bims-mebolo Biomed News
on Metabolomics
Issue of 2026–06–14
fifty-two papers selected by
Daniel Méndez Rodríguez, Vbi-Ugent
  1. Host-Dependent Metabolomic Variations in Viscum coloratum Revealed by UHPLC-Q-TOF-MS/MS and Integrated Annotation Strategies.
  2. Chemical Profiling of Aboveground and Underground Parts of Pterocephalus hookeri by Integrated FBMN, Untargeted LC-MS Metabolomics, and PAD-DESI-MSI.
  3. Untargeted metabolomics suggests a trade-off between phenolamides and flavonoids in the pollen of Petunia hybrida with contrasting flower colours.
  4. Phytochemical Characterization and Comparative Tissue-Specific Metabolomics of Coffea arabica Using LC-HRMS.
  5. Integrated UHPLC-MS/MS and HRMS Strategy for Screening Potentially Hepatotoxic Furan-Containing Compounds in Dioscorea bulbifera L.
  6. Integrated MALDI-MSI and UHPLC-OE-MS for Spatial Visualization and Biosynthetic Pathway Elucidation of Bioactive Metabolites in Lilium lancifolium Thunb.
  7. Systematic profiling of chemical components in Tribulus terrestris L. extract and its absorbed parent compounds/metabolites in rat plasma utilizing UPLC-Q-TOF/MS.
  8. eMZed 3: flexible and interactive development of scalable LC-MS/MS data analysis workflows in python.
  9. MultiMS2: A curated multi-modal, multi-energy spectral library for metabolomics.
  10. ToxBase: A Multidimensional ToxCast Reference Database for High-Throughput Human Exposome Analysis.
  11. Untargeted and Targeted Cerebrospinal Fluid Neurometabolomics via Chromatography-Mass Spectrometry-Based Methods.
  12. Metabolic alterations and potential biomarkers in unstable angina investigated by lipidomic analysis.
  13. A Standardized Metabolomics Protocol For Analyzing Exercise-Induced Metabolic Shifts In Elite Boxers By Liquid Chromatography-Mass Spectrometry.
  14. Metabolomic profiling reveals dynamic lipid reprogramming during adipogenesis in 3T3-L1 cells.
  15. The Language of Elution: Autoregressive Prediction of the Next Feature in Untargeted LC-HRMS Lipidomics.
  16. When aging becomes degeneration: Evidence from Metabolomics of Healthy Age-stratified organ-donor and degenerated lumbar discs.
  17. Polyphenol oxidase mediated substrate selectivity underlies enzymatic browning in the edible insect Antheraea pernyi pupa.
  18. Novel Ion Suppression-Based MS Approach (IS-MS) for Straightforward, Robust, and Rapid Identification and/or Differentiation of Liquid Samples: Wine as a Case Study.
  19. Development and application of a simple LC-MS/MS method for therapeutic drug monitoring to guide colistin dosing in critically Ill patients.
  20. LC-MS/MS-based dereplication and molecular networking of Ficus superba with identification and quantification of furanocoumarins.
  21. Robust analytical methods for bis(monoacylglycero)phosphate profiling in health and disease.
  22. Differences among canine serum, plasma and urine metabolite profiles from samples that were collected at the same time.
  23. Analysis of the effect of variable frequency fan on tobacco metabolites during curing via untargeted metabolomics.
  24. Isocratic zwitterionic HILIC-ESI-MS method for assay of ropinirole and quantitation of degradation impurities in extended-release tablets.
  25. Tandem Mass Spectrometry Imaging of Low-Abundance Ether Phospholipids Guided by Bulk Lipidomics Analysis.
  26. Metabolomics of Andrographis paniculata Burm. F. green extracts by an integrated LC-MS and NMR approach and evaluation of acetylcholinesterase and tyrosinase inhibitory activity of extracts and isolated compounds.
  27. Chemical characterization and antinociceptive and anti-inflammatory activities of the aqueous extract of Myrciaria floribunda (H. West ex Willd.) O. Berg fruit peel.
  28. Development of a Dispersive µSPE Method for the Determination of Pesticide Residues in Water Samples by LC-MS/MS.
  29. Validation of qualitative and quantitative methods for 40 benzodiazepines and Z-drugs in hair by UPLC-MS/MS.
  30. Metabolomics analysis identifies differential metabolites and potential diagnostic biomarkers among pediatric sepsis subtypes.
  31. Untargeted Metabolomics Reveals Major Patterns of Metabolic Shifts in Potato Seed Tubers during Storage.
  32. Comprehensive metabolic profiling of Dictamni Cortex in normal and rheumatoid arthritis rats using UHPLC-Q/TOF-MS/MS coupled with polygonal mass defect filtering.
  33. Solvent-Dependent GC-MS and LC-MS/MS Phytochemical Profiling of Echium italicum L. Aerial Part Extracts and In Vitro Enzyme Inhibitory and DNA-Protective Effects.
  34. A Step-by-Step Protocol From METASPACE to Biological Interpretation.
  35. Comprehensive profiling of chemical composition, plasma-absorbed prototypes, metabolites, and pharmacokinetics of Jiannao Bushen Pill in rats using UHPLC-MS and microdialysis.
  36. Comprehensive Metabolomic Analysis of Saliva Using SWATH-DIA Reveals Systemic Metabolic Adaptations to Exercise.
  37. An integrated multitarget screening strategy combining network pharmacology with cell membrane chromatography coupled to high-performance liquid chromatography for identifying candidate antidepressant compounds from Cyperus rotundus L.
  38. Authenticity Assessment of Five Monofloral Honeys Based on Phytochemical Profiles.
  39. Olive oil-derived monounsaturated fat influences metabolic signatures in serotonergic regions of the brain in broiler chicken.
  40. Analysis of suzetrigine, a novel non-opioid pain medication, in whole blood and urine using liquid chromatography-tandem mass spectrometry.
  41. Suspected and Non-Targeted Screening of Non-Edible Substances in Food by UPLC-Q-TOF-MS.
  42. Medicinal Amazonian Oleoresins: An Eco-Friendly Chemical Fingerprinting Method.
  43. Metabolomics reveals the chemical basis of quality formation in GABA-enriched fermented green tea by lactic acid bacteria.
  44. Evaluation of the Therapeutic Potential and Safety Profile of Six Salvia Species Native to Türkiye.
  45. Molecular Transformation of Dissolved Organic Matter at the Coal-Water Interface: Insights from High-Resolution Mass Spectrometry.
  46. Metabolomics coupled with machine learning highlights metabolite profile differences of Scutellaria baicalensis Georgi from different specifications and geographical origins.
  47. Exploring the Chemical Profile and Biological Activities of Eryngium dichotomum: UHPLC-MS/NMR Characterization, and In Vitro Antioxidant Activity Along with the Antitumor Effect of Falcarinol.
  48. Rapid Determination of 13 Insecticides and Acaricides in Human Blood Using Dispersive Liquid-Liquid Microextraction Coupled With DART-MS/MS.
  49. Effects of extraction solvents on the recovery of bioactive compounds from Platycladus orientalis Leaves of different tree ages: a comparative metabolomics approach.
  50. UPLC-QTOF-MS/MS and Antifungal Activity of a Fraction Enriched in Saponins From Sarcomphalus joazeiro Against Candida spp.
  51. From Conventional Methods to Innovation: Caffeine and Chlorogenic Acid Extraction and Quantification in the Rise of Smart and Green Techniques-A Systematic Review.
  52. Chemical Composition, Antioxidant, and Anti-Inflammatory Activity of the Antarctic Lichen Leptogium puberulum: A Combination of Metabolomic, In Vitro, and In Silico Approaches.
  1. J Sep Sci. 2026 Jun;49(6): e70464
    Host-Dependent Metabolomic Variations in Viscum coloratum Revealed by UHPLC-Q-TOF-MS/MS and Integrated Annotation Strategies.
    Jiajie Wang, Jiong Chen, Xiaoyu Ju, Fendi Liu, Weili Yu, Lingling Huang, Jiexin Qiu, Shan Jia, Chunying Wang, Congkun Xiang, Qian Sun.
      Viscum coloratum (Kom.) Nakai, a widely distributed hemiparasitic medicinal plant in Asia, exhibits host-influenced metabolic traits that remain inadequately characterized. To further investigate this issue, an untargeted metabolomics approach was employed to comprehensively delineate host-dependent metabolic variation. Ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS/MS), a powerful analytical platform for untargeted metabolomics, was used to examine samples collected from two host species in Hubei Province, China. Differential metabolites were screened through multivariate chemometric analysis and characterized using an integrated annotation strategy to ensure systematic and reliable identification. Sixty-eight differential constituents were annotated and mainly assigned to phenylpropanoids, polyketides, lipids and lipid-like compounds, and organic acids and derivatives. These classes represented the major metabolic groups associated with host-dependent variation among the samples analyzed in this study. Furthermore, an exploratory 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay revealed significant differences in antioxidant capacity between the two groups, suggesting that host plants may influence the pharmacologically relevant properties of V. coloratum. These findings highlight the importance of considering host factors in its quality control, provide a comprehensive framework for differential metabolite identification, and enhance the understanding of the metabolic characteristics of V. coloratum.
    Keywords:  Viscum coloratum; antioxidation; chemometric analysis; host plant; molecular networking; plant metabolomics
    DOI:  https://doi.org/10.1002/jssc.70464
  2. Molecules. 2026 May 29. pii: 1868. [Epub ahead of print]31(11):
    Chemical Profiling of Aboveground and Underground Parts of Pterocephalus hookeri by Integrated FBMN, Untargeted LC-MS Metabolomics, and PAD-DESI-MSI.
    Jiaxing Luo, Lanlan Fang, Muze Yu, Di Yang, Jing Zhang, Jia Yu, Ce Tang, Tingting Kuang.
      Pterocephalus hookeri (C.B.Clarke) Höeck is a classic traditional Tibetan medicinal herb with multiple pharmacological activities. The inconsistent usage of its medicinal parts (whole herb, aboveground part (AP), and underground part (UP)) in commercial circulation severely restricts its clinical safety and quality stability. Currently, most existing chemical investigations focus on the whole herb, whereas the intraspecific chemical discrepancies between AP and UP remain poorly clarified. Herein, an integrated analytical strategy combining ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS)-based untargeted metabolomics, feature-based molecular networking (FBMN), and paper-based analytical device desorption electrospray ionization mass spectrometry imaging (PAD-DESI-MSI) was established to characterize differential metabolites and their spatial distribution in P. hookeri. A total of 101 compounds were annotated, and 12 vital differential metabolites were further screened with variable importance in projection (VIP) values > 1. The visualized distribution differences of these biomarkers were validated via heatmap and PAD-DESI-MSI analysis. Obvious differences in chemical accumulation characteristics were confirmed between AP and UP, which can guide reasonable clinical medication and rational dosage regulation referring to metabolite abundance. Moreover, optimized data filtering thresholds effectively eliminated metabolomic false positives, and FBMN exhibited excellent capacity for differential biomarker screening. This study provides a solid chemical basis for the quality evaluation and rational medicinal application of P. hookeri.
    Keywords:  DESI-MSI; Pterocephalus hookeri; UPLC-Q-TOF/MS; different medicinal parts; differential markers; feature-based molecular networking; untargeted metabolomics
    DOI:  https://doi.org/10.3390/molecules31111868
  3. BMC Plant Biol. 2026 Jun 11.
    Untargeted metabolomics suggests a trade-off between phenolamides and flavonoids in the pollen of Petunia hybrida with contrasting flower colours.
    Matheus Fernandes Alves, Abigail Moreno-Pedraza, Paula Carolina Pires Bueno, Khabat Vahabi, Nicole M van Dam.
       BACKGROUND: Pollen is vital for reproduction of flowering plants. Several nutritional and biological benefits for humans and pollinators are described and related to its richness and diversity of metabolites. However, the chemical composition of pollen from several horticulturally significant plant species, such as those in the genus Petunia, has not been thoroughly characterised. Here, we present the first comprehensive description of the chemical profile of two distinct P. hybrida lines: V26 (violet flowers) and W115 (white flowers) using untargeted metabolomics. Our workflow started from pollen sampling and isolation, followed by a 3-in-1 liquid phase extraction for wide-range metabolite recovery, and data acquisition by ultra-high-performance liquid chromatography coupled to high-resolution tandem mass spectrometry (UHPLC-HRMS/MS) using three chromatographic columns (C8, C18, and HILIC) with positive and negative ionisation. For data analysis, we implemented a user-friendly and reproducible data processing pipeline based on open-source computational tools.
    RESULTS: The P. hybrida pollen is rich in glycosylated flavonoids, phenolamides, and lipids, which were detected mostly in non-polar phase extracts analysed by reversed-phase chromatography. Simple phenylpropanoids, fatty acids, amino acids, and terpenoids were annotated to a lesser extent. Statistical analyses integrated with molecular networking demonstrated a distinct metabolic dichotomy: phenolamide derivatives are predominantly present in the pollen of the V26 line, while flavonoids are accumulated in the pollen of W115. This finding suggests different regulation of the phenylpropanoid metabolism in pollen of P. hybrida lines with differing flower colours and sheds light on hypotheses of ecological roles of pollen secondary metabolites, for example, in plant-pollinator interactions.
    CONCLUSIONS: Our findings suggest a metabolic trade-off in the phenylpropanoid pathway in the two studied P. hybrida lines. These cultivar-specific chemical signatures may have significant implications for pollen viability and interactions with pollinators. Furthermore, our analytical and computational workflow serves as a robust template for the deep metabolic profiling of other under-characterised and complex natural matrices.
    Keywords:   Petunia hybrida ; Liquid chromatography - mass spectrometry; Metabolic profiling; Phenylpropanoids; Pollen analysis; Pollen metabolomics; Specialised metabolites
    DOI:  https://doi.org/10.1186/s12870-026-08968-y
  4. Chem Biodivers. 2026 Jun;23(6): e03403
    Phytochemical Characterization and Comparative Tissue-Specific Metabolomics of Coffea arabica Using LC-HRMS.
    Mega Karina Putri, Anastasia Wheni Indrianingsih, Anjar Windarsih, Yuniar Khasanah, Ade Erma Suryani, Eka Noviana, Ayu Septi Anggraeni, Indrawati Dian Utami, Rina Astuti Br Sebayang, Lusty Istiqomah, Lucky Prabowo Miftachul Alam.
