Progress on the Link between Nutrient Availability and Toxin Production by Ostreopsis cf. ovata: Field and Laboratory Experiments.

This study aimed to improve the understanding of the nutrient modulation of Ostreopsis cf. ovata toxin content. During the 2018 natural bloom in the NW Mediterranean, the total toxin content (up to ca. 57.6 ± 7.0 pg toxin cell-1) varied markedly. The highest values often coincided with elevated O. cf. ovata cell abundance and with low inorganic nutrient concentrations. The first culture experiment with a strain isolated from that bloom showed that cell toxin content was higher in the stationary than in the exponential phase of the cultures; phosphate- and nitrate-deficient cells exhibited similar cell toxin variability patterns. The second experiment with different conditions of nitrogen concentration and source (nitrate, urea, ammonium, and fertilizer) presented the highest cellular toxin content in the high-nitrogen cultures; among these, urea induced a significantly lower cellular toxin content than the other nutrient sources. Under both high- and low-nitrogen concentrations, cell toxin content was also higher in the stationary than in the exponential phase. The toxin profile of the field and cultured cells included ovatoxin (OVTX) analogues -a to -g and isobaric PLTX (isoPLTX). OVTX-a and -b were dominant while OVTX-f, -g, and isoPLTX contributed less than 1-2%. Overall, the data suggest that although nutrients determine the intensity of the O. cf. ovata bloom, the relationship of major nutrient concentrations, sources and stoichiometry with cellular toxin production is not straightforward.

Multiply charged ion profiles in the UHPLC-HRMS analysis of palytoxin analogues from Ostreopsis cf. ovata blooms.

Analogues of palytoxin (PLTX), one of the most potent marine biotoxins, are produced by some species of the marine dinoflagellates of the genus Ostreopsis. The proliferation of these species in different coastal zones represents a potential threat of seafood poisoning in humans because the produced toxins can be transferred through marine food webs. Thus, the determination of the concentration of PLTX analogues (ovatoxins-OVTXs, ostreocins-OSTs and isobaric PLTX) in different matrices (seawater, marine fauna, etc.) is necessary to protect human health. This study is addressed to overcome some of the challenges that the chemical complexity of these molecules poses to their quantification by ultra-high-performance liquid chromatography high-resolution mass spectrometry-based techniques (UHPLC-HRMS). In particular, the mass spectra of the palytoxin analogues show the presence of a large number of ions (including mono- and multiply charged ions) whose nature, relative abundances and behavior can lead to quantitation errors if the correct ions are not selected. In this work, the variability of the PLTX and OVTX profiles under different instrument conditions, including the use of diverse electrospray generation sources and different quantitation methods, is studied. Moreover, the extraction protocol in seawater containing Ostreopsis sp. ovata cells is also evaluated. The use of a heated electrospray operating at 350 °C and a quantitative method including ions from different multiply charged species provides a more robust and reliable method for overcoming the problems due to the variability in the toxin's mass spectrum profile. A single MeOH : H2O (80 : 20, v/v) extraction is proposed as the best and reliable procedure. The overall method proposed was applied to quantify OVTXs (-a to -g) and iso-PLTX along the 2019 Ostreopsis cf. ovata bloom. The cells contained a total toxin concentration of up to 20.39 pg per cell.

Potential of desorption electrospray ionization and paper spray ionization with high-resolution mass spectrometry for the screening of sports doping agents in urine.

In this work, desorption electrospray ionization and paper spray ionization both with high-resolution mass spectrometry (DESI-HRMS and PSI-HRMS) were explored for the fast and direct analysis of stimulants and diuretics in urine samples. The analysis was performed at a resolution of 70 000 FWHM (m/z 200) using a quadrupole-Orbitrap mass spectrometer in full scan acquisition mode, detecting stimulants and diuretics in positive and negative ion modes, respectively. The most critical parameters affecting the desorption and ionization efficiencies of compounds were optimized, paying particular attention to the optimization of the spray solvent for PSI-HRMS analysis and to the selection of the DESI sample substrate. For stimulants, the PSI-HRMS method performed better than DESI-HRMS, allowing the direct analysis of raw urine samples with better signal-to-noise ratios than DESI. However, results obtained for diuretics were not as satisfactory as we expected. The PSI-HRMS method was applied to the screening of 52 stimulants for doping control purposes, providing satisfactory detectability for most of them at the Minimum Reporting Level (MRL) in less than 2 minutes for each single analysis. Despite the advantages offered by the PSI-HRMS method, in this study is also included a discussion on the limitations observed because of the presence of interference for some compounds.

