Ultrahigh Resolution Mass Spectrometry Research Articles

Rapid Generation of Microplastics and Plastic-Derived Dissolved Organic Matter from Food Packaging Films under Simulated Aging Conditions.

In this study, we show that low-density polyethylene films, a prevalent choice for food packaging in everyday life, generated high numbers of microplastics (MPs) and hundreds to thousands of plastic-derived dissolved organic matter (DOM) substances under simulated food preparation and storage conditions. Specifically, the plastic film generated 66-2034 MPs/cm2 (size range 10-5000 μm) under simulated aging conditions involving microwave irradiation, heating, steaming, UV irradiation, refrigeration, freezing, and freeze-thaw cycling alongside contact with water, which were 15-453 times that of the control (plastic film immersed in water without aging). We also noticed a substantial release of plastic-derived DOM. Using ultrahigh-resolution mass spectrometry, we identified 321-1414 analytes with molecular weights ranging from 200 to 800 Da, representing plastic-derived DOM containing C, H, and O. The DOM substances included both degradation products of polyethylene (including oxidized forms of oligomers) and toxic plastic additives. Interestingly, although no apparent oxidation was observed for the plastic film under aging conditions, plastic-derived DOM was more oxidized (average O/C increased by 27-46%) following aging with a higher state of carbon saturation and higher polarity. These findings highlight the future need to assess risks associated with MP and DOM release from plastic wraps.

Characterisation of dissolved organic matter in two contrasting arsenic-prone sites in Kandal Province, Cambodia

Aquifers throughout Asia are impacted by the release of geogenic arsenic (As) into groundwater by microbial reduction of As-bearing Fe(III) (oxy)hydroxide minerals, severely impacting water quality. Groundwater dissolved organic matter (DOM) is likely key to As release, mainly as electron donor or electron shuttles. This study used optical analyses and ultra-high resolution mass spectrometry to examine the sources and composition of groundwater DOM in the As-prone aquifers of Kandal Province, Cambodia, at boreholes with differing host lithology (clay- and sand-dominated). Groundwater and surface water DOM composition were related to As concentrations, to infer the potential role of DOM in promoting As release. Optical and molecular-level analyses indicated an overall dominance of terrestrial-derived DOM in the groundwater samples, with higher freshness index and relative abundance (RA) of aliphatic compounds in clay compared to sand-dominated lithology. Compared to surface water, groundwater DOM had relatively lower O/C ratios and nominal oxidation state of carbon (−0.19 to −0.13 compared to 0.04 for ground and surface water, respectively), with a lower %RA of aliphatic compounds and higher %RA of carboxyl-rich alicyclic molecules, suggesting microbial processing of DOM since percolation into the aquifer. Concentrations of As across both sites were negatively correlated with DOM tryptophan:fulvic-like fluorescence and the %RA of aliphatics, potentially indicating microbial degradation of biolabile DOM in connection with As release, which is consistent with its role as an electron donor source. Together these data support DOM composition as an important control on microbial mediated As release.

Annotation of DOM metabolomes with an ultrahigh resolution mass spectrometry molecular formula library

Increased accessibility of liquid chromatography mass spectrometry (LC-MS) metabolomics instrumentation and software have expanded their use in studies of dissolved organic matter (DOM) and exometabolites released by microbes. Current strategies to annotate metabolomes generally rely on matching tandem MS/MS spectra to databases of authentic standards. However, spectral matching approaches typically have low annotation rates for DOM. An alternative approach is to annotate molecular formula based on accurate mass and isotopic fine structure measurements that can be obtained from state-of-the-art ultrahigh resolution Fourier Transform Ion Cyclotron Resonance mass spectrometry (FT-ICR MS), but instrument accessibility for large metabolomic studies is generally limited. Here, we describe a strategy to annotate exometabolomes obtained from lower resolution LC-MS systems by matching metabolomic features to a molecular formula library generated for a representative sample analyzed by LC 21T- FT-ICR MS. The molecular formula library approach successfully annotated 53% of exometabolome features of the marine diatom Phaeodactylum tricornutum – a nearly ten-fold increase over the 6% annotation rate achieved using a conventional MS/MS approach. There was 94% agreement between assigned formula that were annotated with both approaches, and mass error analysis of the discrepancies suggested that the FT-ICR MS formula assignments were more reliable. Differences in the exometabolome of P. tricornutum grown under iron replete and iron limited conditions revealed 668 significant metabolites, including a suite of peptide-like molecules released by P. tricornutum in response to iron deficiency. These findings demonstrate the utility of FT-ICR MS formula libraries for extending the accuracy and comprehensiveness of metabolome annotations.

