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This study investigates agro-industrial by-products as sources of bioactive compounds, particularly focusing on phenolic compounds known for their antioxidant properties. With growing interest in natural alternatives to synthetic antioxidants due to safety concerns, this study highlights the health benefits of plant-derived phenolic compounds in food preservation and healthcare products. Traditional and advanced analytical techniques were used to obtain phytochemical profiles of various residue extracts, including espresso (SCG) and cold-brew spent coffee grounds (CBCG), pineapple peel (PP), beetroot pomace (BP), apple pomace (AP), black carrot pomace (BCP), and garlic peel (GP). Assessments of total phenolic content (TPC), total flavonoid content (TFC), and antioxidant capacity (AC) supported their revalorization. CBCG showed the highest TPC, TFC, and AC. TPC content in by-products decreased in the order CBCG > SCG > GP > BCP > PP > AP > BP, with a similar trend for TFC and AC. Phytochemical profiling via FI-ESI-FTICR-MS enabled the preliminary putative identification of a range of compounds, with polyphenols and terpenes being the most abundant. Univariate and multivariate analyses revealed key patterns among samples. Strong positive correlations (Pearson's R > 0.8) indicated significant contribution of polyphenols to antioxidant capacities. These findings highlight the potential of agro-industrial residues as natural antioxidants, advocating for their sustainable utilization. [ABSTRACT FROM AUTHOR]

Dissolved organic matter (DOM) assemblages in freshwater rivers are formed from mixtures of simple to complex compounds that are highly variable across time and space. These mixtures largely form due to the environmental heterogeneity of river networks and the contribution of diverse allochthonous and autochthonous DOM sources. Most studies are, however, confined to local and regional scales, which precludes an understanding of how these mixtures arise at large, e.g., continental, spatial scales. The processes contributing to these mixtures are also difficult to study because of the complex interactions between various environmental factors and DOM. Here we propose the use of machine learning (ML) approaches to identify ecological processes contributing toward mixtures of DOM at a continental-scale. We related a dataset that characterized the molecular composition of DOM from river water and sediment with Fourier-transform ion cyclotron resonance mass spectrometry to explanatory physicochemical variables such as nutrient concentrations and stable water isotopes (2H and 18O). Using unsupervised ML, distinctive clusters for sediment and water samples were identified, with unique molecular compositions influenced by environmental factors like terrestrial input and microbial activity. Sediment clusters showed a higher proportion of protein-like and unclassified compounds than water clusters, while water clusters exhibited a more diversified chemical composition. We then applied a supervised ML approach, involving a two-stage use of SHapley Additive exPlanations (SHAP) values. In the first stage, SHAP values were obtained and used to identify key physicochemical variables. These parameters were employed to train models using both the default and subsequently tuned hyperparameters of the Histogram-based Gradient Boosting (HGB) algorithm. The supervised ML approach, using HGB and SHAP values, highlighted complex relationships between environmental factors and DOM diversity, in particular the existence of dams upstream, precipitation events, and other watershed characteristics were important in predicting higher chemical diversity in DOM. Our data-driven approach can now be used more generally to reveal the interplay between physical, chemical, and biological factors in determining the diversity of DOM in other ecosystems. [ABSTRACT FROM AUTHOR]

While significant progress has been made in understanding global carbon (C) cycling, the mechanisms regulating belowground C fluxes and storage are still uncertain. New molecular technologies have the power to elucidate these processes, yet we have no widespread standardized implementation of molecular techniques. To address this gap, we introduce the Molecular Observation Network (MONet), a decadal vision from the Environmental Molecular Sciences Laboratory (EMSL), to develop a national network for understanding the molecular composition, physical structure, and hydraulic and biological properties of soil and water. These data are essential for advancing the next generation of multiscale Earth systems models. In this paper, we discuss the 1000 Soils Pilot for MONet, including a description of standardized sampling materials and protocols and a use case to highlight the utility of molecular-level and microstructural measurements for assessing the impacts of wildfire on soil. While the 1000 Soils Pilot generated a plethora of data, we focus on assessments of soil organic matter (SOM) chemistry via Fouriertransform ion cyclotron resonance-mass spectrometry and microstructural properties via X-ray computed tomography to highlight the effects of recent fire history in forested ecosystems on belowground C cycling. We observed decreases in soil respiration, microbial biomass, and potential enzyme activity in soils with high frequency burns. Additionally, the nominal oxidation state of carbon in SOM increased with burn frequency in surface soils. This results in a quantifiable shift in the molecular signature of SOM and shows that wildfire may result in oxidation of SOM and structural changes to soil pore networks that persist into deeper soils. [ABSTRACT FROM AUTHOR]

Prostate cancer (PCa) is one of the most common male malignancies worldwide. In the current study, we evaluated the effects of a natural deep eutectic solvent (NADES) extract of Pueraria lobata roots rich in isoflavones (ISF) and Phaffia rhodozyma extract rich in astaxanthin (ASX) on an N-methyl-N-nitrosourea plus testosterone PCa model in rats. ISF consisted of puerarin, daidzein, genistein, formononetin and other polyphenols, while ASX contained lipids and unsaturated species in addition to astaxanthin. Extracts were administered through a whole promotion period in daily doses shown by our group to successfully inhibit benign prostate hyperplasia (BPH) development — 200 mg/kg for ISF and 25 mg/kg for ASX. Though a similar effect was found for BPH processes accompanying PCa induction, the incidence of PCa in animals treated with placebo, ISF and ASX was 37%, 37% and 41%, respectively, showing no chemopreventive activity of ISF and ASX. PCa development was associated with a decrease in the Ca/Mg ratio in serum and an increase in prostate tissue. Treatment with both extracts produced a normalization effect on Ca balance in serum, which, combined with a decrease in the prostatic index, suggests some positive health effects of ISF and ASX. [ABSTRACT FROM AUTHOR]

