Dissolved Organic Matter Samples Research Articles

Post column infusion of an internal standard into LC-FT-ICR MS enables semi-quantitative comparison of dissolved organic matter in original samples.

Ultrahigh resolution mass spectrometry hyphenated with liquid chromatography (LC) is an emerging tool to explore the isomeric composition of dissolved organic matter (DOM). However, matrix effects limit the potential for semi-quantitative comparison of DOM molecule abundances across samples. We introduce a post-column infused internal standard (PCI-IS) for reversed-phase LC-FT-ICR MS measurements of DOM and systematically evaluate matrix effects, detector linearity and the precision of mass peak intensities. Matrix effects for model compounds spiked into freshwater DOM samples ranging from a headwater stream to a major river were reduced by 5-10% for PCI-IS corrected mass peak intensities as compared to raw (i.e., untransformed) intensities. A linear regression of PCI-IS corrected DOM mass peak intensities across a typical DOM concentration range (2-15 mg dissolved organic carbon L-1) in original, non-extracted freshwater samples demonstrates excellent linearity of the detector response (r2 > 0.9 for 98% of detected molecular formulas across retention times). Importantly, PCI-IS could compensate for 80% of matrix effects across an environmental gradient of DOM composition from groundwater to surface water. This enabled studying the ionization efficiency of DOM isomers and linking the observed differences to the biogeochemical sources. With PCI-IS original, non-extracted DOM samples can be analysed by LC-FT-ICR MS without carbon load adjustment, and mass peak intensities can be reliably used to semi-quantitatively compare isomer abundances between compositionally similar DOM samples.

Characteristics of DOM and Their Relationships with Potentially Toxic Elements in the Inner Mongolia Section of the Yellow River, China.

The characterization of dissolved organic matter (DOM) is important for better understanding of the migration and transformation mechanisms of DOM in water bodies and its interaction with other contaminants. In this work, fluorescence characteristics and molecular compositions of the DOM samples collected from the mainstream, tributary, and sewage outfall of the Inner Mongolia section of the Yellow River (IMYR) were determined by using fluorescence spectroscopy and Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). In addition, concentrations of potentially toxic elements (PTEs) in the relevant surface water and their potential relationships with DOM were investigated. The results showed that the abundance of tyrosine-like components increased significantly in downstream waters impacted by outfall effluents and was negatively correlated with the humification index (HIX). Compared to the mainstream, outfall and tributaries have a high number of molecular formulas and a higher proportion of CHOS molecular formulas. In particular, the O5S class has a relative intensity of 41.6% and the O5-7S class has more than 70%. Thirty-eight PTEs were measured in the surface water samples, and 12 found above their detective levels at all sampling sites. Protein-like components are positively correlated with Cu, which is likely indicating the source of Cu in the aquatic environment of the IMYR. Our results demonstrated that urban wastewater discharges significantly alter characteristics and compositions of DOM in the mainstream of IMYR with strongly anthropogenic features. These results and conclusions are important for understanding the role and sources of DOM in the Yellow River aquatic environment.

Spectroscopic studies of the role of dissolved organic matter in acenaphthene photodegradation in liquid water and ice

The effect of concentration and origin of dissolved organic matter (DOM) on acenaphthene (Ace) photodegradation in liquid water and ice was investigated, and the components in DOM which were involved in Ace photodegradation were identified. The DOM samples included Suwannee River fulvic acid (SRFA), Elliott soil humic acid (ESHA), and an effluent organic matter (EfOM) sample. Due to the production of hydroxyl radical (•OH) and triplet excited-state DOM (3DOM*) which react with Ace, DOM had promotion effects on Ace photodegradation. However, the promotion effects of DOM were prevailed over by their suppressing effect of DOM including screening light effect, intermediates reducing effect and RS quenching effect, and thus, the photodegradation rates of Ace decreased in the presence of the three DOM with concentrations of 0.5–7.5 mg C/L in liquid water and ice. ESHA had higher light absorption and thus had higher screening light effect on Ace photodegradation in liquid water than SRFA and EfOM. At each DOM concentration, ESHA exhibited higher promotion effect on Ace photodegradation than SRFA and EfOM, in liquid water and ice. The binding of Ace with DOM was indicated by decreases in fluorescence intensity of Ace when coexisted with DOM. However, the binding of Ace to DOM played an unimportant role in suppressing Ace photodegradation. The photodegradation behavior of fluorophores in Ace with DOM present in ice was not similar to that in liquid water. C–O, C═O, carboxyl groups O–H and aliphatic C–H functional groups in DOM were involved in the interaction of DOM with Ace. The presence of Ace seemed to have no influence on the photodegradation behavior of functional groups in DOM.