Photoinduced Evolutions of Permafrost-Derived Carbon in Subarctic Thermokarst Pond Surface Waters.

In subarctic regions, rising temperature and permafrost thaw lead to the formation of thermokarst ponds, where organics from eroding permafrost accumulate. Despite its environmental significance, limited knowledge exists regarding the photosensitivity of permafrost-derived carbon in these ponds. In this study, laboratory experiments were conducted to explore the photochemical transformations of organic matter in surface water samples from thermokarst ponds from different environments in northern Quebec, Canada. One pond near Kuujjuarapik is characterized by the presence of a collapsing palsa and is therefore organically rich, while the other pond near Umiujaq is adjacent to a collapsing lithalsa and thus contains fewer organic matters. Photobleaching occurred in the Umiujaq sample upon irradiation, whereas the Kuujjuarapik sample exhibited an increase in light absorbance at wavelength related to aromatic functionalities, indicating different photochemical aging processes. Ultrahigh-resolution mass spectrometry analysis reveals that the Kuujjuarapik sample preferentially photoproduced highly unsaturated CHO compounds with great aromaticity, while the irradiated Umiujaq sample produced a higher proportion of CHON aromatics with reduced nitrogen functionalities. Overall, this study illustrates that the photochemical reactivity of thermokarst pond water varies with the source of organic matter. The observed differences in reactivity contribute to an improved understanding of the photochemical emission of volatile organic compounds discovered earlier. Further insights into the photoinduced evolutions in thermokarst ponds may require the classification of permafrost-derived carbon therein.

Arsenic redox disequilibrium in geogenic contaminated groundwater: Bioenergetic insights from organic molecular characterization and gene-informed modeling

Biotransformation of arsenic (As) influences its speciation and mobility, obscuring mechanistic comprehension on spatiotemporal variation of As concentration in geogenic contaminated groundwater. In particular, unresolved processes underlying As redox disequilibrium in comparison to major redox couples discourage practical efforts to rehabilitate the As-contaminated groundwater. Here, quantitative metagenomic sequencing and ultrahigh-resolution mass spectrometry (FT-ICR-MS) were jointly applied to reveal the links between vertical distribution of As metabolic gene assemblages and that of free energy density of dissolved organic matter (DOM) in As-contaminated groundwater of Datong Basin. Observed small excess of Gibbs free energy available by DOM relative to that required for As(V)-to-As(III) reduction exerts thermodynamic constraint on metabolism-mediated redox transformation of As. Accordingly, the vertical distribution of dissolved As(V)/As(III) ratio correlated significantly with that of ars+acr3 and arr encoding As(V) reduction and aio encoding As(III) oxidation in the moderately/strongly reducing groundwater. Further gene-informed biogeochemical modeling suggests that a net effect of these kinetics-restricted bidirectional metabolic pathways leads to co-preservation of As(V) and As(III) even at relatively high rates of ars+acr3 encoded As(V) reduction. This study therefore provides new insights into bioenergetic constraints on As hydrobiogeochemical behavior, with implications for other redox-sensitive contaminants in the groundwater systems.