      This study presents a non-targeted metabolomic of different parts of Coffea arabica (bean, flower, leaf, cascara, and parchment) using liquid chromatography Orbitrap high-resolution mass spectrometry (LC-Orbitrap HRMS) and chemometrics. The total phenolic content (TPC), total flavonoid content (TFC), total terpenoid content (TTeC), and antioxidant activity were also determined. Leaf extract exhibited the highest TPC (129.54 ± 3.58 mg GAE/g) and TFC (28.47 ± 1.12 mg QE/g), whereas flower extract demonstrated the highest TTeC (581.67 ± 6.11 mg UAE/g). The highest antioxidant activity (measured using the DPPH assay) was found in leaf extract (IC50 34.93 ± 3.30 ppm), followed by bean extract (75.54 ± 4.35 ppm). Metabolite profiling using LC-HRMS revealed distinct chemical signatures across C. arabica parts, highlighting tissue-specific accumulation of primary and secondary metabolites. Partial least squares regression with discriminant analysis (PLS-DA) and hierarchical cluster analysis (HCA) effectively discriminated sample groups and identified key metabolites contributing to their variation. Variable importance for projection (VIP) analysis highlighted metabolites, such as caffeine, D-(-)-quinic acid, chlorogenic acid, flavonoids, and lipids, as key discriminating compounds. In conclusion, the integration of LC-HRMS metabolomics underscores the distinct metabolite profiles of Coffea arabica plants as potential sources of bioactive compounds and provides a foundation for future studies aimed at evaluating their bioavailability, safety, and potential functional applications.
    Keywords:  C. arabica; LC‐HRMS; chemometrics; non‐targeted metabolomics; phytochemical characterization
    DOI:  https://doi.org/10.1002/cbdv.202503403
  5. J Vis Exp. 2026 May 19.
    Integrated UHPLC-MS/MS and HRMS Strategy for Screening Potentially Hepatotoxic Furan-Containing Compounds in Dioscorea bulbifera L.
    Rong Tan, Ying Liu, Yunfang Jiang, Xin Zha, Jiamin Xin, Cong Wang, Jinghua Ruan, Weiwei Li, Shiyu Zhang.
      Dioscorea bulbifera L. (DBL), a traditional herbal medicine used to treat thyroid disorders and tumors, has raised considerable safety concerns due to its potential hepatotoxic effects. Previous research suggests that this liver injury may be associated with the metabolic activation of furan-containing compounds (FCCs) present in DBL. Here, we systematically characterized FCCs and their reactive metabolites in DBL extract through an integrated analytical approach combining ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) and ultra-high performance liquid chromatography coupled with tandem high-resolution mass spectrometry (UHPLC-HRMS). Employing both targeted and untargeted mass spectrometric analyses, we identified a total of 849 constituents in the aqueous extract of DBL, including 10 furanoditerpenoids and 17 additional FCCs. To elucidate the metabolic activation pathways, we used glutathione (GSH) and 4-bromobenzylamine (BBA) as dual trapping agents within a mouse liver microsomal (MLM) incubation system. Through P450-mediated metabolism, seven corresponding adducts derived from diosbulbin B (DSB), C (DSC), D (DSD), E (DSE), L (DSL), N (DSN), and 2-pentylfuran were successfully trapped and characterized. This study establishes a sensitive and specific approach for the comprehensive profiling of potentially hepatotoxic furan compounds in DBL, and provides valuable insights into the formation of reactive metabolites and toxicity mechanisms related to furan-containing herbal medicine.
    DOI:  https://doi.org/10.3791/70865
  6. Molecules. 2026 May 25. pii: 1820. [Epub ahead of print]31(11):
    Integrated MALDI-MSI and UHPLC-OE-MS for Spatial Visualization and Biosynthetic Pathway Elucidation of Bioactive Metabolites in Lilium lancifolium Thunb.
    Qibo Deng, Zhihui Wang, Jiajia Ji, Minsi Xie, Qiaozhen Tong, Kunlai Sun, Qinghua Peng, Zhiying Yuan.
      Lilium lancifolium Thunb. is an important economic crop widely cultivated and traded across Asia and has significant pharmacological activity. Despite decades of research on their chemical composition, the spatial distribution patterns of characteristic secondary metabolites within the bulbs remain poorly understood. In this study, we used matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) technology to characterize and spatially visualize multiple metabolites within the bulb for the first time. Additionally, ultra-high-performance liquid chromatography-Orbitrap Exploris mass spectrometry (UHPLC-OE-MS) was used to obtain comprehensive metabolite information from the bulbs. Using spatial metabolomics, we successfully identified nine steroidal saponins, three phenolic acid glycerides, and six other metabolites. Subsequently, we analyzed the spatial distribution of steroidal saponins and phenolic acid glycerides, which are key bioactive components. The analysis revealed that most of the steroidal saponins and phenolic acid glycerides, such as deacylbrownioside and regaloside A, exhibited a similar distribution pattern, mainly being enriched in the outer regions (A2, B2) and basal regions (B1, B2) on an individual scale. Further metabolomic and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses indicated that 11 substances detected in the bulbs, including diosgenin, phenylalanine, and acetyl-CoA, were jointly associated with 39 metabolic pathways, including "phenylpropanoid biosynthesis" and "terpenoid backbone biosynthesis". Based on the above findings, we propose biosynthetic pathways and accumulation patterns of steroidal saponins and phenolic acid glycerides in bulbs. This study provides a basis for precise resource utilization of L. lancifolium bulbs and a methodology to elucidate the biosynthesis of plant metabolites.
    Keywords:  Lilium lancifolium Thunb.; MALDI-MSI; UHPLC-OE-MS; biosynthetic pathway; phenolic acid glycerides; steroidal saponins
    DOI:  https://doi.org/10.3390/molecules31111820
  7. J Chromatogr B Analyt Technol Biomed Life Sci. 2026 Jun 09. pii: S1570-0232(26)00267-9. [Epub ahead of print]1281 125178
    Systematic profiling of chemical components in Tribulus terrestris L. extract and its absorbed parent compounds/metabolites in rat plasma utilizing UPLC-Q-TOF/MS.
    Yu Han, Xuanran Cong, Lijian Cai, Guohua Zhao, Xin Sun.
      Ischemic stroke (IS) often results in neuronal injury and persistent neurological impairment, underscoring the necessity for effective therapeutic approaches during the recovery phase. Tribulus terrestris L. has a historical application in promoting blood circulation and eliminating blood stasis, showing potential relevance to IS-related pathological processes. This study systematically characterized the chemical constituents of Tribulus terrestris L. extract using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry, identifying 67 compounds. In vivo analysis of absorbed prototype constituents and metabolites revealed 73 compounds, comprising 40 prototype constituents and 33 metabolites. Based on these absorbed constituents, network pharmacology analysis predicted that TP53, SRC, AKT1, and PIK3R1 could be key targets associated with IS-related biological processes. Additionally, integrated analysis of the protein-protein interaction network and pathway enrichment indicated that the PI3K/AKT signaling pathway might be involved in the potential actions of Tribulus terrestris L. extract. These results systematically elucidated the chemical composition of Tribulus terrestris L. extract and offered a network pharmacology-based prediction of potential mechanisms related to IS, without direct experimental evidence of a therapeutic mechanism. Given the absence of ischemic stroke animal models, pharmacodynamic evaluations, neuroprotection assays, target validations, or pathway validations in this study, the proposed targets and pathways should be considered hypotheses requiring further experimental validation.
    Keywords:  Ischemic stroke; Network pharmacology; Tribulus terrestris L.; UPLC-Q-TOF-MS; Untargeted metabolomics
    DOI:  https://doi.org/10.1016/j.jchromb.2026.125178
  8. Bioinform Adv. 2026 ;6(1): vbag138
    eMZed 3: flexible and interactive development of scalable LC-MS/MS data analysis workflows in python.
    Uwe Schmitt, Jethro Hemmann, Nicola Zamboni, Julia A Vorholt, Patrick Kiefer.
       Summary: Liquid chromatography-mass spectrometry (LC-MS/MS) data analysis requires adaptable software solutions to meet diverse analytical needs. We present eMZed 3, a modern Python framework for flexible and interactive analysis of LC-MS/MS data. eMZed 3 enables users to develop scalable workflows tailored to their specific requirements while leveraging Python's extensive ecosystem of libraries. Building on its predecessor, eMZed 3 is now Python 3-based and includes substantial enhancements, including support for chromatogram-based LC-MS data, a new SQLite-based backend supporting optional out-of-memory processing, and rich interactive visualization tools. Compared to the previous version, eMZed 3 is now split into three packages: emzed (core functionalities), emzed-gui (interactive data visualization), and emzed-spyder (an integrated development environment). This modular architecture allows straightforward integration of the emzed core library into headless Python environments, including computational notebooks (such as Jupyter) or high-performance computing clusters. eMZed 3 incorporates well-established libraries such as OpenMS, and is suited for both targeted and untargeted metabolomics. Overall, eMZed 3 supports the efficient development of scalable and reproducible LC-MS data analysis and is accessible to both novice and advanced programmers.
    Availability and implementation: eMZed 3 and its documentation are freely available at https://emzed.ethz.ch, the source code is hosted at https://gitlab.com/groups/emzed3.An online-executable example workflow is available on Binder at: https://mybinder.org/v2/gl/emzed3%2Femzed-example-workflow/HEAD?_labpath=example.ipynb.
    DOI:  https://doi.org/10.1093/bioadv/vbag138
  9. Gigascience. 2026 Jun 10. pii: giag069. [Epub ahead of print]
    MultiMS2: A curated multi-modal, multi-energy spectral library for metabolomics.
    Adriano Rutz, Mario S P Correia, Nicola Zamboni.
       BACKGROUND: Spectral libraries are essential for mass spectrometry-based metabolomics, enabling accurate metabolite annotation. Collision-induced dissociation (CID) dominates existing public libraries, but is rarely sufficient for structural elucidation. Electron-activated dissociation (EAD) provides complementary, radical-driven fragmentation, but remains sparsely represented. The lack of datasets spanning multiple dissociation mechanisms, energies, and ionization modes limits both analytical workflows and the development of robust machine learning models.
    FINDINGS: We present MultiMS2, a curated metabolomics spectral library comprising 43,728 MS/MS spectra from 2,899 unique compounds. Spectra were acquired using both CID and EAD at three energies each, in positive and negative ionization modes. The dataset substantially expands publicly available EAD coverage while preserving matched acquisition conditions across energies and dissociation types.
    CONCLUSIONS: By systematically combining CID and EAD across multiple energies and polarities, MultiMS2 provides a unique resource for metabolite annotation, benchmarking, and machine learning. The library supports energy-aware and dissociation-aware analysis, enabling methodological innovation and improved generalization in computational metabolomics.
    Keywords:  Collision-induced dissociation; Electron-activated dissociation; Metabolomics; Spectral library
    DOI:  https://doi.org/10.1093/gigascience/giag069
  10. Environ Sci Technol. 2026 Jun 09.
    ToxBase: A Multidimensional ToxCast Reference Database for High-Throughput Human Exposome Analysis.
    Ryan Nguyen, Griffin Rangel, Xingzhu Liu, Reuben A Santoso, Amogh Bantwal, Dylan H Ross, Ryan P Seguin, Jennifer Liem, Yvonne S Lin, Brian Pratt, Brendan X MacLean, Michael J MacCoss, Libin Xu.
      High-resolution mass spectrometry (HRMS) is the gold-standard technique for comprehensively profiling chemical exposures in complex human matrices, making it a powerful analytical tool for advancing human exposome research. Yet the scarcity of HRMS reference data, including collision cross-section (CCS) measurements from ion mobility-mass spectrometry (IM-MS) and MS/MS fragmentation spectra, hinders confident structural annotation of chemical exposure agents across laboratories. We therefore developed ToxBase, a multidimensional (m/z, retention time, CCS, MS/MS) reference database for over 2,000 chemicals sourced from the U.S. Environmental Protection Agency's ToxCast chemical library. Built via high-throughput liquid chromatography-ion mobility-tandem mass spectrometry (LC-IM-MS/MS), the ToxBase database comprises 3,598 precursor ions spanning 2,075 unique compounds with excellent precision (98.5% of compounds display interday CCS RSDs < 1%) and strong cross-platform agreement. A high-quality MS/MS reference library of the fragmented precursors was assembled using targeted data-dependent acquisition and DDARawProcessor, a novel data extraction algorithm. When applied to LC-IM-MS/MS data obtained from human plasma, urine, and fecal samples (n = 20 per matrix), ToxBase rapidly enabled 42 high-confidence (Level 1) identifications. The ToxBase database is freely available and compatible with the open-source MS data processing platform Skyline for vendor-agnostic suspect screening workflows, providing a valuable resource for standardized, large-scale exposome analysis.
    Keywords:  MS/MS; ToxCast; collision cross section; exposome; ion mobility-mass spectrometry; suspect screening
    DOI:  https://doi.org/10.1021/acs.est.5c18068
  11. Molecules. 2026 May 25. pii: 1822. [Epub ahead of print]31(11):
    Untargeted and Targeted Cerebrospinal Fluid Neurometabolomics via Chromatography-Mass Spectrometry-Based Methods.
    Alisa K Pautova.
      Neuroscience is a rapidly advancing field; however, a comprehensive understanding of brain function at the molecular, cellular, and systems levels remains incomplete. Neurological and psychiatric disorders represent a major global health burden, highlighting the need for improved diagnostic and therapeutic strategies. Cerebrospinal fluid (CSF) is one of the most informative biofluids for investigating central nervous system (CNS) pathology due to its close biochemical relationship with brain tissue. Recent advances in neurometabolomics, defined as the comprehensive analysis of small-molecule metabolites in CSF, have been driven by the development of highly sensitive and informative mass spectrometry-based techniques. These approaches enable the identification of disease-associated metabolic signatures. This review summarizes current chromatography-mass spectrometry-based methods used in both untargeted and targeted CSF metabolomics, with particular emphasis on their analytical performance, reproducibility, and limitations. Special attention is given to method standardization and validation, as well as to the identification of reliable metabolic biomarkers for the diagnosis and monitoring of neurological disorders, including neurodegenerative, psychiatric, oncological, and neuroinflammatory diseases.
    Keywords:  biomarkers; central nervous system; diagnosis; gas chromatography–mass spectrometry; liquid chromatography–mass spectrometry; metabolites; standardization; targeted analysis; untargeted analysis; validation
    DOI:  https://doi.org/10.3390/molecules31111822
  12. Front Mol Biosci. 2026 ;13 1833340
    Metabolic alterations and potential biomarkers in unstable angina investigated by lipidomic analysis.
    Nan Feng, Linhe Wang, Guoan Zhao, Yanlin Zhou.
       Introduction: Unstable angina (UA) represents a critical condition within the broader context of acute coronary syndromes, characterized by episodes of sudden chest pain resulting from insufficient blood flow to the myocardium. The pathophysiological mechanisms underlying UA remain complex and poorly understood, necessitating further investigation into the metabolic alterations associated with this condition. Identifying specific biomarkers for UA is crucial for improving diagnostic accuracy and facilitating timely therapeutic interventions.