Biochar mediates nitrogen investment strategy involved in Chinese fir growth as revealed by molecular information on soil dissolved organic matter

Soil dissolved organic matter (DOM) is the most reactive pool of soil organic matter. Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) is a powerful tool for obtaining molecular-level DOM information. The ongoing atmospheric nitrogen (N) deposition could cause some negative effects on tropical and subtropical forest ecosystems, and biochar is considered to be an effective soil ameliorator. However, few studies have evaluated the effects of biochar and N deposition, either separately or in combination, on the molecular composition of soil DOM. In this study, we conducted a pot experiment using Chinese fir seedlings amended with biochar (0, 12, and 36 t biochar ha−1, prepared from corn stover at 450 °C) and inorganic N (NH4NO3: 0, 40, and 80 kg N ha−1 y-1) in a full factorial design (four replications per treatment) and characterized soil DOM using FT-ICR-MS. A total of 6084 types of formulas were identified in this study. All 36 samples had 2109 types of formulas in common, which had much lower N/C and P/C ratios than the bulk DOM samples. Permutation-based multivariate analysis of variance revealed that both N addition and biochar application had a significant impact on DOM chemodiversity. N addition increased the relative abundance of lignins and lipids, which may be a way for Chinese fir to acclimate to an acidic environment by expanding roots and fixing N. Biochar amendment increased the relative abundance of carbohydrates and tannins in soil DOM at each N application level. Soil pH is a key soil variable controlling DOM composition, and an increase in pH after biochar amendment contributed greatly to the occurrence of DOM compounds with an O/C > 0.5, which are easily degradable. In particular, lignins associated with low N/C were more abundant at higher pH. Biochar application also improved the efficiency of roots in acquiring nutrients, reduced the relative biomass of roots, and decreased the relative content of lignins or lipids, which primarily originated from the roots. In summary, biochar amendment effectively alleviated the negative effects of N addition by investing more N into the production of easily degradable DOM compounds and stimulating the biogeochemical cycle of N.

Toward a More Comprehensive Approach for Dissolved Organic Matter Chemical Characterization Using an Orbitrap Fusion Tribrid Mass Spectrometer Coupled with Ion and Liquid Chromatography Techniques.

Dissolved organic matter (DOM) represents one of the largest active organic carbon pools in the global carbon cycle. Although extensively studied, only <10% of DOM has been chemically characterized into individual dissolved compounds due to its molecular complexity. This study introduced a more comprehensive DOM characterization method by coupling both ion chromatography (IC) and liquid chromatography (LC) with high mass accuracy and resolution mass spectrometry. We presented a new on-the-fly mass calibration of the Orbitrap technique by utilizing the "lock mass" function in the Orbitrap Fusion Tribrid mass spectrometer (OT-FTMS), which assures high mass accuracy at every scan by a postcolumn introduction of internal labeled standards. With both IC and LC, tested unlabeled standards of amino acids, small peptides, and organic acids were consistently below 1.0 ppm mass error, giving the OT-FTMS the potential of reaching mass accuracy of the Fourier-transform ion cyclotron resonance mass spectrometer. In addition to mass accuracy, a pooled quality control sample (QC) was used to increase reproducibility by applying systematic error removal using random forest (SERRF). Using an untargeted mass spectrometry approach, estuarine DOM samples were analyzed by OT-FTMS coupled to IC in negative mode and LC in positive mode detection to cover a wide range of highly cationic to highly anionic molecules. As a proof of concept, we focused on elucidating the structures of three distinct DOM compound classes with varied acidities and basicities. In UPLC-OT-FTMS, a total of 915 compounds were detected. We putatively elucidated 44 small peptides and 33 deaminated peptides of these compounds. With IC-OT-FTMS, a total of 1432 compounds were detected. We putatively elucidated 20 peptides, 268 deaminated peptides, and 188 organic acids. Except for five compounds, all putatively elucidated compounds were uniquely detected in their corresponding chromatography technique. These results highlight the need for combining these two techniques to provide a more comprehensive method for DOM characterization. Application of the combined IC and LC techniques is not limited to DOM chemical characterization. It can analyze other complex compound mixtures, such as metabolites, and anthropogenic pollutants, such as pesticides and endocrine-disrupting chemicals, in environmental and biological samples.