Molecular-level insight into the effect of fertilization regimes on the chemodiversity of dissolved organic matter in tropical cropland

Fertilization is a critical agronomic measure for croplands in tropical regions, owing to their low fertility. However, the effects of fertilization on the quantity and chemodiversity of latosolic dissolved organic matter (DOM) in tropical regions remain largely unknown. Therefore, in this study, the variations in latosol DOM concentrations and chemodiversity induced by inorganic fertilization and the co-application of inorganic fertilization with straw return, sheep manure, biochar, and vermicompost fertilizers at a molecular level were systematically investigated using multispectral techniques and ultrahigh-resolution mass spectrometry. In line with our expectations, the results showed that combined inorganic-organic fertilization improved soil quality by increasing soil organic carbon content compared to that under inorganic fertilization. However, as the most active and bioavailable organic carbon pool, dissolved organic carbonconcentrations between the fertilization treatments were not significantly different (p = 0.07). However, the dissolved organic carbon concentrations under combined inorganic-organic fertilization treatment (NPK plus straw return, 263.45 ± 37.51 mg/kg) were lower than those under inorganic fertilization treatment (282.10 ± 18.57 mg/kg). Spectral analysis showed that the DOM in the combined inorganic-organic fertilization treatments had a higher degree of humification and lower autogenetic contributions. Furthermore, Fourier transform ion cyclotron resonance mass spectrometry analysis indicated that the combined inorganic-organic fertilization increased the chemodiversity of latosolic DOM and promoted the production of large, oxidized, and stable molecules, including lignin, aromatic, and tannin compounds, which potentially benefits soil carbon sequestration in tropical regions. This study could provide a theoretical basis for elucidating on the potentially relevant ecological functions and environmental effects of DOM under fertilization regimes.

Husk Separation (Kubessa Method) Impacts the Aging Chemistry of Beer.

The removal of husks before the mashing process, also known as the Kubessa method, is an established brewing practice often positively associated with smoothness and better flavor-stability of beer. Empirical evidence on the effect of the Kubessa method on beer, however, has been lacking. Similarly, our study's comprehensive analysis of established brewing attributes revealed that traditional methods do not fully capture the impact of husk separation in beer brewing. Conclusive evidence of the Kubessa method's impact on beer aging chemistry was obtained through ultrahigh resolution mass spectrometry (FT-ICR-MS), revealing intricate molecular details inaccessible to conventional analytical techniques. The compositional information on thousands of molecules in Kubessa beer was resolved and compared to whole malt mashing. Machine learning algorithms applied to aging experiments identified over 500 aging-related compounds inhibited by husk separation. Complementary Time of flight mass spectrometry (ToF-MS) coupled with chromatography further confirmed that the mashing of husks introduces sulfur-containing lipid compounds. These significant differences in the beer composition provide valuable insights for further investigation into the staling protective effect of husk-separation (Kubessa process) during beer production, as empirically demonstrated in this work.

Direct analysis by ultra-high-resolution mass spectrometry of lipid A and phospholipids from Acinetobacter baumannii cells

Acinetobacter baumannii, classified as priority number one by the World Health Organization (WHO), is an opportunistic pathogen responsible for infection and is able to develop antibiotic resistance easily. Membranes are bacteria's first line of defense against external aggression, such as antibiotics. A chemical modification of a lipid family or a change in lipid composition can lead to resistance to antibiotics. In this work, we analyzed different A. baumannii strains from various environments with different antibiotic resistance profiles, using matrix-assisted laser desorption ionization-Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FT-ICR MS). This study shows that it is possible to describe the main lipidome (phospholipids and lipid A) from the simple preparation of lysed cells, and that despite the complexity of the mixture. This ultra-high resolution mass spectrometry technique enables the separation of isobaric ion, to report a new class of lipids. Given its performance, this technique can be used to quickly and reliably characterize the lipidome of clinical strains from different environments.