    Methods: The present study was designed to elucidate the metabolic profile of UA by enrolling 60 patients diagnosed with UA, alongside 60 healthy controls. Participants were stratified into discovery and validation cohorts, with each group comprising 30 UA patients and 30 controls. The diagnosis of UA was performed in accordance with the 2020 European Society of Cardiology guidelines and the 2019 Chinese clinical pathway for UA management. Blood samples were meticulously collected following an overnight fast, processed, and subsequently stored at -80 °C. A comprehensive lipidomic analysis was executed utilizing ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), with compound identification referenced from the Metware Database. Data analysis involved advanced multivariate statistical techniques, including principal component analysis and orthogonal partial least squares-discriminant analysis.
    Results: The analysis revealed significant metabolic discrepancies between the UA and control groups, identifying a total of 193 differential metabolites, of which 67 were notably upregulated in the UA cohort. Pathway enrichment analysis highlighted critical alterations in glycerophospholipid metabolism and necroptosis pathways. Furthermore, the application of machine learning algorithms, specifically neural networks and random forests, enabled the identification of key metabolic biomarkers, including TxB3, LPC(16:0/0:0), and DL-Carnitine, which exhibited robust diagnostic potential.
    Conclusion: In summary, our findings indicate that these metabolites may serve as promising candidate diagnostic biomarkers for UA, providing valuable insights into its pathophysiology.
    Keywords:  UPLC-MS/MS; biomarkers; lipidomic analysis; machine learning; unstable angina
    DOI:  https://doi.org/10.3389/fmolb.2026.1833340
  13. J Vis Exp. 2026 May 22.
    A Standardized Metabolomics Protocol For Analyzing Exercise-Induced Metabolic Shifts In Elite Boxers By Liquid Chromatography-Mass Spectrometry.
    Peng Sun, Dong Zhang, Qinyi Chen, Mengyang He, Biao Li.
      Elite boxing induces rapid metabolic changes that are not fully captured by conventional physiological measurements. A standardized untargeted serum metabolomics workflow based on liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (LC-QTOF-MS/MS) was applied to samples collected before sparring, immediately after sparring, and 24 h after sparring in seven elite male boxers. The workflow included standardized sample collection, pooled quality-control monitoring, metabolite profiling, multivariate statistical analysis, and pathway interpretation. Acute sparring was associated with changes in metabolites related to glycolysis and gluconeogenesis, whereas the 24-h timepoint was associated with sulfur metabolism. Phosphatidylinositol PI(16:0/18:2(9Z,12Z)) showed strong discrimination of the immediate post-sparring state, and thiosulfate was associated with the 24-h recovery state. These findings support the use of this workflow for reproducible profiling of exercise-related metabolic changes in this cohort of seven elite male boxers. The protocol is intended for controlled small-cohort studies of acute exercise and short-term recovery and emphasizes reproducible sample handling, pooled quality-control monitoring, and interpretable downstream analysis.
    DOI:  https://doi.org/10.3791/70719
  14. Front Mol Biosci. 2026 ;13 1821798
    Metabolomic profiling reveals dynamic lipid reprogramming during adipogenesis in 3T3-L1 cells.
    Nanping Shi, Zeyi Huang, Jianan Liu, Wuping Sun, Jian Li.
       Objective: Adipogenesis, the process of adipocyte differentiation, plays a central role in obesity development. However, the molecular mechanisms underlying adipogenesis and its regulation remain incompletely understood. This study aimed to investigate metabolomic changes during adipocyte differentiation in 3T3-L1 cells and elucidate the associated molecular mechanisms, with potential implications for obesity-related metabolic diseases.
    Methods: Murine 3T3-L1 pre-adipocytes were cultured and induced to differentiate into mature adipocytes over an 8-day period. Samples were collected at pre-differentiation (Pre), 4-day differentiation (Middle) and 8-day differentiation (Mature) stages. Metabolites were extracted using hydrophilic and hydrophobic methods and analyzed by ultra-performance liquid chromatography coupled with mass spectrometry. Data analysis employed kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis to identify differential metabolites and their associated metabolic pathways.
    Results: Significant differences in metabolite profiles were observed among Pre, Middle and Mature. We identified 170 differential metabolites between Pre and Middle (164 increased, 6 decreased), 246 differential metabolites between Pre and Mature (223 increased, 23 decreased), and 124 differential metabolites between Middle and Mature (102 increased, 22 decreased). The most pronounced changes occurred in lipid metabolites, particularly triglycerides and phosphatidylcholines. KEGG enrichment analysis revealed that these differential metabolites were mainly involved in glycerolipid metabolism, glycerophospholipid metabolism, and insulin resistance pathways. Correlation network analysis further identified key genes associated with these metabolites, highlighting the interplay between lipid and amino acid metabolism during adipogenesis.
    Conclusion: This study provides a comprehensive metabolomic profile of adipocyte differentiation in 3T3-L1 cells, revealing significant alterations in lipid metabolism and key metabolic pathways. These findings enhance our understanding of the molecular mechanisms underlying adipogenesis and may contribute to the development of novel diagnostic tools and therapeutic strategies for obesity-related metabolic diseases.
    Keywords:  3T3-L1 cells; adipocyte differentiation; adipogenesis; metabolomic; obesity
    DOI:  https://doi.org/10.3389/fmolb.2026.1821798
  15. ArXiv. 2026 Jun 02. pii: arXiv:2606.05225v1. [Epub ahead of print]
    The Language of Elution: Autoregressive Prediction of the Next Feature in Untargeted LC-HRMS Lipidomics.
    Dayanjan S Wijesinghe.
      Untargeted liquid chromatography-high-resolution mass spectrometry (LC-HRMS) detects thousands of molecular features per sample, yet only 2-20% receive confident structural annotations. A root cause of this "dark metabolome" is that tandem MS/MS acquisition is reactive: instruments select precursors only after ions appear, blind to what elutes next. We reframe chromatographic elution as an autoregressive sequence prediction task. Because reversed-phase elution order is governed by hydrophobicity, successive features form a physically constrained sequence, like tokens in language. We discretize the mass-to-charge (m/z) axis into 110 bins and train long short-term memory (LSTM) and Transformer models to predict the next eluting m/z bin from five annotation-free per-token features: m/z bin, mass defect, retention-time gap, polarity, and intensity rank. Trained on 15,242 features from four clinical lipidomics cohorts (342 plasma samples; SCIEX TripleTOF 6600+, Waters CSH C18), the LSTM reaches 98.4% top-1 accuracy (99.99% top-5; mean absolute error 3.6 Da) and the Transformer 98.0%. Ablation shows autoregressive context accounts for 55.5 percentage points while no single feature contributes more than 0.2 pp: the sequential pattern, not molecular properties, drives prediction. Models transfer across instruments sharing the method (r=0.999 on an independent Agilent 6530 dataset) but fail under a different column chemistry (5.1% top-1) or polarity mode (2.6%), confirming method- and mode-specificity. Fine-tuning on as few as two to five quality-control injections recovers held-out accuracy from 2.6% to nearly 50%, so cross-condition deployment needs minimal calibration. These results establish that elution sequences are highly predictable and lay the groundwork for predictive MS/MS acquisition to improve annotation coverage in untargeted metabolomics.
  16. Spine J. 2026 Jun 09. pii: S1529-9430(26)00175-0. [Epub ahead of print]
    When aging becomes degeneration: Evidence from Metabolomics of Healthy Age-stratified organ-donor and degenerated lumbar discs.
    Shanmuganathan Rajasekaran, Chitraa Tangavel, Divya Arunachalam, Gnanaprakash Gurusamy, Sharon Miracle Nayagam, Arun Meena, Sri Vijay Anand.
       BACKGROUND: Aging and degeneration are biologically distinct processes in intervertebral discs, but are difficult to differentiate radiologically. Metabolomics reflects real-time biochemical activity, and age-stratified metabolomic profiling of normal and degenerated discs may identify preclinical degeneration and reveal molecular signatures distinguishing normal aging from degeneration.
    PURPOSE: To characterize the metabolomic changes in the MRI-normal healthy lumbar intervertebral disc and compare them with degenerated discs to identify metabolomic signatures that differentiate normal aging from degeneration.
    STUDY DESIGN AND SETTING: Comparative metabolomics study using human nucleus pulposus tissue from organ donors and surgical specimens, conducted at a tertiary spine care center and an affiliated research laboratory.
    PATIENT SAMPLE: Nucleus pulposus tissue from 21 healthy organ donors (Pfirrmann grade I) was stratified by age: young (20-30 years), middle-aged (31-50 years), and old (>50 years), and compared with 40 degenerated discs (grades III-V) from surgical specimens.
    OUTCOME MEASURES: Primary outcome measures included differential metabolite abundance (metabolites with Variable Importance in Projection [VIP] scores>1), pathway enrichment profiles, and identification of age-specific versus degeneration-specific metabolic signatures.
    METHODS: Untargeted ultra-high performance liquid chromatography- tandem mass spectrometry (UHPLC-MS/MS) was performed in both positive and negative ionization modes. Metabolites were identified using Compound Discoverer v3.7, with reference to HMDB and KEGG. Statistical and pathway enrichment analyses were performed using MetaboAnalyst 6.0.
    RESULTS: Untargeted UHPLC-MS/MS analysis revealed 831 significant metabolites (VIP >1), contributing to group separation in the partial least squares discriminant analysis (PLS-DA) model. Lipids and lipid-like molecules, especially sphingolipids, fatty acyls, and steroids, constituted 39%. MRI normal organ donor discs in progressive age periods demonstrated four distinct metabolic trends: (i) progressive decline of antioxidants (ubiquinone, glutathione, N-acetyl seretonin); (ii) increased oxidative/inflammatory markers (4-HNE, prostaglandin E₂ ethanolamide, N1-acetylspermidine); (iii) transient midlife antioxidant elevation; (4OH benozoic acid, 4OH phenylpyruvic acid); (iv) partial recovery in older discs (Hypoxanthine, Paraxanthine, CerP(d18:1/18:0)). In old-aged discs, accumulation of sphingolipids (sphingosine, ceramides) and redox drift indicated enhanced senescence and energy imbalance. Degenerated discs exhibited a different profile, characterized by the suppression of bioactive lipids, particularly resolvins, PGE2, and SOFAs, accompanied by disrupted sphingolipid metabolism and reduced redox capacity.
    CONCLUSION: Distinct metabolomic signatures differentiate physiological aging from disc degeneration. Degeneration is characterized by disrupted sphingolipid and redox homeostasis, suggesting a pathobiological process beyond normal senescence. These data provide a metabolic framework for future translational studies to identify disc degeneration even at the preclinical stage and in therapeutic stratification.
    Keywords:  Aging vs degeneration; Intervertebral disc; Metabolomics; Molecular signatures; Redox imbalance; Sphingolipids
    DOI:  https://doi.org/10.1016/j.spinee.2026.05.010
  17. Food Res Int. 2026 Sep 01. pii: S0963-9969(26)01198-1. [Epub ahead of print]239 119515
    Polyphenol oxidase mediated substrate selectivity underlies enzymatic browning in the edible insect Antheraea pernyi pupa.
    Yuyang She, Jinghan Zhang, Yanqun Liu, Jun-Hua Shao.
      Antheraea pernyi pupa is highly susceptible to enzymatic browning during processing, yet the endogenous phenolic substrates involved and the substrate selectivity of PPO remain poorly understood. In this study, LC-MS/MS, initial rate analysis, and untargeted metabolomics were integrated to investigate browning mediated by PPO and dependent on substrate type in A. pernyi pupa. Fourteen endogenous compounds related to phenolic metabolism and browning reactions were detected, among which taxifolin, caffeic acid, and chlorogenic acid showed clear browning potential mediated by PPO. Taxifolin exhibited the strongest catalytic response across the tested concentration range, indicating that PPO activity depended on substrate type. It also showed the highest browning activity across the tested pH range and reached its maximum at pH 6, whereas the caffeic acid and chlorogenic acid systems showed their highest activity at pH 5 and declined markedly as pH increased. After tissue disruption, visible browning developed rapidly, while total phenolic content decreased from 0.01032 to 0.00768 mg GAE/g within 2 h, corresponding to an overall reduction of 25.6%; the most pronounced decrease (18.3%) occurred within the first 0.5 h. Untargeted metabolomics further revealed clear substrate-dependent metabolic divergence and identified key oxidation-associated metabolites, including hydroquinone, pyrocatechol, 3,4-dihydroxy-L-phenylalanine, and 5,6-dihydroxyindole-2-carboxylic acid. The browning process was also associated with perturbation of pathways related to amino acid metabolism. These results indicate that browning in A. pernyi pupa is initiated by selective oxidation of susceptible endogenous substrates and then extended through continued chemical evolution centered on quinones. This work provides a mechanistic basis for targeted browning control during the processing of A. pernyi pupa.
    Keywords:  Antheraea pernyi pupa; Edible insect; Endogenous phenolics; Enzymatic browning; Metabolomics; Polyphenol oxidase; Substrate selectivity
    DOI:  https://doi.org/10.1016/j.foodres.2026.119515
  18. Anal Chem. 2026 Jun 08.
    Novel Ion Suppression-Based MS Approach (IS-MS) for Straightforward, Robust, and Rapid Identification and/or Differentiation of Liquid Samples: Wine as a Case Study.
    Maria Bikaki, Lu Hostettler, Matthias Kleiner, David Zollikofer, Götz Schlotterbeck, Markus Ehrat.
      A rapid and robust mass spectrometric approach based on ion suppression effects was developed for the identification and authentication of liquid food samples. Wine was used as a model system to evaluate sample discrimination according to vintage, country of origin, producer and name. A defined mixture of 25 chemical marker compounds was added to each sample, followed by dilution and direct analysis using electrospray ionization mass spectrometry (ESI-MS) operated in selected ion monitoring (SIM) mode. Sample-specific matrix effects on the ionization efficiencies of the marker compounds produced characteristic mass spectral response patterns, which were acquired within approximately 1 min per sample. These patterns were compared with reference data derived from a comprehensive wine database. Multivariate statistical analysis was performed using linear discriminant analysis (LDA) combined with complementary distance-based metrics. The approach was validated using five vintages of several premium wines. High authentication scores were obtained for all samples, demonstrating the suitability of the approach for rapid wine authentication and verification of origin. The results indicate that ion suppression-based MS fingerprinting represents a promising tool for fast, minimal-preparation assessment of authenticity in food analysis.
    Keywords:  ion-suppression; machine learning; mass-spectrometry; wine authenticity
    DOI:  https://doi.org/10.1021/acs.analchem.6c01501
  19. BMC Pharmacol Toxicol. 2026 Jun 09.
    Development and application of a simple LC-MS/MS method for therapeutic drug monitoring to guide colistin dosing in critically Ill patients.