Understanding the mechanisms underlying the assembly of communities has long been the goal of many ecological studies. While several studies have evaluated community wide ecological assembly, fewer have focused on investigating the impacts of individual members within a community or assemblage on ecological assembly. Here, we adapted a previous null model β-nearest taxon index (βNTI) to measure the contribution of individual features within an ecological community to overall assembly. This new metric, called feature-level βNTI (βNTIfeat), enables researchers to determine whether ecological features (e.g., individual microbial taxa) contribute to divergence, convergence, or have insignificant impacts across spatiotemporally resolved metacommunities or meta-assemblages. Using βNTIfeat, we revealed that unclassified microbial lineages often contributed to community divergence while diverse groups (e.g., Crenarchaeota, Alphaproteobacteria, and Gammaproteobacteria) contributed to convergence. We also demonstrate that βNTIfeat can be extended to other ecological assemblages such as organic molecules comprising organic matter (OM) pools. OM had more inconsistent trends compared to the microbial community though CHO-containing molecular formulas often contributed to convergence, while nitrogen and phosphorus-containing formulas contributed to both convergence and divergence. A network analysis was used to relate βNTIfeat values from the putatively active microbial community and the OM assemblage and examine potentially common contributions to ecological assembly across different communities/assemblages. This analysis revealed that P-containing formulas often contributed to convergence/divergence separately from other ecological features and N-containing formulas often contributed to assembly in coordination with microorganisms. Additionally, members of Family Geobacteraceae were often observed to contribute to convergence/divergence in conjunction with both N- and P-containing formulas, suggesting a coordinated ecological role for family members and the nitrogen/phosphorus cycle. Overall, we show that βNTIfeat offers opportunities to investigate the community or assemblage members, which shape the phylogenetic or functional landscape, and demonstrate the potential to evaluate potential points of coordination across various community types. [ABSTRACT FROM AUTHOR]

Micomeria fruticosa and Teucrium polium are Lamiaceae plants found throughout the Mediterranean, including Lebanon. Aerial parts of both plants were taken from a rocky mountain in Lebanon's Bekaa region and alcoholic extraction and chromatographic separation were performed. Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS) was used to examine crude extracts of both plants A and B, as well as their fractions. The mass spectra of both plants revealed great chemical complexity with a total of 6352 ions (peaks) for crude extract of plant A and 5147 ions (peaks) for extract B. The majority of chemicals found in both plants are condensed aromatics, particularly polyphenols which could explain the previously documented anticancer effects of both plants. [ABSTRACT FROM AUTHOR]

We characterized the dissolved organic matter (DOM) under baseflow conditions from a set of rivers in the Mohawk and Hudson River watersheds. The rivers in this study drain a range of bedrock geologies and land cover. We identify how those factors influence riverine DOM reactivity and the source, age, and composition of the biolabile DOM. We performed laboratory incubation experiments to characterize each river's reactive and non‐reactive DOM pools. Measurements of dissolved organic carbon concentration, radiocarbon, Ultraviolet‐visible spectroscopy absorbance, and Fourier‐transform ion cyclotron resonance mass spectrometry (FTICR‐MS) analysis were performed at each incubation start and end, allowing us to determine the quantity, age, and composition of the reactive and nonreactive DOM pools. We find that lithology controls bulk DOM ages, with watersheds underlain by shale/limestone having the most aged DOM and crystalline/metasedimentary watersheds having the youngest DOM. We observe that for a given lithology, bulk DOM age increases with the proportion of agricultural land in the watershed–suggesting agricultural practices mobilize aged DOM. FTICR‐MS analysis reveals that both lithology and land cover influence DOM composition. Shale/limestone watersheds showed DOM compositions distinct from other watershed lithologies, and the percentage of nitrogen‐containing DOM correlated with agricultural influence. In two of the studied rivers we find that the biolabile DOM fraction is older than the bulk DOM (upwards of 7 kyr) revealing that aged DOM may be preferentially consumed in these rivers. Our findings provide insight into how riverine carbon cycles may respond to watershed disturbances that influence DOM inputs to rivers. Plain Language Summary: As rivers drain land areas around the globe they pick up organic matter (OM) which is then transported and transformed as water flows downstream. Through the degradation of some of this OM to carbon dioxide (CO2), rivers play a critical role in the global carbon cycle, with CO2 emissions from rivers estimated to be equivalent to >20% of the CO2 emissions from the burning of fossil fuels. Despite the importance of rivers in the global carbon cycle, many questions remain as to what controls the composition and reactivity of OM within rivers. We investigated how watershed geology and land cover (e.g., forested vs. agricultural land) influence the chemical composition, age, and reactivity of the OM in rivers. We found that rivers draining shale/limestone watersheds had chemically distinct OM that is much older and more reactive than OM from other types of watersheds. We also found that across all watersheds the age of OM increased with the proportion of the watershed that is devoted to agricultural activity and that in some watersheds the older OM was preferentially consumed. Our findings suggest that disturbances to land (e.g., agriculture) may release aged, previously buried OM that can then be transformed to CO2. Key Points: Watershed geology and land cover influence river dissolved organic matter composition and biolability under baseflow conditionsDissolved organic carbon (DOC) bulk ages are strongly influenced by watershed geology and the proportion of agricultural land coverUnder baseflow, high molecular weight compounds are preferentially consumed and aged DOC is preferentially consumed in smaller streams [ABSTRACT FROM AUTHOR]