Assessment of Accelerated Aging Effect of Bio-Oil Fractions Utilizing Ultrahigh-Resolution Mass Spectrometry and k-Means Clustering of van Krevelen Compositional Space.

Bio-oils contain a substantial number of highly oxygenated hydrocarbons, which often exhibit low thermal stability during storage, handling, and refining. The primary objectives of this study are to characterize the hydroxyl group in bio-oil fractions and to investigate the relationship between the type of hydroxyl group and accelerated aging behavior. A bio-oil was fractionated into five solubility-based fractions, classified in two main groups: water-soluble and water-insoluble fractions. These fractions were then subjected to chemoselective reactions to tag molecules containing hydroxyl groups and analyzed by negative-ion electrospray ionization 21 T Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). The fractions were also subjected to accelerated aging experiments and characterized by FT-ICR MS and bulk viscosity measurements. Extracting insightful information from ultrahigh-resolution data to aid in predicting upgrading methodologies and instability behaviors of bio-oils is challenging due to the complexity of the data. To address this, an unsupervised learning technique, k-means clustering analysis, was used to semiquantify molecular compositions with a close Euclidean distance within the (O/C, H/C) chemical space. The combination of k-means analysis with findings from chemoselective reactions allowed the distinctive hydroxyl functionalities across the samples to be inferred. Our results indicate that the hexane-soluble fraction contained numerous molecules containing primary and secondary alcohols, while the water-soluble fraction displayed diverse groups of oxygenated compounds, clustered near to carbohydrate-like and pyrolytic humin-like materials. Despite its high oxygen content, the water-soluble fraction showed minimal changes in viscosity during aging. In contrast, a significant increase in viscosity was observed in the water-insoluble materials, specifically, the low- and high-molecular-weight lignin fractions (LMWL and HMWL, respectively). Among these two fractions, the HMWL exhibited the highest increase in viscosity after only 4 h of accelerated aging. Our results indicate that this aging behavior is attributed to an increased number of molecular compositions containing phenolic groups. Thus, the chemical compositions within the HMWL are the major contributors to the viscosity changes in the bio-oil under accelerated aging conditions. This highlights the crucial role of oxygen functionality in bio-oil aging, suggesting that a high oxygen content alone does not necessarily correlate with an increase of viscosity. Unlike other bio-oil categorization methods based on constrained molecule locations within the van Krevelen compositional space, k-means clustering can identify patterns within ultrahigh-resolution data inherent to the unique chemical fingerprint of each sample.

A trapped ion mobility enabled Fourier transform ion cyclotron resonance mass spectrometer for infrared ion spectroscopy at FELIX

We report the installation of a new infrared ion spectroscopy platform at the free-electron laser facility FELIX, based on a Bruker SolariX Fourier Transform ion cyclotron resonance (FT-ICR) mass spectrometer equipped with a trapped ion mobility spectrometry (TIMS) stage. The instrument allows one to record infrared multiple-photon dissociation (IRMPD) spectra for mass and mobility selected ions, produced by a range of ion sources. We describe two strategies to achieve consistent overlap between the laser beam and the ion packet. Removing the original ECD cathode significantly enhanced the IR transmission and the overlap with the ion cloud, resulting in improved IRMPD yield per pulse. Enhancement of the photodissociation yield is observed when multiple pulses are used, as there is negligible collisional deactivation in the ultra-high vacuum trapping region of the ICR cell. We compare IRMPD spectra recorded on the new platform with IRMPD spectra of the same species recorded on one of our 3D-quadrupole ion trap platforms. We demonstrate the instrument’s performance using a sample containing a mixture of two trisaccharides. Mobility selection allows us to record individual IR spectra for the two isomeric species. This multi-modal platform, encompassing liquid chromatography, ion mobility spectrometry, ultra-high resolution (tandem) mass spectrometry, infrared ion spectroscopy (and soon also mass spectrometry imaging), is available to users at the FELIX facility.