Dissolved organic matter promoted hydroxyl radical formation and phenanthrene attenuation during oxygenation of iron-pillared montmorillonites

The oxygenation of Fe(II)-bearing minerals for hydroxyl radicals (HO•) formation and contaminant attenuation receives increasing attentions. However, information on dissolved organic matter (DOM) with different types, concentrations, and molecular weights (MWs) in manipulating HO• formation and contaminant attenuation during mineral oxygenation remain unclear. In this study, four iron-pillared montmorillonites (IPMs) and two DOM samples [e.g., humic acids (HA) and fulvic acids (FA)] were prepared to explore the HO• formation and phenanthrene attenuation during the oxygenation of IPMs in the presence or absence of DOMs. Results showed that iron-pillared and high-temperature calcination procedures extended the interlayer domain of IPMs, which provided favorable conditions for a high HO• production from 1293 to 14537 μmol kg−1. The surface-absorbed/low crystalline Fe(Ⅱ) was the predominant Fe(Ⅱ) fractionations for HO• production, and presence of DOMs significantly enhanced the HO• production and phenanthrene attenuation. Moreover, regardless of the types and concentrations, the low MW (LMW, <1 kDa) fraction within DOM pool contributed highest to HO• production and phenanthrene attenuation, followed by the bulk and high MW (HMW-, 1 kDa∼0.45 μm) fractions, and FA exhibited more efficient effects in promoting HO• production and phenanthrene attenuation than HA. The fluorescent spectral analysis further revealed that phenolic-like fluorophores in LMW-fraction were the main substances responsible for the enhanced HO• production and phenanthrene attenuation. The results deepen our understandings toward the behaviors and fate of aquatic HO• and contaminants, and also provide technical guidance for the remediation of contaminated environments.

Stratification of dissolved organic matter in the upper 5000 m water column in the western Pacific Ocean

Marine dissolved organic matter (DOM) is a principal reservoir involved in biogeochemical cycles and exerts a pivotal influence on global carbon flux dynamics. In this study, excitation-emission matrix fluorescence spectroscopy combined with parallel factor analysis (EEM-PARAFAC) was conducted on 230 DOM samples collected from 21 sites between February and April 2022 in the Western Pacific Ocean (WPO). We identified five distinct fluorescence peaks (peaks B, T, A, C, and M), predominantly protein-like and humic-like components. These findings, marked by significant differences (p < 0.01) in fluorescence intensities and spectral indices, characterized the transformation of DOM with ocean depth, illustrating a transition from active to recalcitrant forms. Additionally, random forest analysis (RFA) identified depth as a key factor influencing marine dissolved organic carbon (DOC), with a 32.59% importance value. Correlations between hydrological and fluorescent parameters underscored the complexity of DOM sources and influencing processes. Overall, this work broadens our understanding of DOM variability in the upper 5000 m of the WPO, enhancing our knowledge of the marine environment's role in the global carbon cycle.

Molecular formula assignment of dissolved organic matter by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry using two non-targeted data processing approaches: A case study from recirculating aquaculture systems

BackgroundThe accumulation of dissolved organic matter (DOM) poses an issue in the management of the water quality from recirculating aquaculture systems (RAS), but its characterization is often not detailed enough to understand the DOM transformations in RAS. In this study, we investigated the application of two distinct non-targeted data processing approaches using ultra-performance liquid chromatography (UPLC) with quadrupole time-of-flight mass spectrometry (QTOF-MS) and two software with different algorithmic designs: PetroOrg and Progenesis QI to accurately characterize the molecular composition of DOM in RAS by UPLC-QTOF-MS. ResultsThe UPLC-QTOF-MS resolution in combination with PetroOrg and Progenesis QI software successfully assigned 912 and 106 unique elemental compositions, respectively, including compounds containing carbon, hydrogen, and oxygen (CHO) and nitrogen-containing CHO compounds (CHON), in the DOM samples from RAS. The results of these two distinct data processing approaches were consistent with the list of DOM formulas from RAS identified by higher resolution mass spectrometry techniques confirming their reliability. PetroOrg approach revealed only compositional information in the DOM samples from RAS, while Progenesis QI in addition to identifying new elemental compositions, increased their chemical space by giving information about their polarity and their possible key structures. DOM samples from RAS were found to be rich in unsaturated CHO compounds, with tentatively key structures of terpenoids with medium polarity indicating natural origins in their composition. The analysis also revealed probable structures of sucrose fatty acid esters and polyethylene glycol, indicating anthropogenic sources. Significance and NoveltyThe combination of these two non-targeted data processing approaches significantly improves the characterization of the complex mixture of DOM from RAS by UPLC-QTOF-MS reporting for the first time accurate DOM results in terms of its composition, while proposing its key structures. The presented methods can also be used to analyze different DOM samples with other HRMS techniques and software.