Soil redox conditions exert substantial influence on biogeochemical processes in terrestrial ecosystems. Humid tropical forest soils are often characterized by fluctuating redox, yet how these dynamics affect patterns of organic matter decomposition and associated CO2 fluxes remains poorly understood. We used a 13C-label incubation experiment in a humid tropical forest soil to follow the decomposition of plant litter and soil organic matter (SOM) in response to four redox regimes—static oxic or anoxic, and two oscillating treatments. We used high-resolution mass spectrometry to characterize the relative composition of organic compound classes in the water extractable OM. CO2 production from litter and SOM showed different responses to redox treatments. While cumulative production of SOM-derived CO2 was positively correlated with the length of oxic exposure (r = 0.89, n = 20), cumulative 13C-litter-derived CO2 production was not linked to oxygen availability. Litter-derived CO2 production was highest under static anoxic conditions in the first half of the experiment, and later dropped to the lowest rate amongst the treatments. In anoxic soils, we observed depletion of more oxidized water-extractable OM (especially amino sugar-, carbohydrate-, and protein-like compounds) over the second half of the experiment, which likely served as substrates for anaerobic CO2 production. Results from two-pool kinetic modeling showed that more frequent anoxic exposure limited decomposition of a slow-cycling C pool, but not a fast-cycling pool. These results suggest that aerobic and anaerobic heterotrophs were equally effective at degrading labile substrates released from fresh plant litter in this humid tropical forest soil, while aerobic decomposers were more effective in breaking down the potentially refractory compounds found in SOM. [ABSTRACT FROM AUTHOR]

Most soil carbon (C) is in the form of soil organic matter (SOM), the composition of which is controlled by the plant–microbe–soil continuum. The extent to which plant and microbial inputs contribute to persistent SOM has been linked to edaphic properties such as mineralogy and aggregation. However, it is unknown how variation in plant inputs, microbial community structure, and soil physical and chemical attributes interact to influence the chemical classes that comprise SOM pools. We used two long‐term biofuel feedstock field experiments to test the influence of cropping systems (corn and switchgrass) and soil characteristics (sandy and silty loams) on microbial selection and SOM chemistry. Cropping system had a strong influence on water‐extractable organic C chemistry with perennial switchgrass generally having a higher chemical richness than the annual corn cropping system. Nonetheless, cropping system was a less influential driver of soil microbial community structure and overall C chemistry than soil type. Soil type was especially influential on fungal community structure and the chemical composition of the chloroform‐extractable C. Although plant inputs strongly influence the substrates available for decomposition and SOM formation, total C and nitrogen (N) did not differ between cropping systems within either site. We conclude this is likely due to enhanced microbial activity under the perennial cropping system. Silty soils also had a higher activity of phosphate and C liberating enzymes. After 8 years, silty loams still contained twice the total C and N as sandy loams, with no significant response to biofuel cropping system inputs. Together, these results demonstrate that initial site selection is critical to plant–microbe interactions and substantially impacts the potential for long‐term C accrual in soils under biofuel feedstock production. [ABSTRACT FROM AUTHOR]

Predictive biogeochemical modeling requires data-model integration that enables explicit representation of the sophisticated roles of microbial processes that transform substrates. Data from high-resolution organic matter (OM) characterization are increasingly available and can serve as a critical resource for this purpose, but their incorporation into biogeochemical models is often prohibited due to an over-simplified description of reaction networks. To fill this gap, we proposed a new concept of biogeochemical modeling—termed substrate-explicit modeling —that enables parameterizing OM-specific oxidative degradation pathways and reaction rates based on the thermodynamic properties of OM pools. Based on previous developments in the literature, we characterized the resulting kinetic models by only two parameters regardless of the complexity of OM profiles, which can greatly facilitate the integration with reactive transport models for ecosystem simulations by alleviating the difficulty in parameter identification. The two parameters include maximal growth rate (μmax) and harvest volume (Vh) (i.e., the volume that a microbe can access for harvesting energy). For every detected organic molecule in a given sample, our approach provides a systematic way to formulate reaction kinetics from chemical formula, which enables the evaluation of the impact of OM character on biogeochemical processes across conditions. In a case study of two sites with distinct OM thermodynamics using ultra high-resolution metabolomics datasets derived from Fourier transform ion cyclotron resonance mass spectrometry analyses, our method predicted how oxidative degradation is primarily driven by thermodynamic efficiency of OM consistent with experimental rate measurements (as shown by correlation coefficients of up to 0.61), and how biogeochemical reactions can vary in response to carbon and/or oxygen limitations. Lastly, we showed that incorporation of enzymatic regulations into substrate-explicit models is critical for more reasonable predictions. This result led us to present integrative biogeochemical modeling as a unifying framework that can ideally describe the dynamic interplay among microbes, enzymes, and substrates to address advanced questions and hypotheses in future studies. Altogether, the new modeling concept we propose in this work provides a foundational platform for unprecedented predictions of biogeochemical and ecosystem dynamics through enhanced integration with diverse experimental data and extant modeling approaches. [ABSTRACT FROM AUTHOR]

Abstract: The path of carbon (C) from plant litter to soil organic matter (SOM) is key to understanding how soil C stocks and microbial decomposition will respond to climate change and whether soil C sinks can be enhanced. Long‐term ecosystem‐scale litter manipulations and molecular characterization of SOM provide a unique opportunity to explore these issues. We incubated soils from a 20‐year litter input experiment for 525 days and asked how litter quantity and source (i.e., roots versus aboveground litter) affected C cycling, microbial function, and the size and molecular composition of C pools. Input exclusion led to a 30% loss of soil C, attributable largely to the nonmineral‐associated C fraction, and to declines in soil C decomposition. The absence of roots caused a shift in the microbial catabolic profile, though there was little evidence that root litter was preferentially stabilized. Although C pool size did not change with litter additions, Fourier transform ion cyclotron resonance mass spectrometry analysis of the finest mineral fraction revealed dramatic changes to the chemical composition of carbon. Lipid content increased proportionally with input addition and was subsequently mineralized during incubation, indicating that this fraction was metabolically active. Moreover, nonmetric dimensional scaling showed that both litter treatments and incubation caused the molecular composition of SOM to change. We conclude that the path of C from litter to soil may involve labile pools and root‐driven microbial activity associated directly with SOM in the soil mineral matrix otherwise previously hypothesized to be stable. [ABSTRACT FROM AUTHOR]