An automated and high-throughput data processing workflow for PFAS identification in biota by direct infusion ultra-high resolution mass spectrometry

The increasing recognition of the health impacts from human exposure to per- and polyfluorinated alkyl substances (PFAS) has surged the need for sophisticated analytical techniques and advanced data analyses, especially for assessing exposure by food of animal origin. Despite the existence of nearly 15,000 PFAS listed in the CompTox chemicals dashboard by the US Environmental Protection Agency, conventional monitoring and suspect screening methods often fall short, covering only a fraction of these substances. This study introduces an innovative automated data processing workflow, named PFlow, for identifying PFAS in environmental samples using direct infusion Fourier transform ion cyclotron resonance mass spectrometry (DI-FT-ICR MS). PFlow’s validation on a bream liver sample, representative of low-concentration biota, involves data pre-processing, annotation of PFAS based on their precursor masses, and verification through isotopologues. Notably, PFlow annotated 17 PFAS absent in the comprehensive targeted approach and tentatively identified an additional 53 compounds, thereby demonstrating its efficiency in enhancing PFAS detection coverage. From an initial dataset of 30,332 distinct m/z values, PFlow thoroughly narrowed down the candidates to 84 potential PFAS compounds, utilizing precise mass measurements and chemical logic criteria, underscoring its potential in advancing our understanding of PFAS prevalence and of human exposure.Graphical abstract

Comparison of dissolved organic matter composition from various sorbents using ultra-high resolution mass spectrometry

Solid phase extraction (SPE) of a variety of diverse dissolved organic matter (DOM) endmembers through eight commercially available sorbents was examined (ENV, PLEXA, PPL, HLB, Isolute 101, C18/ENV+, C18, EnvirElut) representing styrene divinylbenzene polymer (SDVB) and silica-based sorbents. We assessed dissolved organic carbon (DOC) recovery and DOM composition via 21T Fourier transform-ion cyclotron resonance mass spectrometry (FT-ICR MS). DOC recoveries and SPE-DOM composition differed more by endmember type than by sorbent. Silica-based sorbents retained DOM with many N-containing formulae, while SDVB-based sorbents retained DOM with more S-containing formulae. Extraction pH exerted a greater influence on DOM composition, notably through the presence of strong groupings composed of saturated and lowly oxygenated formulae at basic pH, and of aromatic and highly oxygenated formulae at pH 2, irrespective of endmember or sorbent. There was above 25% DOC recovery, regardless of sorbent or endmember; >90% of the relative abundance (RA) of molecular formulae were shared with PPL, which is currently the most commonly utilized sorbent for DOM. This clearly highlights the ability of the selected sorbents to retain representative DOM across diverse endmembers. Such findings may be useful for future targeted DOM studies (e.g., bioincubations, wastewater and drinking water applications) interested in focusing on specific compositional changes and will provide a better understanding of how organic carbon cycling is impacted by anthropogenic processes.

Molecular features of uranium-binding natural organic matter in a riparian wetland determined by ultrahigh resolution mass spectrometry

