bims-mebolo Biomed News
on Metabolomics
Issue of 2026–06–21
fifty-one papers selected by
Daniel Méndez Rodríguez, Vbi-Ugent
  1. Integrated metabolomics and antioxidant activity assessment of Sphagneticola trilobata (L.) Pruski.
  2. Determination of Coblopasvir in Mouse Plasma by Liquid Chromatography-Tandem Mass Spectrometry and Its Application to Pharmacokinetic Study.
  3. BAGO: A Self-Optimizing Tool for LC-MS Gradient Design in Metabolomics.
  4. An improved method for quantification of cannabidiol, Δ9-tetrahydrocannabinol, and their metabolites in human plasma and urine.
  5. Development of new liquid chromatography-tandem mass spectrometry methods for the quantification of seven metabolites in three related breast cancer cell lines.
  6. Fragmentation property identity card-assisted reliable structural annotation of calycosin metabolites in vivo: A strategy for isomer discrimination using mass spectrometry.
  7. Siderophore screening in marine sponge extracts using LC-HRMS and an R-based metabolomics workflow.
  8. Benchmarking MS/MS Featurization Strategies for Machine Learning-Driven Metabolite Structure Annotation.
  9. Online solid-phase extraction coupled to liquid chromatography-tandem mass spectrometry for fungicide translocation studies in soybean plants.
  10. MS2-SMILES AlignNet: A cross-modal contrastive learning framework for direct alignment of tandem mass spectra and molecular structures.
  11. An Ultra-High performance liquid chromatography-tandem mass spectrometry method for simultaneous determination of etomidate, analogues, and metabolites in human nail: Unveiling a unique accumulation profile.
  12. Rapid and targeted HILIC-MS/MS quantification of urinary metabolites reveals metabolic alterations in COVID-19 patients.
  13. Comparative transcriptomic and metabolomic profiling of forest tea and terrace tea in Sandu, Guizhou, China.
  14. A multi-dimensional approach combining HPLC, ion mobility and high resolution mass spectrometry for investigating transition metal complexes.
  15. Lipidome and Targeted Metabolome Analysis of Locational Picrorhiza Kurroa Royle. Ex Benth., Using UPLC-PDA and UHPLC-QTOF-IMS.
  16. Two Flavones as Markers for the Distinction of Natural Acacia Honey Based on the UHPLC-QE-MS Method and Research on Their Potential Immunomodulatory Activity.
  17. Metabolomic characterization and temporal dynamics of Burkholderia cepacia for polyhydroxyalkanoates production from oleic acid.
  18. Urinary organic acid levels and their associations with clinical characteristics in patients with schizophrenia.
  19. A cost-efficient internal standard approach for busulfan quantification: MS-based investigation of adduct formation in therapeutic drug monitoring.
  20. Microbial biotransformation of Syzygium australe modifies metabolomic profile assessed with multivariate analysis and molecular networking: In vitro and computational studies.
  21. Genome-guided discovery of antibiotic activity in Streptomyces virginiae THA-960 against multidrug resistant Staphylococcus aureus.
  22. Untargeted metabolomics for chemotaxonomy of blow fly pupae (Diptera: Calliphoridae) supported by DNA barcoding.
  23. Mechanisms of Shengxian Quyu Decoction In Heart Failure Using an Untargeted Metabolomics and Network Pharmacology Approach In Rats.
  24. Novel Strongly Basic Molecularly Imprinted Solid-Phase Extraction Sorbent for Simultaneous Determination of Catecholamines and Their Metabolites in Urine.
  25. Integrated metabolomics and machine learning identify predictive biomarkers via SHAP analysis for sintilimab-induced rash in lung cancer patients.
  26. Simultaneous Determination and Enantiomeric Separation of Five DL-Thiol Compounds Using UHPLC-HRMS With Chiral Derivatization in Sake: Dynamic Monitoring of Human Urine After Drinking.
  27. Real-time metered dose inhalers (MDIs) characterization by secondary electrospray ionization (SESI)-mass spectrometry: a proof-of-principle study.
  28. Comprehensive profiling of synthetic cannabinoids fragment ions using UPLC-HRMS in conjunction with online databases.
  29. Integrative metabolomics and molecular networking reveal the progressive metabolic continuum of resin formation in Dracaena cochinchinensis wood.
  30. Corona: A Virtual Mass Spectrometer for the Development of Real-Time Mass Spectrometry Software.
  31. Protocol for high-throughput, automated, solid-phase extraction of sphingolipids from human urine.
  32. UPLC-MS/MS profiling, antioxidant and hemorheological effects of D101 resin-based purified flavonoids from Allium polyrhizum.
  33. Comparative Phytochemical Analysis of Five South African Bulbine Species Using Chromatography-Based Methods.
  34. UPLC-ESI-QToF-MS/MS profiling and anxiolytic-like effects of Simira paraensis (Baill.) Steyerm bark.
  35. Integrating Aptasensor With Mass-Tag Signal Amplification Strategy for the Detection of Three Steroid Hormones in Serum.
  36. Forensic Tracing of Unknown Compounds at the Human-Environment Interface: An All-in-One LC-HRMS Strategy Guided by a Curated Methodological Database.
  37. Seasonal Phytochemical Profiling and Chemometric Assessment of Crataegus songarica K. Koch: Implications for Bioactivity and Resource Utilization.
  38. Enantioselective analysis of synthetic cathinones and amphetamine-type stimulants in biological fluids via LC-MS/MS.
  39. Bio-guided identification of novel antiparasitic natural products from Gardenia imperialis K. Schum (Rubiaceae).
  40. Bioanalysis of Ametryn by UHPLC-MS3 Coupled With Multiple Fragmentation to Decrease Interference and Enhance Sensitivity.
  41. An integrated multi-technique strategy for in vitro metabolite identification and profiling of gymnodimine A using LC-HRMS/MS.
  42. Rapid spatial distribution analysis of lipids in wheat grain using DESI/PI mass spectrometry imaging.
  43. Bioactive potential of Rhododendron lowndesii: Extraction, antioxidant analysis, antimicrobial assay, and Liquid Chromatography-mass Spectrometry profiling.
  44. Evaluation of FASP for Mass Spectrometry-Based Untargeted Metabolomics Analysis of Urine Samples.
  45. FNA-based lipidomics reveals coordinated lipid metabolic remodeling in pancreatic cancer.
  46. Chemical Composition and Anti-Inflammatory Activity of Selaginella tamariscina.
  47. Comparative Phytochemical Profiling, Antioxidant, and Enzyme Inhibitory Activities of Different Parts of Aspilia africana Extracted by Maceration and Soxhlet Methods.
  48. Cyanobacterial Secondary Metabolites: genotoxicity and carcinogenicity in silico evaluation.
  49. Metabolic Stability and Metabolic Profile of Schisandrol B in Rat and Human Liver Microsomes Employing HPLC-MS/MS Analysis.
  50. Development and Validation of a Stability-Indicating RS HPLC Method for Leuprolide Acetate and Its Related Substances: Characterization of Degradation Products With LC-MS.
  51. An ultra-performance convergence chromatography tandem mass spectrometry strategy for quantification of monodisperse polyethylene glycol polymers coupled with multiple fragmentation to enhance sensitivity.
  1. Sci Rep. 2026 06 13. pii: 18361. [Epub ahead of print]16(1):
    Integrated metabolomics and antioxidant activity assessment of Sphagneticola trilobata (L.) Pruski.
    Manar T Ali, Muhammad A Alsherbiny, Dalia A Al-Mahdy, Ahlam M El Fishawy, Asmaa M Otify.
      Sphagneticola trilobata (L.) Pruski (Asteraceae), formerly known as Wedelia trilobata, is a widely distributed ornamental plant utilized in traditional medicine for various ailments. This study aims to provide the first comprehensive comparative analysis of the metabolomic profiles of Sphagneticola trilobata flowerheads and leaves to elucidate the chemical basis for their differential antioxidant capacities. Ultra-high-performance liquid chromatography-electrospray ionization-quadrupole time-of-flight mass spectrometry was employed in both positive and negative ionization modes. The resulting datasets were subjected to multivariate chemometric analyses, including Principal Component Analysis and Orthogonal Partial Least Squares Analysis. Antioxidant potential was assessed using in vitro DPPH, FRAP, and ABTS assays. A total of 86 metabolites were tentatively annotated, spanning classes of terpenoids, flavonoids, phenolic acids, lipids, and amino acids. Notably, 18 of these compounds were observed in this plant for the first time. Chemometric analysis revealed distinct metabolic variances: flowerheads were enriched with flavonoids and phenolic acids, whereas leaves were characterized by a predominance of sesquiterpenoids and diterpenoids. Biological assays demonstrated that flowerheads possess significantly higher antioxidant activity compared to leaves. Moreover, correlation analysis identified key biomarkers responsible for this activity, including 3',4',7-trihydroxy-flavanone hexoside, luteolin, myricetin hexoside, and 4,5-dicaffeoyl-quinic acid. The study establishes that the superior antioxidant potential of Sphagneticola trilobata flowerheads is driven by their high flavonoid and phenolic acid content, distinguishing them from the terpenoid-rich leaves. These findings validate the utility of metabolomics in identifying bioactive markers and suggest the flowerheads as a promising source of natural antioxidants for pharmaceutical or cosmeceutical applications.
    Keywords:   Sphagneticola trilobata ; Antioxidant; Chemometrics; Flowerheads; Leaves; UPLC-MS/MS
    DOI:  https://doi.org/10.1038/s41598-026-54096-w
  2. Biomed Chromatogr. 2026 Jul;40(7): e70525
    Determination of Coblopasvir in Mouse Plasma by Liquid Chromatography-Tandem Mass Spectrometry and Its Application to Pharmacokinetic Study.
    Zhongjie Shi, Yunhui Chen, Jinfang Jiang, Pan Deng.
      This study aims to develop and validate a highly sensitive and high-throughput liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of coblopasvir in mouse plasma and to apply it to a pharmacokinetic study. Plasma samples (10.0 μL) were deproteinized by protein precipitation using acetonitrile. Chromatographic separation is achieved on a Waters CORTECS C8 column with a total run time of 3.0 min. Mass spectrometric detection employed electrospray positive ionization (ESI+) and multiple reaction monitoring (MRM) modes, with quantitative ion transitions of m/z 783.5 → 609.2 (coblopasvir) and m/z 455.2 → 164.9 (internal standard verapamil). The method was fully validated according to the bioanalytical guidelines and subsequently applied to a pharmacokinetic study in ICR mice following oral administration of coblopasvir at a dose of 10 mg/kg. The method exhibited excellent linearity over the concentration range of 2.00-5000 ng/mL. Intraday and interday accuracy (relative error [RE]) ranges from -9.3% to 9.3%. This study presents the first dedicated LC-MS/MS method for the determination of coblopasvir, offering distinct advantages including an extremely low plasma volume requirement (10 μL), high throughput (3 min per sample), and operational simplicity. The validated method provides a reliable and efficient analytical tool for nonclinical and future clinical studies of coblopasvir.
    Keywords:  LC–MS/MS; bioanalysis; coblopasvir; pharmacokinetics
    DOI:  https://doi.org/10.1002/bmc.70525
  3. Anal Chem. 2026 Jun 15.
    BAGO: A Self-Optimizing Tool for LC-MS Gradient Design in Metabolomics.
    Huaxu Yu, Puja Biswas, Elizabeth Rideout, Yankai Cao, Oliver Fiehn, Tao Huan.
      A self-driving metabolomics laboratory has long been envisioned but remains largely unrealized due to the complexity of analytical method design. As an initial step toward this goal, we developed BAGO, a self-optimizing framework for automated liquid chromatography (LC) gradient design in mass spectrometry-based untargeted metabolomics. BAGO aims to enhance global metabolite detection by improving the separation of all compounds, regardless of whether their identities are known or unknown. It operates through a data-driven Bayesian optimization process that iteratively learns from acquired MS data to propose improved gradients. To support this, we propose a global separation index that quantifies coelution among both annotated and unannotated features, enabling robust and structure-agnostic optimization across diverse sample types. Benchmarking across four metabolomics assays involving diverse sample matrices, column chemistries, and gradient durations, BAGO achieved substantial improvements within only 10 optimization iterations by balancing exploration and exploitation. The optimized gradients led to increased numbers of Gaussian-shaped peaks, higher MS/MS acquisition rates, and more annotated metabolites using both identity and analog search approaches. We further applied BAGO to a sex-differentiated metabolomics study of Drosophila abdominal carcasses, completing the workflow in parallel under both initial and optimized gradients. The optimized method resulted in a 41.9% increase in Gaussian-shaped peaks, a 36.8% increase in MS/MS-acquired peaks, and the identification of 18 additional biologically significant metabolites, including sex-associated compounds such as octopamine and pyroglutamic acid. BAGO (https://github.com/HuanLab/bago) is freely available as an open-source tool and represents a generalizable step toward fully automated, self-optimizing experimental workflows in untargeted metabolomics.