Competition between Dissolved Organic Matter and Freshwater Plankton Control Methylmercury Isotope Fractionation during Uptake and Photochemical Demethylation.

Isotope fractionation related to photochemical reactions and planktonic uptake at the base of the food web is a major uncertainty in the biological application of mercury (Hg) stable isotopes. In freshwater systems, it is unclear how competitive interactions among methylmercury (MeHg), dissolved organic matter (DOM), and phytoplankton govern the magnitude of mass-dependent and mass-independent fractionation. This study investigated how DOM alters rates of planktonic MeHg uptake and photodegradation and corresponding Hg isotope fractionation in the presence of freshwater phytoplankton species, Raphidocelis subcapitata. Outdoor sunlight exposure experiments utilizing R. subcapitata were performed in the presence of different DOM samples using environmentally relevant ratios of MeHg-DOM thiol groups. The extent of Δ199Hg in phytoplankton incubations (2.99‰ St. Louis River HPOA, 1.88‰ Lake Erie HPOA) was lower compared to paired abiotic control experiments (4.29 and 2.86‰, respectively) after ∼30 h of irradiation, resulting from cell shading or other limiting factors reducing the extent of photodemethylation. Although the Δ199Hg/Δ201Hg ratio was uniform across experiments (∼1.4), Δ199Hg/δ202Hg slopes varied dramatically (from -0.96 to 15.4) across incubations with R. subcapitata and DOM. In addition, no evidence of Hg isotope fractionation was observed within R. subcapitata cells. This study provides a refined examination of Hg isotope fractionation markers for key processes occurring in the lower food web prior to bioaccumulation, critical for accurately accounting for the photochemical processing of Hg isotopes across a wide spectrum of freshwater systems.

Proton Binding Characteristics of Dissolved Organic Matter Extracted from the North Atlantic.

Marine dissolved organic matter (DOM) presents key thermodynamic properties that are not yet fully constrained. Here, we report the distribution of binding sites occupied by protons (i.e., proton affinity spectra) and parametrize the median intrinsic proton binding affinities (log K̅H) and heterogeneities (m), for DOM samples extracted from the North Atlantic. We estimate that 11.4 ± 0.6% of C atoms in the extracted marine DOM have a functional group with a binding site for ionic species. The log K̅H of the most acidic groups was larger (4.01-4.02 ± 0.02) than that observed in DOM from coastal waters (3.82 ± 0.02), while the chemical binding heterogeneity parameter increased with depth to values (m1= 0.666 ± 0.009) ca. 10% higher than those observed in surface open ocean or coastal samples. On the contrary, the log K̅H for the less acidic groups shows a difference between the surface (10.01 ± 0.08) and deep (9.22 ± 0.35) samples. The latter chemical groups were more heterogeneous for marine than for terrestrial DOM, and m2 decreased with depth to values of 0.28 ± 0.03. Binding heterogeneity reflects aromatic carbon compounds' persistence and accumulation in diverse, low-abundance chemical forms, while easily degradable low-affinity groups accumulate more uniformly in the deep ocean.

The effect and spectral response mechanism of dissolved organic matter (DOM) in Pb(II) adsorption onto biochar

This study investigated the interaction of biochar-derived dissolved organic matter (DOM) and metal pollutant Pb(II) during adsorption. Research data showed that the adsorption capacity of biochar and residual biochar (after removal of DOM) was 277.62 and 204.47 mg g−1, respectively, indicating that DOM played a significant role in enhancing the adsorption efficiency of Pb(II). The results indicated that the adsorption process of Pb(II) on biochar and residual biochar followed the pseudo-second-order kinetic equation (R2 > 0.99), and the adsorption isotherm was much closer Freundlich model. A combination of three-dimensional excitation-emission matrix (3D-EEM), synchronous fluorescence, Fourier transform infrared spectroscopy (FT-IR), and two-dimensional correlation spectroscopy (2D-COS) was employed to investigate the interaction between DOM and Pb(II). 3D-EEM indicated the presence of four major components of DOM, namely two protein-like substances, one humic acid-like substance, and one fulvic acid-like substance. The synchronous fluorescence revealed a gradual quenching of the fluorescence of the DOM sample with an increase in Pb(II) concentration, indicating a static quenching mechanism between DOM and Pb(II). FTIR-2D-COS analysis showed a specific sequence of DOM binding with Pb(II) is as follows: 1500 > 1010 > 880 > 1390 > 1260 cm−1. This findings have significant implications for the role of DOM in environmental science and have potential applications in various fields.

Expanding the Limits of Structural Characterization of Marine Dissolved Organic Matter Using Nonuniform Sampling Frequency-Reversed Edited HSQC NMR.

