Combination Of High-resolution Mass Spectrometry Research Articles

Chemical profiling and mechanistic studies of Zhi-Shang-Feng granules against influenza virus by high-performance liquid chromatography coupled with Q exactive focus hybrid quadrupole orbitrap high-resolution mass spectrometry in combination with network pharmacology analysis.

Zhi-Shang-Feng Granules are used in the clinical treatment of influenza to relieve headaches, chills and fever, bronchitis, nasal congestion, neuralgia and other symptoms. To decipher the components responsible for therapeutic effects of Zhi-Shang-Feng g ranules against influenza virus, an analytical method based on high-performance liquid chromatography coupled with Q exactive focus hybrid quadrupole orbitrap high resolution mass spectrometry was developed and the chemical profile of Zhi-Shang-Feng granules was characterized. Then, the identified components were used to conduct network pharmacological analysis and determine the potential mechanism of Zhi-Shang-Feng Granules. As a result, 177 compounds were putatively identified through comprehensive analysis by liquid chromatography coupled with high-resolution mass spectrometry, of which 23 compounds were unambiguously confirmed with reference standards. Components in Zhi-Shang-Feng Granules were found to specifically act on different enzymes, G-protein-coupled receptors, ion channels and transporters in the immune, endocrine, nervous, and circulatory systems. The potential mechanism was related to several biological processes, including cell growth and death, pattern recognition receptor signalling, signalling by interleukins, and lipid metabolism. The combination of chemical profile characterization and network construction provided useful insight into the overall chemical composition of Zhi-Shang-Feng granules and revealed their potential anti-infection, anti-inflammatory and immunoregulatory mechanisms against influenza virus infected disease.

Characterization of the recently detected cathinone N-cyclohexyl butylone: From structure elucidation to in silico supported pharmacological/toxicological considerations

One of the most widely consumed new psychoactive substances (NPS) families in Europe are synthetic cathinones. Cathinone structure can be easily modified resulting in new derivatives that rapidly reach drug markets. In this work, the recently detected synthetic cathinone N-cyclohexyl butylone has been characterized by gas chromatography coupled to mass spectrometry (GC–MS), liquid chromatography coupled to high-resolution mass spectrometry (HRMS), nuclear magnetic resonance (NMR) and Fourier-transformed infrared (FTIR) spectroscopy, using research chemicals samples collected by the drug analysis service Energy Control from anonymous users. Compound identification was performed by the combination of HRMS and NMR data. The elemental composition and putative moieties of the compound were determined based on the accurate-mass ions observed by HRMS. Then, different NMR experiments, including bidimensional, allowed the establishment of the chemical structure and confirmation of compound identity. Furthermore, FTIR spectrum was also acquired in order to provide a complete analytical characterization of the novel cathinone. Finally, pharmacological/toxicological characterization was attempted using in silico methods. Based on these predictions, N-cyclohexyl butylone probably has similar effects to stimulants like MDMA.

A model study on the site-specificity of (−)-epicatechin-induced reactions in β-lactoglobulin by high-resolution mass spectrometry in combination with bioinformatics

Polyphenol–protein reactions in model solutions of β-lactoglobulin (β-LG) incubated with (−)-epicatechin at 37 °C and 60 °C were monitored by microLC–timsTOF Pro-MS/MS combined with bioinformatics strategies. The addition of (−)-epicatechin to the model solutions resulted in changes in tryptic peptide profiles. Covalent bond formation between (−)-epicatechin o-quinones and β-LG was identified for the residues S27, S30, K60, C66, K69, and C160, with C160 being the predominant binding site. Furthermore, the incubation of β-LG with (−)-epicatechin significantly promoted oxidation, especially for the residues M7 and M24. The reaction of monomeric (−)-epicatechino-quinone at C160 was also identified in the milk chocolate sample. The adaptation of this study by extending the scope of the reaction products offers significant potential for comprehensive food profiling strategies.

CCS Predictor 2.0: An Open-Source Jupyter Notebook Tool for Filtering Out False Positives in Metabolomics.