    DOI:  https://doi.org/10.1021/acs.analchem.6c01208
  4. J Pharm Biomed Anal. 2026 Jun 02. pii: S0731-7085(26)00267-0. [Epub ahead of print]280 117599
    An improved method for quantification of cannabidiol, Δ9-tetrahydrocannabinol, and their metabolites in human plasma and urine.
    Ayat Zagzoog, Deborah Michel, Nini Ha, Keren Bachi, Christopher Kudrich, Jasper van Oort, Yasmin L Hurd, Jane Alcorn, Robert B Laprairie.
      Cannabidiol (CBD), the second most prevalent phytocannabinoid in Cannabis sativa, potentially offers numerous therapeutic benefits without psychoactive effects. Research into CBD's therapeutic applications must be supported by an understanding of the time course of CBD concentrations in the body and how these concentrations relate to CBD's safety and efficacy. This study aimed to develop a simplified protocol for quantifying CBD, Δ9-Tetrahydrocannabinol (THC) and its metabolites in human plasma and urine, eliminating complex and time-consuming sample preparation, especially for urine samples, while maintaining accuracy comparable to traditional methods. This protocol was used to assess the influence of an oral capsule of CBD (BSPG CBD BRAINS Bioceutical laboratories) on the pharmacokinetics of CBD and its metabolites in plasma and urine relative to an FDA approved CBD formulation (Epidiolex®). The method development focused on optimizing mass spectrometry (MS) conditions with an ultra-high performance liquid chromatography (UHPLC) C18 column. Plasma and urine were collected at fourteen and eight time points, respectively, across 24 h and analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) to quantify CBD and its metabolites. Additionally, (THC) and its metabolites were examined, with no detectable levels found in either plasma or urine. This newly developed analytical method successfully quantified CBD and its metabolites in both plasma and urine, contributing valuable data to the understanding of CBD pharmacokinetics in different oral products. This study's findings have important implications for optimizing CBD administration and assessing therapeutic outcomes.
    Keywords:  Cannabidiol (CBD); LC-MS/MS; Metabolites; Plasma; Quantification; Urine; Δ(9)-tetrahydrocannabinol (THC)
    DOI:  https://doi.org/10.1016/j.jpba.2026.117599
  5. J Pharm Biomed Anal. 2026 Jun 13. pii: S0731-7085(26)00280-3. [Epub ahead of print]280 117612
    Development of new liquid chromatography-tandem mass spectrometry methods for the quantification of seven metabolites in three related breast cancer cell lines.
    Marziyeh Ghanbarian, Deborah Michel, Maryam Alyari, Amir Khajavinia, Oleg Y Dmitriev, Anas El-Aneed.
      Metabolites of the tricarboxylic acid (TCA) cycle play crucial roles in cancer biology, and their accurate quantification is essential for understanding energy metabolism, signaling dynamics, and identifying metabolic vulnerabilities in cancer cells. However, traditional liquid chromatograph-tandem mass spectrometry (LC-MS/MS) methods for these polar metabolites often encounter challenges, such as limited retention on reversed-phase columns and ion suppression. This study developed and validated two LC-MS/MS methods for the accurate quantification of seven key TCA cycle metabolites in MDA-MB-231, M67-2 (MEMO1 knockdown), and M67-9 (MEMO1 knockout) breast cancer cell lines. For five metabolites, namely citrate (CA), L-malate (MA), fumarate (FA), α-ketoglutarate (AKG), and glutamate (GA), an isotope-coded derivatization approach utilizing 12C/13C-labeled dimethylaminophenacyl (DmPA) bromide was employed to develop a targeted high-performance liquid chromatography (HPLC)-MS/MS method. Inefficient DmPA derivatization in aqueous matrices was addressed by optimizing sample preparation in non-aqueous conditions, and the presence of multiple peaks of AKG was resolved by selecting triethanolamine (TEOA) as the reaction base to improve specificity. Conversely, due to persistent interferences with DmPA derivatization, pyruvic acid (PA) and succinic acid (SA) were quantified using another novel hydrophilic interaction liquid chromatography (HILIC)-MS/MS method in their native underivatized forms. Both methods were validated according to regulatory bodies, ensuring linearity, accuracy, precision, selectivity, and stability. The methods ensured the utilization of two multiple reaction monitoring (MRM) transitions to enhance specificity. The validation approach was adjusted to fit tissue culture studies. The validated methods were successfully used to measure the TCA metabolites in tested cell lines, providing valuable tools for investigating metabolic dynamics in cancer research.
    Keywords:  Cancer cells; DmPA derivatization; HILIC-MS/MS; HPLC-MS/MS; LC-MS/MS; TCA cycle metabolites; Targeted metabolomics
    DOI:  https://doi.org/10.1016/j.jpba.2026.117612
  6. J Chromatogr A. 2026 Jun 15. pii: S0021-9673(26)00514-5. [Epub ahead of print]1783 467185
    Fragmentation property identity card-assisted reliable structural annotation of calycosin metabolites in vivo: A strategy for isomer discrimination using mass spectrometry.
    Rui Wang, Yang Liu, Wenhan Song, Chenxi Yu, Jing Lei, Jiading Weng, Lin Lin, Yunan Zhang, Yaolei Ma, Wei Zhou, Xin Shen, Yanxu Chang, Yue Gao.
      Drug metabolites play a pivotal role in the assessment of pharmacological efficacy and safety. The reliable structural annotation of drug metabolites is challenging due to their inherent complexity. Calycosin (Cal) is a bioactive compound derived from Astragali Radix, exhibiting diverse pharmacological activities, including anti-inflammatory, antioxidant and cardioprotective properties; however, its metabolic profile has not yet been comprehensively elucidated. Ultra-high pressure liquid chromatography quadrupole time of flight tandem mass spectrometry (UHPLC-Q-TOF-MS) and ultra-high pressure liquid chromatography triple quadrupole mass spectrometry (UHPLC-QqQ-MS) were integratively employed for the identification of Cal metabolites. A total of 22 metabolites of Cal were detected in biological matrices, including plasma, urine, and feces. Based on the empirical mass fragmentation rules, the chemical structures of certain metabolites were elucidated. For the metabolites exhibiting isomeric forms with highly similar MS/MS spectra, the fragmentation property identity card (FPIC), a descriptor of compound mass spectrometric fragmentation patterns, was developed and its rapid visualization was implemented using Python. The compound FPIC was constructed by fitting, normalization, and transposition of ion transition response intensities under gradient collision energies, enabling the identification of fragment ions with isomer discriminative characteristics. Subsequently, the integration of FPIC with quantum chemical calculations enabled reliable structural elucidation of the isomers. Glucuronidation, sulfation, methylation, and dehydroxylation constituted the predominant in vivo metabolic pathways of Cal. Collectively, the present study comprehensively delineated the in vivo metabolic profile of Cal and proposed a reliable and broadly applicable strategy for the structural characterization of compounds metabolites.
    Keywords:  Bond dissociation energy; Calycosin; Fragmentation property identity card; Isomer; Metabolite
    DOI:  https://doi.org/10.1016/j.chroma.2026.467185
  7. PLoS One. 2026 ;21(6): e0343544
    Siderophore screening in marine sponge extracts using LC-HRMS and an R-based metabolomics workflow.
    Alejandro García Ríos, Massuo Jorge Kato, Lydia Fumiko Yamaguchi, Breno Pannia Espósito, Ailan Farid Arenas-Soto.
      Siderophores are pivotal ‌‌iron-acquisition biomolecules integral to microbial survival, pathogenicity, and ecology. Elucidating these compounds offers critical insights into the microbial dynamics of marine holobionts and potential therapeutic applications. In this study, we present a culture-independent, data-centric strategy to annotate siderophores from the body mass of three marine sponge species: Dragmacidon reticulatum, Aplysina fulva, and Amphimedon viridis. Utilizing Liquid Chromatography-High Resolution Mass Spectrometry (LC-HRMS) coupled with a custom R-based analytical workflow (XCMS and MetaboAnnotation), we putatively annotated 59 siderophores. We employed a validation pipeline, utilizing iron-adduct calculations [M-2H + Fe]+, [M-H + Fe]2+, [2M-2H + Fe]+, mass accuracy thresholds (<3 ppm), retention time deviation (Coefficient of variation < 2%), and chromatograph peak analysis. According to the Metabolomics Standards Initiative (MSI), these annotations correspond to Level 2 (putatively annotated compounds) because they are based on accurate mass matching without chemical standard confirmation. Notably, iron supplementation during extraction did not significantly alter siderophore detection, suggesting constitutive production or environmental saturation. This workflow bypasses the limitations of traditional cultivation, revealing a diverse landscape of iron-chelating metabolites, including Ferricrocin, Aeruginic acid, and Madurastatin directly within the sponge's body.
    DOI:  https://doi.org/10.1371/journal.pone.0343544
  8. J Am Soc Mass Spectrom. 2026 Jun 15.
    Benchmarking MS/MS Featurization Strategies for Machine Learning-Driven Metabolite Structure Annotation.
    Roger Giné, Ivan Pérez-López, Josep M Badia, Jordi Capellades, Oscar Yanes.
      Reference MS/MS libraries remain incomplete due to the vast chemical diversity of metabolites, leaving many spectra from untargeted metabolomics experiments unannotated─the "dark matter" of metabolomics. Machine learning can extend metabolite annotation beyond direct library matches, but its success depends critically on how MS/MS spectra are converted into numerical representations that capture chemically meaningful features while reducing sparsity. Although numerous spectral representations exist, they have not been systematically compared. Using over 71,000 unique compounds with merged-energy MS/MS spectra, we benchmarked a broad set of spectral featurization methods, including fixed and adaptive binning, global-quantile variable-width bins, frequent-peaks representations, spectrum hashing, and learned embeddings such as Spec2Vec, MS2DeepScore, DreaMS, and SpecEmbedding. We further evaluated how vector dimensionality affects performance. A total of 105 neural network models were trained under 5-fold cross-validation to predict Mol2Vec molecular embeddings and retrieve correct structures from a 0.6-million-compound database. Retrieval was assessed at 0.1, 3, and 10 ppm mass tolerances, and a null ranking model was generated to determine expected Top-N accuracy under random candidate ordering. Adaptive binning, frequent-peaks, and DreaMS produced the most accurate embedding predictions. On the test data set, Top-1 retrieval reached 46%, 44%, and 38% for 0.1, 3, and 10 ppm, respectively, with Top-5 accuracies up to 77%. In the CASMI2022 data set, Top-1 performance remained similar at 0.1 ppm but dropped markedly at wider tolerances, reaching only 26% at 3 ppm and 23% at 10 ppm. To ensure reproducibility and broad community applicability, results were further validated on two fully open benchmark data sets, MassSpecGym and Spectraverse, with findings consistent across all three resources. These results underscore clear performance differences among featurization strategies, the strong dependence of retrieval accuracy on mass precision, and the need for evaluation metrics aligned with structure-level annotation tasks.
    Keywords:  machine learning; metabolite annotation; spectral featurization; structure retrieval; tandem mass spectrometry; untargeted metabolomics
    DOI:  https://doi.org/10.1021/jasms.5c00428
  9. Talanta. 2026 Jun 13. pii: S0039-9140(26)00809-X. [Epub ahead of print]310 130153
    Online solid-phase extraction coupled to liquid chromatography-tandem mass spectrometry for fungicide translocation studies in soybean plants.
    Eliézer de Oliveira, Carla Beatriz Grespan Bottoli.
      This study describes the development and validation of an online solid-phase extraction coupled to liquid chromatography-tandem mass spectrometry (online SPE-LC-MS/MS) method for evaluating the translocation of fluxapyroxad, bixafen, pyraclostrobin, trifloxystrobin and prothioconazole (determined as prothioconazole-desthio) in soybean leaves following foliar application, either alone or combined with a copper-based fertilizer. The method was optimized and validated after comparison with the conventional QuEChERS approach, highlighting important methodological advancements such as reduced sample handling, lower analysis time, and partial integration of sample preparation with instrumental analysis. Validation results demonstrated significant matrix effects for all analytes, with ionization suppression ranging from -60 to -27%; however procedural standard calibration efficiently compensated for these effects, resulting in satisfactory trueness and precision. Limits of detection and quantification were ≤1 μg L-1 and ≤5 μg L-1, respectively, while calibration curves showed good linearity (r > 0.997). Mean recoveries ranged from 90 to 120%, with intra- and inter-day precision below 19 and 14%, respectively, in accordance with the SANTE guidelines. The method was successfully applied to greenhouse translocation experiments, revealing enhanced fungicide redistribution in treatments combined with copper-based fertilizer. The results highlight the analytical and operational advantages of automation in plant-tissue pesticide analysis.
    Keywords:  LC-MS/MS analysis; Online SPE approach; Pesticide translocation; Soybean matrices
    DOI:  https://doi.org/10.1016/j.talanta.2026.130153
  10. Talanta. 2026 Jun 11. pii: S0039-9140(26)00777-0. [Epub ahead of print]310 130121
    MS2-SMILES AlignNet: A cross-modal contrastive learning framework for direct alignment of tandem mass spectra and molecular structures.
    Xuemeng Geng, Ruxi Gao, Chang Wang, Xinning Li, Jixuan Song, Haixue Kuang, Liu Yang, Hai Jiang.