Tims Branch riparian wetland located in South Carolina, USA has immobilized 94 % of the U released >50 years ago from a nuclear fuel fabrication facility. Sediment organic matter (OM) has been shown to play an important role in immobilizing U. Yet, uranium-OM-mineral interactions at the molecular scale have never been investigated at ambient concentrations. The objectives of this study were to extract, purify, and concentrate U-bound sediment OM along the stream water pathway and perform molecular characterization using Fourier transform ion cyclotron resonance mass spectrometry (FTICRMS). Out of 9614 identified formulas, 715 contained U. These U-containing formulas were enriched with Fe, N, and/or S compared to the total OM. Lignin-like and protein-like molecules accounted for 40 % and 19 % of the U-containing formulas, respectively. Phosphorus-containing formulas were found to exert an insignificant influence on complexing U. U-containing formulas in the ‘mobile’ (groundwater extractable) OM fraction had lower (reduced) nominal oxidation states of carbon (NOSC); and less aromatic moieties than OM recovered from the ‘immobile’ (sodium pyrophosphate extractable) OM fraction. U-containing formulas in the redox interfacial zones (stream banks) compared to those in nearby up-slope zones tended to have smaller molecular weights; lower NOSC; higher contents of COO and/or CONO functional groups; and higher abundance of Fe-containing formulas. Fe was present in 38 % of the U-containing formulas but only 20 % of the total OM formulas. It is postulated that Fe played an important role in stabilizing the structure of sedimentary OM, especially U-containing compounds. The identification for the first time of hundreds of Fe-U-OM formulas demonstrates the complexity of such system is much greater than commonly believed and numerically predicting U binding behavior in OM-rich systems may require greater use of statistical or artificial intelligence approaches rather than deterministic approaches limited to measuring metal complexation with well-defined individual analogue organic ligands.

Aqueous Photolysis of Water-Soluble Brown Carbon from Simulated Prescribed and Wildfire Biomass Burning.

This work, as part of the Georgia Wildland fire Simulation Experiment (G-WISE) campaign, explores the aqueous photolysis of water-soluble brown carbon (W-BrC) in biomass burning aerosols from the combustion of fuel beds collected from three distinct ecoregions in Georgia: Piedmont, Coastal Plain, and Blue Ridge. Burns were conducted under conditions representative of wildfires, which are common unplanned occurrences in Southeastern forests (low fuel moisture content), and prescribed fires, which are commonly used in forest management (higher fuel moisture content). Upon exposure to radiation from UV lamps equivalent to approximately 5 h in the atmosphere, the absorption spectra of all six samples exhibited up to 40% photobleaching in the UV range (280-400 nm) and as much as 30% photo-enhancement in the visible range (400-500 nm). Together, these two effects reduced the absorption Ångström exponent (AAE), a measure of the wavelength dependence of the spectrum, from 6.0-7.9 before photolysis to 5.0-5.7 after. Electrospray ionization ultrahigh-resolution mass spectrometry analysis shows the potential formation of oligomeric chromophores due to aqueous photolysis. This work provides insight into the impacts that aqueous photolysis has on W-BrC in biomass burning aerosols and its dependence on fuel bed composition and moisture content.

A Gated TIMS FTICR MS Instrument to Decipher Isomeric Content of Complex Organic Mixtures.

Modern research faces increasingly complex materials with a constant need for new analytical strategies that can provide deeper levels of chemical insight. Ultrahigh resolution mass spectrometry (MS), particularly Fourier transform ion cyclotron resonance (FTICR) MS, has provided a robust analytical foundation. However, MS alone offers limited structural information. Here, we present the first implementation and results from an FTICR MS with fully integrated dual accumulation analysis with gated trapped ion mobility spectrometry (gTIMS) capability. The drastically extended charge capacity and parallel accumulation facilitate the analysis of complex mixtures. We achieved a high dynamic range of 4 orders of magnitude within a single FTICR acquisition event. Simultaneously, the valuable linear relationship between the TIMS elution voltage and reduced mobility was retained over a wide mobility range. Benchmarking the instrument performance with Suwannee River fulvic acid (SRFA) by variable ramp gTIMS analysis allowed separation and unambiguous assignment of different charge state distributions. Application to bio-oils has proven the capability to distinguish the isomeric diversity in these ultracomplex samples, while maintaining the expected FTICR MS resolving power and mass accuracy. Valuable information about the molecular distribution, isomeric diversity, and main molecular differences could directly be extracted within the analysis time of a classical "dilute and shoot" direct infusion experiment. The development of this fully integrated and flexible gTIMS with FTICR MS analysis possesses the potential to significantly change the current landscape of high-resolution mass spectrometric analysis of complex mixtures through the added insight of isomeric complexity afforded by TIMS. The exploration of the added IMS dimension promises transformative effects across diverse fields including energy transition, environmental studies, and biological research.