    Yueyuan Wang, Yani Gu, Kai Fan, Chen Feng, XinYun Zhang, Aiming Shi, Yunli Yu.
       OBJECTIVES: This study aims to develop a simple, rapid, and cost-effective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for determining the concentration of colistin in human serum, including its two active components, colistin A and colistin B, to support routine therapeutic drug monitoring (TDM).
    METHODS: Protein precipitation was performed using pure acetonitrile, with polymyxin B1 as the internal standard (IS). Chromatographic separation was achieved using a Phenomenex KINETEX XB-C18 column (2.6 μm, 3 × 50 mm) with a gradient elution system consisting of 0.1% formic acid (FA) in water and pure acetonitrile as the mobile phase. Detection was carried out using an electrospray ionization (ESI) source in positive ion mode with multiple reaction monitoring (MRM). After applying the validated method to critically ill patients for approximately one year, we used the receiver operating characteristic (ROC) curve to identify the preliminary cut-off value of colistin trough concentration for predicting acute kidney injury (AKI).
    RESULTS: The total runtime for chromatographic separation and mass spectrometric detection was 5 min. Excellent linearity was achieved over the concentration range of 0.27-8.69 µg/mL for colistin A and 0.08-2.56 µg/mL for colistin B, with R2 greater than 0.99 for both analytes. The method demonstrated acceptable matrix effects and recovery rates. In addition‌, the ROC curve analysis of colistin TDM in critically ill patients determined a preliminary cut-off value for AKI at a trough concentration of 2.22 µg/mL.
    CONCLUSION: The validated method enables simultaneous and precise quantification of the two active substances, with a simple procedure and minimal analytical cost. The utility of TDM in critically ill patients is paramount for individualizing dosing regimens to optimize outcomes and safety. Specifically, the correlation between trough concentration and AKI risk highlights the importance for clinicians to monitor renal function during colistin treatment to prevent drug-induced AKI.
    CLINICAL TRIAL NUMBER: Not applicable.
    Keywords:  Acute kidney injury; Colistin; Colistin methanesulfonate; Liquid chromatography-tandem mass spectrometry; Therapeutic drug monitoring
    DOI:  https://doi.org/10.1186/s40360-026-01162-8
  20. Nat Prod Res. 2026 Jun 08. 1-10
    LC-MS/MS-based dereplication and molecular networking of Ficus superba with identification and quantification of furanocoumarins.
    Muhammad Afiffikri Ahmad, Sarah Sofea Mohd Sali, Nur Hanis Syuhada Abdul Rauf, Nur Ain Sofia Mohd Khairi, Shamsul Khamis, Nurunajah Ab Ghani, Juliana Yusoff, Nurulfazlina Edayah Rasol.
      Ficus superba (Moraceae) is a limestone-associated plant species from Malaysia that remains chemically unexplored. This study aimed to establish the phytochemical profile of its bark, leaves, and twigs using an untargeted mass spectrometry-based approach for rapid dereplication of known constituents. Feature-based molecular networking (FBMN) revealed distinct clusters of metabolites, leading to the annotation of 23 compounds distributed across major classes, including stilbenoids, coumarins, flavonoids, terpenoids and others. Comparative profiling demonstrated variation in metabolite composition between plant parts, but consistently highlighted two furanocoumarins, oxypeucedanin hydrate and oxypeucedanin hydrate-3'-O-β-D-glucopyranoside, as organ-conserved metabolites and candidate marker compounds. Their levels were quantified using a validated UHPLC-UV method, confirming their presence in bark, leaves, and twigs at concentrations ranging from 10 to 118 µg/mg of dry extract. This integrated approach provides a focused dereplication-guided chemical overview of F. superba and identifies candidate marker compounds that may support future chemotaxonomic and pharmacological studies.
    Keywords:  FBMN; Ficus superba; dereplication; furanocoumarin; quantification
    DOI:  https://doi.org/10.1080/14786419.2026.2683920
  21. Nat Protoc. 2026 Jun 10.
    Robust analytical methods for bis(monoacylglycero)phosphate profiling in health and disease.
    Wentao Dong, Kwamina Nyame, Eshaan S Rawat, Uche N Medoh, Jian Xiong, Caio C Bonin, Hisham N Alsohybe, Hunter Y Liu, Sara Gomes, Tammy Shalit, Marianna Arnold, Frank Hsieh, Esther Sammler, Monther Abu-Remaileh.
      Bis(monoacylglycero)phosphates (BMPs), a distinct class of anionic phospholipids predominantly found in late endosomes and lysosomes, plays a pivotal role in supporting lysosomal functions and maintaining metabolic homeostasis. Dysregulation of BMPs is associated with an array of disorders, notably neurodegenerative diseases. However, the identification and quantitation of BMP remains difficult because of its structural similarity to its isomer, phosphatidylglycerol (PG), thus necessitating robust analytical methods for accurate and reliable BMP profiling. In this study, we present comprehensive liquid chromatography (LC)-tandem mass spectrometry (MS2) methodologies for the precise and systematic analysis of BMP species in biological samples. We detail LC/MS methods for both an untargeted Orbitrap mass spectrometer and a targeted triple quadrupole mass spectrometer. We use differences in hydrophobicity and structure to annotate BMPs and PGs on the basis of retention time and positive-mode MS2 fragmentation patterns, respectively. Because genetic ablation of the BMP synthase CLN5 leads to specific depletion of BMPs but not PGs, lipid extracts from CLN5 knockout and wild-type cells can be compared to confidently annotate BMPs when MS2 data are incomplete. Lipid extraction and preparation of samples for LC/MS takes ~4 h, unattended LC/MS instrument time depends on the number of samples and computer-based data analysis takes ~1 d. Altogether, this approach constitutes a robust method for BMP profiling and annotation, furthering research into health and disease.
    DOI:  https://doi.org/10.1038/s41596-026-01379-1
  22. Front Vet Sci. 2026 ;13 1837846
    Differences among canine serum, plasma and urine metabolite profiles from samples that were collected at the same time.
    Robin Moore, Johanna Anturaniemi, Vidya Velagapudi, Anna Hielm-Björkman.
       Background: Biospecimen choice can substantially influence metabolite measurements, yet matched comparisons of serum, EDTA plasma, lithium-heparin plasma, and urine remain limited in canine targeted metabolomics.
    Objectives: To evaluate cross-matrix comparability of targeted small polar metabolite profiles in dogs and to assess how different biospecimens capture diet- and health-associated variation.
    Animals: Client-owned dogs from a previously reported feeding trial.
    Methods: Targeted ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was performed on matched biospecimens from a baseline blood-matrix cohort (serum, EDTA plasma, lithium-heparin plasma) and an end-of-trial serum-urine cohort. Metabolites with >20% missing values within a matrix and samples with >20% missing metabolite values within a biofluid were excluded. Cross-matrix agreement was assessed using Pearson correlation and mean paired log10 concentration differences. PCA and PERMANOVA were used to compare global matrix effects and diet- and health-associated separation.
    Results: After filtering, 88 metabolites remained for serum, 85 for EDTA plasma, and 88 for lithium-heparin plasma in the blood-matrix cohort. Globally, EDTA plasma separated clearly from both serum and lithium-heparin plasma, whereas serum and lithium-heparin plasma overlapped substantially. At the metabolite level, 10/84 (11.9%) serum-EDTA plasma, 16/88 (18.2%) serum-lithium-heparin plasma, 20/84 (23.8%) lithium-heparin plasma-EDTA plasma, and 4/75 (5.3%) serum-urine comparisons were comparable to varying degrees. Serum showed the clearest diet-associated separation at baseline (PERMANOVA pseudo-R 2 = 0.13, p = 0.013) and end-of-trial (pseudo-R 2 = 0.22, p = 0.021). Health-associated separation was weak and exploratory.
    Conclusions and Clinical Importance: Biospecimen choice materially affects targeted metabolite profiling in dogs. Serum and lithium-heparin plasma were globally more similar to each other than either was to EDTA plasma, whereas urine was a distinct complementary matrix rather than a blood surrogate. Serum was the most robust matrix for detecting diet-associated variation.
    Keywords:  canine; metabolomics; plasma; serum; urine
    DOI:  https://doi.org/10.3389/fvets.2026.1837846
  23. BMC Plant Biol. 2026 Jun 08.
    Analysis of the effect of variable frequency fan on tobacco metabolites during curing via untargeted metabolomics.
    Di Shi, Xiaoqiang Zhang, Guanhui Li, Hongfeng Liu, Zhengfa Shi, Jiawei Luo, Xin Jin, Shengjiang Wu, Ning Huang, ZeYu Zhao, Lingli Xie, Kesu Wei.
      The Variable-frequency (VF) fan regulation is a promising technique for flue-curing of tobacco (Nicotiana tabacum L.), and exploring its mechanism is of great significance for improving the curing quality. The middle leaves of "Yunyan 87" were used as experimental materials for exploring the changes of metabolic components by untargeted metabolomics and monitoring dynamic changes of key physiological indicators during flue-curing, VF curing (dynamic frequency: 35-40-45-40 Hz) and constant-frequency curing (50 Hz, CK). VF treatment resulted in a more uniform yellowing process, reduced leaf browning, and improved color retention compared with the control. Leaves under VF treatment showed enhanced superoxide dismutase (SOD), peroxidase (POD), and polyphenol oxidase (PPO) activities, accompanied by lower malondialdehyde (MDA) accumulation. At 48 h, Pathway enrichment analysis revealed the glyoxylate and dicarboxylate metabolism pathway was most affected, with significantly enriched metabolites including isocitrate, citric acid, and glyoxylate. At 120 and 168 h, the most significant metabolic pathways affected by variable frequency fan were flavonoid and flavanol biosynthesis, accompanied significantly enrichment of kaempferol, quercetin, and quercetin. Notably, total aroma substances were reduced under VF (Neophytadiene - 42%, benzaldehyde - 28%). This study provided a metabolomics-based framework for improving intensive curing processes.
    Keywords:  Aroma components; Fan frequency; Metabolic pathway; Physiological changes; Untargeted metabolome
    DOI:  https://doi.org/10.1186/s12870-026-09101-9
  24. Talanta. 2026 Jun 08. pii: S0039-9140(26)00739-3. [Epub ahead of print]310 130083
    Isocratic zwitterionic HILIC-ESI-MS method for assay of ropinirole and quantitation of degradation impurities in extended-release tablets.
    Vasiliki Brakoulia, Ariadni Geballa-Koukoula, Dimitrios Bogeas, Andreas Kakouris, Irene Panderi.
      Given the therapeutic importance of ropinirole in Parkinson's disease and its long-term use, reliable quality control of this drug is required, with particular emphasis on impurity control in the finished pharmaceutical product. A zwitterionic hydrophilic interaction liquid chromatography-electrospray ionization mass spectrometry (HILIC-ESI-MS) method was developed and validated for the determination of ropinirole and three degradation impurities in pharmaceutical tablets. Chromatographic separation was achieved on a ZIC®-HILIC column (150 × 2.1 mm, 3.5 μm particle size, 200 Å) under isocratic elution with a mobile phase of 18.8 mM aqueous ammonium formate solution at pH 6.20 and acetonitrile in an 8:92 (v/v) ratio, delivered at a flow rate of 0.25 mL min-1. Detection was performed in positive electrospray ionization mode using selected ion monitoring and the target analytes were determined within 10 min for assay and 16 min for impurity testing. The method was validated for linearity, accuracy, precision, sensitivity, robustness, and selectivity. Calibration curves showed good linearity over the concentration range of 21 to 122 ng mL-1 with correlation coefficients of at least 0.997. Intra-day precision (% CV) did not exceed 6.7 %, while total precision ranged between 2.5 and 8.8 % for all compounds. The method was successfully applied to the analysis of commercially available ropinirole extended-release tablets. To the best of our knowledge, this is the first reported HILIC-ESI-MS method for the assay of ropinirole and the quantitation of three degradation impurities in pharmaceutical tablets.
    Keywords:  HILIC; Hydrophilic interaction liquid chromatography; Impurities; LC-MS; Method validation; Ropinirole
    DOI:  https://doi.org/10.1016/j.talanta.2026.130083
  25. Anal Chem. 2026 Jun 12.
    Tandem Mass Spectrometry Imaging of Low-Abundance Ether Phospholipids Guided by Bulk Lipidomics Analysis.
    Tommy Zhang, Sara Amer, Julia Laskin.
      A central challenge in mass spectrometry imaging (MSI) of lipids is detecting and identifying species overshadowed by higher abundance isobaric lipids. Although tandem mass spectrometry can differentiate many lipid isobars and some isomers, the diversity and heterogeneity of lipids observed in MSI experiments necessitate the development of targeted approaches to differentiate and localize species with an overlapping m/z. To address this challenge, we leverage the fast acquisition speed of multiple reaction monitoring (MRM) to identify and localize hundreds of lipids in a single MSI experiment. We present a workflow to generate comprehensive, system-specific, and information-rich MRM transition lists for MSI and use them to examine the spatial localization of specific lipid classes with nanospray desorption electrospray ionization (nano-DESI) MSI in MRM mode. The lists are generated by integrating information from the LIPID MAPS database with data-dependent acquisition (DDA) and filtering by examining MRM signals of the corresponding tissue lipid extract. Using 165 MRM transitions in a single nano-DESI MSI experiment, we examined the localization of plasmalogen species, their isomers, and corresponding isobars in mouse brain tissue with sn-chain-level annotation. This workflow, which can be readily adapted to other lipid classes and tissue types, establishes MRM-MSI as a powerful strategy for mapping lipid targets in complex tissues.
    DOI:  https://doi.org/10.1021/acs.analchem.6c01209
  26. Fitoterapia. 2026 Jun 06. pii: S0367-326X(26)00243-1. [Epub ahead of print]192 107324
    Metabolomics of Andrographis paniculata Burm. F. green extracts by an integrated LC-MS and NMR approach and evaluation of acetylcholinesterase and tyrosinase inhibitory activity of extracts and isolated compounds.
    Luciana Maria Polcaro, Roberta Cuozzo, Sonia Piacente, Milena Masullo.