      Structural annotation of metabolites via tandem mass spectrometry (MS2) remains a long-standing core challenge in analytical chemistry. To address this issue, we introduce MS2-SMILES AlignNet (MSAN), a cross-modal contrastive learning framework tailored for direct alignment between MS2 spectra and molecular structures. Its key innovations lie in a dual-branch Transformer-based MS2 encoder and a hybrid loss function, which jointly enable effective cross-modal alignment while preserving molecular structural similarity. Trained on more than 1.6 million high-quality spectrum-structure pairs, MSAN achieves state-of-the-art performance on the unified test subset of the CASMI 2022 benchmark, attaining a Recall@1 of 54.23% in positive ion mode and 45.37% in negative ion mode. Notably, it outperforms CSU-MS2 by a substantial 14.81 percentage points in the more challenging negative ion mode. Furthermore, its generalization capacity and structural isomer discrimination ability are validated on the CASMI 2016 dataset. Meanwhile, MSAN exhibits favorable robustness across diverse experimental conditions. Collectively, this work provides a precise and robust tool for high-throughput metabolite annotation in untargeted metabolomics research.
    Keywords:  Contrastive learning; Deep learning; Metabolite identification; Metabolomics; Molecular fingerprinting; Tandem mass spectrometry
    DOI:  https://doi.org/10.1016/j.talanta.2026.130121
  11. Forensic Toxicol. 2026 Jun 19.
    An Ultra-High performance liquid chromatography-tandem mass spectrometry method for simultaneous determination of etomidate, analogues, and metabolites in human nail: Unveiling a unique accumulation profile.
    Pinting Zhang, Kaiyu Wang, Xinyi Yan, Dan Zhang, Jiale Bai, Zixia Shao, Yihan Liu, Zhenjiang Liao, Yunde Tang, Hao Zeng, Na Jing, Qiuping Huang, Hongxian Shen, Bin Di, Fang Yan, Tao Lu.
       PURPOSE: In response to the increasing abuse of etomidate-related substances in China, this study developed and validated an ultra-high performance liquid chromatography-tandem mass spectrometry method for the simultaneous quantification of etomidate (ET), metomidate (MET), propoxate (PRO), isopropoxate (I-PRO), CF3-etomidate (CF3-ET), and their metabolite etomidate acid (ETA) in human nails. This method enabled the first investigation into their accumulation patterns in authentic samples, offering scientific support for forensic monitoring.
    METHODS: A 20 mg portion of nail was cryogenically ground and extracted with methanol containing the internal standard. After vortex mixing, ultrasonication, and filtration, the extract was concentrated under nitrogen flow and analyzed by ultra-high performance liquid chromatography-tandem mass spectrometry.
    RESULTS: The limits of quantification (LOQs) for the target substances in nail samples ranged from 1 to 10 pg/mg. The calibration curves exhibited good linearity over the concentration range from the LOQ to 1000 pg/mg. All other evaluated validation parameters proved satisfactory. The validated method was applied to authentic nail samples to investigate the accumulation profiles of ET and its analogs. Analysis of the samples revealed that 52 tested positive. These positive cases showed a distinct accumulation pattern dominated by the parent drugs, with ETA as the sole metabolite detected.
    CONCLUSION: This study developed and validated a reliable method for the simultaneous quantification of ET, its analogues, and its metabolite in human nails. Notably, it is the first to be applied to authentic samples to investigate their accumulation profiles, supporting the use of nail analysis in forensic toxicology and drug abuse monitoring.
    Keywords:  Drug accumulation; Etomidate; Etomidate acid; Etomidate analogues; Nail; Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry
    DOI:  https://doi.org/10.1007/s11419-026-00776-1
  12. Anal Methods. 2026 Jun 11.
    Rapid and targeted HILIC-MS/MS quantification of urinary metabolites reveals metabolic alterations in COVID-19 patients.
    Ondrej Hodek, Anna Edman, Christoffer Granvik, Alicia Lind, Anna K Överby, Mareike Gutensohn, Annika I Johansson.
      Urinary metabolites and their concentrations serve as biomarkers for identification of metabolic pathways that relate to specific diseases; therefore, fast and accurate quantification of the metabolites in urine is essential in health assessment and diagnosis. As many urinary metabolites are of polar nature, hydrophilic interaction liquid chromatography (HILIC) has been used over the last several years because it offers faster and more reproducible analyses compared to traditional techniques such as reversed-phase chromatography or capillary electrophoresis. In our study, we developed a HILIC method by using a 3 cm analytical column in connection with tandem mass spectrometry detection for quantification of 10 urinary metabolites including creatinine as the reference for normalization. As all tested metabolites contain ionizable functional groups, pH of the mobile phase was optimized to achieve baseline separation of 2 isomeric pairs (1-methyl-4-imidazoleacetic acid/1-methyl-5-imidazoleacetic acid and 1-methylhistidine/3-methylhistidine) and to obtain overall better separation efficiency resulting in a 7 min analysis. The developed method was validated in terms of sensitivity, carry-over, linearity, matrix effects, accuracy, and precision. The metabolite concentrations in healthy subjects determined by the developed method correspond well with the normal reference values found in the literature. Moreover, the method was tested on a small cohort of COVID-19 patients, where it enabled identification of differences in metabolite levels. Thus, the developed method has potential to be used routinely in a diagnostic field for high-throughput analysis of urine samples.
    DOI:  https://doi.org/10.1039/d6ay00400h
  13. Food Chem (Oxf). 2026 Dec;13 100429
    Comparative transcriptomic and metabolomic profiling of forest tea and terrace tea in Sandu, Guizhou, China.
    Dani Wei, Huan Wu, Mengxue Yang, Chaya Sarathchandra, Yanfei Geng.
      Ancient forest tea is locally valued in Guizhou, but the transcriptomic and metabolomic differences associated with forest and terrace tea cultivation systems remain unclear. We conducted an exploratory integrated transcriptomic and untargeted metabolomic comparison across three shoot types (single buds, one-bud-one-leaf, and one-bud-two-leaves). Transcriptomic analysis showed the largest number of DEGs at the one-bud-two-leaves stage, suggesting substantial transcriptional variation under the present sampling conditions. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses indicated that phenylpropanoid and flavonoid biosynthesis pathways were over-represented. Untargeted liquid chromatography-mass spectrometry (LC-MS) detected 864 differential metabolite features with Metabolomics Standards Initiative (MSI) Level 2 putative annotations, including 374 features with higher relative abundance in forest tea. The largest number of differential metabolite features occurred at the one-bud-one-leaf stage, and these features were enriched in flavonoid biosynthesis. Variable importance in projection (VIP) ranking prioritized discriminant features from false discovery rate (FDR)-filtered data, including four MSI Level 2 features putatively annotated as polyphenol-related compounds that showed higher relative abundance in forest tea. In addition, the expression of genes encoding phenylalanine ammonia-lyase (PAL), a key enzyme in the phenylpropanoid biosynthesis pathway, differed significantly between forest tea and terrace tea. These results suggest that secondary-metabolism-related pathways are associated with molecular differentiation between forest and terrace tea. Because this study lacked sensory evaluation, targeted metabolite quantification, and functional validation, these findings require future validation. This work provides candidate pathways and metabolite features for future validation of tea quality differences.
    Keywords:  Camellia sinensis; Flavonoid biosynthesis; Metabolite profiling; Secondary metabolism; Tea quality
    DOI:  https://doi.org/10.1016/j.fochms.2026.100429
  14. Sci Rep. 2026 Jun 16.
    A multi-dimensional approach combining HPLC, ion mobility and high resolution mass spectrometry for investigating transition metal complexes.
    Laura Zellner, Julian Süß, Samuel Vorbach, Uwe Monkowius, Markus Himmelsbach, Stephan Hann, Christian W Klampfl.
      Organometallic complexes play a central role in catalysis and medicinal chemistry, yet their analytical characterization via mass spectrometry remains challenging due to their reactivity, susceptibility to fragmentation and the resulting overlapping isotopologue patterns, especially in complex reaction mixtures. Herein, we present a high-performance liquid chromatography ion mobility mass spectrometry workflow for the separation and analysis of group-six metal complexes. Chromatographic separation was combined with variation of electrospray ionization source conditions, allowing fragmentation behavior to be tuned according to specific analytical requirements. Ion mobility spectrometry provided an additional separation dimension that enabled the determination of collision cross sections as molecular descriptors for all investigated complexes. Analysis of the ion mobility data revealed systematic, metal-dependent trends consistent with expected structural differences. These correlations facilitated the assignment of the central metal even for species of low abundance or those lacking characteristic isotopologue patterns. Overall, this work demonstrates that the combination of chromatographic separation and ion mobility-based trend analysis provides a versatile platform for the characterization of organometallic species.
    Keywords:  Collision cross sections; Drift-tube ion-mobility mass spectrometry; HPLC; Organometallics; Transition metal complexes
    DOI:  https://doi.org/10.1038/s41598-026-57597-w
  15. Rapid Commun Mass Spectrom. 2026 Sep 15. 40(17): e70108
    Lipidome and Targeted Metabolome Analysis of Locational Picrorhiza Kurroa Royle. Ex Benth., Using UPLC-PDA and UHPLC-QTOF-IMS.
    Bindu Rawat, Robin Joshi, Dinesh Kumar.
       INTRODUCTION: Lipids are a fundamental class of biomolecules essential for membrane structure, energy storage, and cellular signaling, whereas lipidomics is an advanced dedicated way to understand the lipids, including biological lipids.
    OBJECTIVE: Keeping in mind the importance of lipid makeup, the current study was focused to understand the lipid diversity present within Picrorhiza kurroa (leaves, rhizomes, and roots) at three different localities of Himachal Pradesh, India: Bhatori (Pangi), Bharmaur (Chamba), and Gumna (Shimla).
    METHOD: UHPLC-QTOF-IMS was used for lipid profiling of P. kurroa locational samples. The data were analyzed for each extraction parameter. Moreover, targeted specialized metabolites were also analyzed using UPLC-PDA. Further, multivariate analysis including supervised orthogonal projections to latent structures discriminant analysis and unsupervised principal component analysis were employed to understand the lipid makeup, its dissemination, and similarity traits among the samples.
    RESULTS: Q-TOF/MS revealed the comprehensive lipidome of P. kurroa that includes the important presence of Cer, FA, (L) PE, TG, PA, PI, PS, MGDG, and DGDG. Leaves of P. kurroa collected from the Pangi region showed higher lipid content than other targeted parts and localities, whereas targeted metabolites including picrosides were present higher in the rhizomes of the Pangi region. Further, statistical analysis showed clear dissemination and similarity traits among the samples. The current study suggested variability among the chemo-profile of different locational samples.
    CONCLUSION: We concluded that leaves of P. kurroa collected from the Pangi region constitute higher lipid content than other parts and P. kurroa plants collected from another area. Also, the findings revealed that the plant P. kurroa contains Cer, FA, (L)PE, TG, DG, PG, PA, PI, PS, MGDG, and DGDG. The variation in picroside contents was also observed through UPLC-DAD analysis; rhizomes collected from the Bhatori (Pangi) region exhibited the highest picroside content.
    Keywords:   Picrorhiza kurroa ; Lipidomics; Picrosides; ultraperformance liquid chromatography coupled with ESI Q‐TOF MS
    DOI:  https://doi.org/10.1002/rcm.70108
  16. Food Sci Nutr. 2026 Jun;14(6): e72021
    Two Flavones as Markers for the Distinction of Natural Acacia Honey Based on the UHPLC-QE-MS Method and Research on Their Potential Immunomodulatory Activity.
    Bingkang Wang, Min Hua, Yuecheng Liu, Meng Li, Ning Zhang, Yi Wu, Qi Gao, Hongfu Sun, Yanpeng Dai, Meng Zhou, Qian Zhou.
      Honey is a natural sweet substance produced by bees from nectar and plant secretions, which is abundant in nutrients and exhibits excellent biological activities. However, owing to deficiencies in the quality control index system, prominent issues such as honey adulteration and improper processing are prevalent in the current market, resulting in a severe decline in honey quality. Therefore, this study focuses on acacia honey, one of the four major honey varieties in China, by screening its characteristic components and analyzing the correlation between these components and immunomodulatory activity, aiming to provide targeted theoretical basis and indicator support for resolving the aforementioned practical issues. The results demonstrated that natural acacia honey (NAH) had significant advantages over commercial acacia honey (CAH) in key quality indices (total flavonoids, total phenols, proline content, amylase activity, etc.) and quality consistency, with significantly higher immunomodulatory activity. Combined with machine learning and chemometric analyses, hesperetin and pinocembrin were confirmed as the characteristic components of NAH from the 50 components identified via ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap mass spectrometry (UPLC-QE-MS) analysis. In vitro experiments demonstrated that both components at 50 μM effectively promoted mouse splenic lymphocyte proliferation, reduced NO and TNF-α secretion in LPS-induced RAW264.7 macrophages, and showed no obvious cytotoxicity, indicating their potential as characteristic markers for evaluating NAH quality and functional activity. Further network pharmacology and molecular docking studies indicated that hesperetin and pinocembrin could mediate immunomodulatory effects by regulating 10 core targets including MMP9, MMP2, BCL2, and SRC, as well as the PI3K-AKT pathway, with the markers showing the strongest binding activity toward MMP2. This study established a characteristic marker identification system for NAH, and provided scientific indicator support for formulating acacia honey quality standards and improvement of processing technologies.