Purpose: Dissolved organic matter (DOM) composition is influenced by and modulates biogeochemical processes, yet DOM characterization techniques are challenged by its heterogeneous properties and structures. In this paper, ultrahigh electrospray ionization Fourier transform ion cyclotron resonance (ESI-FTICR) mass spectrometry is used to characterize previously isolated and well-characterized four DOM fractions from a water reservoir in the southwestern part of the USA.Materials and methods: About 50-60 L water samples were collected from Lake Pleasant (a water reservoir of Colorado River) and isolated and fractionated using a sequential isolation and fractionation procedure. DOM was firstly fractionated to the colloids and soluble permeate, then hydrophobic acids (HPO-A) and neutrals (HPO-N) plus amphiphilic acids (AMP-A) and neutrals (AMP-N) were fractionated using the resin-in-tandem procedure. Approximately 0.7 mg sample was dissolved in 250 μL methanol/water mixture (1:1, v/v) and diluted twice with 0.1 % (by volume) NH4OH (in 1:1 methanol/water, v/v) and measured by ESI-FTICR in negative ion mode.Results and discussion: In the m/z range of <700, among the sum of 4107 peaks, HPO-N contained the most assigned 1763 molecules and HPO-A, AMP-N, and AMP-A contained 971, 990, and 293, respectively. Acid fractions were relatively more oxygenated than the corresponding neutrals. Except for the major assigned CHO molecules, a high portion of heteroatom-containing molecules were assigned in the neutral fractions. Van Krevelen and Venn diagrams analyses showed significant molecular differentiation among the fractions. Among the assigned formulas, the Venn diagram indicated only a small portion (<4 %) of distinct formulas overlapped. S-containing molecules were efficiently ionized and identified in the neutrals. Some of the S1-containing compounds with high signal magnitudes are tentatively assigned to alkylbenzene sulfonates and their biodegradation and biotransformation derivatives indicating anthropogenic input on site.Conclusions: By using the XAD resin-in-tandem approach and FTICR-MS, numbers of DOM molecules in fractions were distinctly separated, accordingly the complexity of DOM was reduced. Furthermore, to a great extent, the compositions and structures of each fraction in the molecular level are remarkably different as evidenced by only small portion of molecules overlapping. [ABSTRACT FROM AUTHOR]

Soyasaponins are oleanene-type triterpenoid saponins, naturally occurring in many edible plants that have attracted a great deal of attention for their role in preventing chronic diseases. The aim of this study was to establish the distribution and the content of soyasaponins in 21 ecotypes of Fagioli di Sarconi beans (Phaseolus vulgaris, Leguminosae). High-performance reversed-phase liquid chromatography (RPLC) with positive electrospray ionization (ESI(+)) and Fourier transform ion cyclotron resonance (FTICR) mass spectrometry (MS) in conjunction with infrared multiphoton dissociation (IRMPD) was applied for the unambiguous identification of soyasaponins Ba (m/z 959.5213, [C48H79O19]+), Bb (m/z 943.5273, [C48H79O18]+), Bd (m/z 957.5122, [C48H77O19]+), and Be (m/z 941.5166, [C48H77O18]+), which are the only commercially available reference standards. In addition, the several diagnostic product ions generated by IRMPD in the ICR-MS cell allowed us the putative identification of soyasaponins Bb' (m/z 797.4680, [C42H69O14]+), αg (m/z 1085.5544, [C54H85O22]+), βg (m/z 1069.5600, [C54H85O21]+), and γg (m/z 923.5009, [C48H75O17]+), establishing thus their membership in the soyasaponin group. Quantitative and semiquantitative analysis of identified soyasaponins were also performed by RPLC-ESI(+) FTICR-MS; the total concentration levels were found ranging from 83.6 ± 9.3 to 767 ± 37 mg/kg. In vitro hypoglycemic outcomes of four soyasaponin standards were evaluated; significant inhibitory activities were obtained with IC50 values ranging from 1.5 ± 0.1 to 2.3 ± 0.2 μg/mL and 12.0 ± 1.1 to 29.4 ± 1.4 μg/mL for α-glucosidase and α-amylase, respectively. This study represents the first detailed investigation on the antidiabetic activity of bioactive constituents found in Fagioli di Sarconi beans.The first detailed RPLC-ESI(+) FTICR-MS investigation of the qualitative and semiquantitative profile of soyasaponins, occurring in 21 ecotypes of Fagioli di Sarconi beans (P. vulgaris L.). [ABSTRACT FROM AUTHOR]

Abstract: In light of increasing terrestrial carbon (C) transport across aquatic boundaries, the mechanisms governing organic carbon (OC) oxidation along terrestrial‐aquatic interfaces are crucial to future climate predictions. Here we investigate the biochemistry, metabolic pathways, and thermodynamics corresponding to OC oxidation in the Columbia River corridor using ultrahigh‐resolution C characterization. We leverage natural vegetative differences to encompass variation in terrestrial C inputs. Our results suggest that decreases in terrestrial C deposition associated with diminished riparian vegetation induce oxidation of physically bound OC. We also find that contrasting metabolic pathways oxidize OC in the presence and absence of vegetation and—in direct conflict with the “priming” concept—that inputs of water‐soluble and thermodynamically favorable terrestrial OC protect bound‐OC from oxidation. In both environments, the most thermodynamically favorable compounds appear to be preferentially oxidized regardless of which OC pool microbiomes metabolize. In turn, we suggest that the extent of riparian vegetation causes sediment microbiomes to locally adapt to oxidize a particular pool of OC but that common thermodynamic principles govern the oxidation of each pool (i.e., water‐soluble or physically bound). Finally, we propose a mechanistic conceptualization of OC oxidation along terrestrial‐aquatic interfaces that can be used to model heterogeneous patterns of OC loss under changing land cover distributions. [ABSTRACT FROM AUTHOR]

Tandem mass spectrometry (MS/MS) provides detailed information for structural characterization of biomolecules. The combination of electron capture dissociation (ECD) techniques with Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS) often provides unique ion-electron reactions and fragmentation channels in MS/MS. ECD is often a complimentary, sometimes even a superior tool to conventional MS/MS techniques. This article is aimed at providing a short overview of ECD-based fragmentation techniques (ExD) and optimization of ECD experiments for FTICR mass analyzers. Most importantly, it is meant to pique the interest of potential users for this exciting research field. © 2015 Wiley Periodicals, Inc. Mass Spec Rev 36:4-15, 2017. [ABSTRACT FROM AUTHOR]