      Andrographis paniculata Burm. F. (Acanthaceae) is receiving significant interest from the scientific and medical communities due to its immunological, anti-inflammatory, antibacterial, and neuroprotective properties. This study aimed at comparing green extraction methods for A. paniculata in order to identify an environmentally friendly process capable of yielding a product rich in bioactive compounds. Andrographis paniculata aerial parts were submitted to extraction methods (maceration, Ultrasound Assisted Extraction UAE, and Solid Liquid Dynamic Extraction SLDE) using different solvent mixtures (100% EtOH, 75% EtOH/H2O, and 50% EtOH/H2O). Chemical profiling of the extracts was performed using an integrated approach based on high-resolution mass spectrometry (HRMS) via liquid chromatography-tandem mass spectrometry (LC-MS) and Nuclear Magnetic Resonance (NMR) spectroscopy, facilitating the identification of metabolites. A comparative analysis, conducted using multivariate statistical techniques, revealed that the extracts obtained by SLDE were the richest in metabolites. A detailed phytochemical investigation of the 75% EtOH/H2O extract obtained by SLDE was carried out to isolate and unambiguously characterize compounds putatively identified by LC-MS analysis. Among diterpenes, 7-hydroxy-14-deoxyandrographolide (15) is here reported for the first time from a natural source. Furthermore, the ability of extracts and isolated compounds to inhibit tyrosinase and acetylcholinesterase (AChE) enzymes was tested by spectrophotometric assays. Flavonoids (IC50 = 54.8-97.0 μM) inhibited tyrosinase enzyme slightly better than diterpenoids (IC50 = 98.8-173.8 μM), while the diterpenes 7-hydroxy-14-deoxyandrographolide (IC50 = 87.1 μM) and andrographolide (IC50 = 97.1 μM) showed the highest AChE inhibitory activity.
    Keywords:  AChE inhibitory activity; Andrographis paniculata; Diterpenes; Green extracts; HRMS; NMR; Tyrosinase inhibitory activity
    DOI:  https://doi.org/10.1016/j.fitote.2026.107324
  27. Inflammopharmacology. 2026 Jun 12.
    Chemical characterization and antinociceptive and anti-inflammatory activities of the aqueous extract of Myrciaria floribunda (H. West ex Willd.) O. Berg fruit peel.
    Simone Patricia De Freitas Rosa, Izabelly Bianca da Silva Santos, Wendel César eSilva Pereira, Thaís Vitória Freitas de Souza, Paulo Henrique Eloi Fernandes, Gabriela Ribeiro de Sousa, José Fernandes da Silva Cardoso, Alisson Macário de Oliveira, Márcia Vanusa da Silva, Wêndeo Kennedy Costa, Maria Tereza Dos Santos Correia.
      Natural products derived from medicinal plants represent an important source of bioactive compounds with potential therapeutic applications in the management of pain and inflammatory disorders. In this study, the chemical profile and pharmacological activities of the aqueous extract obtained from the fruit peel of Myrciaria floribunda (AeMf) were investigated. Chemical characterization was performed using high-performance liquid chromatography coupled with electrospray ionization mass spectrometry (HPLC-ESI-MSⁿ). The antinociceptive activity of AeMf was evaluated in mice using acetic acid-induced abdominal writhing, formalin, and tail immersion tests, while the anti-inflammatory potential was assessed through carrageenan-induced paw edema and peritonitis models. Chromatographic analysis revealed a chemical profile predominantly composed of organic acids and related metabolites, including quinic acid, citrate, maleate, and a fatty acid hexoside. In nociceptive models, AeMf produced a significant and dose-dependent reduction in acetic acid-induced writhing and markedly decreased paw-licking behavior in both phases of the formalin test. In addition, the extract increased tail withdrawal latency in the thermal nociception assay, indicating the involvement of both peripheral and central analgesic mechanisms. In inflammatory models, AeMf significantly inhibited carrageenan-induced paw edema, reduced leukocyte and neutrophil migration to the peritoneal cavity, and markedly suppressed the production of pro-inflammatory cytokines, including TNF-α and IL-1β. Collectively, these findings demonstrate that AeMf exerts potent antinociceptive and anti-inflammatory effects, likely mediated through the modulation of inflammatory mediators and cellular recruitment. The results highlight the therapeutic potential of Myrciaria floribunda as a promising natural source of bioactive compounds for the development of novel strategies for pain and inflammation management.
    Keywords:  Caatinga; Cytokines; Inflammation; Myrtaceae; Pain
    DOI:  https://doi.org/10.1007/s10787-026-02294-3
  28. Molecules. 2026 May 26. pii: 1826. [Epub ahead of print]31(11):
    Development of a Dispersive µSPE Method for the Determination of Pesticide Residues in Water Samples by LC-MS/MS.
    Gabrielle D Pereira, Igor F de Souza, Luana Floriano, Osmar D Prestes, Renato Zanella.
      The increasing occurrence of pesticides in aquatic environments has raised concern due to their potential impact on human health and ecosystems. In this context, the development of sensitive, reliable, and environmentally sustainable analytical methods is essential for monitoring these contaminants. Therefore, the aim of this study was to develop and validate a miniaturized dispersive solid-phase extraction (DµSPE) method for the determination of current-use multiclass pesticides in water samples using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Initially, a simple and rapid sample preparation procedure was developed, in which different experimental variables were evaluated to obtain suitable extraction efficiency. The validated method has a quantification limit of 0.01 µg L-1 and was applied to the determination of pesticides in surface water from different regions in Rio Grande do Sul State, Brazil. In addition, the environmental sustainability of the method was evaluated using the AGREEprep tool, allowing a quantitative and visual assessment of its compliance with the principles of Green Analytical Chemistry. The results demonstrated that the proposed method provides adequate analytical performance for the determination of 28 compounds in water matrices while offering a simple sample preparation procedure with reduced solvent consumption and waste generation.
    Keywords:  DµSPE; LC-MS/MS; green analytical chemistry; miniaturization; pesticides; water
    DOI:  https://doi.org/10.3390/molecules31111826
  29. J Pharm Biomed Anal. 2026 Jun 05. pii: S0731-7085(26)00269-4. [Epub ahead of print]280 117601
    Validation of qualitative and quantitative methods for 40 benzodiazepines and Z-drugs in hair by UPLC-MS/MS.
    Miao Zhang, Junbo Zhao, Shujie Yu, Ping Xiang, Fang Yan, Hui Yan.
      In recent years, the analysis and identification of benzodiazepines and Z-drugs have presented significant challenges in forensic medicine and pharmaceutical analysis due to their widespread use. Notably, some of these substances are classified as new psychoactive substances (NPS) by the UNODC. Hair was chosen as a biological matrix for monitoring long-term drug intake in this study. We aimed to develop and validate a rapid and sensitive method for simultaneous determination of 40 benzodiazepines and Z-drugs in hair using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). All the analytes were detected within 11 min. The method was validated with linear calibration curves over the range of 5-500 pg mg-1 (coefficient of determination, r2 > 0.99). The limits of detection (LOD) were 1-3 pg mg-1, and the limits of quantification (LOQ) were 5 pg mg-1. The method demonstrated acceptable selectivity, accuracy, and precision (< 20%). Matrix effects ranged from -29.8-23.5%, and recoveries ranged from 77.9% to 119.9%. The method was successfully applied to six authentic hair samples, detecting nine substances: Nitrazepam, Temazepam, Lorazepam, 7-Aminoflunitrazepam, Clobazam, Lormetazepam, α-Hydroxyalprazolam, 7-Aminoclonazepam, and Zolpidem. Benzodiazepines concentrations ranged from 5.6 to 436.1 pg mg-1, while zolpidem concentrations ranged from 11.1 to 113.8 pg mg-1. This method provided a reliable tool for detecting benzodiazepines and Z-drugs in hair and addresses the requirements of routine forensic practice.
    Keywords:  Benzodiazepine; Hair analysis; New psychoactive substances; UPLC-MS/MS; Z-drugs
    DOI:  https://doi.org/10.1016/j.jpba.2026.117601
  30. PLoS One. 2026 ;21(6): e0351295
    Metabolomics analysis identifies differential metabolites and potential diagnostic biomarkers among pediatric sepsis subtypes.
    Sisi Zhuang, Lili Zuo.
       BACKGROUND: Sepsis in children can be caused by a variety of pathogens, with bacteria and viruses being the most common. This study used metabolomics to identify differences in metabolic profiles and potential biomarkers among pathogens causing pediatric sepsis.
    METHODS: Serum metabolomic profiles of pediatric bacterial and viral sepsis were obtained from the MetaboLights database (MTBLS563). Principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), and orthogonal PLS-DA were employed to explore metabolic distinctions. Differential expression metabolites (DEMs) were identified using the Wilcoxon rank-sum test and variable importance in projection (VIP) scores. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, receiver operating characteristic (ROC) analysis, Extreme Gradient Boosting (XGBoost) modeling, and Shapley Additive exPlanations (SHAP) analysis were conducted to determine diagnostic metabolites and evaluate model performance.
    RESULTS: PCA and PLS-DA revealed distinct metabolic profiles among bacterial pediatric sepsis (PBID_PS), viral pediatric sepsis (VID_PS), and healthy controls. Fourteen differential metabolites were identified, primarily enriched in nitrogen metabolism, arginine biosynthesis, and the metabolism of alanine, aspartate, and glutamate. Among them, choline, glutamate, and glutamine exhibited strong discriminatory ability between PBID_PS and VID_PS. XGBoost and SHAP analyses confirmed these metabolites as key diagnostic indicators, achieving excellent predictive performance and revealing distinct metabolic reprogramming underlying different etiologies of pediatric sepsis.
    CONCLUSION: Metabolomic profiling revealed distinct metabolic signatures between bacterial and viral pediatric sepsis, with glutamate, glutamine, and choline serving as potential biomarkers.
    DOI:  https://doi.org/10.1371/journal.pone.0351295
  31. Potato Res. 2026 ;69(4): 131
    Untargeted Metabolomics Reveals Major Patterns of Metabolic Shifts in Potato Seed Tubers during Storage.
    Chunmei Zou, Ric C H de Vos, Roland Mumm, Henriëtte D L M van Eekelen, Aurin M Vos, Alejandro Thérèse Navarro, Robert D Hall, Martin K van Ittersum, Willemien J M Lommen, Paul C Struik.
      To obtain comprehensive insight into metabolic changes in potato seed tubers during storage and to identify patterns associated with physiological ageing, we analysed tuber metabolite composition using untargeted metabolomics based on gas and liquid chromatography-mass spectrometry (GC-MS and LC-MS). During two seasons, seed tubers of four contrasting cultivars (Agria, Festien, Innovator, and Lady Claire) were produced in a single field and after harvest, stored at different temperatures. During storage, the number of detected secondary metabolites increased progressively, particularly at higher temperatures (7-17 ˚C). Principal component analysis (PCA) revealed clear cultivar-specific metabolic profiles, with the starch cultivar Festien distinctly separated from the consumption cultivars, primarily caused by different amino acid composition. PCA further highlighted the impact of cold storage (4 ˚C) on primary metabolism, especially the accumulation of reducing sugars, as well as the combined effects of storage duration and elevated temperature on secondary metabolites, notably glycoalkaloids. Factor analysis (FA) supported these findings, with most metabolites strongly associated with the factor distinguishing cv. Festien from the other cultivars. Additionally, the leading factors captured cultivar-specific patterns and diverse trajectories reflecting the effects of storage duration and temperature. Together, these results provide a comprehensive overview of metabolic dynamics during storage and contribute to understanding the functional roles of metabolites in tuber physiological ageing.
    Supplementary Information: The online version contains supplementary material available at 10.1007/s11540-026-10083-2.
    Keywords:  Cultivar; GC–MS; LC–MS; Physiological age; Solanum tuberosum; Storage
    DOI:  https://doi.org/10.1007/s11540-026-10083-2
  32. J Pharm Biomed Anal. 2026 Jun 03. pii: S0731-7085(26)00264-5. [Epub ahead of print]280 117596
    Comprehensive metabolic profiling of Dictamni Cortex in normal and rheumatoid arthritis rats using UHPLC-Q/TOF-MS/MS coupled with polygonal mass defect filtering.
    Yanying Li, Wei Guan, Tiantian Wen, Shiqi Xu, Zhichao Hao, Qingshan Chen, Lili Zhang, Shu Liu, Haixue Kuang, Xiaochi Ma, Lili Huang, Yan Liu.
      Due to the low efficiency and time-consuming nature of detecting multiple metabolites from complex backgrounds-especially those with unpredictable metabolic types-as well as significant methodological limitations and data processing complexities, the in vivo metabolite profile analysis of herbs remains a major challenge. Therefore, this study developed an integrated strategy combining VBA background subtraction, polygonal mass defect filter (p-MDF), and untargeted metabolomics for identifying metabolites of quinoline alkaloids and limonoids. Through applying VBA background subtraction and p-MDF, the filtration ranges achieved for the biological samples were 19.30-24.34% and 78.32-85.46%, respectively. Finally, 108 compounds were efficiently and accurately identified from the water extract of Dictamni Cortex (DDW), including 84 quinoline alkaloids and 24 limonoids, among which 24 previously undescribed and 11 first-identified compounds were revealed. And a total of 158 compounds were detected in the plasma, urine, and feces of rheumatoid arthritis (RA) and normal rat following oral administration with DDW, including 69 prototype compounds and 89 metabolites. Untargeted metabolomics was performed to analyse plasma, urine, and feces samples from RA and normal rat. A total of 51, 28, and 9 differential parent compounds and metabolites were identified in the three sample types, respectively. It was speculated that the impact on the hepatointestinal environment and bioavailability under the RA state caused metabolic abnormalities in rats. In conclusion, this study provides a conceptual framework and methodological reference for the comprehensive analysis of herbal metabolites in complex biological matrices.
    Keywords:  Dictamni Cortex; Metabolites; Multivariate statistical analysis; Polygonal Mass defect filtering; Rheumatoid arthritis
    DOI:  https://doi.org/10.1016/j.jpba.2026.117596
  33. Biomed Chromatogr. 2026 Jul;40(7): e70512
    Solvent-Dependent GC-MS and LC-MS/MS Phytochemical Profiling of Echium italicum L. Aerial Part Extracts and In Vitro Enzyme Inhibitory and DNA-Protective Effects.
    İçim Gökkaya, Gökçe Seyhan, Elçin Güney, Ceren Boğuşlu, Mustafa Abdullah Yılmaz, Burak Barut, Gülin Renda.
      Echium italicum is a medicinal plant traditionally used in Türkiye for wound healing and inflammatory conditions. In this study, chloroform, ethyl acetate, ethanol, and 70% ethanol extracts prepared from the aerial parts were comparatively evaluated by gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS), and their in vitro antioxidant, enzyme inhibitory, and DNA interaction properties were investigated. LC-MS/MS profiling of the ethyl acetate, ethanol, and 70% ethanol extracts revealed marked solvent-dependent differences in phenolic composition, with the 70% ethanol extract containing the highest amounts of rosmarinic acid (8.858 mg/g), astragalin (4.551 mg/g), and nicotiflorin (3.874 mg/g). The GC-MS analysis showed that the primary constituents of the chloroform extract were 5-eicosene (25.30%), 1-octadecene (19.82%), and octacosanol (14.85%). The ethyl acetate extract mainly included 9-octadecenoic acid methyl ester (35.11%), methyl palmitate (25.05%), and methyl stearate (11.35%). Among all tested extracts, the 70% ethanol extract exhibited the strongest 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activity (78.10% at 200 μg/mL) and the highest α-glucosidase inhibition (61.45% at 200 μg/mL), whereas effects against α-amylase, tyrosinase, and elastase were weak. In agarose gel electrophoresis assays, the 70% ethanol extract did not induce DNA strand breaks and protected pBR322 plasmid DNA against hydroxyl radical-induced oxidative damage. These results suggest that hydroethanolic extraction more effectively recovers phenolics linked to antioxidant, α-glucosidase inhibitory, and DNA-protective effects in E. italicum.