    Keywords:  SPE‐UPLC‐Q‐Exactive Orbitrap MS; acacia honey; immune regulation; quality evaluation; quality markers
    DOI:  https://doi.org/10.1002/fsn3.72021
  17. Metabolomics. 2026 Jun 16. pii: 95. [Epub ahead of print]22(4):
    Metabolomic characterization and temporal dynamics of Burkholderia cepacia for polyhydroxyalkanoates production from oleic acid.
    Alfonso E Alarcón, Nubia C Moreno, Daniel Pardo-Rodriguez, Mónica P Cala, Carlos A M Riascos.
       BACKGROUND: Polyhydroxyalkanoates (PHA) have become biodegradable alternatives for replacing chemical polymers. However, high production costs remain the primary obstacle to the commercial application of this bioprocess, prompting the use of metabolic engineering approaches to optimize PHA quality and increase productivity.
    OBJECTIVES: This study aimed to identify the endometabolome at different stages during a dynamic fermentation of Burkholderia cepaciausing oleic acid as the carbon source.
    METHODOLOGY: Untargeted metabolomic analysis was conducted using gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) techniques. Univariate (UVA) and multivariate (MVA) statistical analyses were performed to determine significant differences between metabolomic profiles.
    RESULTS: A total of 24 significant metabolites were identified through GC-MS analysis and 223 through LC-MS, highlighting organic compounds and lipids associated with pathways such as ß-oxidation, the tricarboxylic acid cycle, and the pentose phosphate pathway. Acetyl-CoA emerged as a critical common intermediate in both central biosynthetic and PHA-producing pathways.
    CONCLUSION: Acetyl-CoA plays a fundamental role in the complex regulatory system activated by nutrient fluctuations in the culture medium, suggesting that the microorganism undergoes metabolic reorganization to optimize carbon source utilization for cellular maintenance.
    Keywords:   Burkholderia cepacia ; Dynamic analysis; Polyhydroxyalkanoates; Untargeted metabolomics analysis
    DOI:  https://doi.org/10.1007/s11306-026-02473-x
  18. Metabolomics. 2026 Jun 16. pii: 96. [Epub ahead of print]22(4):
    Urinary organic acid levels and their associations with clinical characteristics in patients with schizophrenia.
    Yavuz Yılmaz, Halef Okan Doğan, Ayşegül Murat, Gökmen Zararsız.
       INTRODUCTION: Schizophrenia is a chronic psychiatric disorder characterized by substantial biological and clinical heterogeneity. Beyond classical neurotransmitter-based models, increasing evidence suggests that systemic metabolic alterations may contribute to its pathophysiology.
    OBJECTIVES: This study aimed to characterize urinary organic acid profiles in patients with schizophrenia and investigate their associations with clinical characteristics and pathway-level metabolic alterations.
    METHODS: In this cross-sectional study, urinary organic acids were quantified using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in 55 patients with schizophrenia and 30 age- and sex-matched healthy controls. Organic acid concentrations were normalized to urinary creatinine levels. Clinical severity was evaluated using the Positive and Negative Syndrome Scale and the Clinical Global Impressions-Severity scale. Differential metabolite analysis, subgroup comparisons, principal component analysis, correlation analyses, and pathway enrichment analyses were performed.
    RESULTS: Patients with schizophrenia demonstrated widespread alterations in urinary organic acid profiles compared with healthy controls, with 40 metabolites remaining significantly different after false discovery rate correction. Subgroup analyses identified additional metabolomic variation according to symptom severity, treatment adherence, family history, and current treatment status. Principal component analysis demonstrated partial separation between patients and controls, whereas subgroup distributions showed substantial overlap. Correlation analyses revealed predominantly weak-to-moderate associations between clinical variables and urinary metabolite concentrations. Pathway enrichment analysis identified propanoate metabolism as the only pathway that remained statistically significant after multiple testing correction, while several additional pathways demonstrated nominal enrichment.
    CONCLUSION: These findings suggest that schizophrenia is associated with broad alterations in urinary metabolomic profiles and support the possibility that intermediary metabolic pathways may contribute to the biological complexity and heterogeneity of the disorder. Further longitudinal and validation studies are needed to clarify the biological and clinical relevance of these observations.
    Keywords:  Intermediary metabolism; Metabolomics; Propanoate metabolism; Schizophrenia; Urinary organic acids
    DOI:  https://doi.org/10.1007/s11306-026-02479-5
  19. Daru. 2026 Jun 15. pii: 36. [Epub ahead of print]34(2):
    A cost-efficient internal standard approach for busulfan quantification: MS-based investigation of adduct formation in therapeutic drug monitoring.
    Erfan Jahan Mehmani, Niloofar Rastegarpanah, Issa Zarei, Massoud Amanlou.
       BACKGROUND: Busulfan plays a central role in conditioning regimens for hematopoietic stem cell transplantation (HSCT). However, its narrow therapeutic index and substantial interpatient pharmacokinetic variability necessitate precise therapeutic drug monitoring (TDM). Because busulfan is highly lipophilic, many LC-MS/MS assays rely on costly stable-isotope-labeled internal standards (e.g., busulfan-d8), which limits their routine application in resource-constrained settings.
    METHODS: This study aimed to synthesize cost-effective structural analogs of busulfan and evaluate their chromatographic behavior and cation-adduct formation as potential alternative internal standards. Four dimethanesulfonate analogs were synthesized from symmetrical n-diols (n = 3-6) under alkaline conditions (47-55% yields). The synthesized analogs were evaluated as candidate internal standards by investigating their cation-adduct formation behavior using liquid chromatography-electrospray ionization-quadrupole time-of-flight high-resolution mass spectrometry (LC-ESI-qTOF-HRMS).
    RESULTS: Each analog formed distinct cation adducts (H+, Li+, Na+, K+, and NH4+), with sodium adducts demonstrating the greatest stability in mixed-ion environments. Among the synthesized compounds, 1,6-hexanediol dimethanesulfonate (compound 4) exhibited the most favorable chromatographic retention and adduct stability, supporting its suitability as a structural internal-standard candidate.
    CONCLUSION: These findings provide mechanistic and analytical evidence to support the selection of a cost-efficient internal standard for busulfan quantification. However, full bioanalytical validation in biological matrices will be required before routine clinical implementation.
    Keywords:  Busulfan; Hematopoietic stem cell transplantation; Internal standard; LC-MS; Mass spectrometry; Therapeutic drug monitoring
    DOI:  https://doi.org/10.1007/s40199-026-00615-z
  20. Sci Rep. 2026 06 14. pii: 18372. [Epub ahead of print]16(1):
    Microbial biotransformation of Syzygium australe modifies metabolomic profile assessed with multivariate analysis and molecular networking: In vitro and computational studies.
    Heba H Elzayat, Noha A Seif-Eldein, Lina J M Abdel-Hafez, Hussein N Ghanem, Alaadin E El-Haddad, Shaza A Mohamed.
      Syzygium australe, a comparatively less studied species within the Syzygium genus, is emerging as a prospective source of bioactive phytochemicals. In this study, the impact of microbial biotransformation by Aspergillus niger on the metabolomic and bioactivity profiles of S. australe leaves extract (SAE) was evaluated. UPLC-T-TOF-MS/MS and molecular networking enabled the tentative identification of 80 metabolites in SAE, with flavonoids emerging as the dominant phytoconstituents. After biotransformation, sulfated flavonoids are the main metabolites in S. australe biotransformed extract (SABE), suggesting that enzymatic sulfonation is mediated by fungal sulfotransferase enzymes. Molecular networking revealed two key clusters: cluster A, which is primarily composed of quercetin derivatives, and cluster B, which corresponded to syringetin. Notably, the biotransformed metabolites in SABE were predominantly observed as self-looped nodes, indicating the formation of structurally unique compounds. Multivariate chemometric analyses revealed a significant metabolomic modulation and a clear discrimination between SAE and SABE. Compared with SABE, SAE significantly increased the free radical scavenging capacity, as evidenced by lower IC₅₀ values in DPPH and ABTS assays (36.96 ± 1.20 and 19.80 ± 0.85 µg/mL respectively), which is likely a consequence of tannin degradation during microbial biotransformation. The bioactivity of SABE, particularly against pancreatic lipase, was enhanced, with an inhibition rate of 74.49 ± 4.80% at 100 µg/mL. Molecular docking further supported these findings, highlighting isorhamnetin-3-O-sulfate as a key bioactive constituent with the highest binding affinity to pancreatic lipase (ΔG = - 12.47 kcal/mol). These findings highlight a significant potential and warrant further investigation using alternative microbial strains aiming to develop novel therapeutic agents.
    Keywords:  Antioxidant; Docking; GNPS; LC-MS/MS; Pancreatic lipase; Sulfated flavonoids; α-amylase; α-glucosidase
    DOI:  https://doi.org/10.1038/s41598-026-55423-x
  21. iScience. 2026 Jun 19. 29(6): 116311
    Genome-guided discovery of antibiotic activity in Streptomyces virginiae THA-960 against multidrug resistant Staphylococcus aureus.
    Trang Thi Minh Nguyen, Jeyong Jung, Xiangji Jin, Qiwen Zheng, Jaeyoung Choi, Tae-Hoo Yi.
      The emergence of multidrug-resistant Staphylococcus aureus (MDR-S. aureus) demands innovative strategies to identify robust microbial producers of potent antibiotics. This study characterizes Streptomyces virginiae THA-960, a soil-derived actinomycete, as a producer of the known anti-MDR-S. aureus antibiotic. Phenotypic screening showed THA-960's efficacy against clinical MDR-S. aureus isolates, with MICs as low as 0.08 mg/mL, outperforming conventional antibiotic-producing Streptomyces. Time-kill assays and SEM confirmed rapid bactericidal action via cell wall disruption. Complete genome sequencing revealed a rich biosynthetic potential, housing 31 specialized metabolite gene clusters. Phylogenomic analysis of 521 Streptomyces genomes delineated S. virginiae into distinct groups and showed that the amycomicin biosynthetic gene cluster is conserved within a specific taxonomic group, providing a genomic roadmap for targeted strain selection. Crucially, Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Qtof-MS) analysis detected a metabolite feature putatively assigned as amycomicin in THA-960. This integrated multi-omics study provides a phylogenomic framework for the targeted exploitation of Streptomyces strains against antimicrobial resistance.
    Keywords:  health sciences; medical microbiology; microbial genomics
    DOI:  https://doi.org/10.1016/j.isci.2026.116311
  22. J Med Entomol. 2026 May 05. pii: tjag075. [Epub ahead of print]63(3):
    Untargeted metabolomics for chemotaxonomy of blow fly pupae (Diptera: Calliphoridae) supported by DNA barcoding.
    Alexa Figueroa, Stephen M Baca, Jennifer Y Rosati, Rosalia Hernandez, Rabi A Musah.
      In forensics, accurate species identification of necrophagous insects retrieved from decomposing remains is critical for estimating the postmortem interval (PMI). Blow flies (Diptera: Calliphoridae) are typically the earliest colonizers to arrive on carrion. Because juvenile life stages are morphologically similar across species, forensic entomologists traditionally rear the eggs, larvae, and pupae to adulthood to ensure a definitive identification based on features of the adults, in a process that is time-consuming and relies heavily on specimen viability. This study presents a rapid, high-throughput identification method for forensically significant blow fly pupae utilizing direct analysis in real time-high-resolution mass spectrometry (DART-HRMS) coupled with chemometrics. A prediction model was developed for 11 species of necrophagous blow fly pupae across seven genera of Calliphoridae: Calliphora, Chrysomya, Cochliomyia, Cynomya, Lucilia, Phormia, and Protophormia. It was built using 220 species-validated test samples, and tested with 115 external validation samples. Leave-one-out cross validation accuracies of 97.27% and 98.26%, respectively, were observed for the test and validation datasets. Species identities were independently verified via DNA barcoding with cytochrome c oxidase I. These results demonstrate that DART-HRMS can successfully identify pupae-a historically underutilized developmental stage in forensic casework-based upon their unique metabolome signatures. This work provides critical forensic resources for the utilization of pupae that may facilitate PMI estimations, even for non-viable specimens.
    Keywords:  Calliphoridae pupa species identification; DART-HRMS; DNA barcoding; forensic entomology; metabolome profiling
    DOI:  https://doi.org/10.1093/jme/tjag075
  23. J Vis Exp. 2026 May 29.
    Mechanisms of Shengxian Quyu Decoction In Heart Failure Using an Untargeted Metabolomics and Network Pharmacology Approach In Rats.
    Dongxia Li, Xinyi Zhou, Shiyi Tao, HaoZhe Xiong, Xuefei Gao, Xiang Xiao, Li Huang, Mingjing Shao, Deshuang Yang.