Lignin is the second most abundant natural polymer and a promising alternative energy source for conventional fossil fuels. In this study, we investigated transformations of lignin compounds under artificial UV light conditions at the molecular level. Such light-induced changes of composition profiles in nature after sun exposure have been studied for crude oil in the petroleomics field. We applied a similar high resolution mass spectrometry experimental strategy to lignin and demonstrated various data processing methods to reveal the characteristic differences between the extremely complex data sets of two sample sets, one native control before and one sample after photo-irradiation, using Fourier transform ion cyclotron resonance-mass spectrometry. [ABSTRACT FROM AUTHOR]

Introduction: Sulfur-containing metabolites (S-metabolites) in organisms including plants have unique benefits to humans. So far, few analytical methods have explored such metabolites. Objectives: We aimed to develop an automatic chemically assigning platform by metabolomics and chemoinformatics with S labeling to identify the molecular formula of S-metabolites. Methods: Direct infusion analysis using Fourier transform ion cyclotron resonance-mass spectrometry provided ultra-high-resolution data including clearly separated isotopic ions-N, S, O, and C-in the flower, silique, leaf, stem, and root of non-labeled and S-labeled Arabidopsis thaliana. Chemoinformatic analysis assigned several elemental compositions of S-metabolites to the acquired S-containing monoisotopic ions using mass accuracy and peak resolution in the non-labeled metabolome data. Possible elemental compositions were characterized on the basis of diagnostic scores of the exact mass and isotopic ion pattern, and a database search. By comparing elemental compositions assigned to the S-labeled data with those assigned to the non-labeled data, the elemental composition of S-metabolites were determined. The determined elemental compositions were surveyed using the in-house database, which stores molecular formulae downloaded from metabolome databases. Results: We identified 35 molecular formulae for known S-metabolites and characterized 72 for unknown. Chemoinformatics required around 1.5 min to analyze a pair of the non-labeled and S-labeled data of the organ. Conclusion: In this study, we developed an automation platform for automatically identifying the presence of S-metabolites. We identified the molecular formula of known S-metabolites, which are accessible in free databases, together with that of unknown. This analytical method did not focus on identifying the structure of S-metabolites, but on the automatic identification of their molecular formula. [ABSTRACT FROM AUTHOR]

Lignin is the second most abundant natural biopolymer, and lignin wastes are therefore potentially significant sources for renewable chemicals such as fuel compounds, as alternatives to fossil fuels. Waste valorisation of lignin is currently limited to a few applications such as in the pulp industry, however, because of the lack of effective extraction and characterisation methods for the chemically highly complex mixtures after decomposition. Here, we have implemented high resolution mass spectrometry and developed two-dimensional mass defect matrix plots as a data visualisation tool, similar to the Kendrick mass defect plots implemented in fields such as petroleomics. These 2D matrix plots greatly simplified the highly convoluted lignin mass spectral data acquired from Fourier transform ion cyclotron resonance (FTICR)-mass spectrometry, and the derived metrics provided confident peak assignments and strongly improved structural mapping of lignin decomposition product series from the various linkages within the lignin polymer after electrochemical decomposition. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

Transthyretin amyloidosis (ATTR) belongs to a class of disorders caused by protein misfolding and aggregation. ATTR is a disabling disorder of autosomal dominant trait, where transthyretin (TTR) forms amyloid deposits in different organs, causing dysfunction of the peripheral nervous system. We previously discovered that amyloid fibrils from ATTR patients are glycated by methylglyoxal. Even though no consensus has been reached about the actual role of methylglyoxal-derived advanced glycation end-products in amyloid diseases, evidence collected so far points to a role for protein glycation in conformational abnormalities, being ubiquitously found in amyloid deposits in Alzheimer's disease, dialysis-related amyloidosis and Parkinson's diseases. Human fibrinogen, an extracellular chaperone, was reported to specifically interact with a wide spectrum of stressed proteins and suppress their aggregation, being an interacting protein with TTR. Fibrinogen is differentially glycated in ATTR, leading to its chaperone activity loss. Here we show the existence of a proteostasis imbalance inATTRlinked to fibrinogen glycation by methylglyoxal. [ABSTRACT FROM AUTHOR]

Pyoverdines (PVD) are a group of siderophores produced by fluorescent Pseudomonads. Identification of PVD variants mostly relies on liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID). Here, both CID and the novel dissociation technique electron-capture dissociation (ECD) were applied to characterize PVD succinamide and its Fe(III)-chelated complex. The results clearly showed that ECD produced diagnostic side chain fragmentation of the PVD peptide chain and preserved the labile Fe(III) binding to the chromophore in contrast to CID. The ECD technique is therefore expected to support the understanding of strain-specific Fe(III) transport processes of PVDs. [ABSTRACT FROM AUTHOR]

Fourier transform mass spectrometry (FTMS) is the method of choice for measurements that require ultra-high resolution. The establishment of Fourier transform ion cyclotron resonance (FTICR) MS, the availability of biomolecular ionization techniques and the introduction of the OrbitrapΓ mass spectrometer have widened the number of FTMS-applications enormously. One recent example involves clinical proteomics using FTICR-MS to discover and validate protein biomarker signatures in body fluids such as serum or plasma. These biological samples are highly complex in terms of the type and number of components, their concentration range, and the structural identity of each species, and thus require extensive sample cleanup and chromatographic separation procedures. Clearly, such an elaborate and multi-step sample preparation process hampers high-throughput analysis of large clinical cohorts. A final MS read-out at ultra-high resolution enables the analysis of a more complex sample and can thus simplify upfront fractionations. To this end, FTICR-MS offers superior ultra-high resolving power with accurate and precise mass-to-charge ratio (m/z) measurement of a high number of peptides and small proteins (up to 20 kDa) at isotopic resolution over a wide mass range, and furthermore includes a wide variety of fragmentation strategies to characterize protein sequence and structure, including post-translational modifications (PTMs). In our laboratory, we have successfully applied FTICR "next-generation" peptide profiles with the purpose of cancer disease classifications. Here we will review a number of developments and innovations in FTICR-MS that have resulted in robust and routine procedures aiming for ultra-high resolution signatures of clinical samples, exemplified with state-of-the-art examples for serum and saliva. [ABSTRACT FROM AUTHOR]