Plasma metabolomic and lipidomic profiles accurately classify mothers of children with congenital heart disease: an observational study

IntroductionCongenital heart disease (CHD) is the most common congenital anomaly, representing a significant global disease burden. Limitations exist in our understanding of aetiology, diagnostic methodology and screening, with metabolomics offering promise in addressing these.ObjectiveTo evaluate maternal metabolomics and lipidomics in prediction and risk factor identification for childhood CHD.MethodsWe performed an observational study in mothers of children with CHD following pregnancy, using untargeted plasma metabolomics and lipidomics by ultrahigh performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS). 190 cases (157 mothers of children with structural CHD (sCHD); 33 mothers of children with genetic CHD (gCHD)) from the children OMACp cohort and 162 controls from the ALSPAC cohort were analysed. CHD diagnoses were stratified by severity and clinical classifications. Univariate, exploratory and supervised chemometric methods were used to identify metabolites and lipids distinguishing cases and controls, alongside predictive modelling.Results499 metabolites and lipids were annotated and used to build PLS-DA and SO-CovSel-LDA predictive models to accurately distinguish sCHD and control groups. The best performing model had an sCHD test set mean accuracy of 94.74% (sCHD test group sensitivity 93.33%; specificity 96.00%) utilising only 11 analytes. Similar test performances were seen for gCHD. Across best performing models, 37 analytes contributed to performance including amino acids, lipids, and nucleotides.ConclusionsHere, maternal metabolomic and lipidomic analysis has facilitated the development of sensitive risk prediction models classifying mothers of children with CHD. Metabolites and lipids identified offer promise for maternal risk factor profiling, and understanding of CHD pathogenesis in the future.

Accessing the Low-Polar Molecular Composition of Boreal and Arctic Peat-Burning Organic Aerosol via Thermal Analysis and Ultrahigh-Resolution Mass Spectrometry: Structural Motifs and Their Formation.

Peatland fires emit organic carbon-rich particulate matter into the atmosphere. Boreal and Arctic peatlands are becoming more vulnerable to wildfires, resulting in a need for better understanding of the emissions of these special fires. Extractable, nonpolar, and low-polar organic aerosol species emitted from laboratory-based boreal and Arctic peat-burning experiments are analyzed by direct-infusion atmospheric pressure photoionization (APPI) ultrahigh-resolution mass spectrometry (UHRMS) and compared to time-resolved APPI UHRMS evolved gas analysis from the thermal analysis of peat under inert nitrogen (pyrolysis) and oxidative atmosphere. The chemical composition is characterized on a molecular level, revealing abundant aromatic compounds that partially contain oxygen, nitrogen, or sulfur and are formed at characteristic temperatures. Two main structural motifs are identified, single core and multicore, and their temperature-dependent formation is assigned to the thermal degradation of the lignocellulose building blocks and other parts of peat.

Revealing Molecular Structures of Nitrogen-Containing Compounds in Dissolved Black Carbon Using Ultrahigh-Resolution Mass Spectrometry Combined with Thermodynamic Calculations.