      Shengxian Quyu decoction (SXQY) has been suggested as a potential therapeutic strategy for heart failure (HF), but its therapeutic mechanisms remain unclear. This study investigated the therapeutic effects and underlying mechanisms of SXQY in HF. A rat model of HF was induced by transverse aortic constriction (TAC) and treated with SXQY. Cardiac function was assessed by transthoracic echocardiography, and myocardial structure and fibrosis were evaluated using hematoxylin and eosin and Masson's trichrome staining. Serum samples were analyzed by untargeted metabolomics using ultra-performance liquid chromatography-tandem mass spectrometry. Blood-entering components were mapped to targets, and intersecting HF-related targets were analyzed using protein-protein interaction (PPI) network analysis, followed by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. SXQY significantly improved cardiac function in TAC rats, as evidenced by decreased left ventricular internal diameter in diastole and left ventricular posterior wall thickness in diastole and increased left ventricular ejection fraction, and alleviated myocardial hypertrophy, inflammation, and fibrosis. A total of 563 blood-entry compounds were identified (367 prototypes and 196 metabolites), with 5 key active compounds identified in the Traditional Chinese Medicine Systems Pharmacology database corresponding to 63 targets. Network analysis revealed 44 overlapping genes, with Formononetin, Timosaponin BII, and Sinensetin as core components, and PTGS2, PPARG, and HSP90AA1 as hub targets. PPI analysis further identified key genes including ESR1, PTGS2, PPARG, HSP90AA1, JUN, and 15 additional genes. KEGG analysis indicated that SXQY mainly acts via Ca2+, phosphatidylinositol 3-kinase-protein kinase B, cyclic adenosine monophosphate, and inflammation- and hormone-related pathways. In conclusion, SXQY exerts protective effects against HF by improving cardiac function and attenuating myocardial remodeling through multicomponent, multitarget, and multipathway mechanisms.
    DOI:  https://doi.org/10.3791/70989
  24. J Sep Sci. 2026 Jun;49(6): e70467
    Novel Strongly Basic Molecularly Imprinted Solid-Phase Extraction Sorbent for Simultaneous Determination of Catecholamines and Their Metabolites in Urine.
    Artūrs Šilaks, Antons Podjava, Laura Bernāte, Vladlens Grebnevs, Artur Maciej.
      Catecholamines are important hormones and neuromediators in the human body. Simultaneous determination of both catecholamines and catecholamine metabolites in bodily fluids can help accurately diagnose dangerous health conditions like adrenal tumors. However, the biggest obstacle is the selective separation of these compounds from the biological matrix. In this work, we propose a novel, dual-recognition, non-covalent molecularly-imprinted polymer that utilizes strong anion exchange as the source of sorbent-analyte interaction. (4-Vinylbenzyl)trimethylammonium-homovanillyl alcohol anion salt and (4-vinylbenzyl)trimethylammonium-homoveratric acid anion salt served as template/functional monomer complexes for catecholamines and acidic metabolites, respectively. The sorbent was synthesized using precipitation polymerization and studied with batch adsorption experiments, scanning electron microscopy, Fourier-transform infrared spectroscopy, and Brunauer-Emmett-Teller surface area and pore size analysis. The polymer was loaded into cartridges and tested with acidified urine samples. The analytes are deprotonated and adsorbed via sorbent's tetraalkylammonium moiety in hydroxide form, which also neutralizes excess acid, removing the need for pH readjustment (which is often necessary for urine analysis). The imprinted sorbent can also be reused at least four times without performance deterioration. Norepinephrine, epinephrine, dopamine, normetanephrine, metanephrine, vanillylmandelic acid, and homovanillic acid were separated and analyzed in a single run using molecularly imprinted solid-phase extraction combined with liquid chromatography-tandem mass spectrometry (MISPE-LC-MS/MS), with recoveries ranging from 69% (epinephrine) to 97% (homovanillic acid). Method's limits of detection, limits of quantitation, linearity, repeatability, trueness, and intermediate precision were evaluated. Limits of quantitation ranged from 0.7 to 6.5 µg/L (for catecholamines and metanephrines) and from 0.12 to 0.2 mg/L (for acidic metabolites). Compared to commercially available weak-cation exchange and hydrophilic-lipophilic balance cartridges, the imprinted sorbent produced stronger catecholamine signals with minimal volume of urine (25 µL). This study successfully demonstrated molecularly imprinted solid-phase extraction workflow for simultaneous separation and quantitation of urinary catecholamines and their basic and acidic metabolites, proving its compatibility with bioanalysis.
    Keywords:  catecholamines; molecularly‐imprinted polymers; non‐covalent imprinting; solid‐phase extraction; strong anion exchange
    DOI:  https://doi.org/10.1002/jssc.70467
  25. Front Pharmacol. 2026 ;17 1846667
    Integrated metabolomics and machine learning identify predictive biomarkers via SHAP analysis for sintilimab-induced rash in lung cancer patients.
    Wenxiu Tian, Hehe Tang, Xiaoyuan Liu, Juan Lv, Yunyun Xu, Yujiao Hua, Fen Xie, Yongjuan Ding.
       Background: Sintilimab-induced rash is a significant clinical challenge in lung cancer treatment, often necessitating therapy interruption or discontinuation and thereby compromising patient outcomes. The underlying mechanisms of this adverse event remain poorly understood. This study aimed to investigate potential predictive biomarkers and mechanisms of sintilimab-induced rash through metabolomic profiling.
    Methods: A total of 55 patients with lung cancer who received sintilimab were enrolled, including 32 who developed rash and 23 matched controls without rash. Blood samples were collected before sintilimab infusion and at rash onset. Comprehensive clinical data were recorded. Untargeted metabolomic analysis of plasma was performed using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Differential metabolites were identified and analyzed using pathway enrichment, univariate analysis (AUC ≥0.800), and SHAP analysis.
    Results: No significant differences were observed between groups in demographic characteristics or most clinical parameters. However, the rash group exhibited significantly elevated total bile acids glucose (GLU), and basophil percentage (BAS%), along with reduced AST/ALT ratio, alkaline phosphatase lactate dehydrogenase phosphorus (P), neutrophil count (NEU), and high-sensitivity C-reactive protein (hsCRP) (P < 0.05). Metabolomic analysis identified 92 differentially expressed metabolites. Pathway enrichment revealed alterations in oxytocin signaling, GnRH signaling, platelet activation, FcγR-mediated phagocytosis, retrograde endocannabinoid signaling, pantothenate and CoA biosynthesis, FcεRI signaling, and aldosterone synthesis and secretion. Univariate analysis identified 25 metabolites with high predictive value (AUC ≥0.800), and SHAP analysis highlighted 20 metabolites. Cross-comparison identified five overlapping metabolites: N,N,N-trimethyl-L-histidine, laurolactam, 2-naphthalenesulfonic acid, limonenecarboxylic acid, and N-lauroylsarcosine.
    Conclusion: Distinct clinical and metabolomic alterations are associated with sintilimab-induced rash in lung cancer patients. The identified differential metabolites may serve as predictive biomarkers and potential therapeutic targets, providing new insights for clinical management and mechanistic research into immune-related adverse events.
    Keywords:  differentially expressed metabolites; lung cancer; sintilimab; skin rash; untargeted metabolomics
    DOI:  https://doi.org/10.3389/fphar.2026.1846667
  26. Chirality. 2026 Jul;38(7): e70112
    Simultaneous Determination and Enantiomeric Separation of Five DL-Thiol Compounds Using UHPLC-HRMS With Chiral Derivatization in Sake: Dynamic Monitoring of Human Urine After Drinking.
    Shuyun Xiao, Hang Shi, Shuyi Song, Jun Zhe Min.
      Sake, a fermented alcoholic beverage made from rice and Aspergillus oryzae through microbial fermentation, contains thiol compounds that are the primary source of its distinctive sweet aroma. However, to date, there has been no report on the detection of chiral thiol compounds in sake. This study introduces a novel UHPLC-HRMS method utilizing the (R)-(5-(3-isothiocyanatopyrrolidin-1-yl)-5-oxopentyl) triphenylphosphonium (NCS-OTPP) chiral mass spectrometry probe for the simultaneous detection of five DL-thiol compounds in sake. Separation was achieved using a YMC Triart C18 column (2.0 × 150 mm, 1.9 μm), employing an isocratic elution method to isolate DD/LL-GSH, γ-L-Glu-L-Cys, DL-Cys, DL-Hcy, and DL-Ac-Cys. The method demonstrated excellent linearity (R2 ≥ 0.9992), intraday precision (0.43%-13.18%), and an average recovery rate of 91.62%-110.40%. A comparative analysis of DL-thiol compound content in seven different types of sake from various countries and manufacturers revealed the presence of five chiral thiol compounds-LL-GSH, γ-L-Glu-L-Cys, DL-Cys, DL-Hcy, and L-Ac-Cys-across Japanese and Korean sake, with L-Cys being the most abundant and D-Ac-Cys the least. Notably, γ-L-Glu-L-Cys was detected only in two types of Korean sake and absent in Japanese sake. Additionally, the study investigated the metabolic kinetics of chiral thiol compounds in human urinary specimens after alcohol ingestion, constructing a metabolic fitting curve. The peak concentrations of DL-Cys, DL-Hcy, and LL-GSH were attained 15 min following consumption, with slow clearance, returning to baseline at 60 min. The metabolic fitting curve effectively captures the dynamic changes in urinary metabolism. This research presents a new method for detecting chiral thiol compounds in sake and monitoring urinary metabolism after alcohol consumption.
    Keywords:  DL‐thiol compound; UHPLC–HRMS; chiral resolution; sake; urine
    DOI:  https://doi.org/10.1002/chir.70112
  27. Anal Methods. 2026 Jun 19.
    Real-time metered dose inhalers (MDIs) characterization by secondary electrospray ionization (SESI)-mass spectrometry: a proof-of-principle study.
    Ana María Casas-Ferreira, Miguel Del Nogal Sánchez, Eliseo Herrero-Hernández, Diego García-Gómez, Encarnación Rodríguez-Gonzalo, José Luis Pérez Pavón.
      This study explores the application of secondary electrospray ionization mass spectrometry (SESI-MS) for the real-time characterization of pharmaceutical aerosols from metered-dose inhalers (MDIs). The analysis includes one of the most widely used inhalers worldwide, Ventolin®, a simple formulation containing salbutamol and norflurane for which salbutamol was reliably detected, while norflurane remained undetectable due to its low proton affinity; and Ventoduo®, a multicomponent formulation also including beclomethasone dipropionate, ethanol, and oleic acid for which all major components were successfully detected. The coupling of SESI with high-resolution MS enabled unambiguous identification of all analytes, with excellent mass accuracy, confirming the identity of both active ingredients through characteristic fragmentation patterns. These results demonstrate that SESI-MS, particularly when coupled with high-resolution instrumentation (SESI-HRMS), is a powerful and reliable tool for the comprehensive and real-time analysis of pharmaceutical aerosols, including those with multiple active ingredients and co-solvents.
    DOI:  https://doi.org/10.1039/d6ay00870d
  28. J Chromatogr B Analyt Technol Biomed Life Sci. 2026 Jun 12. pii: S1570-0232(26)00275-8. [Epub ahead of print]1281 125186
    Comprehensive profiling of synthetic cannabinoids fragment ions using UPLC-HRMS in conjunction with online databases.
    Shihao Zhong, Huan Li, Shiyang Qin, Jinghui Zhou, Hanchen Liu, Xiaoyi Wang, Yixuan Han, Yuanfeng Wang.
      This study utilizes Ultra-Performance Liquid Chromatography-High-Resolution Mass Spectrometry (UPLC-HRMS) and integrates open-access online MS databases (e.g., mzCloud) along with analytical software (e.g., Mass Frontier, ChemDraw). By leveraging a well-established structural classification framework, we systematically analyzed the high-resolution mass spectrometry (HRMS) fragmentation pathway and key m/z of known synthetic cannabinoids (SCs). Based on the four common components of synthetic cannabinoids (core, linked group, side chain, and linker), integrate the structural information of known SCs, and obtain HRMS for representative SCs through LC-HRMS analysis. The elemental compositions and chemical properties of recurring or structurally related m/z were individually analyzed with the aid of open-access MS databases and supporting software. The study revealed that the core structures of SCs (such as indole, indazole, and carbazole) produce stable m/z (e.g., m/z 116.050, 144.044) during fragmentation. Additionally, substituents (such as phenyl rings, pyridine rings, and halogenated naphthalenes) and sidechains (such as aliphatic chains and benzyl structures) also generate stable m/z during fragmentation. Moreover, the study uncovered the relationship between the sidechain structures of SCs and neutral losses. The findings could provide significant theoretical and technical support for the metabolic analysis, non-targeted detection, structural identification, and other aspects of drug control research on SCs, and also fill the gap in the comprehensive HRMS characterization of SCs in existing studies.