The multiplicity-edited heteronuclear single quantum correlation (ME-HSQC) NMR method is widely used for the structural characterization of marine dissolved organic matter (DOM), which is a complex molecular mixture comprising millions of individual compounds. However, the standard ME-HSQC suffers from significant signal cancellation and subsequent loss of crucial structural information due to the overlap between CH3/CH (positive) and CH2 (negative) cross-peaks in overcrowded regions. This study introduces nonuniform sampling in frequency-reversed ME-HSQC (NUS FR-ME-HSQC), highlighting its remarkable potential for the comprehensive structural characterization of marine DOM. By reversing the frequency of CH2 cross-peaks into an empty region, the FR-ME-HSQC method effectively simplifies the spectra and eliminates signal cancellation. We demonstrate that nonuniform sampling enables the acquisition of comparable spectra in half the time or significantly enhances the sensitivity in time-equivalent spectra. Comparative analysis also identifies vulnerable CH2 cross-peaks in the standard ME-HSQC that coincide with CH3 and CH cross-peaks, resulting in the loss of critical structural details. In contrast, the NUS FR-ME-HSQC retains these missing correlations, enabling in-depth characterization of marine DOM. These findings highlight the potential of NUS FR-ME-HSQC as an advanced NMR technique that effectively addresses challenges such as signal overcrowding and prolonged experimental times, enabling the thorough investigation of complex mixtures with implications in several fields, including chemistry, metabolomics, and environmental sciences. The advantages of NUS FR-ME-HSQC are experimentally demonstrated on two solid-phase-extracted DOM (SPE-DOM) samples from the surface and deep ocean. With this new technology, differences in the composition of DOM from various aquatic environments can be assigned to individual molecules.

Bioactivity profile of dissolved organic matter and its relation to molecular composition

Dissolved organic matter (DOM) occupies a huge and uncharted molecular space. Given its properties, DOM can be presented as a promising biotechnological resource. However, research into bioactivities of DOM is still in early stages. In this study, the biotechnological potential of terrestrial and marine DOM, its molecular composition and their relationships are investigated. Samples were screened for their in vitro antibacterial, antifungal, anticancer and antioxidant activities. Antibacterial activity was detected against Staphylococcusaureus in almost all DOM samples, with freshwater DOM showing the lowest IC50 values. Most samples also inhibited Staphylococcusepidermidis, and four DOM extracts showed up to fourfold higher potency than the reference drug. Antifungal activity was limited to only porewater DOM towards human dermatophyte Trichophytonrubrum. No significant in vitro anticancer activity was observed. Low antioxidant potential was exerted. The molecular characterization by FT-ICR MS allowed a broad compositional overview. Three main distinguished groups have been identified by PCoA analyses. Antibacterial activities are related to high aromaticity content and highly-unsaturated molecular formulae (O-poor). Antifungal effect is correlated with highly-unsaturated molecular formulae (O-rich). Antioxidant activity is positively related to the presence of double bonds and polyphenols. This study evidenced for the first time antibacterial and antifungal activity in DOM with potential applications in cosmeceutical, pharmaceutical and aquaculture industry. The lack of cytotoxicity and the almost unlimited presence of this organic material may open new avenues in future marine bioprospecting efforts.Graphical abstract

Source, Distribution and Transformation of Organic Matter in a Subtropical Karst Reservoir

In order to improve the understanding of the global carbon cycle and the stability of karst carbon sinks, it is necessary to better understand the source, distribution and transformation characteristics of organic matter (OM) in aquatic ecosystems. Here, stable isotope ratios (δ13C and δ15N), elemental analysis (C/N ratios), and lipid biomarkers were analyzed for dissolved organic matter (DOM) (&lt;0.7 μm), particulate organic matter (POM) (&gt;0.7 μm) of water, and organic matter from sediment cores (SCOM) to identify the sources, distribution, and transformation of OM in a subtropical karst reservoir. The results showed that short-chain (C14–20) n-alkyl lipids were more abundant than long-chain (C21–34) n-alkyl lipids in both the DOM and SCOM samples, indicating that bacteria were the primary sources of these lipids, while terrestrial organic matter (OM) made only a minor contribution to the n-alkyl lipid pool, and aquatic plants (macrophytes) OM contributed major contribution to the n-alkyl lipid pool in POM. Microbial activity and lipid degradation were more pronounced in the DOM. Furthermore, terrigenous and macrophyte-derived lipids were found to be more abundant in POM than in DOM and SCOM, suggesting that they are relatively resistant to degradation compared with phytoplankton-derived OM.