Ultra-high-performance liquid chromatography-atmospheric pressure ionization-tandem mass spectrometry method for the migration studies of primary aromatic amines from food contact materials.

This work describes the development of an ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for the determination of 23 primary aromatic amines (PAAs) that can potentially migrate from food contact materials. The chromatographic separation was performed in a pentafluorophenylpropyl (PFPP) column achieving the separation of all PAAs in less than 6.5 min using water to acetonitrile (0.1% acetic acid in both solvents) as mobile phase and a gradient elution. The feasibility of atmospheric pressure chemical ionization (APCI) and atmospheric pressure photoionization (APPI) was evaluated as alternative to electrospray ionization (ESI) for the analysis of PAAs. Results showed that for most of the compounds, better responses were obtained with APCI, which shows the advantage of being less susceptible to matrix effects. Tandem mass spectrometry (MS/MS) fragmentation studies of [M + H]+ allowed for the selection of the two most characteristic and abundant product ions of the 23 PAAs which led to the development of a selective and sensitive UHPLC-APCI-MS/MS method with limits of detection ranging from 0.2 to 2 µg kg-1. Moreover, intra-day and inter-day precisions of the method in terms of relative standard deviation (RSD%) were lower than 10% and 15%, while trueness as relative error was <15% for most of the compounds. The UHPLC-APCI-MS/MS method was applied to the analysis of twenty black Nylon kitchenware samples that were submitted to migration tests using food simulant B (3% acetic acid, w/v), and the presence of PAAs were detected in eighteen samples at concentrations above the legislated limit (2 µg kg-1 of food or food simulants).

Analysis of Dechlorane Plus and related compounds in gull eggs by GC-HRMS using a novel atmospheric pressure photoionization source.

Here, a new gas chromatography-atmospheric pressure photoionization-high-resolution mass spectrometry (GC-APPI-HRMS) method combined with selective pressurized liquid extraction (sPLE) has been developed for the selective determination of Dechlorane Plus (DP) and its related compounds in gull egg samples used as a bioindicator of contamination. To the best of our knowledge, this is the first time these compounds have been analyzed by GC-MS using atmospheric pressure photoionization (APPI). Negative ion dopant-assisted APPI using vapors of diethyl ether and a source temperature of 250 °C provided high ionization efficiencies and mass spectra characterized by intense in-source fragment ions as well as the presence of molecular ion and characteristic cluster ions containing oxygen atoms in their chemical structure. This made it possible to improve the selectivity in the determination of these compounds compared to that obtained with traditional GC-MS ion sources. Under optimized conditions, the sPLE GC-APPI-HRMS (Orbitrap) method provided high recoveries (> 91%), good precisions (RSD% < 12%), and low method limits of detection (0.1-3.5 pg g-1 wet weight). The developed methodology has been applied to the determination of DP and related compounds in eggs of two gull species (L. michahellis and L. audouinii) from several Spanish protected areas. The results obtained showed significant differences in the DP concentration profiles in eggs from different gull breeding locations and between gull species of the same protected area. These results demonstrated the good performance of the GC-APPI-HRMS system to achieve a selective and sensitive determination of DP and related compounds in complex environmental samples.

Aerosol Toxins Emitted by Harmful Algal Blooms Susceptible to Complex Air-Sea Interactions.