Landscape wildfires generate a substantial amount of dissolved black carbon (DBC) annually, yet the molecular nitrogen (N) structures in DBC are poorly understood. Here, we systematically compared the chemodiversity of N-containing molecules among three different DBC samples from rice straw biochar pyrolyzed at 300, 400, and 500 °C, one leached dissolved organic carbon (LDOC) sample from composted rice straw, and one fire-affected soil dissolved organic matter (SDOMFire) sample using Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR MS). N-Containing molecules contributed 20.0%, 36.1%, and 43.7% of total compounds in Combined DBC (pooling together the three DBC), LDOC, and SDOMFire, respectively, and molecules with fewer N atoms had higher proportions (i.e., N1 > N2 > N3). The N-containing molecules in Combined DBC were dominated by polycyclic aromatic (62.2%) and aromatic (14.4%) components, while those in LDOC were dominated by lignin-like (50.4%) and aromatic (30.1%) components. The composition and structures of N-containing molecules in SDOMFire were more similar to those in DBC than in LDOC. As the temperature rose, the proportion of the nitrogenous polycyclic aromatic component in DBC significantly increased with concurrent enhanced oxidation and unsaturation of N. As indicated by density functional theory (DFT)-based thermodynamic calculations, the proportion of aliphatic amide N decreased from 23.2% to 7.9%, whereas that of nitroaromatic N increased from 10.0% to 39.5% as the temperature increased from 300 to 500 °C; alternatively, the proportion of aromatic N in the 5/6 membered ring remained relatively stable (∼31%) and that of aromatic amide N peaked at 400 °C (32.7%). Our work first provides a comprehensive and thorough description of molecular N structures of DBC, which helps to better understand and predict their fate and biogeochemical behavior.

The Hunt for Chemical Dark Matter across a River-to-Ocean Continuum.

Thousands of mass peaks emerge during molecular characterization of natural dissolved organic matter (DOM) using ultrahigh-resolution mass spectrometry. While mass peaks assigned to certain molecular formulas have been extensively studied, the uncharacterized mass peaks that represent a significant fraction of organic matter and convey biogenic elements and energy have been previously ignored. In this study, we introduce the term dark DOM (DDOM) for unassigned mass peaks and have explored its characteristics and environmental behaviors using a data set of 38 DOM extracts covering the Yangtze River-to-ocean continuum. We identified a total of 9141 DDOM molecules, which exhibited higher molecular weight and greater diversity than the DOM subset with assigned DOM formulas. Although DDOM contributed a smaller fraction of relative abundance, it significantly impacted the molecular weight and molecular composition of bulk DOM. A portion of DDOM with higher molecular weight was found to increase molecular abundance across the river-to-ocean continuum. These compounds could contain halogenated organic molecules and might have a high potential to contribute to the refractory organic carbon pool. With this study, we underline the contribution of dark matter to the total DOM pool and emphasize that more DDOM research is needed to understand its contribution to global biogeochemical cycles and carbon sequestration.

Detection and Exclusion of False-Positive Molecular Formula Assignments via Mass Error Distributions in UHR Mass Spectra of Natural Organic Matter.

Ultrahigh resolution mass spectrometry (UHRMS) routinely detects and identifies thousands of mass peaks in complex mixtures, such as natural organic matter (NOM) and petroleum. The assignment of several chemically plausible molecular formulas (MFs) for a single accurate mass still poses a major problem for the reliable interpretation of NOM composition in a biogeochemical context. Applying sensible chemical rules for MF validation is often insufficient to eliminate multiple assignments (MultiAs)─especially for mass peaks with low abundance or if ample heteroatoms or isotopes are included - and requires manual inspection or expert judgment. Here, we present a new approach based on mass error distributions for the identification of true and false assignments among MultiAs. To this end, we used the mass error in millidalton (mDa), which was superior to the commonly used relative mass error in ppm. We developed an automatic workflow to group MultiAs based on their shared formula units and Kendrick mass defect values and to evaluate the mass error distribution. In this way, the number of valid assignments of chlorinated disinfection byproducts was increased by 8-fold as compared to only applying 37Cl/35Cl isotope ratio filters. Likewise, phosphorus-containing MFs can be differentiated against chlorine-containing MFs with high confidence. Further, false assignments of highly aromatic sulfur-containing MFs ("black sulfur") to sodium adducts in negative ionization mode can be excluded by applying our approach. Overall, MFs for mass peaks that are close to the detection limit or where naturally occurring isotopes are rare (e.g., 15N) or absent (e.g., P and F) can now be validated, substantially increasing the reliability of MF assignments and broadening the applicability of UHRMS analysis to even more complex samples and processes.