Applying fluorescence spectroscopy and DNA pyrosequencing with 2D-COS and co-occurrence network to deconstruct dynamical DOM degradation of air-land-water sources in an urban river

In an urban river, comprehending the interplay between dissolved organic matter (DOM) and atmospheric, terrestrial, and aquatic sources is crucial. This encompassed investigating temporal variations in DOM and its association with the bacterioplankton community to gain profound insights into the biogeochemical dynamics and biodegradability of DOM. DOM was extracted from PM2.5, soil, sediment, bait, and terrestrial/aquatic plant residuals collected along the Wenyuhe River in Beijing, China - a region predominantly supplied with reclaimed water. Subsequently, mixed microbial communities from the river were introduced into DOM samples originating from each source and incubated for 10 days. Principal component analysis (PCA) applied to reassembled excitation-emission matrix (EEM) data revealed two distinct clusters: cluster 1 comprising soil, sediment, and PM2.5 samples; and cluster 2 consisting of bait as well as terrestrial/aquatic plant residuals. According to parallel factor analysis, C1 (microbial humic-like) and C2-C3 (fulvic-like) dominated the DOM from soil, sediment, and PM2.5. These components were continuously degraded during incubation, except for PM2.5. DOM from bait and terrestrial/aquatic plants contained representative components of C6 (phenolic-like) and C7 (tryptophan-like), which underwent extensive decomposition. Interestingly, DOM in PM2.5 contained aliphatic compounds and polycyclic aromatic hydrocarbons (PAHs) but exhibited weak degradation with the complete disappearance of C6 and C7. Rhodococcus was a unique species capable of degrading PAHs, which might be particularly important considering the specificity of PM2.5 pollution. Based on two-dimensional correlation spectroscopy (2D-COS), variations in DOM components such as C6, and C7 were significantly larger compared to those of C1, C2, C3, and C5 (terrestrial humic-like) from bait samples, sediments, and residual terrestrial plants. MW-2D-COS analysis revealed that DOM from bait samples and terrestrial/aquatic plants experienced substantial degradation by the second day while DOM from soil or sediment decomposed mainly on the fourth day. Notably, the decomposition of DOM fractions in PM2.5 occurred throughout the entire four-day period. Co-occurrence network analysis classified sources of DOM into two clusters similar to PCA results: cluster 1 showed significant microbial degradation of fulvic-like compounds while cluster 2 demonstrated deep microbial decomposition of tyrosine-like and phenolic compounds. Therefore, the artificial loading of DOM into rivers not only expands the chemical diversity within DOM but also perturbs bacterioplankton diversities.

Chemical diversity of dissolved organic matter revealed by ultra performance liquid chromatography-Fourier transform ion cyclotron resonance mass spectrometry