Mass spectrometry (MS)-based profiling of serum peptides for biomarker discovery purposes and comparative analyses of clinical cohorts has been widely applied on surface-enhanced laser desorption/ionization- and matrix-assisted laser desorption/ionization (MALDI)-platforms. Most commonly, these strategies involve a single-step sample clean-up procedure and time-of-flight MS readout. Previously, we have developed fully automated high-throughput magnetic bead-based solid-phase extraction (SPE) procedures to enable peptide and protein profiling studies. In this study, it will be shown that the combination of SPE with MALDI and ultrahigh resolution Fourier transform ion cyclotron resonance MS provides a next-generation platform that allows such profiling up to molecular weights of 15 kDa with full isotopic resolution for each species. Beneficial aspects of using ultrahigh resolution MS for MALDI profiling are mass precision and spectral alignment, resolving overlapping signals, and identification (structure characterization) of differentiating features. [ABSTRACT FROM AUTHOR]

Marine microorganisms adapt to their habitat by structural modification of their membrane lipids. This concept is the basis of numerous molecular proxies used for paleoenvironmental reconstruction. Archaeal tetraether lipids from ubiquitous marine planktonic archaea are particularly abundant, well preserved in the sedimentary record and used in several molecular proxies. We here introduce the direct, extraction-free analysis of these compounds in intact sediment core sections using laser desorption ionization (LDI) coupled to Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS). LDI FTICR-MS can detect the target lipids in single submillimeter-sized spots on sediment sections, equivalent to a sample mass in the nanogram range, and could thus pave the way for biomarker-based reconstruction of past environments and ecosystems at subannual to decadal resolution. We demonstrate that ratios of selected archaeal tetraethers acquired by LDI FTICR-MS are highly correlated with values obtained by conventional liquid chromatography/MS protocols. The ratio of the major archaeal lipids, caldarchaeol and crenarchaeol, analyzed in a 6.2-cm intact section of Mediterranean sapropel S1 at 250-μm resolution (∼4-y temporal resolution), provides an unprecedented view of the fine-scale patchiness of sedimentary biomarker distributions and the processes involved in proxy signal formation. Temporal variations of this lipid ratio indicate a strong influence of the ∼200-y de Vries solar cycle on reconstructed sea surface temperatures with possible amplitudes of several degrees, and suggest signal amplification by a complex interplay of ecological and environmental factors. Laser-based biomarker analysis of geological samples has the potential to revolutionize molecular stratigraphic studies of paleoenvironments. [ABSTRACT FROM AUTHOR]

Natural populations of marine invertebrates often exhibit measureable morphologic variation resulting in taxonomic confusion. This potentially has severe consequences for experimental design and data management. Species of the sponge genus Ianthella embody a number of different morphologies and a diverse range of secondary metabolites. Among them, Ianthella basta (Pallas, 1776), a common sponge in Papua New Guinea and the Great Barrier Reef (GBR), exhibits two dominant colour morphotypes: yellow and purple. Specimens collected from Orpheus Island on the GBR were investigated using phylogenetic (CO1, ITS-2 sequence analysis), chemical (mass spectrometry) and microbial (DGGE and 16S rRNA clone library) techniques in an effort to fully characterise the two colour morphs. Phylogenetic analyses indicated sharp genetic discontinuities within I. basta sensu lato independent of colour variation. The two morphotypes did, however, correspond to distinct DGGE profiles largely due to the presence of additional bands in the purple morpho-group. Further comparison of the microbial communities by 16S rRNA gene sequencing revealed that whilst both colour morphs were dominated by only two bacterial symbionts (residing within the Gamma and Alphaproteobacteria), the purple morph also contained minor representatives of the Cyanobacteria, Chloroflexi and Verrucomicrobia. Untargeted metabolic profiling by Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS) indicated two distinct clusters corresponding to the different sponge colours. A clear association was found between the araplysillin class of compounds and the purple morphotype of I. basta, indicating the utility of a metabolomic approach to assess differences between colour morphs. These results have important implications for ecological investigations in sponges and other invertebrate taxa whose morphology is fundamentally dynamic, stressing the need for precise taxonomic, chemical and microbial descriptions. [ABSTRACT FROM AUTHOR]

Redox cofactors like NADH and NADPH are essential for the catalytic activity of several oxidoreductases. Here, we describe a comparative study of the thermal degradation products of both cofactors in the dry and liquid states. The degradation products were first separated, detected, and quantified by high-performance liquid chromatography (HPLC). Subsequently, selected main fractions were investigated by nanoelectrospray ionization-Fourier transform ion cyclotron resonance mass spectrometry (MS). Additionally, HPLC-MS was used to elucidate the structure of all degradation products. From these data, degradation pathways for both the liquid and the solid states were elucidated. Thermal degradation in water is significantly faster compared to degradation in the solid state. Hydrolysis and oxidative ring opening of the reduced nicotinamide adenine dinucleotide (phosphate) were shown to be the main reaction paths. Surprisingly, no significant differences were observed between the degradation of both cofactors in solution and in the solid state. Our results demonstrate that the stability of both cofactors is not limiting at moderate temperatures if they are used in the dry state (e.g., solid/gas catalysis). Significant degradation of dry cofactors was only observed under conditions, which are usually not appropriate for biocatalysis (>95 °C). Besides, the situation is completely different in solution where degradation is already observed at moderate temperatures. [ABSTRACT FROM AUTHOR]