Metabolite annotation continues to be the widely accepted bottleneck in nontargeted metabolomics workflows. Annotation of metabolites typically relies on a combination of high-resolution mass spectrometry (MS) with parent and tandem measurements, isotope cluster evaluations, and Kendrick mass defect (KMD) analysis. Chromatographic retention time matching with standards is often used at the later stages of the process, which can also be followed by metabolite isolation and structure confirmation utilizing nuclear magnetic resonance (NMR) spectroscopy. The measurement of gas-phase collision cross-section (CCS) values by ion mobility (IM) spectrometry also adds an important dimension to this workflow by generating an additional molecular parameter that can be used for filtering unlikely structures. The millisecond timescale of IM spectrometry allows the rapid measurement of CCS values and allows easy pairing with existing MS workflows. Here, we report on a highly accurate machine learning algorithm (CCSP 2.0) in an open-source Jupyter Notebook format to predict CCS values based on linear support vector regression models. This tool allows customization of the training set to the needs of the user, enabling the production of models for new adducts or previously unexplored molecular classes. CCSP produces predictions with accuracy equal to or greater than existing machine learning approaches such as CCSbase, DeepCCS, and AllCCS, while being better aligned with FAIR (Findable, Accessible, Interoperable, and Reusable) data principles. Another unique aspect of CCSP 2.0 is its inclusion of a large library of 1613 molecular descriptors via the Mordred Python package, further encoding the fine aspects of isomeric molecular structures. CCS prediction accuracy was tested using CCS values in the McLean CCS Compendium with median relative errors of 1.25, 1.73, and 1.87% for the 170 [M - H]-, 155 [M + H]+, and 138 [M + Na]+ adducts tested. For superclass-matched data sets, CCS predictions via CCSP allowed filtering of 36.1% of incorrect structures while retaining a total of 100% of the correct annotations using a ΔCCS threshold of 2.8% and a mass error of 10 ppm.

Solvation of Large Polycyclic Aromatic Hydrocarbons in Helium: Cationic and Anionic Hexabenzocoronene.

The adsorption of helium on charged hexabenzocoronene (Hbc, C42H18), a planar polycyclic aromatic hydrocarbon (PAH) molecule of D6h symmetry, was investigated by a combination of high-resolution mass spectrometry and classical and quantum computational methods. The ion abundance of HenHbc+ complexes versus size n features prominent local anomalies at n = 14, 38, 68, 82, and a weak one at 26, indicating that for these “magic” sizes, the helium evaporation energies are relatively large. Surprisingly, the mass spectra of anionic HenHbc− complexes feature a different set of anomalies, namely at n = 14, 26, 60, and 62, suggesting that the preferred arrangement of the adsorbate atoms depends on the charge of the substrate. The results of our quantum calculations show that the adsorbate layer grows by successive filling of concentric rings that surround the central benzene ring, which is occupied by one helium atom each on either side of the substrate. The helium atoms are fairly localized in filled rings and they approximately preserve the D6h symmetry of the substrate, but helium atoms in partially filled rings are rather delocalized. The first three rings contain six atoms each; they account for magic numbers at n = 14, 26, and 38. The size of the first ring shrinks as atoms are filled into the second ring, and the position of atoms in the second ring changes from hollow sites to bridge sites as atoms are filled into the third ring. Beyond n = 38, however, the arrangement of helium atoms in the first three rings remains essentially frozen. Presumably, another ring is filled at n = 68 for cations and n = 62 for anions. The calculated structures and energies do not account for the difference between charge states, although they agree with the measurements for the cations and show that the first solvation shell of Hbc± is complete at n = 68. Beyond that size, the adsorbate layer becomes three-dimensional, and the circular arrangement of helium changes to hexagonal.

Adsorption of Helium and Hydrogen on Triphenylene and 1,3,5-Triphenylbenzene.

