Lignin is a promising renewable source of valuable organic compounds and environmentally benign materials. However, its involvement in economic circulation and the creation of new biorefining technologies require an understanding of its chemical composition and structure. This problem can be overcome by applying mass spectrometry analytical techniques in combination with advanced chemometric methods for mass spectra processing. The present study is aimed at the development of mass defect filtering to characterize the chemical composition of lignin at the molecular level. This study introduces a novel approach involving resolution-enhanced Kendrick mass defect (REKMD) analysis for the processing of atmospheric pressure photoionization Orbitrap mass spectra of lignin. The set of priority Kendrick fractional base units was predefined in model experiments and provided a substantially expanding available mass defect range for the informative visualization of lignin mass spectra. The developed REKMD analysis strategy allowed to obtain the most complete data on all the homologous series typical of lignin and thus facilitated the interpretation and assignment of elemental compositions and structural formulas to oligomers detected in extremely complex mass spectra, including tandem ones. For the first time, the minor modifications (sulfation) of lignin obtained in ionic liquid-based biorefining processes were revealed.

With significant advancements in high-resolution mass spectrometry, there has been a substantial increase in the amount of chemical component data acquired from natural products. Therefore, the rapid and efficient extraction of valuable mass spectral information from large volumes of high-resolution mass spectrometry data holds crucial significance. This study illustrates a targeted annotation of the metabolic products of alkaloid and sesquiterpene components from Dendrobium nobile (D. nobile) aqueous extract in mice serum through the integration of an in-houses database, R programming, a virtual metabolic product library, polygonal mass defect filtering, and Kendrick mass defect strategies. The research process involved initially establishing a library of alkaloids and sesquiterpenes components and simulating 71 potential metabolic reactions within the organism using R programming, thus creating a virtual metabolic product database. Subsequently, employing the virtual metabolic product library allowed for polygonal mass defect filtering, rapidly screening 1705 potential metabolites of alkaloids and 3044 potential metabolites of sesquiterpenes in the serum. Furthermore, based on the chemical composition database of D. nobile and online mass spectrometry databases, 95 compounds, including alkaloids, sesquiterpenes, and endogenous components, were characterized. Finally, utilizing Kendrick mass defect analysis in conjunction with known alkaloids and sesquiterpenes targeted screening of 209 demethylation, methylation, and oxidation products in phase I metabolism, and 146 glucuronidation and glutathione conjugation products in phase II metabolism. This study provides valuable insights for the rapid and accurate annotation of chemical components and their metabolites in vivo within natural products.

Capturing the breadth of chemical exposures in utero is critical in understanding their long-term health effects for mother and child. We explored methodological adaptations in a Non-Targeted Analysis (NTA) pipeline and evaluated the effects on chemical annotation and discovery for maternal and infant exposure. We focus on lesser-known/underreported chemicals in maternal and umbilical cord serum analyzed with liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-QTOF/MS). The samples were collected from a demographically diverse cohort of 296 maternal-cord pairs (n = 592) recruited in San Francisco Bay area. We developed and evaluated two data processing pipelines, primarily differing by detection frequency cut-off, to extract chemical features from non-targeted analysis (NTA). We annotated the detected chemical features by matching with EPA CompTox Chemicals Dashboard (n = 860,000 chemicals) and Human Metabolome Database (n = 3140 chemicals) and applied a Kendrick Mass Defect filter to detect homologous series. We collected fragmentation spectra (MS/MS) on a subset of serum samples and matched to an experimental MS/MS database within the MS-Dial website and other experimental MS/MS spectra collected from standards in our lab. We annotated ~72 % of the features (total features = 32,197, levels 1-4). We confirmed 22 compounds with analytical standards, tentatively identified 88 compounds with MS/MS spectra, and annotated 4862 exogenous chemicals with an in-house developed annotation algorithm. We detected 36 chemicals that appear to not have been previously reported in human blood and 9 chemicals that were reported in less than five studies. Our findings underline the importance of NTA in the discovery of lesser-known/unreported chemicals important to characterize human exposures.