Dissolved organic matter (DOM) is a highly complex and heterogeneous mixture that exists in various environments, including rivers, oceans, soils, and atmospheric aerosols. DOM plays a crucial role in biogeochemical cycles and significantly influences the environment by regulating water quality, changing the climate, and transporting pollutants. Therefore, clarifying the detailed molecular composition of DOM is essential to obtain a better understanding of its physical and chemical properties, thereby enabling further elucidation of its biogeochemical behavior. In this study, Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) combined with quadrupole detection (QPD) was used to conduct the online ultra performance liquid chromatography (UPLC)-MS analysis of DOM in water, aerosol, and soil samples collected in Tianjin, China. The samples were extracted with pure water and filtered through a glass fiber membrane (0.45 μm). The DOM in the samples was then enriched by solid-phase extraction (SPE) and redissolved in water-acetonitrile (1∶1, v/v) at mass concentration of 200 mg/L for the LC-MS experiments. The mobile phases used for UPLC were water containing 0.1% (v/v) formic acid (A) and acetonitrile containing 0.1% (v/v) formic acid (B). The gradient elution procedure was as follows: 0-5 min, 0B; 5-11 min, 0B-95%B; 11-25 min, 95%B; 25-28 min, 95%B-0B; 28-30 min, 0B. The flow rate was 0.1 mL/min, and the injection volume was 10 μL. The UV wavelength was set at 274 nm. MS detection was performed in negative electrospray ionization (ESI(-)) mode with a capillary voltage of 5.0 kV, and the MS data were collected in broadband (m/z 150-1000) and QPD modes. The transient data size was set to 2M, the free induction decay signal length was 0.74 s, and the ion accumulation time was 0.030 s. Four chromatographic peaks were observed in the chromatograms. The first peak was identified as salt adduct compounds containing sodium formate. The three other peaks contained complex components, such as oxygen-rich, unsaturated tannin-like compounds, as well as low-oxygen, highly saturated lignin-like and protein/amino-like compounds. UPLC-FT-ICR MS was suitable for assigning the detailed elemental compositions of the DOM samples. UPLC effectively improved the ionization efficiency of difficult-to-ionize compounds and enhanced the detection accuracy of MS. Indeed, MS peaks with a mass difference of as small as 3.4 mDa were well identified. A total of 12027, 15593, and 8029 peaks in the mass spectra of the water, aerosol, and soil samples, respectively, were assigned to known elemental formulae. Peaks Ⅱ and Ⅲ were hydrophilic components mainly including CHNO and CHO compounds. Compared with peak Ⅱ, peak Ⅲ exhibited a significant increase in CHNOS and CHOS, indicating that UPLC exerted a certain separation effect on these compounds. Furthermore, the aerosol samples contained a higher concentration of sulfur-containing compounds than the water and soil samples, primarily because of the abundance of organic sulfates present in atmospheric and cloud water. Data processing and graphic visualization revealed that the unique components in the water samples mainly appeared in the area of 0.1<O/C<0.5 and 1.0 <H/C<1.7. The compounds detected were low-oxygen and highly condensed lignin-like compounds. The unique components in the aerosol samples appeared in the area of 0.4<O/C<1.0 and 1.5<H/C<2.0, and were classified as carbohydrates. The unique components in the hydrophilic fraction of the soil samples were found in the area of 0.6<O/C<1.0 and 0.5<H/C<1.0, and were determined to be tannin-like compounds. By contrast, the components in the hydrophobic fraction were similar to those found in the water samples and appeared in the region containing lignin-like compounds. In summary, this study proposed a novel analytical protocol to characterize DOM from different ecosystems using UPLC-FT-ICR MS. This method could separate DOM components using UPLC with eluents of different polarities and analyze them using high-resolution FT-ICR MS to reveal their molecular compositions and possible chemical types. This protocol offers solid technical support for the comprehensive profiling of DOM at the molecular level.

Linkages between fluorescent dissolved organic matter and lignin phenols in the Canada Basin, Arctic Ocean

Lignin is one of the more refractory components of dissolved organic matter (DOM) and its concentration and composition are used to trace terrigenous DOM in the ocean. In this study, lignin oxidation products (LOP), and chromophoric and fluorescent DOM were measured in Polar Mixed Layer (PML), Atlantic water (AW), and Canada Basin Deep Water (CBDW) layers in the Canada Basin to elucidate the composition and source of terrestrial DOM. Significant differences in total lignin phenol concentration (Σ11) and cinnamyl to vanillyl ratio (C/V) measured with CuO oxidation in PML, AW, and CBDW samples were found. The fluorescence intensities of humic-like C1 were correlated with Σ11 in PML, congruent with the humic-like terrestrial origin. However, no significant correlation was found when all layers are considered (p > 0.05), suggesting a change in the molecular composition of C1. The protein-like C3 and C5 intensities were strongly correlated with Σ11 (rs = 0.55–0.64) corroborating that lignins can contribute to the protein-like fluorescence in the water column. The principal component analysis (PCA) confirmed the origin and composition of DOM vary between the different water masses, especially in terms of C/V, DOC, and fluorescent components. Significant relationships between humic-like fluorescence intensities and C/V reveal the significant contribution of woody sources to humic material in the Canada Basin. Elevated S/V and humic-like C6 levels were associated with a cold eddy DOM sample but further studies are required to validate the use of these DOM characteristics to trace deep eddy.

Assessment of salinity and heat impacts on natural organic matter composition in bentonite for used nuclear fuel storage