    Keywords:  Characteristic fragment ions; Drug control; LC-HRMS; Non-targeted analysis; Synthetic cannabinoids
    DOI:  https://doi.org/10.1016/j.jchromb.2026.125186
  29. Front Mol Biosci. 2026 ;13 1795393
    Integrative metabolomics and molecular networking reveal the progressive metabolic continuum of resin formation in Dracaena cochinchinensis wood.
    Meijia Chen, Yunfei Cui, Jinhua Su, Jiahui Ren, Yu Zhu, Jinhui Wang, Guang Li.
       Introduction: The resinification of Dracaena cochinchinensis wood represents a sophisticated physiological reprogramming triggered by environmental stress, culminating in the production of the high-value "dragon's blood" resin.
    Methods: In this study, we systematically characterized the chemical space of three representative wood morphotypes-hollow cork cambium (P), whole-body resin-containing (MZ), and resin-secreting aggregated (LZ)-to decipher the molecular mechanisms underpinning resinogenesis within the xylem matrix. Integrating UPLC-Q-TOF-MS/MS with Feature-Based Molecular Networking (FBMN), we overcame annotation bottlenecks inherent in complex woody tissues and successfully annotated 299 specialized metabolites, including flavonoids, phenylpropanoids, steroids, and lipids.
    Results: Multivariate statistical analysis revealed significant metabolic shifts across these wood morphotypes, reflecting distinct biological strategies: the P form prioritized phenylpropanoid biosynthesis (e.g., sinapyl alcohol and coniferaldehyde) for structural barrier reinforcement of the wood cell walls; the MZ morphotype accumulated steroids and fatty acids to maintain membrane integrity during chronic adaptation; and the LZ morphotype exhibited a defensive burst of flavonoids and terpenoids, typical of acute stress responses.
    Discussion: Collectively, these findings support a putative "progressive metabolic continuum" model (P→MZ→LZ), illustrating a hypothesized metabolic gradient and a dynamic shift in resource allocation from physical repair to chronic adaptation and finally to acute chemical defense. This study provides a comprehensive phytochemical framework for understanding specialized metabolite mobilization in resinous wood and establishes an efficient metabolomic workflow for the quality evaluation and sustainable utilization of woody medicinal resources.
    Keywords:  Dracaena cochinchinensis; Feature-Based Molecular Networking (FBMN); metabolic reprogramming; phytochemical mobilization; stress response; therapeutic compounds; wood morphotypes
    DOI:  https://doi.org/10.3389/fmolb.2026.1795393
  30. Anal Chem. 2026 Jun 17.
    Corona: A Virtual Mass Spectrometer for the Development of Real-Time Mass Spectrometry Software.
    Michael R Hoopmann, Christopher D McGann, Jesse D Canterbury, William D Barshop, Qing Yu, Meagan Gadzuk-Shea, Alexander Hogrebe, Judit Villén, Devin K Schweppe.
      Real-time mass spectrometry (RTMS), the process by which mass spectral data are analyzed during acquisition on a mass spectrometer, is an integral part of mass spectrometer development. Particularly within the fields of proteomics and metabolomics, RTMS has evolved to include the creation of third-party software that interfaces with a mass spectrometer to provide novel acquisition methodologies, such as applications that use the IAPI from Thermo Fisher Scientific. Developing and testing RTMS applications with the use of a mass spectrometer is a slow and expensive process that creates a bottleneck in the laboratory. Here, we present Corona, a virtual mass spectrometer for use in RTMS application development independent of a mass spectrometer. RTMS applications developed with IAPI connect to Corona seamlessly and operate exactly the same as if connected to a physical instrument. In this manner, it is possible to rapidly create and test RTMS applications prior to deployment on a mass spectrometer.
    DOI:  https://doi.org/10.1021/acs.analchem.6c01637
  31. STAR Protoc. 2026 Jun 18. pii: S2666-1667(26)00297-2. [Epub ahead of print]7(3): 104644
    Protocol for high-throughput, automated, solid-phase extraction of sphingolipids from human urine.
    Rebekah J Nicholson, J Alan Maschek, Trevor Lonergan, Scott A Summers.
      Urinary sphingolipids may serve as biomarkers of renal diseases. Here, we present a high-throughput, automated protocol for solid-phase extraction and tandem mass spectrometry (LC-MS/MS) quantification of lipids from urine. We describe steps for sample preparation and lipid extraction, and then detail procedures for targeted LC-MS/MS analysis of urinary sphingolipids. In this protocol, we focus on the analysis of urine samples; however, this platform can be optimized for alternative biospecimens. Additionally, this method allows for tandem preparation of metabolomic and lipidomic samples. For complete details on the use and execution of this protocol, please refer to Nicholson et al.1.
    Keywords:  Health Sciences; High Throughput Screening; Metabolism; Metabolomics; Protocols in Metabolomics and Lipidomics
    DOI:  https://doi.org/10.1016/j.xpro.2026.104644
  32. Front Pharmacol. 2026 ;17 1755360
    UPLC-MS/MS profiling, antioxidant and hemorheological effects of D101 resin-based purified flavonoids from Allium polyrhizum.
    Caiquan Zhao, Lige Bai, Jian Gao, Jie Liu, Ling Lu, Meng Wang, Jiahua Guo, Peng Zhao.
       Introduction: Oxidative stress and blood stasis represent critical pathological drivers underlying the progression of cardiovascular and cerebrovascular diseases. Total flavonoids derived from Allium polyrhizum Turcz. ex Regel (APTF) have been reported to possess potential antioxidant and blood-activating properties. However, the purification process and systematic pharmacological characteristics of APTF remain insufficiently elucidated, which substantially limits its further development and practical application.
    Methods: In this study, orthogonal experiments were performed to optimize the purification conditions of APTF using D101 macroporous resin. Chemic antioxidant activity of purified APTF was evaluated via multiple free radical scavenging assays. A rat model of blood stasis was established to verify the in vivo efficacy of APTF, with hemorheological, vascular endothelial, and blood coagulation indices determined for comprehensive assessment. Additionally, ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) was employed to systematically identify the metabolic components of APTF.
    Results: The optimized purification protocol yielded a high APTF recovery rate of 95.112±1.507%. Purified APTF exhibited potent free radical scavenging activity. In vivo experimental results demonstrated that APTF effectively ameliorated hemorheological abnormalities, restored vascular endothelial functional homeostasis, and exerted remarkable anticoagulant effects in blood stasis model rats. A total of 309 flavonoid metabolites were successfully identified, which constituted the potential material basis for the pharmacological activities of APTF.
    Discussion: This study established a stable and reliable purification protocol for APTF using D101 macroporous resin. The purified APTF displays prominent antioxidant and blood-activating bioactivities, conferring great application potential for the development of vascular protective functional products and therapeutic agents. Future investigations will focus on screening the active monomer components of APTF and clarifying its underlying molecular mechanisms of action.
    Keywords:  Allium polyrhizum Turcz. ex Regel; antioxidant; blood stasis; flavonoids; macroporous resins
    DOI:  https://doi.org/10.3389/fphar.2026.1755360
  33. Chem Biodivers. 2026 Jun;23(6): e71385
    Comparative Phytochemical Analysis of Five South African Bulbine Species Using Chromatography-Based Methods.
    Ramoagi T Segone, Maxleene Sandasi, Efficient Ncube, Weiyang Chen, Bharathi Avula, Kumar Katragunta, Ikhlas A Khan, Alvaro M Viljoen.
      Bulbine Wolf (Asphodelaceae) comprises over 80 species, with five holding significant medicinal value in South Africa for treating skin and gastrointestinal ailments. While roots are traditionally prioritised, the chemical profiles of leaves and rhizomes remain underexplored although they are also medicinally important. This study used chromatographic and mass spectrometry-based analytical techniques (HPTLC, LC-QToF-MS, and UPLC-MS) combined with chemometric analysis to chemically profile the metabolites of five Bulbine species from the Eastern Cape. Chemical profiling tentatively identified 73 compounds, including flavonoids, phenolic acids, and anthraquinones. Phenylanthraquinones, specifically knipholone and knipholone 6'-methyl ether, were ubiquitous across all taxa. Species-specific markers were identified, such as bulbine-knipholone in B. abyssinica, gaboroquinones in B. frutescens, and bulbnatalonosides in B. latifolia. Chemometric modelling revealed that while rhizome chemistry is relatively conserved, root profiles provide the most robust inter-species differentiation. Bulbine frutescens and B. latifolia displayed the highest chemical diversity, whereas B. asphodeloides was notably rich in anthocyanins. The significant overlap in bioactive constituents between leaves, roots and rhizomes suggests that aerial parts could serve as viable substitutes in traditional remedies. These findings establish a chemical basis for species authentication and support sustainable harvesting practices by reducing the destructive exploitation of rhizomes and roots.
    Keywords:  Bulbine species; HPTLC; LC‐QToF‐MS; UPLC‐MS; chemometrics
    DOI:  https://doi.org/10.1002/cbdv.71385
  34. J Pharm Biomed Anal. 2026 Jun 05. pii: S0731-7085(26)00270-0. [Epub ahead of print]280 117602
    UPLC-ESI-QToF-MS/MS profiling and anxiolytic-like effects of Simira paraensis (Baill.) Steyerm bark.
    Larissa Rodrigues Jales Martins, Nadia Eligia Nunes Pinto Paracampo, Paulo Riceli Vasconcelos Ribeiro, Francisco Oiram Filho, Amanda Maria Barros Alves, Hélcio Silva Dos Santos, Jane Eire Silva Alencar de Menezes, Márcia Machado Marinho, Matheus Nunes da Rocha, Emmanuel Silva Marinho, Kirley Marques Canuto.
      Simira paraensis (Baill.) Steyerm (Rubiaceae), popularly known as araribá-rosa, is a small tree found mainly in the Amazon. Studies on the genus Simira report a chemical composition rich in indole alkaloids of the β-carboline class, which are associated with effects on the central nervous system. The present study aimed to characterize the chemical profile and evaluate the anxiolytic activity of S. paraensis bark through in vivo and in silico assays. The powdered bark was extracted with a hydromethanolic solution, yielding a red-colored extract (SP-EMB), which was analyzed by Ultra High Performance Liquid Chromatography coupled to Electrospray Ionization Mass Spectrometry and Quadrupole-Time-of-Flight Analyzer (UPLC-ESI-QToF-MS/MS). The neuropharmacological activity was evaluated in zebrafish (Danio rerio) using locomotor activity, light/dark, novel tank, GABAergic neuromodulation, and PTZ-induced seizure assays. UPLC-ESI-QToF-MS/MS analysis revealed 15 β-carboline alkaloids, 4 phenolic acid derivatives, and 1 phenolic glycoside. Harmane, the major constituent, was isolated by preparative HPLC and identified by UPLC-ESI-QToF-MS/MS and NMR analyses. SP-EMB exhibited anxiolytic-like effects in both light/dark and novel tank tests, with low acute toxicity (LD₅₀ > 400 mg·kg⁻¹) and reduced sedative effects compared to diazepam (DZP, 4 mg·kg⁻¹). Molecular docking showed that harmane and chlorogenic acid have high affinity for the GABAA receptor, indicating their influence on the anxiolytic effect. Both compounds showed low toxicity (LD₅₀ > 40 mg·kg⁻¹) and dose-dependent anxiolytic-like activity without locomotor impairment. Furthermore, chlorogenic acid exhibited anxiolytic effects mediated by GABAA receptors. These findings may suggest that S. paraensis is a promising alternative source of anxiolytic agents with lower sedative effects.
    Keywords:  Anxiety; Mass spectrometry; Rubiaceae; Simira paraensis; β-carboline alkaloids
    DOI:  https://doi.org/10.1016/j.jpba.2026.117602
  35. Rapid Commun Mass Spectrom. 2026 Sep 15. 40(17): e70121
    Integrating Aptasensor With Mass-Tag Signal Amplification Strategy for the Detection of Three Steroid Hormones in Serum.
    Jia Zhao, Huimin Liu, Liujuan Zhan, Wei Shao, Guangming Huang.
       RATIONALE: Steroid hormone testing is critical for assessing endocrine function, diagnosing related disorders, and monitoring therapeutic efficacy. However, current mainstream detection methods have limitations. Although liquid chromatography-tandem mass spectrometry (LC-MS/MS), regarded as the gold standard, offers high sensitivity and specificity, it involves complex and time-consuming procedures. Immunoassays such as enzyme-linked immunosorbent assay (ELISA) are simple and fast but are limited by poor throughput for multitarget detection. Therefore, it is crucial to develop an analytical method that streamlines procedures and enables efficient parallel detection of multiple targets.
    METHODS: We developed an integrated signal-enhanced aptasensor platform for pooled MS detection of three steroid hormones in serum. It combines aptamer recognition with mass-tag amplification. This approach uses biorecognition instead of chromatography, while mass spectrometry enables simultaneous readout of multiple mass tags from combined samples.
    RESULTS: The method was evaluated by detecting three steroid hormones (vitamin D, cortisol, and testosterone) in simulated serum samples and human serum samples. The limits of detection (LODs) for these analytes ranged from 0.411 to 6.796 nM, which are below the established clinical cut-off values for each steroid, demonstrating the requisite sensitivity for detection.