Lignin is the second most abundant biopolymer in nature and a promising renewable feedstock for the production of aromatic compounds, composite materials, sorbents, etc. Being a complex mixture of oligomeric molecules with an irregular structure, natural lignin is an extremely difficult object to study. Its molecular level characterization requires advanced analytical techniques among which atmospheric pressure photoionization Orbitrap mass spectrometry holds a promising place. In the present study, Kendrick mass defect (KMD) analysis was proposed to facilitate the visualization and interpretation of Orbitrap mass spectra of the biopolymer on an example of Siberian pine dioxane lignin preparation. The use of the typical guaiacylpropane structure C10H12O4 as a Kendrick base unit made it possible to effectively identify oligomer series with different polymerization degrees and structurally related compounds, as well as to reliably determine the elemental compositions and structures of oligomers with high molecular weights (> 1 kDa). For the first time, KMD analysis was applied to the interpretation of the complex tandem mass spectra of lignin oligomers, rapid discrimination of the product ion series, and the establishment of the main collision-induced dissociation pathways. It was demonstrated that especially promising was the use of KMD filtering in the study of broadband fragmentation tandem mass spectra, which allows for the structural characterization of all oligomers with a particular degree of polymerization.

Soil contaminations with per- and polyfluoroalkyl substances (PFAS) are of great concern due to their persistence, leading to continuous, long-term groundwater contamination. A composite sample from contaminated agricultural soil from northwestern Germany (Brilon-Scharfenberg, North Rhine-Westphalia) was investigated in depth with nontarget screening (NTS) (Kendrick mass defect and MS2 fragment mass differences with FindPFΔS). Several years ago, selected PFCAs and PFSAs were identified on this site by detection in nearby surface and drinking water. We identified 10 further PFAS classes and 7 C8-based PFAS (73 single PFAS) previously unknown in this soil including some novel PFAS. All PFAS classes except for one class comprised sulfonic acid groups and were semi-quantified with PFSA standards from which ∼97% were perfluorinated and are not expected to be degradable. New identifications made up >75% of the prior known PFAS concentration, which was estimated to >30 μg/g. Pentafluorosulfanyl (-SF5) PFSAs are the dominant class (∼40%). Finally, the soil was oxidized with the direct TOP (dTOP) assay, revealing PFAA precursors that were covered to a large extent by identified H-containing PFAS and additional TPs (perfluoroalkyl diacids) were detected after dTOP. In this soil, however, dTOP + target analysis covers <23% of the occurring PFAS, highlighting the importance of NTS to characterize PFAS contaminations more comprehensively.

Only a few dozens of the several thousand existing per- and polyfluoroalkyl substances (PFAS) are monitored using conventional target analysis. This study employed suspect screening to examine patterns of emerging and novel PFAS in German and Chinese river water affected by industrial point sources. In total, 86 PFAS were (tentatively) identified and grouped into 18 structure categories. Homologue patterns revealed distinct differences between fluoropolymer production sites of the two countries. In the Chinese Xiaoqing River Basin, the C8 homologue was the most prevalent compound of the emerging series of chlorinated perfluoroalkyl carboxylic acids (Cl-PFCAs) and perfluoroalkylether carboxylic acids (PFECAs). In contrast, C6 and shorter homologues were dominant in the German Alz River. This indicates that the phaseout of long-chain compounds in Europe and their ongoing production in Asian countries also apply to unregulated emerging PFAS classes. Additional characteristics to differentiate the point sources were the peak area ratio of perfluorobutane sulfonic acid (PFBS) versus the emerging compound hydro-substituted PFBS (H-PFBS) as well as the occurrence of byproducts of the sulfonated tetrafluoroethylene-based polymer Nafion. The large number of identified unregulated PFAS underlines the importance of a grouping approach on a regulatory level, whereas the revealed contamination patterns can be used to estimate, prioritize, and minimize contributions of specific sources.

Per- and polyfluoroalkyl substances (PFASs) are a large family of thousands of chemicals, many of which have been identified using nontargeted time-of-flight and Orbitrap mass spectrometry methods. Comprehensive characterization of complex PFAS mixtures is critical to assess their environmental transport, transformation, exposure, and uptake. Because 21 tesla (T) Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR MS) offers the highest available mass resolving power and sub-ppm mass errors across a wide molecular weight range, we developed a nontargeted 21 T FT-ICR MS method to screen for PFASs in an aqueous film-forming foam (AFFF) using suspect screening, a targeted formula database (C, H, Cl, F, N, O, P, S; ≤865 Da), isotopologues, and Kendrick-analogous mass difference networks (KAMDNs). False-positive PFAS identifications in a natural organic matter (NOM) sample, which served as the negative control, suggested that a minimum length of 3 should be imposed when annotating CF2-homologous series with positive mass defects. We putatively identified 163 known PFASs during suspect screening, as well as 134 novel PFASs during nontargeted screening, including a suspected polyethoxylated perfluoroalkane sulfonamide series. This study shows that 21 T FT-ICR MS analysis can provide unique insights into complex PFAS composition and expand our understanding of PFAS chemistries in impacted matrices.