Critical research is needed regarding harmful algal blooms threatening ecosystem and human health, especially through respiratory routes. Additional complexity comes from the poorly understood factors involved in the physical production of marine aerosols coupled with complex biogeochemical processes at ocean surfaces. Here-by using a marine aerosol generation tank-five bubble-bursting experiments (with contrasting incubation times and, likely, physiological microalgal states) were run to investigate simultaneously the concentrations of the toxins, synthesized by a natural Ostreopsis cf. ovata bloom, in suspension in the water and in the atmosphere. The first two experiments (EXP1-2) were run with moderate levels of O. cf. ovata cell numbers (ca. 105 cells·L-1) and total toxin in suspension (4 × 106 pg·Lwater-1) obtained at an early phase of the bloom. After 0.75-4 h incubation, toxin concentration in the aerosols accounted for 49-69 pg·Lair-1. By striking contrast, three experiments (EXP3-5)-conducted with samples collected two weeks later with higher cell abundances and higher toxin concentration in the seston (respectively, about 1 × 106 cells·L-1 and 2 × 108 pg·Lwater-1) and incubated for 21 h-showed about 15-fold lower atmospheric concentrations (3-4 pg·Lair-1), while important foam accumulation was observed in the water surface in the tank. Offline spectroscopic analysis performed by proton-nuclear magnetic resonance spectroscopy showed that the particulate organic carbon in the water was drastically different from that of bubble-bursting aerosols from the tank experiments-suggesting a selective transfer of organic compounds from seawater into the atmosphere. Overall, the results suggest that aerosol production and diffusion of marine toxins in the atmosphere are regulated by complex interactions between biological processes and air-sea aerosol production dynamics.

Analysis of hydroxylated phenylalkylamine stimulants in urine by GC-APPI-HRMS.

A gas chromatography-atmospheric pressure photoionization-high-resolution mass spectrometry (GC-APPI-HRMS) method was developed for the determination of eight phenylalkylamine stimulants in urine samples. Spiked urine samples were hydrolyzed, processed by solid-phase extraction, and derivatized before analysis. Two derivatization reactions were studied: the formation of trimethylsilyl (TMS) derivatives with N-methyl-N-trimethylsilyl trifluoroacetamide (MSTFA) and trimethylsilyl/trifluoroacetyl (TMS/TFA) derivatives with MSTFA and N-methyl-bis (trifluoroacetamide) (MBTFA) as derivatization reagents. Gas chromatography of both derivatives was performed with a 100% dimethylsiloxane column and a good separation of all isomeric compounds was achieved. To maximize the signal of the protonated molecule [M+H]+, the APPI most critical parameters were optimized. Three solvents were tested as dopant agents, with acetone yielding the lower in-source collision-induced dissociation (CID) fragmentation. The acquisition was performed in full scan and product ion scan (parallel reaction monitoring, PRM) using a quadrupole-Orbitrap mass analyzer (35,000 FWHM at m/z 200) in positive ion detection mode. At the optimal working conditions, the full scan method was evaluated for the fulfillment of identification requirements in doping analysis. Selectivity, limits of detection, matrix effect, and precision were estimated to validate the method for confirmation purposes and its applicability was tested by the analysis of spiked samples as well as by the analysis of samples obtained after the administration of some of the compounds to healthy volunteers. Results were compared with those obtained by GC-electron ionization-MS, demonstrating that the GC-APPI-HRMS method improved selectivity and sensibility, achieving lower limits of detection and satisfactory reproducibility.

Pigment profiles of Spanish extra virgin olive oils by ultra-high-performance liquid chromatography coupled to high-resolution mass spectrometry.

This work studies the natural pigment profiles (chlorophylls and carotenoids) of Spanish Extra Virgin Olive Oils (EVOO) produced in different Spanish regions. The simultaneous qualitative and quantitative analysis of EVOO natural pigments has been performed by ultra-high-performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS) using atmospheric pressure chemical ionisation (APCI). The results showed a similar natural pigment pattern for all the analysed EVOOs, although the total pigments content differed significantly. Moreover, the chlorophyll/carotenoid ratio was close to 1, while the lutein/ß-carotene ratio was higher than 1, showing that lutein is the most abundant carotenoid in the studied Spanish EVOOs. Data from multivariate statistical approach demonstrated that the olive variety does not discriminate between EVOO samples. However, they were classified based on their origin allowing good differentiation of samples from the Basque Country and Canary Islands from the rest of regions. The results of this study show the differences of the nature and pigments concentration of Spanish EVOO samples, parameters that are of significance for reliable characterisation.

Fragmentation studies of neutral per- and polyfluoroalkyl substances by atmospheric pressure ionization-multiple-stage mass spectrometry.