    Keywords:   Echium ; DNA interaction properties; LC–MS; antioxidant; α‐glucosidase
    DOI:  https://doi.org/10.1002/bmc.70512
  34. J Mass Spectrom. 2026 Jul;61(7): e70072
    A Step-by-Step Protocol From METASPACE to Biological Interpretation.
    Abigail Moreno-Pedraza, Brittney Gorman, Marija Velickovic, Max Bentelspacher, Jaime Barros, Dusan Velickovic, Christopher R Anderton.
      Mass spectrometry imaging (MSI) represents an exceptional tool for exploring complex biological systems spatially at the molecular level. However, its multidimensional nature and large data outputs make it challenging to extract meaningful biological insights. Advancements such as the METASPACE platform allow researchers to efficiently process, annotate, and interpret MSI datasets by leveraging machine learning and a cloud-based infrastructure. In this tutorial, we present a detailed and user-friendly R-pipeline designed to help METASPACE users navigate untargeted metabolomic annotations and translate them into practical biological insights, particularly in complex systems. This approach has broad potential applications, including diagnostics, drug discovery, environmental, and ecological research. We envision this pipeline will be particularly useful for newcomers to MSI and encourage experienced users to customize and extend it to meet more advanced analytical needs.
    DOI:  https://doi.org/10.1002/jms.70072
  35. J Pharm Biomed Anal. 2026 Jun 10. pii: S0731-7085(26)00275-X. [Epub ahead of print]280 117607
    Comprehensive profiling of chemical composition, plasma-absorbed prototypes, metabolites, and pharmacokinetics of Jiannao Bushen Pill in rats using UHPLC-MS and microdialysis.
    Xiaojun Yu, Lingmei Kong, Bin Wang, Jing Liu, Xueli Lv, Qingyuan Sun, Ruoxuan Liu, Bonian Zhao, Jing Lv, Yan Gao.
      Jiannao Bushen Pill (JNBSP) is a traditional Chinese medicine (TCM) preparation used for nourishing the brain and kidney, tonifying qi and spleen, calming the mind, and improving memory. Clinically, it has been widely applied for the treatment of amnesia and cognitive decline; however, its pharmacodynamic material basis remains largely unclear. In the present study, an ultra-high-performance liquid chromatography coupled with Q-Exactive Orbitrap high-resolution mass spectrometry (UHPLC-Q-Exactive Orbitrap-HRMS) strategy was established to systematically characterize the chemical constituents of JNBSP. A total of 181 compounds were identified, and their herbal origins were further assigned through comparison with the chemical profiles of the 25 individual medicinal materials comprising the formula. In addition, 40 prototype compounds absorbed into the bloodstream and 41 metabolites were tentatively characterized in rat plasma after oral administration of JNBSP. To further evaluate the in vivo exposure behavior of representative absorbed constituents, microdialysis sampling combined with ultra-high-performance liquid chromatography-triple quadrupole tandem mass spectrometry was employed to monitor the temporal concentration profiles of six prototype compounds. The results revealed comparable Tmax values but different systemic exposure levels and elimination characteristics among these constituents, suggesting potentially different temporal and quantitative contributions to the pharmacological effects of JNBSP. To the best of our knowledge, this is the first systematic study to integrate comprehensive chemical profiling, characterization of absorbed prototypes and metabolites, and dynamic in vivo monitoring of representative constituents for JNBSP. These findings provide an important foundation for subsequent pharmacokinetic-pharmacodynamic studies, target identification, and mechanistic investigations of JNBSP in the treatment of cognitive dysfunction.
    Keywords:  Cognitive dysfunction; Material basis; Microdialysis; Traditional Chinese medicine; UHPLC-Q-Exactive Orbitrap-HRMS
    DOI:  https://doi.org/10.1016/j.jpba.2026.117607
  36. ACS Omega. 2026 Jun 02. 11(21): 30561-30569
    Comprehensive Metabolomic Analysis of Saliva Using SWATH-DIA Reveals Systemic Metabolic Adaptations to Exercise.
    Maroof Athar Hashmi, Diksha Gogia, Nirpendra Singh, Arvind Ramanathan.
      Saliva is an accessible, noninvasive biofluid for real-time monitoring of physiological and metabolic changes. However, its potential to capture physical-exciton-induced biochemical responses has been limited by the metabolite identification depth and reproducibility of conventional metabolomics tools. In this study, we established a Sequential Window Acquisition of All Theoretical Fragment-Ion Spectra (SWATH-DIA)-based untargeted LC-MS metabolomics workflow for comprehensive profiling and relative quantitation of the salivary metabolome before and after physical exercise. Saliva samples were collected from 27 recreational runners before and immediately after a standardized 5 km run to investigate acute metabolic fluctuations in participants. The Zeno SWATH-DIA method enabled the simultaneous acquisition of precursor and fragment ion spectra across the full m/z range (50-800 Da) in positive and negative mode of electrospray ionization (ESI), resulting in detection and validation metabolites spanning lipids, amino acids, organic acids, carbohydrates, and short-chain carnitines. Compared with traditional data-dependent acquisition (DDA) approaches, Zeno SWATH-DIA provided enhanced metabolite coverage, improved reproducibility, and reduced precursor selection bias (a statement of quantitation of how much more). Multivariate analyses (PCA, OPLS-DA) revealed clear separation between pre- and postexercise samples, highlighting metabolic shifts involving carbohydrate metabolism (lactate, pyruvate), fatty acid oxidation (acylcarnitines, glycerol), amino acid turnover (BCAAs, arginine, ornithine), and nitrogen metabolism (urea, spermidine). Collectively, these findings establish Zeno SWATH-DIA saliva metabolomics as a robust, high-coverage analytical approach for noninvasive assessment of acute metabolic responses to the exercise-induced physiological changes. The workflow provides a methodological foundation for future integrative studies linking saliva-based metabolomics with performance, fatigue, and metabolic health monitoring.
    DOI:  https://doi.org/10.1021/acsomega.5c11847
  37. J Ethnopharmacol. 2026 Jun 09. pii: S0378-8741(26)00859-7. [Epub ahead of print] 122005
    An integrated multitarget screening strategy combining network pharmacology with cell membrane chromatography coupled to high-performance liquid chromatography for identifying candidate antidepressant compounds from Cyperus rotundus L.
    Minghui Liu, Wanjiao Wang, Yuting Zhang, Jie Yong, Qingsong Jiang, Rui Sun, Xiao Han, Shengli Han, Langchong He, Heshui Yu, Jiaxin Yin, Zheng Li.
       ETHNOPHARMACOLOGICAL RELEVANCE: Cyperus rotundus L. (CR) is a traditional Chinese medicine (TCM) herb that has been used for centuries to soothe the liver, regulate Qi, and relieve emotional distress, with classical indications for "Yu syndrome" encompassing depressive mood, chest distension, and dysphoria. CR is incorporated into representative TCM formulas targeting depression-related conditions, such as Yueju Pill, substantiating its ethnopharmacological relevance in mood disorder management. However, its complex chemical composition presents considerable challenges for systematic bioactive compound identification, and the specific antidepressant constituents of CR remain poorly characterized, hindering the elucidation of its antidepressant material basis and the development of related therapeutics.
    AIM OF THE STUDY: This study aims to establish a comprehensive multi-method strategy for the rapid screening of multi-target candidate antidepressant compounds from CR and to conduct a preliminary, in vitro evaluation of their antidepressant-relevant activity.
    METHODS: Headspace gas chromatography-ion mobility spectrometry (HS-GC-IMS), headspace-solid phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS), and ultra-performance liquid chromatography-quadrupole Orbitrap mass spectrometry (UPLC-Q-Orbitrap-MS) were employed to comprehensively characterize the volatile and non-volatile chemical constituents of CR. On this basis, a multidimensional screening strategy integrating network pharmacology (NP) with a high-performance liquid chromatography-cell membrane chromatography (HPLC-CMC) online platform was developed to systematically identify candidate antidepressant compounds in CR. Molecular docking and surface plasmon resonance (SPR) were subsequently applied to evaluate the binding affinities between the identified candidate antidepressant compounds and their target receptors. Finally, with fluoxetine hydrochloride as a positive control, the neuroprotective effects of these compounds were assessed in a corticosterone-induced PC12 cell injury model.
    RESULTS: HS-GC-IMS and HS-SPME-GC-MS collectively identified 115 volatile organic compounds (VOCs), with four compounds (1,8-cineole, p-cymene, terpinen-4-ol, and α-pinene) detected by both methods. UPLC-Q-Orbitrap-MS characterized 30 non-volatile organic compounds (NVOCs). A total of seven candidate antidepressant compounds were identified that showed binding affinity for 5-Hydroxytryptamine 1A Receptor (5-HT1A), 5-Hydroxytryptamine 2A Receptor (5-HT2A), Dopamine D1 Receptor (DRD1), and Dopamine D2 Receptor (DRD2). Molecular docking and SPR analyses confirmed direct binding between these compounds and their target receptors. In a corticosterone-induced PC12 cell injury model, most of these compounds exhibited comparable or greater in vitro neuroprotective potency than fluoxetine hydrochloride under the tested conditions. Furthermore, cyperenone and aristolone are reported here for the first time to possess antidepressant potential.
    CONCLUSION: This study developed an integrated NP-HPLC-CMC screening strategy combined with HS-GC-IMS, HS-SPME-GC-MS, and UPLC-Q-Orbitrap-MS to identify seven candidate antidepressant compounds from CR targeting 5-HT1A, 5-HT2A, DRD1, and DRD2 receptors, with cyperenone and aristolone reported for the first time to possess antidepressant potential. These findings clarify the antidepressant material basis of CR and provide a reproducible framework for GPCR-targeted bioactive compound screening from TCMs, with in vivo validation of antidepressant efficacy identified as the essential next step.
    Keywords:  Candidate antidepressant compounds; Cell membrane chromatography; Cyperus rotundus L.; Depression; Network pharmacology
    DOI:  https://doi.org/10.1016/j.jep.2026.122005
  38. Foods. 2026 Jun 01. pii: 1954. [Epub ahead of print]15(11):
    Authenticity Assessment of Five Monofloral Honeys Based on Phytochemical Profiles.
    Yinan Du, Xinyue Du, Yue Wang, Hongcheng Zhang, Jiangtao Qiao, Yuncai Lu.
       BACKGROUND: Ensuring authenticity and verifying the floral origin of honey are persistent and critical issues in the quality control of bee products; in particular, the characteristic components and practical authenticity evaluation standards of several specialty fruit monofloral honeys are still insufficiently defined.
    METHODS: To address this, we conducted a comparative analysis of five fruit monofloral honey (loquat, pomegranate, citrus, apple, and blueberry) phytochemicals using high-performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC-MS), and the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine (TCM).
    RESULTS: Based on the currently available literature and databases, eleven identified phytochemicals appear to be reported in honey for the first time, including quinic acid derivatives, phenolamides, and flavonoid glycosides. Characteristic components with high species dependence were identified in distinct honey samples: anisic acid in loquat honey; methyl syringate in pomegranate honey; caffeine in citrus honey; cinnamic acid and methyl syringate in apple honey; and phaseic acid, methyl syringate, isorhamnetin-3-O-neohesperidoside, and callunene in blueberry honey. Twenty-three commercial samples were collected from the retail market to assess authenticity using HPLC fingerprints and quantitative thresholds for characteristic components. Authenticity was assessed based on both chromatographic fingerprint similarity and the content thresholds of characteristic phytochemicals specific to each monofloral honey type. The results indicated that 19 commercial samples satisfied the proposed authenticity criteria, whereas four commercial samples showed inconsistencies in characteristic phytochemical profiles or fingerprint similarity.
    CONCLUSIONS: This research establishes reliable chemical markers and a quantitative method to assess the authentication of five monofloral honeys, supporting high-value product development.
    Keywords:  HPLC fingerprints; authenticity evaluation; characteristic components; fruit monofloral honeys; phytochemicals
    DOI:  https://doi.org/10.3390/foods15111954
  39. Front Physiol. 2026 ;17 1826795
    Olive oil-derived monounsaturated fat influences metabolic signatures in serotonergic regions of the brain in broiler chicken.
    Odinaka C Iwuozo, Paul C Omaliko, Nathanael I Lichti, Bruce R Cooper, Yewande O Fasina.
       Introduction: Dietary fat modulates the dorsal raphe nucleus (DRN) and hypothalamic (HYP) serotonergic regions of the brain, influencing neurotransmitter activity and behavior in broiler chickens. Identifying metabolites that reach these serotonergic regions in response to dietary fat may support the formulation of diets that enhance neurological function and improve wellbeing. This study aimed to determine how monounsaturated fatty acids (MUFA) from olive oil influence the composition of metabolites that reach the DRN and HYP in broiler chickens.
    Methods: Day-old Ross 708 chicks (n = 160) were weighed and randomly assigned to two dietary treatments consisting of a corn-soybean meal basal diet supplemented with either poultry fat (CON) or olive oil (OLIV) at 3%, with five replicate pens of 16 chicks each. On d 20, plasma, DRN and HYP were aseptically collected and subjected to HPLCMS analysis for serotonin and metabolite profiling. Serotonin concentrations were analyzed using a Student's t-test, and metabolomic data were evaluated using linear modeling and principal component analysis to identify distinct metabolite profiles. Differentially abundant metabolites were annotated using HMDB and KEGG databases, followed by pathway enrichment analysis.
    Results and discussion: Results showed higher (P < 0.05) serotonin concentrations in both the DRN and HYP of OLIVfed birds. A total of 15,084 metabolites were differentially abundant (P < 0.05), of which 617 were annotated. Among these, Ssuccinylcysteine was upregulated in both plasma and DRN, while sphingomyelin was upregulated in DRN but downregulated in plasma. Dimethoxyflavone and cytosine were upregulated in HYP. Gammaglutamylleucine and epigallocatechin metabolites were differentially abundant in plasma and HYP but downregulated in DRN. Enrichment analysis revealed that glutamate metabolism was enriched in plasma and HYP but not significantly in DRN, whereas aspartate metabolism was altered in plasma and DRN only. Metabolic changes associated with purine metabolism across plasma, DRN, and HYP, and alterations in alanine, aspartate, and glutamate metabolism were also noted. Overall, dietary olive oil altered metabolites and pathways in serotonergic brain regions, indicating that MUFA intake modulates their metabolic activity. Higher serotonin concentrations in the DRN and HYP further support enhanced central serotonergic function in OLIV fed broiler chickens.