Establishing the analytical platforms for characterizing human milk oligosaccharides is important to fully assess their specific functionalities. The characterization of these biomolecules, however, is still considered challenging, owing to their overall complexity and diversity. Addressed here are the technical difficulties with an emphasis on the application of mass spectrometry to rapidly profile and quantify human milk oligosaccharides. Fundamental concepts and improvements in instrumentation and an overview of the biological functions and structures of these compounds are also discussed. Results reveal that small-chain oligosaccharides, evident in abundance in the early stage of lactation, are selectively consumed by specific stains of Bifidobacterium longum biovar, infantis. [ABSTRACT FROM AUTHOR]

With the discovery of Na-sulphate minerals on Mars and Europa, recent studies using these minerals have focused on their ability to assist in the detection of bio/organic signatures. This study further investigates the ability of thenardite (Na2SO4) to effectively facilitate the ionization and identification of aromatic amino acids (phenylalanine, tyrosine and tryptophan) using a technique called geomatrix-assisted laser desorption/ionization in conjunction with a Fourier transform ion cyclotron resonance mass spectrometry. This technique is based on the ability of a mineral host to facilitate desorption and ionization of bio/organic molecules for detection. Spectra obtained from each aromatic amino acid alone and in combination with thenardite show differences in ionization mechanism and fragmentation patterns. These differences are due to chemical and structural differences between the aromatic side chains of their respective amino acid. Tyrosine and tryptophan when combined with thenardite were observed to undergo cation-attachment ([M+Na]+), due to the high alkali ion affinity of their aromatic side chains. In addition, substitution of the carboxyl group hydrogen by sodium led to formation of [M-H+Na]Na+ peaks. In contrast, phenylalanine mixed with thenardite showed no evidence of Na+ attachment. Understanding how co-deposition of amino acids with thenardite can affect the observed mass spectra is important for future exploration missions that are likely to use laser desorption mass spectrometry to search for bio/organic compounds in extraterrestrial environments. [ABSTRACT FROM AUTHOR]

Geomatrix-assisted laser desorption/ionization (GALDI) in conjunction with a Fourier transform ion cyclotron resonance mass spectrometer (FTICR-MS) has been employed to determine how effectively bio/organic molecules associated with the mineral thenardite (Na2SO4) can be detected. GALDI is based on the ability of the mineral host to assist desorption and ionization of bio/organic molecules without additional sample preparation. When glycine was mixed with thenardite, glycine was deprotonated to produce C2H4NO2- at m/z 74.025. The combination of stearic acid with thenardite produced a complex cluster ion at m/z 390.258 in the negative mode, which was assigned a composition of C18H39O7Na-. A natural sample of thenardite from Searles Lake in California also produced a peak at m/z 390.260. The bio/organic signatures in both the laboratory-based and natural samples were heterogeneously dispersed as revealed by chemical imaging. The detection limits for the stearic acid and thenardite combination were estimated to be 3 parts per trillion or ∼7 zeptomoles (10- 21) per laser spot. Attempts to improve the signal-to-noise ratio by co-adding FTICR-MS data predetermined to contain the biosignatures of interest revealed problems due to a lack of phase coherence between data sets. [ABSTRACT FROM AUTHOR]

With unmatched mass resolution, mass accuracy, and exceptional detection sensitivity, Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FTICR-MS) has the potential to be a powerful new technique for high-throughput metabolomic analysis. In this study, we examine the properties of an ultrahigh-field 12-Tesla (12T) FTICR-MS for the identification and absolute quantitation of human plasma metabolites, and for the untargeted metabolic fingerprinting of inbred-strain mouse serum by direct infusion (DI). Using internal mass calibration (mass error ≤1 ppm), we determined the rational elemental compositions (incorporating unlimited C, H, N and O, and a maximum of two S, three P, two Na, and one K per formula) of approximately 250 out of 570 metabolite features detected in a 3-min infusion analysis of aqueous extract of human plasma, and were able to identify more than 100 metabolites. Using isotopically-labeled internal standards, we were able to obtain excellent calibration curves for the absolute quantitation of choline with sub-pmol sensitivity, using 500 times less sample than previous LC/MS analyses. Under optimized serum dilution conditions, chemical compounds spiked into mouse serum as metabolite mimics showed a linear response over a 600-fold concentration range. DI/FTICR-MS analysis of serum from 26 mice from 2 inbred strains, with and without acute trichloroethylene (TCE) treatment, gave a relative standard deviation (RSD) of 4.5%. Finally, we extended this method to the metabolomic fingerprinting of serum samples from 49 mice from 5 inbred strains involved in an acute alcohol toxicity study, using both positive and negative electrospray ionization (ESI). Using these samples, we demonstrated the utility of this method for high-throughput metabolomics, with more than 400 metabolites profiled in only 24 h. Our experiments demonstrate that DI/FTICR-MS is well-suited for high-throughput metabolomic analysis. [ABSTRACT FROM AUTHOR]

Since highly sensitive on-line coupling of UPLC with FTICR-MS is technically infeasible due to their different scan rates, at-line coupling of these techniques was developed for rapid analysis. To enable cutting of one peak of the chromatogram into one fraction, several conditions and relationships were investigated, e.g. the optimum volume of the inserted delay loop, the relationship between retention time, loop outlet drop speed, individual drop volume versus mobile phase composition under constant speed, and linear solvent strength gradient elution modes. Good and reproducible results were achieved applying UPLC as an efficient separation and fast fractionation tool before the FTICR-MS measurements. A chip-based nanoelectrospray ionization system was employed which was perfectly suited to handling the small-volume fractions and was thus chosen for the at-line coupling. The method was initially applied to spiked extracts of cell-free bacterial culture supernatants in which bacterial signalling compounds, namely N-acyl homoserine lactones (AHL), were detected. Good reproducibility and high recovery was observed. Afterwards, a culture supernatant of Erwinia sp. JX3.2, a putative AHL producer, was investigated and N-hexanoyl-homoserine lactone was determined as a possible signalling molecule. More reliable assignments were achieved by use of at-line coupling of UPLC and FTICR-MS compared with off-line measurements. [ABSTRACT FROM AUTHOR]