Bentonite clay (MX-80, Wyoming, USA) is a proposed sealing material for the safe and long-term management of Canada's used nuclear fuel in a deep geological repository (DGR). Due to the physiochemical properties of bentonite, this buffer layer can limit radionuclide mobilization, and minimize air and water circulation, and hence suppress microbial activity near the used fuel container. However, natural organic matter (NOM) in mined Wyoming-type bentonite (MX-80) may serve as microbial substrates and cause further corrosion to the used fuel container. In the proposed DGR, bentonite will be exposed to high salinity due to saline groundwater and heat released from radioactive decay of radionuclide inventory of the used nuclear fuel. There is limited knowledge about how NOM quantity and quality in MX-80 may change after being exposed to the anticipated levels of salinity and heat in a DGR, especially at the molecular-level. To explore this, organic carbon contents and NOM composition (both soluble- and solid-phase) in MX-80 after exposure to different saline (NaCl) solutions (1 M, 2 M, 3 M, 4 M and 5 M) with and without heat exposure (90 °C) were analyzed. Dissolved organic carbon (DOC) solubility decreased with increasing salinity but these differences were not statistically significant when compared to the control (P < 0.05). DOC solubilization was not significantly altered with heat (P < 0.05). Dissolved organic matter (DOM) composition was analyzed with solution-state 1H nuclear magnetic resonance (NMR) and ultraviolet–visible (UV–Vis) spectroscopy. The NMR results indicated that DOM samples with varied salinities and heat exposure had similar chemical signatures. The results from UV–Vis analysis revealed a small decrease in less unsaturated components in DOM with NaCl solution compared to deionized water. The chemistry of solid-phase NOM that remained in MX-80 after DOM isolation was similar and suggests an insensitivity to either heat and/or salinity. The solubilized DOC contributes less than 1% to the total organic carbon in MX-80, thus, the anticipated level of salinity and heat in a DGR is unlikely to enhance NOM solubilization and change NOM chemistry in the bentonite markedly. As the bentonite in this study was only exposed to the heat and salinity in a short-term experiment, further studies examining NOM composition and reactivity under long-term saline and heat conditions are recommended.

Molecular composition of dissolved organic matter across diverse ecosystems: Preliminary implications for biogeochemical cycling

Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR MS) has been widely applied to characterize the molecular composition of dissolved organic matter (DOM) in different ecosystems. Most previous studies have explored the molecular composition of DOM focused on one or a few ecosystems, which prevents us from tracing the molecular composition of DOM from different sources and further exploring its biogeochemical cycling across ecosystems. In this study, a total of 67 DOM samples, including soil, lake, river, ocean, and groundwater, were analyzed by negative-ion electrospray ionization FT-ICR MS. Results show that molecular composition of DOM varies dramatically among diverse ecosystems. Specifically, the forest soil DOM exhibited the strongest terrestrial signature of molecules, while the seawater DOM showed the most abundant of biologically recalcitrant components, for example, the carboxyl-rich alicyclic molecules were abundant in the deep-sea waters. Terrigenous organic matter is gradually degraded during its transport along the river-estuary-ocean continuum. The saline lake DOM showed similar DOM characteristics with marine DOM, and sequestrated abundant recalcitrant DOM. By comparing these DOM extracts, we found that human activities likely lead to an increase in the content of S and N-containing heteroatoms in DOM, this phenomenon was commonly found in the paddy soil, polluted river, eutrophic lake, and acid mine drainage DOM samples. Overall, this study compared molecular composition of DOM extracted from various ecosystems, providing a preliminary comparison on the DOM fingerprint and an angle of view into biogeochemical cycling across different ecosystems. We thus advocate for the development of a comprehensive molecular fingerprint database of DOM using FT-ICR MS across a wider range of ecosystems. This will enable us to better understand the generalizability of the distinct features among ecosystems.

Theoretical modeling of molecular fractionation of dissolved organic matter on ferrihydrite and its impact on proton and metal binding properties

Molecular fractionation of dissolved organic matter (DOM) at the mineral-liquid interfaces in soil changes its molecular composition, thus altering its reactivity, such as proton and metal binding properties. Therefore, a quantitative understanding of compositional change of DOM molecules after adsorptive fractionation by minerals is of great environmental significance for predicting the cycling of organic carbon (C) and metals in the ecosystem. In this study, we conducted adsorption experiments to investigate the adsorption behaviors of DOM molecules on ferrihydrite. The molecular compositions of the original and fractionated DOM samples were analyzed with Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS). For all DOM molecules, three molecular groups with significantly different chemical properties were identified through Spearman correlation analysis between the relative intensities of DOM molecules and organic C concentrations in solutions after adsorptive fractionation. Three corresponding molecular models for the three molecular groups were constructed based on Vienna Soil-Organic-Matter Modeler and FT-ICR-MS results, which were used as base units to construct molecular models for the original or fractionated DOM samples (model(DOM)). The models well described the chemical properties of the original or fractionated DOM as compared with the experimental data. Furthermore, based on model(DOM), the proton and metal binding constants of DOM molecules were quantified by SPARC chemical reactivity calculations and linear free energy relationships. We found the density of binding sites of the fractionated DOM samples was negatively correlated with the adsorption percentage. Our modeling results suggested that adsorption of DOM on ferrihydrite gradually removed acidic functional groups from the solution, dominated by the adsorption of both carboxyl and phenol groups. This study proposed a new modeling approach to quantify the molecular fractionation processes of DOM on Fe oxides and their impact on proton and metal binding properties, which is expected to be applicable to DOM from different environments.