The establishment of fragmentation pathways has a great interest in the identification of new or unknown related compounds present in complex samples. On that way, tentative fragmentation pathways for the ions generated by atmospheric pressure ionization of neutral per- and polyfluorinated alkyl substances (PFASs) have been proposed in this work. Electrospray (ESI), atmospheric pressure chemical ionization (APCI) and photoionization (APPI) were evaluated using mobile phases and source conditions that enhance the ionization efficiency of ions generated. A hybrid mass spectrometer consisting of a linear ion trap and an Orbitrap was used to combine the information of both multiple-stage mass spectrometry (MSn) and mass accuracy measurements to characterize and establish the genealogical relationship between the product ions observed. The ionization mechanisms to generate ions such as [M-H]-, [M]-•, and [M+O2]-• or the in-source collision-induced dissociation (CID) fragment ions in each API source are discussed in this study. In general, fluorotelomer olefins (FTOs) ionized in negative-ion APCI and APPI generated the molecular ion, while fluorotelomer alcohols (FTOHs) also provided the deprotonated molecule. Besides, fluorooctane sulfonamides (FOSAs) and sulfonamido-ethanols (FOSEs) led to the deprotonated molecule and in-source CID fragment ions, respectively. The fragmentation pathways from these precursor ions mainly involved initial α,ß-eliminations of HF units and successive losses of CF2 units coming from the perfluorinated alkyl chain. Moreover, FTOHs and FOSEs showed a high tendency to generate adduct ions under negative-ion ESI and APPI conditions. The fragmentation study of these adduct ions has demonstrated a strong interaction with the attached moiety. Graphical abstract.

Formation of new disinfection by-products of priority substances (Directive 2013/39/UE and Watch List) in drinking water treatment.

The degradation of priority substances (Directive 2013/39/UE and Watch List) by chlorine dioxide (ClO2) and the formation of disinfection by-products (DBPs) in a drinking water treatment plant (DWTP) located near Barcelona (NE Spain) were investigated. For the first time, the reactivity with ClO2 of several compounds frequently found at the entrance of the DWTP such as erythromycin, clarithromycin, chlorpyrifos, and imidacloprid was evaluated in both simulated and real conditions. To identify potential DBPs, experiments were performed at laboratory scale by simulating the operational disinfection conditions in the DWTP. Liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) working in full scan and target-MS/HRMS modes was used for the identification of the generated DBPs. Several new DBPs were found, three from erythromycin, one from clarithromycin, two from chlorpyrifos, and one from imidacloprid. Then, the presence and behavior through DWTP treatment of priority substances and their DBPs were investigated in order to evaluate their generation in real working conditions. Two of the potential DBPs, anhydroerythromycin, and N-desmethyl clarithromycin were already identified in the raw water of DWTP, but N-desmethyl clarithromycin was also generated after the chlorine dioxide treatment step. Both compounds were eliminated by the treatments applied in the DWTP; anhydroerythromycin was eliminated after ozonation in the upgraded conventional treatment and after reverse osmosis in the advanced treatment while N-desmethyl clarithromycin is recalcitrant in the upgraded conventional treatment, but it was eliminated by reverse osmosis.

Trace analysis of polystyrene microplastics in natural waters.

The development of quantitative and qualitative analytical methods to assess micro-plastics (MPLs) and nano-plastics (NPLs) content in the environment is a central issue for realistic risk assessment studies. However, the quantitative analysis continues being a critical issue, in particular for MPLs from 100 µm down to the nano-sized range in complex environmental samples. This paper evaluates the potential of mass spectrometry for the analysis of MPLs and NPLs. The performance of different techniques including matrix-assisted laser desorption ionisation (MALDI) coupled to time-of-flight mass spectrometry (TOF-MS), liquid chromatography coupled to high-resolution mass spectrometry (LC-HRMS), and the ambient ionisation approaches as desorption electrospray ionisation (DESI) and direct analysis real-time (DART), were assessed for the study of polystyrene (PS) MPLs and NPLs in natural waters. A method based on LC-HRMS, equipped with an atmospheric pressure photoionisation source (APPI), operated in negative conditions for the quantitative analysis of PS MPLs and NPLs in natural waters, was developed. The chromatographic separation was achieved using an advanced polymer chromatographic (APC) column using toluene isocratic as the mobile phase. The optimal analytical method showed an instrumental limit of detection (ILOD) of 20 pg and methods limits of detection and quantification around 30 pg L-1 and 100 pg L-1, respectively. And, recoveries of 60 and 70% in samples from rivers and the marine coast, respectively. The performance of the new method was proved by the analysis of fortified samples and natural seawater samples.