    CONCLUSIONS: This integrated signal-enhanced aptasensor outperforms conventional LC-MS/MS in efficiency and ELISA in throughput, enabling the quantification of multiple serum steroid hormones. Therefore, we believe that this method could be potentially useful in the clinical screening of hormone-related disorders and suitable for the analysis of serum.
    Keywords:  aptasensor; mass spectrometry; multitarget analysis; signal amplification; steroid hormone detection
    DOI:  https://doi.org/10.1002/rcm.70121
  36. ACS Omega. 2026 Jun 09. 11(22): 32435-32451
    Forensic Tracing of Unknown Compounds at the Human-Environment Interface: An All-in-One LC-HRMS Strategy Guided by a Curated Methodological Database.
    Alix Munoz, Jeanne Brotier, Laetitia Barthe, Guénaël Thiault, Joëlle Vinh.
      The detection and identification of unknown organic pollutants in complex environmental and forensic samples remain major analytical challenges. While numerous finely tuned liquid chromatography-mass spectrometry (LC-MS) methods exist for specific compound classes, no transversal strategy has been proposed to date. Here we propose an "all-in-one" LC-MS strategy that is robust enough to handle the unknown and broad enough to encompass multiple chemical families within a single analytical framework. Our approach combines a structured, literature-derived analysis of LC-based methods with experimental validation on model mixtures and real forensic samples. From a systematic review of hundreds of reported conditions, we established a structured database of key LC parameters for selected pesticides, household products, and dyes. This enabled the design of a cross-family LC-MS method validated on representative compounds and successfully applied to authentic casework, including suspicious delivery packages and household cleaning agents in forensic investigations. Rather than relying on novel instrumentation, this work provides the first literature-driven, experimentally validated workflow for broad-spectrum, nontargeted LC-MS detection. This strategy, widely applicable across environmental and forensic applications, is a valuable resource for harmonization, spectral library growth, and automated annotation. Moreover, the compiled reference database of chromatographic conditions serves as a unique tool for method development in the community.
    DOI:  https://doi.org/10.1021/acsomega.6c00585
  37. Chem Biodivers. 2026 Jun;23(6): e71383
    Seasonal Phytochemical Profiling and Chemometric Assessment of Crataegus songarica K. Koch: Implications for Bioactivity and Resource Utilization.
    Zokirova Mashkhura, Narges Samanian, Erkin Botirov, Baratov Kuzijon, Rakhmonova Gulnora, Isaboyeva Dilnoza.
      Crataegus songarica K. Koch is a widely distributed species in Central Asia with recognized ethnobotanical importance; however, its seasonal phytochemical variation remains insufficiently characterized. In this study, leaves and flowers collected at different developmental stages were analyzed to evaluate phenolic profiles, mineral composition, and associated biological responses. High-performance liquid chromatography identified apigenin and rutin as the dominant flavonoids in leaves, while flowers showed pronounced but transient increases in flavonoid content during flowering. Inductively coupled plasma-mass spectrometry revealed high levels of potassium, calcium, and magnesium. Ethanolic extracts exhibited low acute toxicity (LD50 > 5000 mg/kg) and demonstrated measurable biological activity, including attenuation of carbon tetrachloride-induced biochemical alterations and improved tolerance to normobaric hypoxia. Multivariate analysis using principal component analysis revealed clear organ- and season-dependent differentiation of phytochemical profiles, while partial least squares regression indicated model-indicated associations between chemical composition and physiological response parameters. The results highlight pronounced seasonal and organ-specific variation in C. songarica K. Koch and demonstrate that its biological effects are linked to coordinated phytochemical patterns, supporting its value as a chemically rich plant resource for phytochemical and biodiversity-oriented studies.
    Keywords:  Crataegus songarica K. Koch; antihypoxic activity; chemometrics; hepatoprotective activity; phenolic compounds
    DOI:  https://doi.org/10.1002/cbdv.71383
  38. J Chromatogr B Analyt Technol Biomed Life Sci. 2026 Jun 08. pii: S1570-0232(26)00266-7. [Epub ahead of print]1281 125177
    Enantioselective analysis of synthetic cathinones and amphetamine-type stimulants in biological fluids via LC-MS/MS.
    Bartłomiej Feigel, Piotr Adamowicz, Wiktor Kasprzyk, Elisa Ziliani, Sławomir Wybraniec.
      Synthetic cathinones are the second largest group of new psychoactive substances (NPS), with 123 new compounds reported in Europe between 2014 and 2023. Many possess a chiral center and are sold as racemic mixtures. Since enantiomers and other structural isomers may differ in pharmacological and pharmacokinetic properties, developing analytical methods capable of their separation and quantitative analysis is essential, particularly in forensic and clinical toxicology, where accurate determination of therapeutic and toxic concentrations is critical. This study developed a liquid chromatography-tandem mass spectrometry method for separating cathinone-type NPS, including structural isomers and enantiomers. Chromatographic separation was performed at 10 °C using a Lux 3 μm chiral column (Cellulose-2 phase). The method enables both qualitative and quantitative analysis, improving differentiation of structurally similar compounds and supporting toxicological interpretation. The method achieved successful chromatographic separation of the target cathinone enantiomers and numerous structural isomers within a 15-min runtime after a simple liquid-liquid extraction step. Validation demonstrated excellent analytical performance: calibration models were linear (R2 ≥ 0.99), accuracy and precision exceeded predefined acceptance criteria across the tested range, limits of detection and quantification at 1 ng/mL produced signal-to-noise values well above thresholds, and no significant carry-over or cross-interference was observed. Matrix effects were present in the range of -72% to +57%, and stability testing indicated superior preservation at higher concentrations and under frozen or cooled autosampler conditions. Therefore, the developed method constitutes a significant advancement in the analysis of synthetic cathinones, providing precise and reliable tools for forensic and clinical applications.
    Keywords:  Chiral column; Designer drugs; Drug screening; LC-MS/MS; NPS enantiomers; NPS isomers; New psychoactive substances (NPS)
    DOI:  https://doi.org/10.1016/j.jchromb.2026.125177
  39. Daru. 2026 Jun 19. pii: 37. [Epub ahead of print]34(2):
    Bio-guided identification of novel antiparasitic natural products from Gardenia imperialis K. Schum (Rubiaceae).
    Christelle Wayoue Kom, Sylvester Osei Bobbie, Darline Dize, Cyrille Armel Njanpa Ngansop, Mariscal Brice Tchatat Tali, Lauve Rachel Tchokouaha Yamthe, Bill Baker, Fabrice Fekam Boyom.
       BACKGROUND: Gardenia imperialis K. Schum. is a medicinal plant used in traditional medicine to treat malaria and related symptoms. This bio-guided study aimed at identifying antiparasitic compounds from G. imperialis against the causative agents of malaria (Plasmodium falciparum), visceral leishmaniasis (Leishmania donovani), and sleeping sickness (Trypanosoma brucei brucei).
    METHODS: Crude methanolic extracts from the leaf and stem bark of G. imperialis were studied using liquid chromatography-mass spectrometry (LC-MS) and preparative medium/high-performance liquid chromatography (M/HPLC) guided by evidence of antiparasitic activity. The structures of the isolated compounds were elucidated using nuclear magnetic resonance (NMR) and MS. All the extracts, fractions, subfractions, and pure compounds were evaluated for antiparasitic activity. Cytotoxicity was tested against Vero and Raw264.7 mammalian cell lines.
    RESULTS: The leaf methanolic extract (GIlMeOH) exhibited broad-spectrum antiparasitic activity (IC₅₀ < 11 µg/mL) and low cytotoxicity (median cell cytotoxicity-CC50 >100 µg/mL). The dereplication of the extract and isolation yielded fractions, subfractions, and four flavonoids with improved activity (IC₅₀ values ranging from 0.67 to 23.8 µg/mL). Among the isolated flavonoids, gradenin A (3), salvigenin (4), hispidulin (1), and 5,7,3'-trihydroxy-6,4',5'-trimethoxyflavone (2) showed potent antitrypanosomal activity with IC₅₀ values of 2.8, 2.5, 16.6, and 9.7 µg/mL, respectively. The antiparasitic activities of gradenin A, salvigenin, and 5,7,3'-trihydroxy-6,4',5'-trimethoxyflavone are reported here for the first time.
    CONCLUSION: The promising antiparasitic profile of the G. imperialis leaf extract and its isolated constituents, particularly and 5,7,3'-trihydroxy-6,4',5'-trimethoxyflavone (2), gradenin A (3), and salvigenin (4), warrants further investigation into their potential as novel and selective agents against the target parasitic diseases.
    Keywords:   Gardenia imperialis ; Antiparasitic activity; Flavonoids; Malaria; Sleeping sickness; Visceral leishmaniasis
    DOI:  https://doi.org/10.1007/s40199-026-00618-w
  40. Rapid Commun Mass Spectrom. 2026 Sep 15. 40(17): e70124
    Bioanalysis of Ametryn by UHPLC-MS3 Coupled With Multiple Fragmentation to Decrease Interference and Enhance Sensitivity.
    Yue Deng, Hongjie Ju, Fangyu Liu, Jiarui Zhang, Shuang Feng, Jiale Liu, Xinyue Zhou, Lei Yin, Meiyun Shi.
       RATIONALE: Ametryn poses significant risks to aquatic ecosystems, but conventional MRM methods suffer from matrix interference and limited sensitivity. To overcome these limitations, we established a more selective and sensitive UHPLC-MS3 approach that effectively suppresses background noise and enhances signal-to-noise ratio, enabling reliable quantification of ametryn in zebrafish tissues.
    METHODS: A novel UHPLC-MS3 method with electrospray ionization was developed. Quantification employed the MS3 transition m/z 228.0 → 186.1 → 158.0. Five zebrafish tissues (liver, muscle, brain, intestine, gill) were analyzed following sample preparation. Fast chromatographic separation was achieved in 2.8 min.
    RESULTS: Compared with MRM, the MS3 method substantially reduced matrix interference, enhanced signal intensity from 6.4 × 104 to 1.6 × 107 cps, and increased the signal-to-noise ratio from 6.3 to 45. The assay exhibited excellent linearity (10-1000 pg/mL), and validation results for accuracy, precision, recovery, matrix effects, and stability all met FDA bioanalytical requirements.
    CONCLUSIONS: This study presents the first application of UHPLC-MS3 for quantitative analysis of ametryn in multiple zebrafish tissues, providing a reliable high-throughput platform for residue monitoring and mechanistic toxicology research.
    Keywords:  UHPLC‐MS3; ametryn; method validation; tissue distribution; zebrafish
    DOI:  https://doi.org/10.1002/rcm.70124
  41. J Pharm Biomed Anal. 2026 Jun 08. pii: S0731-7085(26)00271-2. [Epub ahead of print]280 117603
    An integrated multi-technique strategy for in vitro metabolite identification and profiling of gymnodimine A using LC-HRMS/MS.
    Jiangang Chen, Shuai Liu, Sudan Liang, Minglei Zhong, Jiaquan Li, Li Ling, Yihou Peng, Liangjiao Qu, Yan Zhang, Wenyan Huang, Shaoping Li, Jie Wu, Jing Zhao.
      Gymnodimine A (GYM-A) is a lipophilic cyclic imine toxin produced by marine dinoflagellates that accumulates in shellfish and poses potential risks to human health. However, regulatory limits have not been established worldwide, largely due to insufficient understanding of its metabolic pathways and toxicokinetic behavior. This study characterized the in vitro metabolism of GYM-A using human liver microsomes (HLM) and human liver S9 fractions (HS9) combined with liquid chromatography-high-resolution tandem mass spectrometry (LC-HRMS/MS). An integrated identification strategy incorporating segmented data-dependent acquisition (DDA), targeted/untargeted screening, MS/MS fingerprint comparison, density functional theory (DFT) calculation, and μElution solid phase extraction (SPE) enabled confident structural elucidation of metabolites derived from low-concentration substrates in complex matrices. Four phase I hydroxylated metabolites were identified: three mono-hydroxylated derivatives (M1-M3) and one di-hydroxylated derivative (M4), with 21-hydroxy-GYM-A (M1) as the highest relative abundance. No phase II glucuronide conjugates were detected under the conditions tested. Metabolic stability studies revealed high intrinsic clearance following first-order kinetics, with elimination primarily mediated by CYP450-dependent phase I oxidation. Enzyme phenotyping identified CYP3A5 as the principal isoform responsible for GYM-A hydroxylation. Detection of metabolite M2 in urine from dosed rats supported the in vitro findings. This work provides the first systematic metabolic profile of GYM-A and advances knowledge of its kinetic behavior, while the analytical workflow may serve as a useful tool for risk assessment and metabolism studies of related marine toxins.
    Keywords:  GYM-A; In vitro metabolism; LC-HRMS/MS; Metabolites identification; Segment DDA; Targeted and untargeted screening
    DOI:  https://doi.org/10.1016/j.jpba.2026.117603
  42. J Chromatogr A. 2026 Jun 11. pii: S0021-9673(26)00510-8. [Epub ahead of print]1783 467181
    Rapid spatial distribution analysis of lipids in wheat grain using DESI/PI mass spectrometry imaging.