    Keywords:  broiler chickens; dorsal raphe nucleus; hypothalamus; metabolomics; olive oil
    DOI:  https://doi.org/10.3389/fphys.2026.1826795
  40. J Anal Toxicol. 2026 Jun 09. pii: bkag041. [Epub ahead of print]
    Analysis of suzetrigine, a novel non-opioid pain medication, in whole blood and urine using liquid chromatography-tandem mass spectrometry.
    Isabelle R Parry, Britni N Skillman.
      Suzetrigine (JOURNAVX®) is a novel non-opioid pain medication that was approved in January 2025. Given the well-documented abuse potential of opioids, suzetrigine may become more widely prescribed as an alternative for acute pain management. However, it could still appear in forensic casework as little is understood about its role in polydrug toxicity, comorbidity with pre-existing health conditions, or abuse potential within larger populations. As such, this research aimed to develop the first forensically focused method to detect suzetrigine in blood and urine. Due to its similar physicochemical properties, suzetrigine was seamlessly integrated into a previously validated liquid-liquid extraction method developed for novel dual orexin receptor antagonists (DORAs). This report focuses on the subsequent method development and validation for suzetrigine detection using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The method was validated in accordance with ANSI/ASB 036 for the quantification of suzetrigine in blood and its qualitative identification in urine. A quadratic calibration model with 1/x weighing was utilized (1.0-1000 ng/mL) with quality controls (QCs) at 1.5, 80, 400, and 800 ng/mL. The limit of detection in blood and urine was 0.25 ng/mL, while the limit of quantitation in blood was 1.0 ng/mL. Post-extraction addition at 5 ng/mL and 500 ng/mL yielded matrix effects less than ± 25%, but increased variability at the low concentration in blood was deemed to have no impact on LOD. No carryover or interferences were observed from the internal standard, matrices, or common drugs. Dilution integrity (5X) was acceptable in blood, and processed samples were stable up to 72 hours in the autosampler. In this report we describe a simple extraction for suzetrigine in blood and urine followed by LC-MS/MS analysis. This analytical workflow will be valuable as the use of suzetrigine becomes more prevalent in casework samples.
    Keywords:  Suzetrigine; blood; liquid chromatography–tandem mass spectrometry; novel analgesic; urine
    DOI:  https://doi.org/10.1093/jat/bkag041
  41. Foods. 2026 Jun 03. pii: 2001. [Epub ahead of print]15(11):
    Suspected and Non-Targeted Screening of Non-Edible Substances in Food by UPLC-Q-TOF-MS.
    Ting Wang, Fuhong Chen, Lirong Pan, Wenxuan Yuan, Jie Pang, Xianliang Li, Cunxian Xi, Dunming Xu.
      A screening method based on dispersive solid-phase extraction (DSPE) coupled with ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was established for the analysis of non-edible substances in food. This method is applicable to a wide range of non-edible substances, including but not limited to antihypertensive, hypoglycemic, weight-loss, antimicrobial, antipyretic-analgesic, sedative-hypnotic, and antifatigue agents. Through systematic optimization of sample pretreatment and UPLC-Q-TOF-MS conditions, ultrasonic extraction with methanol followed by cleanup using 25 mg Primary Secondary Amine (PSA) and 50 mg C18 was identified as the optimal procedure. The methodological validation demonstrated that all 38 quality control compounds exhibited excellent linear correlation coefficients (R2 > 0.99) across a concentration range of 0.005~5.0 mg/kg. At three spiking levels, the mean recoveries and relative standard deviations (RSDs) in four matrices ranged from 67.79% to 110.93% and from 0.23% to 9.37%, respectively. The screening detection limits (SDLs) and limits of quantification (LOQs) were within the range of 0.003~0.5 mg/kg. A screening database comprising 390 substances was constructed. In addition, an identification strategy for the unknown structural analogues was established by summarizing the mass spectrometric fragmentation patterns of the phosphodiesterase-5 (PDE-5) inhibitor analogues. Applied to 110 batches of samples, the method screened 12 known non-edible substances and identified a new PDE-5 inhibitor analogue, phenyl 3-desethyl 3-propyl carbodenafil. The workflow integrates suspected screening using a comprehensive database with a non-targeted identification strategy for unknown analogues. Overall, this strategy is efficient, sensitive and accurate, providing a robust analytical platform for high-throughput screening and discovery of illegally added unknown substances in food.
    Keywords:  PDE-5 inhibitors; dispersive solid-phase extraction; high-resolution mass spectrometry; non-edible substances; suspected screening; unknown structural analogues
    DOI:  https://doi.org/10.3390/foods15112001
  42. Plants (Basel). 2026 May 28. pii: 1651. [Epub ahead of print]15(11):
    Medicinal Amazonian Oleoresins: An Eco-Friendly Chemical Fingerprinting Method.
    Rayssa Ribeiro, Gabriel Reis Alves Carneiro, Henrique Marcelo Gualberto Pereira, Monica Costa Padilha, Valdir F Veiga-Junior.
      Oleoresins are complex natural lipophilic matrices traditionally analyzed using chromatographic techniques that require extensive sample preparation, derivatization, and authentic standards. Amazonian oleoresins from Copaifera and Eperua species (Fabaceae) represent valuable bioresources with recognized pharmacological potential, largely attributed to diterpenoids such as copalic and hardwickiic acids, as well as bioactive sesquiterpenes, including the cannabinoid β-caryophyllene. In this study, we present a proof-of-concept application of Direct Analysis in Real Time coupled with High-Resolution Mass Spectrometry (DART-HRMS) as a rapid, direct, and environmentally friendly approach for chemical fingerprinting and semi-targeted screening of the two most important amazonian oleoresins from these two genera: Eperua oleifera and Copaifera multijuga. Analyses were performed using a Q Exactive Orbitrap coupled to a DART ion source under optimized conditions. Hardwickiic acid was used as a model compound for method optimization, with optimal performance achieved at 200 °C and 100 V, yielding stable signal intensities (CV < 10%) and high mass accuracy (<1 ppm). The method enabled reproducible detection of diterpenic acids in both oleoresins, allowing differentiation of their chemical profiles and assessment of short-term stability under ambient conditions. In addition to diterpenes, free fatty acids were also detected, expanding the compositional characterization of these matrices. Compound annotation was performed based on accurate mass measurements and literature comparison, corresponding to Level 5 confidence according to established metabolomics criteria. Although the absence of chromatographic separation limits isomer discrimination and absolute quantification, DART-HRMS provides a rapid and solvent-free strategy for chemical fingerprinting and preliminary characterization of oleoresins. This approach aligns with Green Chemistry principles and shows strong potential as a screening and triage tool for quality control, chemotaxonomic studies, and sustainable valorization of Amazonian natural products.
    Keywords:  Amazonian bioeconomy; Copaifera multijuga Hayne; DART-MS; Eperua oleifera Ducke; Green Chemistry; oleoresins
    DOI:  https://doi.org/10.3390/plants15111651
  43. Food Res Int. 2026 Sep 01. pii: S0963-9969(26)01199-3. [Epub ahead of print]239 119516
    Metabolomics reveals the chemical basis of quality formation in GABA-enriched fermented green tea by lactic acid bacteria.
    Junjie Zhang, Lu Liu, Yijie Guan, Shengnan Chen, Guojie Liu, Qi Wang, Xueling Gao, Na Xu, Qi Chen.
      Fermenting tea infusions with lactic acid bacteria (LAB) can effectively increase γ-aminobutyric acid (GABA) content and reshape the chemical composition and flavor profile, providing a new strategy for functional tea beverage development. In this study, instant green tea was used as raw material to screen LAB strains with high GABA production and tea infusion tolerance, and the fermentation process was optimized to prepare GABA-enriched fermented tea (GFT). GC-MS, UPLC-MS/MS, quantitative descriptive analysis (QDA), relative odor activity values (rOAV), and multivariate statistics were integrated to reveal the regulatory mechanisms of LAB fermentation on GFT quality formation. The results showed that Lactobacillus strains 2# and 3# exhibited excellent GABA synthesis ability and maintained robust metabolic activity in the tea infusion system. The optimal conditions for strain 2# were: tea-to-water ratio 1:25, inoculum 3%, fermentation 24 h; and for strain 3# were: tea-to-water ratio 1:25, inoculum 1%, fermentation 36 h. After fermentation, the brightness of the tea infusion was significantly improved, the umami and kokumi were enhanced. The aroma shifted from a single chestnut note to a complex aroma with fermented, woody, and citrus characteristic. A total of 146 volatile compounds were detected by GC-MS, and 38 characteristic volatiles were shared by two GFT groups, among which phenethyl alcohol, damascenone, and jasmone formed the core aroma basis. A total of 1763 non-volatile compounds were identified by UPLC-MS/MS, organic acids, flavonoids, and phenolic acids were the main differential metabolites. The chemical composition of the tea infusion underwent significant changes following lactic acid bacteria fermentation, with the levels of tea polyphenols and epicatechins decreasing significantly, while the levels of free phenolic acids, flavonoid aglycones, organic acids, and umami amino acids increased significantly. This pattern of changes is highly consistent with the known functions of esterases, β-glucosidases, and glutamate decarboxylases produced by lactic acid bacteria, suggesting that these enzymes may have been involved in the aforementioned chemical transformation processes. Flavonoid glycosides were reduced and flavonoid aglycones elevated; esterified phenolic acids were hydrolyzed. Lactic acid and fumaric acid accumulated, while oxalic acid, citric acid, and other TCA cycle intermediates were significantly reduced. KEGG pathway enrichment analysis showed that differentially metabolites were mainly enriched in flavonoid biosynthesis, cysteine/methionine metabolism, and citrate cycle. This study reveals the mechanism of LAB fermentation in regulating the chemical quality of GABA tea from sensory, aroma, and metabolic dimensions, and provides a theoretical foundation for process optimization and product development of functional fermented tea.
    Keywords:  Aroma compounds; Fermented tea; GC–MS; Lactic acid bacteria; Metabolomics; Quality formation; UPLC-MS/MS; γ-Aminobutyric acid
    DOI:  https://doi.org/10.1016/j.foodres.2026.119516
  44. Plants (Basel). 2026 Jun 02. pii: 1718. [Epub ahead of print]15(11):
    Evaluation of the Therapeutic Potential and Safety Profile of Six Salvia Species Native to Türkiye.
    Nagehan Saltan, Fatmanur Tunç, Merve Baysal, Gamze Göger, Serkan Levent.
      The genus Salvia L. represents one of the most pharmacologically significant groups within the Lamiaceae family. This study investigates the phytochemical profiles and biological activities of six Salvia species native to Türkiye (S. dorystaechas B.T.Drew, S. sclarea L., S. glutinosa L., S. tomentosa Mill., S. argentea L., and S. aethiopis L.) to scientifically validate their extensive use in Turkish traditional medicine. Phytochemical characterization was performed using Liquid Chromatography-High-Resolution Mass Spectrometry (LC-HRMS), while biological potential was evaluated through antioxidant (DPPH), antimicrobial (MIC), and cytotoxicity (MTT on NIH/3T3 cells) assays. Among the taxa, S. dorystaechas exhibited the most potent antioxidant activity, with IC50 values of 0.090 mg/mL (infusion) and 0.072 mg/mL (ethanolic), which strongly correlated with high total phenolic contents (111.50 and 125.55 mg GAE/g, respectively). This species may also serve as a potential source of bioactive compounds. Antimicrobial screenings against pathogenic bacteria and Candida spp. demonstrated modest inhibitory effects, with MIC values ranging from 625 to >5000 µg/mL. Safety profiling indicated that the ethanolic extract of S. tomentosa showed the lowest cytotoxicity (IC50 562.37 ± 49.50 µg/mL) among the tested samples, which nonetheless indicates a relatively narrow therapeutic window. LC-HRMS profiling revealed the presence of flavonoids and phenolic diterpenes, such as carnosol and rosmanol, providing a chemical rationale for the observed moderate activities. Consequently, rather than direct systemic pharmacological agents, these findings suggest that the studied Salvia species could serve as preliminary botanical sources for the isolation of specific secondary metabolites or for restricted topical applications.
    Keywords:  LC-HRMS; Salvia; antimicrobial activity; antioxidant activity; cytotoxic activity
    DOI:  https://doi.org/10.3390/plants15111718
  45. ACS Omega. 2026 Jun 02. 11(21): 31542-31551
    Molecular Transformation of Dissolved Organic Matter at the Coal-Water Interface: Insights from High-Resolution Mass Spectrometry.
    Min Wu, Kaiquan Deng, Jinli Jiang, Yixiang Bao, Jingfeng Li.
      This study employed untargeted high-resolution mass spectrometry combined with multivariate statistical analysis to elucidate the detailed molecular-geochemical differentiation patterns and underlying driving mechanisms during the migration and transformation of coal-derived dissolved organic matter (DOM) into mine groundwater. Results indicate that the coal DOM molecular library was highly complex and diverse, with a total of 40,158 molecular formulas identified, primarily composed of heteroatom-rich, nitrogen- and sulfur-containing compounds (e.g., CHON, CHONS classes) and condensed aromatic structures. In contrast, the total molecular formulas in mine groundwater DOM markedly decreased to 8,829, and its composition underwent fundamental restructuring, shifting toward a predominance of CHO-type compounds, a notable increase in phosphorus-containing constituents, and overall higher hydrophilicity and oxidation state of molecules. Both Van Krevelen diagrams and principal component analysis statistically confirmed substantial differences and clear separation between the two DOM types in terms of elemental composition and molecular characteristics. Further analysis revealed that this differentiation was driven by stringent environmental screening, potentially influenced by synergistic processes including physicochemical selective dissolution, microbial degradation and transformation, and adsorption at the water-rock interface. The identification of 1,529 shared molecules common to both media, predominantly consisting of aliphatic and lignin-like CHO/CHON compounds, provides putative molecular evidence of coal-water interaction. These findings offer essential molecular-level insights and a scientific basis for accurately tracing organic pollutant sources in mine groundwater, assessing pollutant migration risks, and understanding the transformation and stability of the subsurface carbon pool.
    DOI:  https://doi.org/10.1021/acsomega.6c02127
  46. Talanta. 2026 Jun 09. pii: S0039-9140(26)00784-8. [Epub ahead of print]310 130128
    Metabolomics coupled with machine learning highlights metabolite profile differences of Scutellaria baicalensis Georgi from different specifications and geographical origins.