The adsorption of helium or hydrogen on cationic triphenylene (TPL, C18H12), a planar polycyclic aromatic hydrocarbon (PAH) molecule, and of helium on cationic 1,3,5-triphenylbenzene (TPB, C24H18), a propeller-shaped PAH, is studied by a combination of high-resolution mass spectrometry and classical and quantum computational methods. Mass spectra indicate that HenTPL+ complexes are particularly stable if n = 2 or 6, in good agreement with the quantum calculations that show that for these sizes, the helium atoms are strongly localized on either side of the central carbon ring for n = 2 and on either side of the three outer rings for n = 6. Theory suggests that He14TPL+ is also particularly stable, with the helium atoms strongly localized on either side of the central and outer rings plus the vacancies between the outer rings. For HenTPB+, the mass spectra hint at enhanced stability for n = 2, 4 and, possibly, 11. Here, the agreement with theory is less satisfactory, probably because TPB+ is a highly fluxional molecule. In the global energy minimum, the phenyl groups are rotated in the same direction, but when the zero-point harmonic correction is included, a structure with one phenyl group being rotated opposite to the other two becomes lower in energy. The energy barrier between the two isomers is very small, and TPB+ could be in a mixture of symmetric and antisymmetric states, or possibly even vibrationally delocalized.

Detecting Water Constituents Unique to Septic Tanks as a Wastewater Source in the Environment by Nontarget Analysis: South Florida's Deering Estate Rehydration Project Case Study.

The present study has generated a workflow based on nontarget analysis (NTA) with Compound Discoverer Ver 3.1 to characterize a set of source-discriminating compounds identified in water samples from locations in South Florida (USA), particularly those describing a freshwater environment (Everglades based), urban impacted areas (septic tank driven), and coastal (Biscayne Bay) endmembers in and around the Charles Deering Estate property in the Village of Palmetto Bay. Waters from an interconnected managed canal system were assessed to evaluate the influence of localized emissions. Septic tank effluents influence the water in many Southeast Florida environments due to their diminished onsite treatment capacity based on the limestone-dominated geology and canal systems providing a relatively unobstructed connection pathway. Through a combination of high-resolution mass spectrometry and statistical analyses, a set of tracers and indicators was determined (azelaic acid, decanophenone, galaxolidone, methyl violet, monoolein, metoprolol, and 1-stearoylglycerol). Tentatively identified compounds were generally assigned to various categories such as dyes, personal care products, and pharmaceuticals. The NTA Compound Discoverer Ver 3.1 compound data (presented as principal component analysis and Kendrick mass defect plots) showed apparent differences between wastewater-influenced sites and non-wastewater-influenced sites along with the ranked "Top10" compounds found at each location. Waters from different locations were also compared using the presence of sucralose to further inform the NTA. The most septic-influenced site contained 3594 ± 94 ng/L of sucralose with concentrations declining steadily and reaching the lowest concentrations in Biscayne Bay of 122 ± 94 ng/L. The sucralose concentrations provided further evidence of septic influence on this system. Sucralose was determined to be a conservative tracer between the freshwater and coastal sources and complementary to other probable unique tracers of septic tank effluent identified by the NTA. Environ Toxicol Chem 2022;41:1165-1178. © 2022 SETAC.

Novel chlorinated disinfection byproducts from tannic acid: nontargeted identification, formation pathways, and computationally predicted toxicity

Tannic acid is ubiquitously present in various simulated and real water sources and in wastewater. Various chlorinated disinfection byproducts (Cl-DBPs) are generated via reactions with tannic acid during disinfection with chlorine. We used high-resolution mass spectrometry in combination with our self-developed halogen extraction code to selectively identify Cl-DBPs. Our strategy enabled successful detection of 35 Cl-DBP formulas formed by chlorination of tannic acid, and we identified 26 of these structures. The structures of 17 novel Cl-DBPs are firstly reported here. The reaction pathways of these Cl-DBPs were tentatively proposed. These Cl-DBPs are likely to be generated during chlorination at various chlorine doses, from 0.0 to 10.0 mg-Cl2/L, and 14 of the 26 Cl-DBPs were detected in simulated drinking water, which implies a relatively high probability of incidence. Quantitative structure-activity relationship toxicity analyses predicted that most of these Cl-DBPs would exhibit much higher acute toxicity than the common DBPs trichloromethane and monochloroacetic acid and that some of these compounds would induce developmental toxicity and be mutagenic, which further emphasizes that these Cl-DBPs should raise concerns. This study broadens our understanding of the Cl-DBPs formed from tannic acid and should prompt wider application of our analytical strategy in environmental matrices.