Simultaneous analysis of natural pigments and E-141i in olive oils by liquid chromatography-tandem mass spectrometry.

This work describes the development of an ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for the determination of carotenoids (ß-carotene, lutein, ß-criptoxanthin, neoxanthin, violaxanthin) and chlorophylls, as well as their related compounds (chlorophyll A and B, pheophytin A and B and the banned dyes Cu-pyropheophytin A, Cu-pheophytin A and B) in olive oils. For this purpose, the feasibility of electrospray ionization (ESI), atmospheric pressure chemical ionization (APCI) and atmospheric pressure photoionization (APPI) for the ionization of these compounds was evaluated and compared. Tandem mass spectrometry (MS/MS) fragmentation was discussed for each family of compounds, and the most characteristic and abundant product ions were selected to propose a selective and sensitive UHPLC-MS/MS method. The best results were obtained using APCI and APPI, while ESI provided the worst signal-to-noise ratio (S/N) for all compounds. For the analysis of olive oils, a simple solid-phase extraction (SPE) with silica cartridges was applied before the determination by UHPLC-MS/MS (APCI and APPI) in multiple reaction monitoring (MRM) mode. Method quality parameters were stablished, and the results demonstrate the good performance of the new methods, providing low limits of detection (0.004-0.9 mg L-1), high extraction efficiencies (62-95%) and low matrix effects (< 25%). The developed UHPLC-API-MS/MS (APCI and APPI) methods were applied to the analysis of olive oil samples, and ß-carotene, pheophytin A, pheophytin B and lutein were detected and quantified in all of them at concentrations ranging from 0.1 to 9.5 mg L-1. Graphical abstract.

Native Colombian Fruits and Their by-Products: Phenolic Profile, Antioxidant Activity and Hypoglycaemic Potential.

The phenols and fatty acids profile and in vitro antioxidant and hypoglycaemic activity of seed, peel, pulp or pulp plus seeds of Colombian fruits from Solanaceae and Passifloraceae families were investigated. Ultra-High Performance Liquid Chromatography (UHPLC)-High Resolution Mass Spectrometry (HRMS) revealed the presence of chlorogenic acid as dominant phenolic compound in Solanaceae samples. Based on the Relative Antioxidant Score (RACI) and Global Antioxidant Score (GAS) values, Solanum quitoense peel showed the highest antioxidant potential among Solanaceae samples while Passiflora tripartita fruits exhibited the highest antioxidant effects among Passifloraceae samples. P. ligularis seeds were the most active as hypoglycaemic agent with IC50 values of 22.6 and 24.8 µg/mL against α-amylase and α-glucosidase, respectively. Considering that some of the most promising results were obtained by the processing waste portion, its use as functional ingredients should be considered for the development of nutraceutical products intended for patients with disturbance of glucose metabolism.

Modified distribution in the polyphenolic profile of rosemary leaves induced by plant inoculation with an arbuscular mycorrhizal fungus.

BACKGROUND: Rosemary forms an arbuscular mycorrhizal (AM) symbiosis with a group of soilborne fungi belonging to the phylum Glomeromycota, which can modify the plant metabolome responsible for the antioxidant capacity and other health beneficial properties of rosemary. RESULTS: The effect of inoculating rosemary plants with an AM fungus on their growth via their polyphenolic fingerprinting was evaluated after analyzing leaf extracts from non-inoculated and inoculated rosemary plants by ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS). Plant growth parameters indicated that mycorrhizal inoculation significantly increased plant height and biomass. Chemical modifications in the plant polyphenolic profile distribution were found after a principal components analysis (PCA) loading plots study. Four compounds hosting strong antioxidant properties - ferulic acid, asiatic acid, carnosol, and vanillin - were related to mycorrhizal rosemary plants while caffeic and chlorogenic acids had a higher influence on non-mycorrhizal plants. CONCLUSION: Mycorrhization was found to stimulate growth to obtain a higher biomass of plant leaves in a short time, avoiding chemical fertilization, while analytical results demonstrate that there is an alteration in the distribution of polyphenols in plants colonized by the symbiotic fungus, which can be related to an improvement in nutritional properties with future industrial significance. © 2018 Society of Chemical Industry.

