mass spectrometry was applied for further detailed investigation of the chemical constituents in Buddleja dividii. In this two-dimensional liquid chromatography (2D-LC) method, a new at-column dilution (ACD) modulator was introduced in the 2D-LC system to solve the incompatibility problem of the mobile phase between two dimensions, which resulted in a 2D-LC analysis with high orthogonality. For the root extract, as one of the analyzed samples, the optimization of the 1D and 2D gradients was carried out carefully. With this new modulator, much better peak separation and better peak shape were achieved compared to two-dimensional liquid chromatography system using a traditional standard (TS) modulator. With a similar approach, the other four parts of Buddleja davidii were well separated. Comparing the different analyzed parts, flowers and leaves showed the most complex profiles. MS and MS/MS data were obtained successfully, which demonstrated the potential of the proposed RPLCxHILIC-MS system in the constituents' analysis of herbal medicine. However, due to the lack of reported reference information, 24 compounds could be tentatively identified.

Effect of ozone stress on the intracellular metabolites from Cobetia marina.

A GCxGC-MS system was employed with a non-polar × mid-polar column set for the metabolic non-target analysis of Cobetia marina, the model bacteria for marine biofouling. C. marina was treated with ozone to investigate the intracellular metabolic state change under oxidative stress. A minimal inhibitory concentration test was involved to guarantee that the applied ozone dosages were not lethal for the cells. In this study, non-target analyses were performed to identify the metabolites according to the NIST database. As a result, over 170 signals were detected under normal living conditions including 35 potential metabolites. By the comparison of ozone-treated and non-treated samples, five compounds were selected to describe observed trends of signals in the contour plots. Oleic acid exhibited a slight growth by increasing ozone dosage. In contrast, other metabolites such as the amino acid L-proline showed less abundance after ozone treatment, which was more evident once ozone dosage was raised. Thus, this work could provide a hint for searching for up/downregulating factors in such environmental stress conditions for C. marina. Graphical abstract.

Analysis of volatile metabolites from in vitro biofilms of Pseudomonas aeruginosa with thin-film microextraction by thermal desorption gas chromatography-mass spectrometry.

Cystic fibrosis (CF) is an autosomal recessive inherited disease which leads to a production of thickened mucus in the airways. These conditions are conducive to poly-microbial infections, like chronic lung infection, in which Pseudomonas aeruginosa (P. aeruginosa) is the major pathogenic bacterium colonizing CF lungs at the end of the lifetime of CF patients. This in vitro study uses a P. aeruginosa biofilm model under partly cystic fibrosis conditions, with a sampling of volatile extracellular metabolites. The gas sampling was done with thin-film microextraction (TFME) and commercial polydimethylsiloxane (PDMS) films, whereas the analysis of loaded films was done by gas chromatography coupled to quadrupole mass spectrometry and thermodesorption (TD-GC-qMS). For this purpose, two commercially available films were characterized by means of thermogravimetry coupled to a qMS with atmospheric pressure photo ionization (TG-APPI-qMS), regarding homogeneity and temperature stability. The selected film was cleaned using a method developed in this study. The TD-GC-qMS method was successfully used for standards of volatile metabolites which were known to be produced by P. aeruginosa. Limits of detection and quantification of the method for middle and less polar compounds in low nanomolar range (0.5 nM and 1.5 nM) were achieved. The developed method was finally applied to investigate the extracellular volatile metabolites produced by biofilms of the strain P. aeruginosa DSM 50071 under aerobic and anaerobic conditions. In sum, eleven metabolites could be found under both conditions. Furthermore, it was shown in this study that different oxygen conditions (aerobic and anaerobic) resulted in emitting different extracellular volatile metabolites. Specific metabolites, like 1-undecene (aerobic) and 2-undecanone (anaerobic), could be identified. The results are promising, in that the biofilm model may be applicable for the identification of P. aeruginosa under clinical conditions. Furthermore, the model could be the basis for studying extracellular volatile metabolites from different mono- or co-cultures of various bacteria, as well as the implementation of pulmonary conditions, like these in CF lungs. This possibility allows the development of a non-invasive "at-bedside" breath analysis method for CF patients in focus of various bacterial infections. Graphical abstract.

Non-targeted and targeted analysis of oxylipins in combination with charge-switch derivatization by ion mobility high-resolution mass spectrometry.

Eicosanoids and other oxylipins play an important role in mediating inflammation as well as other biological processes. For the investigation of their biological role(s), comprehensive analytical methods are necessary, which are able to provide reliable identification and quantification of these compounds in biological matrices. Using charge-switch derivatization with AMPP (N-(4-aminomethylphenyl)pyridinium chloride) in combination with liquid chromatography ion mobility quadrupole time-of-flight mass spectrometry (LC-IM-QTOF-MS), we developed a non-target approach to analyze oxylipins in plasma, serum, and cells. The developed workflow makes use of an ion mobility resolved fragmentation to pinpoint derivatized molecules based on the cleavage of AMPP, which yields two specific fragment ions. This allows a reliable identification of known and unknown eicosanoids and other oxylipins. We characterized the workflow using 52 different oxylipins and investigated their fragmentation patterns and ion mobilities. Limits of detection ranged between 0.2 and 10.0 nM (1.0-50 pg on column), which is comparable with other state-of-the-art methods using LC triple quadrupole (QqQ) MS. Moreover, we applied this strategy to analyze oxylipins in different biologically relevant matrices, as cultured cells, human plasma, and serum. Graphical abstract.

Innovative Optical-Sensing Technology for the Online Fouling Characterization of Silicon Carbide Membranes during the Treatment of Oily Water.

The oil and gas industry generates a large volume of contaminated water (produced water) which must be processed to recover oil before discharge. Here, we evaluated the performance and fouling behavior of commercial ceramic silicon carbide membranes in the treatment of oily wastewaters. In this context, microfiltration and ultrafiltration ceramic membranes were used for the separation of oil during the treatment of tank dewatering produced water and oily model solutions, respectively. We also tested a new online oil-in-water sensor (OMD-32) based on the principle of light scattering for the continuous measurement of oil concentrations in order to optimize the main filtration process parameters that determine membrane performance: the transmembrane pressure and cross-flow velocity. Using the OMD-32 sensor, the oil content of the feed, concentrate and permeate streams was measured continuously and fell within the range 0.0-200 parts per million (ppm) with a resolution of 1.0 ppm. The ceramic membranes achieved an oil-recovery efficiency of up to 98% with less than 1.0 ppm residual oil in the permeate stream, meeting environmental regulations for discharge in most areas.