Quantitative proteomics identifies biomarkers to distinguish pulmonary from head and neck squamous cell carcinomas by immunohistochemistry.

The differentiation between a pulmonary metastasis and a newly developed squamous cell carcinoma of the lung in patients with prior head and neck squamous cell carcinoma (HNSCC) is difficult due to a lack of biomarkers but is crucially important for the prognosis and therapy of the affected patient. By using high‐resolution mass spectrometry in combination with stable isotope labelling by amino acids in cell culture, we identified 379 proteins that are differentially expressed in squamous cell carcinomas of the lung and the head and neck. Of those, CAV1, CAV2, LGALS1, LGALS7, CK19, and UGDH were tested by immunohistochemistry on 194 tissue samples (98 lung and 96 HNSCCs). The combination of CAV1 and LGALS7 was able to distinguish the origin of the squamous cell carcinoma with high accuracy (area under the curve 0.876). This biomarker panel was tested on a cohort of 12 clinically classified lung tumours of unknown origin after HNSCC. Nine of those tumours were immunohistochemically classifiable.

First direct determination of the 93Mo half-life

This work presents the first direct measurement of the 93Mo half-life. The measurement is a combination of high-resolution mass spectrometry for the determination of the 93Mo concentration and liquid scintillation counting for determining the specific activity. A 93Mo sample of high purity was obtained from proton irradiated niobium by chemical separation of molybdenum with a decontamination factor larger than 1.6 × 1014 with respect to Nb. The half-life of 93Mo was deduced to be 4839(63) years, which is more than 20% longer than the currently adopted value, whereas the relative uncertainty could be reduced by a factor of 15. The probability that the 93Mo decays to the metastable state 93mNb was determined to be 95.7(16)%. This value is a factor of 8 more precise than previous estimations. Due to the man-made production of 93Mo in nuclear facilities, the result leads to significantly increased precision for modelling the low-level nuclear waste composition. The presented work demonstrates the importance of chemical separations in combination with state-of-the-art analysis techniques, which are inevitable for precise and accurate determinations of nuclear decay data.

High-throughput enzyme inhibition screening of 44 Iranian medicinal plants via piezoelectric spraying of planar cholinesterase assays

A rapid and straightforward approach was developed for screening the acetyl- and butyrylcholinesterase (ChE) inhibitory activity of 44 Iranian medicinal plant extracts at laboratory scale. After a fast ChE inhibitory pre-testing of samples applied as band pattern, 40 out of the 44 Iranian medicinal plant extracts were selected. These were adjusted in the application volume depending on their inhibition activity, applied on both plate sides and simultaneously developed in a horizontal developing chamber. Different mobile phases were studied to achieve maximum separation of ChE inhibitors and minimum co-elution with matrix. Contrary to immersion, the piezoelectric spraying reduced the consumption of assay solutions, prevented zone tailing, zone shift and cross-contamination, and homogeneously covered the entire plate surface with the assay solutions. The ChE inhibitors of the six most bioactive plant extracts were tentatively assigned by high-resolution mass spectrometry in combination with the spectral and chromatographic information obtained.