Bacterial intraspecies and interspecies communication in the rhizosphere is mediated by diffusible signal molecules. Many Gram-negative bacteria use N-acyl-homoserine lactones (AHLs) as autoinducers in the quorum sensing response. While bacterial signalling is well described, the fate of AHLs in contact with plants is much less known. Thus, adsorption, uptake and translocation of N-hexanoyl- (C6-HSL), N-octanoyl- (C8-HSL) and N-decanoyl-homoserine lactone (C10-HSL) were studied in axenic systems with barley ( Hordeum vulgare L.) and the legume yam bean ( Pachyrhizus erosus (L.) Urban) as model plants using ultra-performance liquid chromatography (UPLC), Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS) and tritium-labelled AHLs. Decreases in AHL concentration due to abiotic adsorption or degradation were tolerable under the experimental conditions. The presence of plants enhanced AHL decline in media depending on the compounds’ lipophilicity, whereby the legume caused stronger AHL decrease than barley. All tested AHLs were traceable in root extracts of both plants. While all AHLs except C10-HSL were detectable in barley shoots, only C6-HSL was found in shoots of yam bean. Furthermore, tritium-labelled AHLs were used to determine short-term uptake kinetics. Chiral separation by GC-MS revealed that both plants discriminated D-AHL stereoisomers to different extents. These results indicate substantial differences in uptake and degradation of different AHLs in the plants tested. [ABSTRACT FROM AUTHOR]

Natural products are a source of unique chemical entities with specific biological activities of great value to the pharmaceutical industry. However, the determination of unknown structures is usually time consuming and often becomes a bottleneck in the effort to develop natural products into effective drugs. The high-performance features of high magnetic field FTMS have greatly alleviated the structural elucidation bottleneck to meet increasingly shorter discovery timelines for drug candidates based on natural products. The high-performance features of high field FTMS include unsurpassed mass measurement accuracy for elemental formula determination, ultra-high mass resolution for component separation, the ability to perform multiple levels of tandem mass spectrometry for structural elucidation, and moderate sensitivity for limited supply of isolates. A number of applications utilizing these properties of FTMS have been reported recently for the structural elucidation of novel natural product structures originating from terrestrial and marine microorganisms. In this review, FTMS methods and their applications for the structural elucidation and characterization of natural products will be reviewed. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

Speed of analysis is a key factor in liquid chromatography. One possible way is to decrease the particle size and thus increase the pressure (ultra high pressure liquid chromatography, UHPLC). For the separation of a complex mixture, the application of gradient elution techniques is expedient. From a practical point of view, it has to be questioned how the high pressure influences the separation parameters at gradient mode. In our study we investigated the following chromatographic parameters: peak width versus temperature, peak width versus gradient time, retention time versus temperature, and retention factor versus flow rate. It was shown that even in sample volume overload conditions the gradient system worked in a reproducible way; the maximum possible sample volume (20 µL) was injected to get the lowest possible limit of detection. Increasing the flow rate, the peak width increased only moderately, and the same was found when the temperature was decreased. However, at 60°C the signal to noise ratio decreased in a small compass showing the thermal disequilibrium between the column outlet and the detector cell, which might have caused extra noise. Finally, a practical separation was presented in which the UHPLC can be applied for the determination of N-acylhomoserine lantones (AHLs), which have a relatively low UV absorption at low wavelength. The detection limit was low enough for practical applications because of the low dispersion due to a small diameter stationary phase and the low dead volume in the column. The UPLC method was applied as a first screening for AHLs and the results were confirmed by fourier transformation ion cyclotron resonance mass spectrometry (FTICR-MS), which has a high selectivity. [ABSTRACT FROM AUTHOR]

N-Acylated homoserine lactones (AHLs) are produced by Gram-negative bacteria as communication signals and are frequently studied as mediators of the “quorum sensing” response of bacterial communities. Several reports have recently been published on the identification of AHLs from different species and attempts have been made to study their role in natural habitats, for example the surface of plant roots in the rhizosphere. In this article, different analytical methods, including bacterial biosensors and chromatographic techniques, are reviewed. A concept for assignment of the structures of AHLs is also presented. The retention behaviour of derivatives of AHLs containing β-keto or hydroxyl groups and/or double bonds has been evaluated in relation to the separation behaviour of AHLs with saturated and unsubstituted alkanoyl chains. Samples have also been analysed by high resolution mass spectrometry (Fourier-transform ion-cyclotron-resonance mass spectrometry, FTICR-MS), nano liquid chromatography–electrospray ionization ion trap mass spectrometry (nano-LC–MS) and by the aid of a biosensor. The results obtained from ultra performance liquid chromatography (UPLC), FTICR-MS, nano-LC–MS, and bioassays have been compared to attempt structural characterisation of AHL without chemical synthesis of analytical standards. The method was used to identify the major AHL compound produced by the rhizosphere bacterium Acidovorax sp. N35 as N-(3-hydroxydecanoyl)homoserine lactone. [ABSTRACT FROM AUTHOR]

Structural characterization of macromolecules is currently delivering new insights into the behavior of individual molecules or molecular ensembles. Technological advances have made it possible to examine smaller and smaller amounts (down to single molecules) of larger and larger molecular systems. Mass spectrometry in particular is capable of the detailed study of extremely small quantities (down to a single molecule) of very large (biological) molecules. The advent of new ionization techniques such as electrospray and matrix-assisted laser desorption are mainly responsible for these advances. As a result, mass spectrometry has evolved into an enabling discipline that plays an increasingly important role in combinatorial chemistry, polymer science, biochemistry, medicine, environmental and marine science, and archaeology and conservation science. This paper will review a selection of methodological developments in the field of high-performance Fourier transform ion cyclotron resonance mass spectrometry for structural analysis of these macromolecules. [ABSTRACT FROM AUTHOR]

A letter to the editor is presented in response to the article which discusses a study on the use of electron-capture dissociation (ECD) to investigate host/guest complexes of 18-crown-6-ether and peptides.