Toxic effects of bisphenol A diglycidyl ether and derivatives in human placental cells.

BADGE (bisphenol A diglycidyl ether) is a synthesis product of bisphenol A (BPA), which, like other plasticizers, can cross the human placenta and reach the foetus. However, compared to BPA, there is almost no toxicological information. This work investigates the toxicity, endocrine and lipid disruption potential of BADGE and its hydrolysed and chlorinated derivatives (BADGE·H2O and BADGE·2HCl) in human placental JEG-3 cells. The analysis of culture medium by HPLC-ESI(+)-QqQ evidenced a good bioavailability of BADGE·2HCl and BADGE·H2O, but low stability of BADGE. Regardless, BADGE·2HCl and BADGE showed higher cytotoxicity than BADGE·H2O, which was the only compound that significantly inhibited CYP19 activity (IC50 49 ±â€¯5 µM). JEG-3 cells lipidome analyzed by FIA-ESI(+/-)-Orbitrap was significantly altered by exposure to BADGE·2HCl and BADGE at concentrations at the low µM range. BADGE·2HCl lead to a strong decrease of diacyl- and triacyl-glycerides (DGs,TGs) together with some membrane lipids, while BADGE lead to an accumulation of TGs. The results evidence the ability of BADGE and derivatives to affect placental lipid handling and to modulate placental CYP19 activity (BADGE·H2O) and highlights the need to monitor human exposure to these compounds, at least as intensely as BPA is monitored.

Negative-ion atmospheric pressure ionisation of semi-volatile fluorinated compounds for ultra-high-performance liquid chromatography tandem mass spectrometry analysis.

In this work, the feasibility of negative-ion atmospheric pressure chemical ionisation (APCI) and atmospheric pressure photoionisation (APPI) for ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) determination of fluorotelomer alcohols (FTOHs), fluorinated octanesulfonamides (FOSAs) and fluorinated octanesulfonamido-ethanols (FOSEs) was evaluated. The study of the effect of mobile phase composition on the atmospheric pressure ionisation of these compounds indicated that methanol/water mixtures provided the best responses in APCI, while acetonitrile/water with a post-column addition of toluene as dopant was the most appropriated mixture in APPI. Under the optimal working conditions, most of the target compounds produced the ion [M-H]- as base peak, although in-source collision-induced dissociation fragment ions in APCI and APPI and superoxide adduct ions [M+O2]-• in APPI were also present. These ions proved to be more useful as precursor ions for MS/MS determination than the adduct ions generated in electrospray. Although the UHPLC-APCI-MS/MS method allowed the determination of these semi-volatile compounds at low concentration levels, the analysis by UHPLC-APPI-MS/MS provided the lowest limits of detection and it was applied to the analysis of water samples in combination with solid-phase extraction. Quality parameters demonstrated the good performance of the proposed method, providing low method limits of detection (0.3-6 ng L-1), good precision (RSD % < 5%) and an accurate quantification (relative error % < 14%). Among the river water samples analysed by the developed method, 4:2 FTOH and N-EtFOSA were determined at 30 and 780 ng L-1, respectively.

Metabolomic analysis of the effects of cadmium and copper treatment in Oryza sativa L. using untargeted liquid chromatography coupled to high resolution mass spectrometry and all-ion fragmentation.