Transformation of ammonium to nitrophenolic byproducts by sulfate radical oxidation

Sulfate radical (SO4•−) based oxidation shows great promise in wastewater treatment and subsurface remediation. For the first time, we demonstrated that SO4•− could induce the transformation of ammonium (NH4+) to nitrophenolic byproducts. Using high-resolution mass spectrometry in combination with 15N labeling, mono-nitro and di-nitro phenolic byproducts were identified in a sample containing 1 mM NH4+ and 10 mg/L natural organic matter (NOM) following heat activated peroxydisulfate (PDS) oxidation. At PDS dose of 1 mM, the formation of p-nitrophenol and 5-nitrosalicylic acid reached 0.21 and 0.30 μM, respectively, in 12 h and then decreased; the formation of 2,4-dinitrophenol and 3,5-dinitrosalicylic acid increased monotonically, reaching 0.37 and 0.62 μM, respectively, in 24 h. One-electron oxidation of NH4+ to form aminyl radicals (•NH2) was the first step of the transformation. The reaction of •NH2 with oxygen was a key step in propagating radical chain reactions, leading to nitrogen dioxide radicals (NO2•) as a key nitrating agent. The reactive sites susceptible to nitrating in NOM molecules are not limited to phenolic moieties. We found that aromatic carboxylate moieties could be in situ transformed to phenolics by SO4•−, thus contributed to nitrophenolic byproducts formation as well. Considering the ubiquitous presence of NH4+ in the environment, formation of nitrophenolic byproducts will be widespread when SO4•− is applied for onsite remediation, which should be taken into consideration when evaluating the feasibility of this technology.

The skeletome of the red coral Corallium rubrum indicates an independent evolution of biomineralization process in octocorals

BackgroundThe process of calcium carbonate biomineralization has arisen multiple times during metazoan evolution. In the phylum Cnidaria, biomineralization has mostly been studied in the subclass Hexacorallia (i.e. stony corals) in comparison to the subclass Octocorallia (i.e. red corals); the two diverged approximately 600 million years ago. The precious Mediterranean red coral, Corallium rubrum, is an octocorallian species, which produces two distinct high-magnesium calcite biominerals, the axial skeleton and the sclerites. In order to gain insight into the red coral biomineralization process and cnidarian biomineralization evolution, we studied the protein repertoire forming the organic matrix (OM) of its two biominerals.ResultsWe combined High-Resolution Mass Spectrometry and transcriptome analysis to study the OM composition of the axial skeleton and the sclerites. We identified a total of 102 OM proteins, 52 are found in the two red coral biominerals with scleritin being the most abundant protein in each fraction. Contrary to reef building corals, the red coral organic matrix possesses a large number of collagen-like proteins. Agrin-like glycoproteins and proteins with sugar-binding domains are also predominant. Twenty-seven and 23 proteins were uniquely assigned to the axial skeleton and the sclerites, respectively. The inferred regulatory function of these OM proteins suggests that the difference between the two biominerals is due to the modeling of the matrix network, rather than the presence of specific structural components. At least one OM component could have been horizontally transferred from prokaryotes early during Octocorallia evolution.ConclusionOur results suggest that calcification of the red coral axial skeleton likely represents a secondary calcification of an ancestral gorgonian horny axis. In addition, the comparison with stony coral skeletomes highlighted the low proportion of similar proteins between the biomineral OMs of hexacorallian and octocorallian corals, suggesting an independent acquisition of calcification in anthozoans.

Synthesis and characterization of heteroleptic rare earth double-decker complexes involving tetradiazepinoporphyrazine and phthalocyanine macrocycles.

Reaction of (2,3,9,10,16,17,23,24-octabutylphthalocyaninato)lanthanide(iii) acetylacetonates (BuPcLn(acac), 1a-c, Ln = Lu (a), Eu (b), La (c)) with a tetrakis(5,7-bis(4-tert-butylphenyl)-6H-1,4-diazepino)[2,3-b,g,l,q]porphyrazine ligand (tBuPhDzPzH2, 2) produced sandwich compounds (tBuPhDzPz)Ln(BuPc) (3a-c), which represent the first heteroleptic double-deckers incorporating both Pc and DzPz decks. A combination of high-resolution mass spectrometry, UV-Vis/NIR, MCD, and 1H NMR spectroscopy, and square-wave voltammetry provided unambiguous characterization of target complexes 3 indicating that their spectral and electrochemical properties are generally intermediate with respect to their homoleptic relatives. Based on the data of solution-state 1H-1H NMR (COSY, NOESY) correlation spectroscopy supported by DFT calculations, a dimerization tendency of compounds 3 proportional to the Ln(iii) ion size was found. The spectroelectrochemical study of 3 and the corresponding homoleptic double-deckers revealed a pronounced tendency to aggregation of the one-electron oxidized forms of DzPz-containing double-decker complexes compared to homoleptic Pc2Ln compounds.