Zwitterionic, cationic, and anionic per- and polyfluoroalkyl substances (PFAS) are increasingly reported in terrestrial and aquatic environments, but their inputs to agricultural lands are not fully understood. Here, we characterized PFAS in 47 organic waste products (OWP) applied in agricultural fields of France, including historical and recent materials. Overall, 160 PFAS from 42 classes were detected from target screening and homologue-based nontarget screening. Target PFAS were low in agriculture-derived wastes such as pig slurry, poultry manure, or dairy cattle manure (median ∑46PFAS: 0.66 μg/kg dry matter). Higher PFAS levels were reported in urban and industrial wastes, paper mill sludge, sewage sludge, or residual household waste composts (median ∑46PFAS: 220 μg/kg). Historical municipal biosolids and composts (1976-1998) were dominated by perfluorooctanesulfonate (PFOS), N-ethyl perfluorooctanesulfonamido acetic acid (EtFOSAA), and cationic and zwitterionic electrochemical fluorination precursors to PFOS. Contemporaneous urban OWP (2009-2017) were rather dominated by zwitterionic fluorotelomers, which represented on average 55% of ∑160PFAS (max: 97%). The fluorotelomer sulfonamidopropyl betaines (X:2 FTSA-PrB, median: 110 μg/kg, max: 1300 μg/kg) were the emerging class with the highest occurrence and prevalence in contemporary urban OWP. They were also detected as early as 1985. The study informs for the first time that urban sludges and composts can be a significant repository of zwitterionic and cationic PFAS.

Lipids are biomolecules of crucial importance involved in critical biological functions. Yet, lipid content determination using mass spectrometry is still challenging due to their rich structural diversity. Preferential ionisation of the different lipid species in the positive or negative polarity is common, especially when using soft ionisation mass spectrometry techniques. Here, we demonstrate the potency of a dual-polarity approach using surface-assisted laser desorption/ionisation coupled to Fourier transform-ion cyclotron resonance (SALDI FT-ICR) mass spectrometry imaging (MSI) combined with Kendrick mass defect data filtering to (i) identify the lipids detected in both polarities from the same tissue section and (ii) show the complementarity of the dual-polarity data, both regarding the lipid coverage and the spatial distributions of the various lipids. For this purpose, we imaged the same mouse brain section in the positive and negative ionisation modes, on alternate pixels, in a SALDI FT-ICR MS imaging approach using gold nanoparticles (AuNPs) as dual-polarity nanosubstrates. Our study demonstrates, for the first time, the feasibility of (i) a dual-polarity SALDI-MSI approach on the same tissue section, (ii) using AuNPs as nanosubstrates combined with a FT-ICR mass analyser and (iii) the Kendrick mass defect data filtering applied to SALDI-MSI data. In particular, we show the complementarity in the lipids detected both in a given ionisation mode and in the two different ionisation modes. Graphical abstract.

BACKGROUND: Gloriosa superba L. is a promising antitumoural plant species as a source of colchicinoids. Ethnobotanical applications of G. superba are associated with different plant parts such as leaves, seeds, fruits, tuber and the whole plant.
OBJECTIVE: A comparative phytochemical study of purified extracts from in vitro cultures and native tubers of G. superba was carried out by ultrahigh-performance liquid chromatography-high-resolution mass spectrometry (UHPLC-HR-MS) in combination with the mass defect filtering (MDF) technique.
METHODS: The individual compounds were tentatively annotated using database correlations, retention time (Rt), accurate m/z data obtained by electrospray ionisation (ESI) (+)-HR-MS, proposed elemental composition, ring double bond equivalent (RDBeq) values and HR-MS/MS fragmentation patterns. Moreover, the identification was based on transforming the exact mass ratio (m/z) for the protonated molecular ions [М + Н]+ of the observed metabolites in Kendrick nominal masses (NKMs) and calculation of the Kendrick mass defect (KMD), which made it possible to graphically present the ion peaks in Kendrick plots.
RESULTS: Building Kendrick plots allows easy differentiation of small structural differences such as methylation or demethylation of compounds from the same homologous series. In this way, a wide range of tropolone alkaloids was characterised. A greater variety was observed in in vitro cultures, compared to native sources.
CONCLUSIONS: This LC-MS analysis unambiguously demonstrated the presence of tropolone alkaloids in in vitro cultures of G. superba. This approach of LC-MS data interpretation can be used to understand complex mass spectra such as those of plant extracts.