While the knowledge of plant metabolomes has increased in the last few years, their response to the presence of toxicants is still poorly understood. Here, we analyse the metabolomic changes in Japanese rice (Oryza sativa var. Japonica) upon exposure to heavy metals (Cd(ii) and Cu(ii)) in concentrations from 10 to 1000 µM. After harvesting, rice metabolites were extracted from aerial parts of the plants and analysed by HPLC (HILIC TSK gel amide-80 column) coupled to a mass spectrometer quadrupole-Orbitrap (Q-Exactive). Full scan and all ion fragmentation (AIF) mass spectrometry modes were used during the analysis. The proposed untargeted metabolomics data analysis strategy is based on the application of the multivariate curve resolution alternating least squares (MCR-ALS) method for feature detection, allowing the simultaneous resolution of pure chromatographic profiles and mass spectra of all metabolites present in the analysed rice extracts. All-ion fragmentation data were used to confirm the identification of MCR-ALS resolved metabolites. A total of 112 metabolites were detected, and 97 of them were subsequently identified and confirmed. Pathway analysis of the observed metabolic changes suggested an underlying similarity of the responses of the plant to Cd(ii) and Cu(ii), although the former treatment appeared to be the more severe of the two. In both cases, secondary metabolism and amino acid-, purine-, carbon- and glycerolipid-metabolism pathways were affected, in a pattern consistent with reduction in plant growth and/or photosynthetic capacity and with induction of defence mechanisms to reduce cell damage.

Liquid chromatography coupled to tandem and high resolution mass spectrometry for the characterisation of ofloxacin transformation products after titanium dioxide photocatalysis.

The characterization of pharmaceutical drugs and their transformation products have become an important analytical research field because its presence in the environment could induce bacterial resistance. Despite all efforts made by the scientific community, detection and structure identification of unknown chemicals still remains the most challenging task in non-targeted analytics. Given that, the objective of the present study was to develop an untargeted workflow to detect, quantify, identify and characterize ofloxacin and its transformation products (OFX TPs) after photocatalytic treatments based on TiO2 nanoparticles and TiO2 nanofibers. For the characterization and chemical structure assignment of OFX TPs, mass defect filters, mass accurate measurements (HRMS), tandem mass spectrometry in a q-Orbitrap (MS/HRMS) and the photocatalysis of the isotopically labelled ofloxacin (OFX-d3) were used. Since a large set of data was obtained in each run, data treatment based on statistical analysis and mass defect filtering was used to reduce the number of potential TP candidates from 2497 m/z peaks to 70. Moreover, ions generated by in-source CID and by redox reactions in the electrospray source (ESI) were also detected and discarded from the TP candidate list. Moreover, the whole kinetics evolution of the generated TPs provided a deeper insight into the degradation mechanism and was used to propose a degradation pathway for the OFX in the aqueous phase. The time evolution of the TPs generated during the photocatalytic process using both types of catalysts (NPs and NFs) and different set-ups (suspended and supported conditions) indicated that OFX was completely removed from the aqueous solution in less than 4h. Among the condition tested TiO2 nanoparticles in suspended conditions showed the fastest kinetics (k: 0.161 min(-1)).

Ion-molecule adduct formation in tandem mass spectrometry.

Nowadays most LC-MS methods rely on tandem mass spectrometry not only for quantitation and confirmation of compounds by multiple reaction monitoring (MRM), but also for the identification of unknowns from their product ion spectra. However, gas-phase reactions between charged and neutral species inside the mass analyzer can occur, yielding product ions at m/z values higher than that of the precursor ion, or at m/z values difficult to explain by logical losses, which complicate mass spectral interpretation. In this work, the formation of adduct ions in the mass analyzer was studied using several mass spectrometers with different mass analyzers (ion trap, triple quadrupole, and quadrupole-Orbitrap). Heterocyclic amines (AαC, MeAαC, Trp-P-1, and Trp-P-2), photo-initiators (BP and THBP), and pharmaceuticals (phenacetin and levamisole) were selected as model compounds and infused in LCQ Classic, TSQ Quantum Ultra AM, and Q-Exactive Orbitrap (ThermoFisher Scientific) mass spectrometers using electrospray as ionization method. The generation of ion-molecule adducts depended on the compound and also on the instrument employed. Adducts with neutral organic solvents (methanol and acetonitrile) were only observed in the ion trap instrument (LCQ Classic), because of the ionization source on-axis configuration and the lack of gas-phase barriers, which allowed inertial entrance of the neutrals into the analyzer. Adduct formation (only with water) in the triple quadrupole instruments was less abundant than in the ion trap and quadrupole-Orbitrap mass spectrometers, because of the lower residence time of the reactive product ions in the mass analyzer. The moisture level of the CID and/or damper gas had a great effect in beam-like mass analyzers such as triple quadrupole, but not in trap-like mass analyzers, probably because of the long residence time that allowed adduct formation even with very low concentrations of water inside the mass spectrometer.