Study of the sedge (Cárex) lignin by high-resolution mass spectrometry and NMR spectroscopy

A combination of high-resolution mass spectrometry based on an orbital ion trap and 2D NMR spectroscopy was first applied to the characterization of the structure of poorly studied lignins of herbaceous plants using dioxane lignin from sedge (Carex) as an example. Macromolecules of the Carex lignin are found to be constructed from guaiacyl, syringyl, and p-hydroxyphenyl structural units in a ratio of 57:22:21 bound mainly by ether β-O-4 bonds. The key role of p-hydroxycinnamic acids in the formation of the lignin structure and ester bonds with polysaccharides was shown. A possible structural formula of the macromolecule fragment of the Carex lignin was proposed on the basis of an analysis of the 2D NMR spectra (HSQC).

Material characterization of polyamide 12 and related agents used in the multi-jet fusion process: complementary application of high-resolution mass spectrometry and other advanced instrumental techniques

The powder bed-based 3D printing process called multi-jet fusion from HP is used in various industries for different applications. A polymer powder, the so-called fusing and detailing agents, and an IR source are needed. These novel materials offer some challenges for analytical characterization. High-resolution mass spectrometry in combination with a data base search was used as a tool for comprehensive characterization of the agents, the methanol extract of new polyamide 12 powder, and the methanol extract of a grinded 3D printed part. In addition, thermogravimetric studies were carried out. Besides biocides and substances containing ethoxy units, 2-pyrrolidone and triethylene glycol were identified as organic solvents in the agents. The detailing agent contained 3.7% of 2-pyrrolidone and 11.1% of triethylene glycol. The fusing agent contained 18.7% of 2-pyrrolidone and 8.4% of triethylene glycol. Based on the agents’ different functionalities, the water content of the detailing agent was 18% higher as compared to the fusing agent. In addition, the fusing agent contained a carbon black amount of 5.2%. The methanol extract of new polyamide 12 powder contained remaining monomer and oligomer compounds, as well as the oxidation stabilizer Irganox 1098. In the methanol extract of the grinded 3D printed part the cyclic and linear monomer, oligomers, and oxidation stabilizer were found. Moreover, ingredients of the agents were detected. Residues of 2-pyrrolidone, triethylene glycol, 3-benzisothiazolinone, and substances containing ethoxy units were identified in the methanol extract of the grinded 3D printed part. In contrast, no 2-methyl-4-isothiazolin-3-one was observed.Graphical abstract

Isolation of Trametes hirsuta La-7 with high laccase-productivity and its application in metabolism of 17β-estradiol

Estrogens, which are extensive in the eco-environments, are a category of high-toxic emerging contaminants that induce metabolic disorders and even carcinogenic risks in wildlife and humans. Here we investigate whether fungus-secreted laccase can be used as a green catalyst to eliminate a representative estrogen, 17β-estradiol (E2). A white-rot fungus Trametes hirsuta La-7 with high laccase-productivity, was isolated from pig manure-contaminated soil. Extracellular laccase activity expressed by strain La-7 was 65.4 U·mL−1 for a 3 d inoculation under the optimal fermentation parameters. The concentrated-crude laccase from Trametes hirsuta La-7 (CC-ThLac) was capable of effectively metabolizing E2 at pH 4–6, and the apparent pseudo first-order reaction rate constant and half-life values were respectively 0.027–0.055 min−1 and 25.86–12.67 min (R2 > 0.98). The mass measurement of high-resolution mass spectrometry in combination with 13C-isotope labeling identified that the main by-products of E2 metabolism were dimers, trimers, and tetramers, which are consistent with radical-driven C–C and/or C–O–C covalent coupling pathway, involving the initial enzymatic production of phenoxy radical intermediates and then the successive oxidative-oligomerization of radical intermediates. The formation of oligomers dramatically reduced the estrogenic activity of E2. Additionally, CC-ThLac also exhibited high-efficiency metabolism capability toward E2 in the natural water and pig manure, with more than 94.4% and 91.0% of E2 having been metabolized, respectively. These findings provide a broad prospect for the clean biotechnological applications of Trametes hirsuta La-7 in estrogen-contaminated ecosystems.