    Xiuhuan Wang, Axiang Yu, Juan Qin, Jianling Yao, Yu Cao, Yuqing Yang, Letian Ying, Yongqi Yang, Xinru Wu, Jiaqian Xing, Lingzi Wu, Wanzhu Wang, Gaimei She.
      Scutellaria baicalensis Georgi (SBG) is recognized as a species with both medicinal and nutritional value, exhibiting extensive pharmacological activities and edible value. SBG from multiple origins and in two distinct specifications is widely available in the market at the present time. A rapid and accurate analytical method was developed by combining machine learning with metabolomics strategies to discriminate between two specifications and four sources of SBG, and to identify their respective differential markers (DMs). The establishment of a qualitative identification model to distinguish the geographical origins and specifications of SBG was established by employing cost-sensitive models, feature selection techniques, and machine learning algorithms. Both discriminant models demonstrated satisfactory performance. Nine DMs, including organic acids, isoflavones, fungal metabolites, and sesquiterpenes, were considered as potential chemical markers for discriminating the geographical origin of SBG. In contrast to the former, two SBG specifications (Kuqin and Ziqin), abbreviated to KQ and ZQ, contain a total of 15 DMs, which predominantly belong to the flavonoid components. The strategy proposed in this study provides foundation for constructing qualitative identification models, offers valuable insights into how geographical origins and morphological specifications shape the metabolic profiles of SBG and other food-medicine homologous herbs.
    Keywords:  Differential markers; Geographical origins; Metabolomics; Scutellaria baicalensis Georgi; Specifications
    DOI:  https://doi.org/10.1016/j.talanta.2026.130128
  47. Molecules. 2026 Jun 04. pii: 1959. [Epub ahead of print]31(11):
    Exploring the Chemical Profile and Biological Activities of Eryngium dichotomum: UHPLC-MS/NMR Characterization, and In Vitro Antioxidant Activity Along with the Antitumor Effect of Falcarinol.
    Roufia Mezaache, Habiba Laraoui, Anis Bertella, Verónica Bastos, Helena Oliveira, Patrick Pale, Aurelien Blanc, Stefan Chassaing, Oana-Crina Bujor, Diana C G A Pinto, Liliana Bădulescu, Artur M S Silva, Fatma Bitam.
      Qualitative liquid chromatography-mass spectrometry (UHPLC-MS) and NMR analysis of the diethyl ether extract of the aerial part of Eryngium dichotomum plant belonging to the Apiaceae family led to the putative identification of phenolic acids, flavonoid glycosides, triterpenoid saponins, fatty acids, and oxylipins. The tentative identification of several secondary metabolites by UHPLC-MS analysis was further confirmed by compound isolation and comprehensive spectroscopic characterization using 2D NMR and mass spectrometry, leading to the elucidation of seven compounds, a mixture of two hydroxy fatty acids, namely (Z,E)-13-hydroxyoctadeca-9,11-dienoic acid (1) and (E)-13-hydroxyoctadec-11-enoic acid (2); two C17 polyacetylenes, (E)-heptadeca-1,10-dien-4,6-diyne-3,8,9-triol (3), and falcarinol ((Z)-1,9-heptadeca-1,9-dien-4,6-diyn-3-ol) (4); glycerol monopalmitate (5) and two flavonoid glycosides, kaempferol 3-O-β-D-glucopyranosyl-(1 → 6)-O-β-D-galactopyranoside (6), and quercetin 3-O-β-D-glucopyranosyl-(1 → 6)-O-β-D-galactopyranoside (7). Furthermore, the antioxidant activity of the n-butanol and the diethyl ether extracts of the species were evaluated using the DPPH, FRAP, and ABTS assays. In addition, the anticancer activity of the major falcarinol-type polyacetylene was assessed against A375 human melanoma cells.
    Keywords:  Eryngium; UHPLC–MS; falcarinol; flavonoids; triterpenoid saponins
    DOI:  https://doi.org/10.3390/molecules31111959
  48. J Mass Spectrom. 2026 Jul;61(7): e70070
    Rapid Determination of 13 Insecticides and Acaricides in Human Blood Using Dispersive Liquid-Liquid Microextraction Coupled With DART-MS/MS.
    Yuan Zhou, Ke Hu, Xiaolong Hou, Chenyu Xue, Wenfang Zhang, Ying Zhang, Hua Liu.
      A high-throughput, solvent-minimized analytical method was developed by coupling dispersive liquid-liquid microextraction (DLLME) with direct analysis in real-time tandem mass spectrometry (DART-MS/MS) for the simultaneous quantification of multiple insecticides and acaricides in human blood. This method obviates the need for chromatographic separation, substantially reducing total analysis time. Extraction conditions were systematically optimized to achieve efficient enrichment of target analytes from complex biological matrices. The method exhibited good linearity across compound-specific calibration ranges (R2 = 0.9912-0.9993) with limits of quantification as low as 0.1 ng/mL. Recoveries ranged from 78.2% to 110.7%, while matrix effects remained within ±25%. Both intraday and interday precisions met commonly accepted validation criteria (RSD < 20%). Excellent agreement with HPLC-MS/MS results confirmed the method's analytical reliability. Although tested on a limited number of samples, the method demonstrated robust performance in two real-case blood samples and four spiked samples, highlighting its practical potential for the rapid screening and exposure assessment of insecticides and acaricides in time-sensitive forensic casework.
    Keywords:  direct analysis in real‐time tandem mass spectrometry (DART‐MS/MS); dispersive liquid–liquid microextraction (DLLME); human blood; insecticides and acaricides
    DOI:  https://doi.org/10.1002/jms.70070
  49. BMC Plant Biol. 2026 Jun 10.
    Effects of extraction solvents on the recovery of bioactive compounds from Platycladus orientalis Leaves of different tree ages: a comparative metabolomics approach.
    Huayu Li, Bei Cui, Haoran Zhao, Guopeng Wang, Yuanyuan Wang, Sheng Zhang.
       BACKGROUND: Platycladus orientalis foliage, referring to the dried twigs and leaves of the Cupressaceae plant P. orientalis, is a traditional Chinese medicine known for its anti-inflammatory and sedative properties. The composition and concentration of metabolites in P. orientalis foliage vary depending on extraction solvents and the age of the tree, which makes the selection of efficient solvents and appropriate tree age crucial.
    RESULTS: This study evaluated solvent extraction efficiency for bioactive compounds from P. orientalis foliage and analyzed metabolic variations across trees aged 5-3000 years. Acetic acid (AcOH) achieved the highest extraction yield (14.17%), while Ethanol (EtOH) yielded the lowest (9.60%). EtOH extracts exhibited the highest total phenolic and flavonoid contents, with flavonoids increasing with tree age. Water and EtOH showed stronger DPPH scavenging than AcOH; water excelled in ·OH scavenging, whereas AcOH led in ABTS scavenging and total antioxidant activity. Older trees consistently outperformed younger ones in antioxidant assays. Both EtOH and AcOH inhibited E. coli and S. aureus; AcOH's effect against E. coli strengthened with tree age. Water and essential oil showed no antibacterial effects, though all extracts, especially essential oil, inhibited Aspergillus niger. GC-MS identified 88 metabolites and 14 bioactive compounds. β-pinene and geraniol characterized trees under 1000 years, while caryophyllene oxide and α-terpineol accumulated in super-aged trees. These compounds exhibited multi-target pharmacological synergy via anti-tumor, analgesic, anti-inflammatory, and neuroprotective pathways. UHPLC-QE-MS suggested upregulated phenylpropanoid and flavonoid biosynthesis in 3000-year-old trees, leading to accumulation of key antioxidants like isoeugenol, coniferin, myricetin, and quercetin.
    CONCLUSIONS: These findings confirm that even at 3000 years of age, P. orientalis maintains a robust capacity to synthesize active metabolites, underscoring its considerable potential as a valuable resource for pharmaceutical research and development. This study highlights the synergistic effects of solvent and tree age on the bioactive compounds in P. orientalis foliage, providing a theoretical basis for optimizing extraction processes and developing high-value natural products. It also underscores the unique value of ancient tree resources in pharmaceutical applications, offering new strategies for plant-derived drug development.
    Keywords:   Platycladus orientalis extract; ancient tree; antibacterial activity; antioxidant activity; differential metabolomics; phytochemical composition
    DOI:  https://doi.org/10.1186/s12870-026-09199-x
  50. Biomed Chromatogr. 2026 Jul;40(7): e70509
    UPLC-QTOF-MS/MS and Antifungal Activity of a Fraction Enriched in Saponins From Sarcomphalus joazeiro Against Candida spp.
    Ana Raquel Pereira da Silva, Maria do Socorro Costa, Nara Juliana Santos Araújo, Gabriel Gonçalves Alencar, Talysson Felismino Moura, Victor Juno Alencar Fonseca, Antônia Thassya Lucas Dos Santos, Maria Flaviana Bezerra Morais-Braga, Erlânio Oliveira de Souza, Josean Fechine Tavares, Paulo Riceli Vasconcelos Ribeiro, Jacqueline Cosmo Andrade-Pinheiro, Henrique Douglas Melo Coutinho.
      The development of antifungal resistance is a complex process that involves the interaction between hosts, drugs, and microbial factors, all of which contribute to therapeutic ineffectiveness. For this reason, studies on natural products as possible therapeutic alternatives are increasingly necessary in order to gain a better understanding of plant compounds that may be more effective in treating infections caused by fungal pathogens. In this context, this study aimed to investigate the antifungal potential of the saponin-enriched fraction of the Sarcomphalus joazeiro species against the Candida albicans, Candida tropicalis, and Candida krusei strains. The selection of this species for study is due to its rich phytochemical composition and the vast traditional knowledge of its medicinal properties. The fraction obtained from the stem bark of S. joazeiro was analyzed by UPLC-QTOF-MS/MS, in which compounds such as triterpenoids, flavonoids, acids, and saponins were identified. Therefore, the results obtained in this study contribute to understanding the antifungal potential of the S. joazeiro species fraction against fungal infections, especially those caused by Candida spp.
    Keywords:  antifungal activity; flavonoids; liquid chromatography; saponins
    DOI:  https://doi.org/10.1002/bmc.70509
  51. Molecules. 2026 Jun 01. pii: 1890. [Epub ahead of print]31(11):
    From Conventional Methods to Innovation: Caffeine and Chlorogenic Acid Extraction and Quantification in the Rise of Smart and Green Techniques-A Systematic Review.
    Shady H Awwad, Lara M Nasereddin, Ola Al-Tamimi, Ahmad Q Daraosheh, Ali Elrashidi, Lydia Abu Al-Shayeb, Mais Shannag, Beisan A Mohammad, Reem Issa, Mahmoud S Abu-Samak.
      Caffeine and chlorogenic acid are among the most extensively investigated bioactive compounds in coffee, tea, and other plant-derived products due to their noteworthy physical, nutritional, and industrial relevance. Caffeine is primarily acknowledged for its central nervous system stimulant activity, whereas chlorogenic acid, a phenolic ester, contributes antioxidant, anti-inflammatory, and metabolic health benefits. This review was conducted according to the PRISMA guidelines in order to systematically compile and summarize the extraction and analytical conditions reported for caffeine and CGAs in different matrices and to provide a structured comparison among the reported studies. All studies focusing on the extraction and/or quantification of caffeine and chlorogenic acids in several matrices were considered eligible. Three independent electronic searches were performed using PubMed, Science.gov, and BASE to identify relevant articles. Extraction of data was conducted independently by four authors based on consistent selection and extraction criteria. One hundred and twenty-five studies were identified. The results were summarized in tables including several parameters. Conventional extraction techniques, including aqueous and organic solvent-based methods, have formed the foundation for separating caffeine and chlorogenic acids. However, rising interest in green and sustainable technologies has shifted attention towards advanced approaches such as ultrasound-assisted extraction and microwave-assisted extraction. These methods not only enhance extraction yields and reduce processing times but also align with environmental and safety concerns in the modern food and pharmaceutical industries. For quantification, high-performance liquid chromatography equipped with ultraviolet or mass spectrometric detection remains the benchmark, offering precision and reproducibility in different matrices. This review sheds light on recent advances and ongoing research in the extraction and quantification of caffeine and chlorogenic acid in different types of matrices. Continued innovation in green extraction technologies and robust quantification methods is essential for supporting scientific research applications.
    Keywords:  caffeine; chlorogenic acid; extraction; green chemistry; quantification
    DOI:  https://doi.org/10.3390/molecules31111890
  52. Int J Mol Sci. 2026 May 27. pii: 4822. [Epub ahead of print]27(11):
    Chemical Composition, Antioxidant, and Anti-Inflammatory Activity of the Antarctic Lichen Leptogium puberulum: A Combination of Metabolomic, In Vitro, and In Silico Approaches.
    Alfredo Torres-Benítez, José Erick Ortega-Valencia, Juan Rodrigo Salazar, Katherine Monje, Jaqueline Ley-Martínez, Gabriel Vargas-Arana, Mario J Simirgiotis.
      Lichenized fungi are a source of secondary metabolites with multiple biological potential. The objective of the study was to determine the chemical composition, antioxidant, and anti-inflammatory activity of the hydroalcoholic extract of the Antarctic lichen Leptogium puberulum through metabolomic, in vitro, and in silico analyses. Seventeen compounds were tentatively identified using UHPLC-ESI-QToF-MS. The phenolic composition yielded 6.356 mg GAE/g, and antioxidant activity assays showed IC50 values for DPPH• and ABTS•+ of 1187.149 and 207.00 µg/mL, respectively, along with 15.845 µmol Trolox/g for ORAC and 21.925 µmol Trolox/g for FRAP. The in silico evaluation was performed using OSIRIS Data Warrior, ProTox 3.0, and SwissTargetPrediction, identifying 9,10,12,13,14-pentahydroxytetracosanoic acid (PHTA), 9,10,12,13-tetrahydroxytricosanoic acid (THTA), 9,10,12,13-tetrahydroxyheneicosanoic acid (THHA), and 9,10,12,13-tetrahydroxydocosanoic acid (THDA) as the most promising compounds. These metabolites showed favorable pharmacokinetic properties, with no anticipated toxicological risks. Subsequently, their affinity for the cyclooxygenase-2 (COX-2) enzyme was evaluated by molecular docking with AutoDock Vina software version 1.2.3, and the most stable protein-ligand complexes were analyzed to characterize key interactions within the active site and subjected to molecular dynamics simulations with YASARA software version 19.1.27 for 100 ns. Overall, these results indicate that selected metabolites from L. puberulum may act as potential COX-2 inhibitors, supporting their relevance as lichen-derived anti-inflammatory agents and warranting further pharmacological investigation.
    Keywords:  Antarctica; COX-2 inhibition; bioactive compounds; biological potential; extract; metabolomics; molecular docking; pharmacokinetic properties
    DOI:  https://doi.org/10.3390/ijms27114822
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