Albanitriles A-G: Antiprotozoal Polyacetylene Nitriles from a Mycale Marine Sponge.

Seven new nitrile-bearing polyacetylenes, named albanitriles A-G, were isolated from a marine sponge of the Mycale genus (Order: Poecilosclerida, Family: Mycalidae) collected near Albany, Western Australia. Structural elucidation was achieved using a combination of high-resolution mass spectrometry and ultraviolet/visible, infrared, and nuclear magnetic resonance spectroscopy. The compounds were found to possess moderate activity against Giardia duodenalis when compared to a metronidazole positive control.

Formation of Nitrophenolic Byproducts during Heat-Activated Peroxydisulfate Oxidation in the Presence of Natural Organic Matter and Nitrite.

Sulfate radical (SO4•-)-based advanced oxidation is a viable in situ remediation technology for degrading organic contaminants in the subsurface. In this study, we demonstrated that SO4•- could induce the activation of nitrite, an anion commonly present in the subsurface environment, leading to the formation of nitrophenolic byproducts. Fourier-transform infrared spectroscope and 15N nuclear magnetic resonance analysis revealed that the inorganic nitrite was incorporated into natural organic matter (NOM) to form organic nitrogen upon SO4•- oxidation. Nitrophenolic byproducts, including 2-hydroxy-5-nitrobenzoic acid, 4-nitrophenol, and 2,4-dinitrophenol, were identified using high-resolution mass spectrometry in combination with a 15N labeling technique. Formation of nitrated byproducts was ascribed to the scavenging of SO4•- by nitrite, which not only generated the nitrating agent NO2• but also inhibited the degradation of organic compounds, making them more available to the reactions with NO2•. The phenolic moieties in NOM served as the main reactive sites for NO2• attack. The nitration begins with H abstraction on the phenoxy oxygen, followed by the addition of another NO2• to its ortho or para site. Decarboxylation followed by NO2• addition can also generate nitrophenolic byproducts. To the best of our knowledge, this is the first study reporting the nitration of NOM and formation of toxic nitrophenolic byproducts during SO4•--based oxidation. It sheds light on the potential risks of this technology in subsurface remediation practices.

A Study on Tissue-Specific Metabolite Variations in Polygonum cuspidatum by High-Resolution Mass Spectrometry-Based Metabolic Profiling.

Polygonum cuspidatum Sieb. et Zucc. is a traditional Chinese herbal medicine widely used to treat tussis, hepatitis and arthralgia. This study identified and quantitatively described the bioactive compounds in different P. cuspidatum tissues. Metabolic profiles of root, stem, leaf, flower, rhizome and seed were determined using high-resolution mass spectrometry in combination with multivariate analyses. In total, 53 metabolites, 8 reported for the first time in this species, were putatively identified and classified mainly as stilbenes, anthraquinones and flavonoids. A principal component analysis, cluster analysis and heatmap were used to depict the correlations between specimens and the relative abundance levels of these compounds in different plant tissues. An orthogonal partial least square discriminant analysis found that 13 metabolites showed distinct differences among the six plant tissues, making them potential discriminative tissue-identification markers. This study will provide guidance in comparing, selecting and exploiting the medicinal uses of different P. cuspidatum tissues.