    Yang Pan, Yaya Wang, Zhiwei Wen, Weimin Wang, Na Li, Qin Sun, Chengyuan Liu, Yang Pan.
      Wheat is an important cereal grain. Understanding lipid composition and distribution in wheat grain is valuable for plant, food and nutritional research. Conventional methods can analyze lipid composition and content in wheat grain, while the spatial native distribution of lipids is inevitably lost. In this paper, an effective mass spectrometry imaging (MSI) analysis protocol combining desorption electrospray ionization/post-photoionization (DESI/PI) with high-resolution mass spectrometer was established. To overcome the hardness of wheat grains and fragility of wheat grain sections, a rapid hand plane sectioning (HPS) method was developed. By employing accurate mass spectrometry peak identification, the HPS samples detection results achieved ∼23 % similarity with cryosectioning sample (same compounds). DESI/PI has good sensitivity for both non-polar and polar lipids. A small mass spectrum database for wheat grain was established which may provide strong data support for wheat grain component analysis. The rapid MSI analysis protocol was further used to explore the spatial distribution changes of lipids in wheat grain with different storage years, indicating that DESI/PI was a potential means of studying wheat grain aging. This study provides a convenient sectioning method, a comprehensive identification checklist, and spatial distribution information of lipids in wheat grains, which holds potential significance for guiding improvements in wheat grain processing & storage, slowing down the aging, and ensuring grain quality.
    Keywords:  DESI/PI; Food security; Lipids analysis; Mass spectrometry imaging; Wheat grain
    DOI:  https://doi.org/10.1016/j.chroma.2026.467181
  43. J Adv Pharm Technol Res. 2026 Apr-Jun;17(2):17(2): 121-127
    Bioactive potential of Rhododendron lowndesii: Extraction, antioxidant analysis, antimicrobial assay, and Liquid Chromatography-mass Spectrometry profiling.
    Anup Luitel, Rajan Shrestha, Lalit Mohan Pant, Ashwinee Kumar Shrestha, Niru Thapa, Manika Shrestha, Ronak Shrestha, Ganesh Datt Joshi, Kirankumar Hullatti, Janardan Lamichhane.
      Rhododendron lowndesii Davidian is an endemic medicinal plant of Nepal found in an altitude range of 3000-4100 m. This study investigates the phytochemical composition and bioactivities of its methanolic extract. The aerial part of the plant was extracted through methanol maceration to achieve 31.50% ± 0.92% yield. Qualitative and quantitative phytochemical analysis of the extract revealed the presence of alkaloids, flavonoids, phenols, steroids, terpenoids, coumarins, and carbohydrates, with a total phenolic content of 175.63 ± 1.37 mg GAE/g and total flavonoid content (TFC) of 1090.36 ± 2.29 mgQE/g. Antioxidant activity assay of the extract suggested its strong radical scavenging activity through DPPH (IC50:19.46 ± 1.85 µg/mL) and ABST is 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) assays (IC50: 13.4 ± 0.20 µg/mL). Antibacterial activity as determined by the disc diffusion method, showed significant effect against Bacillus subtilis, Enterococcus faecalis, and Staphylococcus aureus (minimum inhibitory concentration: 312.5-1250 µg/mL) but limited efficacy against Salmonella enterica. High-resolution liquid chromatography-mass spectrometry identified pharmacologically significant compounds, including zapotin and cepharanthine. These findings suggest that R. lowndesii has rich phytochemical profile and bioactivity; this supports its potential as a medicinal plant and need of its pharmacological research.
    Keywords:  Antioxidant activity; Rhododendron lowndesii; high-resolution liquid chromatography-mass spectrometry; phytochemicals
    DOI:  https://doi.org/10.4103/JAPTR.JAPTR_235_25
  44. J Proteome Res. 2026 Jun 18.
    Evaluation of FASP for Mass Spectrometry-Based Untargeted Metabolomics Analysis of Urine Samples.
    Muath Khairi Mousa, Luis B Carvalho, Alexander D Giddey, André Figueiredo, Hamza Al-Hroub, Mohammad H Semreen, Mohammed Uddin, Hugo M Santos, Nelson C Soares.
      Filter-Aided Sample Preparation (FASP) is a well-established method in proteomics, yet its potential for the parallel recovery of metabolites remains largely unexplored. Herein, we evaluate the performance of FASP as a straightforward workflow for the simultaneous isolation of protein and corresponding metabolite fractions from a single urine sample. The FASP-based LC-MS/MS approach for both proteomics and metabolomics analysis identified 3,163 nonredundant peptides corresponding to 957 unique protein groups. The metabolomic profile comparison of three urine fractions, specifically FASP-concentrated, FASP flow-through, and raw samples, resulted in the identification of 176 common metabolites. Next, as a proof-of-concept, the FASP protocol was applied to compare the metabolomic profiles of clinical urine samples from healthy individuals (n = 13) and patients with Ta bladder cancer (n = 12). The metabolomic modulation was consistent with previously reported findings, highlighting perturbations in phenylacetate, purine, and tryptophan metabolism, as reflected by changes in metabolites such as adenosine monophosphate (AMP), phenylacetic acid, glutamine, cytosine, and l-tryptophan. FASP protocol can be effectively adapted for the concurrent profiling of both proteomic and metabolomic fractions from urine samples. Thus, FASP-based workflow represents a viable alternative for single-step sample preparation, facilitating subsequent quantitative multiomics data integration.
    Keywords:  Bladder cancer; FASP; Metabolomics; Multiomics; Proteomics; Sample preparation; Urine
    DOI:  https://doi.org/10.1021/acs.jproteome.6c00173
  45. Front Mol Biosci. 2026 ;13 1823383
    FNA-based lipidomics reveals coordinated lipid metabolic remodeling in pancreatic cancer.
    Shenhao Liu, Lifeng He, Gelin Jiang, Songling Lei, Bin Xu, Jian Xu, Songmei Lou.
       Background: Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive malignancy with poor prognosis. Endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) is routinely used for preoperative tissue confirmation; however, its potential for comprehensive lipidomic profiling in a preoperative diagnostic setting remains insufficiently explored. Given the critical role of lipid metabolic reprogramming in PDAC progression, we investigated whether lipidomic alterations could be reliably captured in EUS-FNA-derived specimens.
    Methods: Paired tumor and adjacent non-tumor tissues obtained via EUS-FNA from 13 PDAC patients were subjected to widely targeted (pseudo-targeted) liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based lipidomic analysis. Differential lipid species were identified through multivariate and univariate analyses. A composite lipid score was constructed based on principal component loadings. Serum samples from an independent cohort of 30 PDAC patients were included for exploratory projection analysis.
    Results: A total of 1822 lipid species across 47 lipid classes were detected in EUS-FNA-derived specimens. Tumor tissues displayed coordinated lipid alterations, including accumulation of storage lipids and structural remodeling of fatty acyl chains characterized by elongation and increased unsaturation. These alterations were readily detectable in EUS-FNA-derived specimens. These patterns showed a tendency to distinguish tumor from adjacent non-tumor samples within the FNA cohort. Exploratory projection suggested directionally consistent lipid changes in serum samples.
    Conclusion: Lipid metabolic remodeling in PDAC can be reliably detected in preoperative EUS-FNA-derived specimens. These findings support the feasibility of lipidomic profiling in minimally invasive diagnostic samples and highlight the translational potential of EUS-FNA-based metabolic assessment.
    Keywords:  EUS-FNA; LC-MS/MS; PDAC; lipidomics; metabolic reprogramming; micro-biopsy
    DOI:  https://doi.org/10.3389/fmolb.2026.1823383
  46. Chem Biodivers. 2026 Jun;23(6): e71422
    Chemical Composition and Anti-Inflammatory Activity of Selaginella tamariscina.
    Haimin Kuai, Xiaowei Wu, Yulin Feng, Jingyi Lai, Wenai Xu, Rong Wang, Hao Wang.
      Selaginella tamariscina is a traditional Chinese medicine with promising anti-inflammatory activity. In this study, a bioactivity-guided strategy was employed to characterize its active constituents. The ethyl acetate fraction significantly reduced LPS-induced nitric oxide production in RAW264.7 macrophages at 100 µg/mL. UPLC-QTOF/MS analysis tentatively identified seven biflavonoids, including amentoflavone (1), 2,3-dihydroamentoflavone (2), robustaflavone (3), 7″-methoxyrobustaflavone (4), hinokiflavone (5), isocryptomerin (6), and neocryptomerin (7). Among them, five compounds exhibited strong inhibitory activities against both cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2). Amentoflavone showed the most potent inhibition, with IC50 values of 2.35 ± 0.05 µM for COX-1 and 5.14 ± 0.16 µM for COX-2. In addition, a validated HPLC method was established for simultaneous quantification. Quantitative analysis of 11 batches revealed pronounced variability in biflavonoid content across different geographical origins, highlighting the necessity of multi-marker quality evaluation. Overall, these results provide insights into the chemical basis, anti-inflammatory activity, and quality evaluation of S. tamariscina.
    Keywords:  Selaginella tamariscina; anti‐inflammatory activity; biflavonoids; multicomponent analysis; quality control
    DOI:  https://doi.org/10.1002/cbdv.71422
  47. Chem Biodivers. 2026 Jun;23(6): e71384
    Comparative Phytochemical Profiling, Antioxidant, and Enzyme Inhibitory Activities of Different Parts of Aspilia africana Extracted by Maceration and Soxhlet Methods.
    Koaudio Ibrahime Sinan, Shakeel Ahmed, Ouattara Katinan Etienne, Cziaky Zoltan, Josef Jeko, Milena Terzic, Gokhan Zengin.
      Aspilia africana is a popular medicinal plant mainly found in Africa, with a variety of ethnobotanical uses. The purpose of this research is to make a comparative assessment of the phytochemical content, antioxidant activity, and enzyme inhibition activity of different parts of A. africana prepared by two conventional methods. Leaves, flowers, and the bark of A. africana were subjected to maceration and Soxhlet extraction, and detailed chemical analysis of each of the extracts was carried out using HPLC-ESI-Q-TOF-MS. Antioxidant activities of the extracts were estimated using the following methods: DPPH, ABTS, FRAP, CUPRAC, and MCA, while the enzyme inhibitor activity of the extracts was determined on AChE, tyrosinase, α-amylase, and α-glucosidase. Results from HPLC-ESI-Q-TOF-MS indicated the existence of phenolics (chlorogenic acid, caffeic acid, p-coumaric acid), flavonoids (rutin, quercetin-3,4'-di-O-glucoside), coumarins (scopoletin-7-O-glucoside, esculetin (6,7-dihydroxycoumarin)), and terpenes glycosides among the main active compounds of A. africana responsible for these activities. The highest antioxidant activities were obtained from the leaf extract obtained by Soxhlet extraction. Maceration extracts of flowers and leaves provide the maximum inhibition of tyrosinase (60.38±0.18 mg KAE/g). These results show A. africana as a promising source of antioxidant compounds with their potential applications in neurodegenerative disorders, managing diabetes, and cosmetic formulations.
    Keywords:  Aspilia africana; HPLC‐ESI‐Q‐TOF‐MS; Soxhlet; antidiabetics; antioxidant; neuroprotective
    DOI:  https://doi.org/10.1002/cbdv.71384
  48. Toxicon. 2026 Jun 17. pii: S0041-0101(26)00213-8. [Epub ahead of print] 109195
    Cyanobacterial Secondary Metabolites: genotoxicity and carcinogenicity in silico evaluation.
    Mylena Lemes Cunha, Cynthia Bomfim Pestana, Daniela Morais Leme.
      Cyanobacterial secondary metabolites are considered to have potential as biologically active compounds (e.g., antibacterial, antifungal) and are attracting great interest in medical applications and industrial innovation. However, it is well established that a large proportion of newly identified molecules, including natural compounds, are ultimately rejected due to adverse effects on human health. In silico tools could be a rapid and efficient way to screen the toxicity properties of secondary metabolites with potential biotechnological applications. In this study, 25 cyanobacterial secondary metabolites with known bioactivity were selected from CyanoMetDB (Cyanobacterial Secondary Metabolites Database) - a comprehensive and publicly available database - and screened for the toxicological endpoints genotoxicity and carcinogenicity using different in silico models and/or tools (VEGA, OECD QSAR Toolbox, T.E.S.T. (Toxicity Estimation Software Tool), ADMETlab, Carcinogenicity Predictor). A majority-consensus approach was applied to integrate the predictions. Seven secondary metabolites were predicted as non-genotoxic and non-carcinogenic, suggesting they could be promising candidates for biocidal and/or other biotechnological applications. The results support integrating in silico approaches for screening and prioritizing cyanobacterial secondary metabolites, while highlighting limitations related to the applicability domain and the underrepresentation of complex natural products in model training sets. In the future, NAMs-based data generated for microbial secondary metabolites could improve the representation of these compounds in the in silico model datasets, thereby increasing the robustness of the predictions.
    Keywords:  carcinogenicity; cyanobacteria; genotoxicity; in silico predictions; secondary metabolites
    DOI:  https://doi.org/10.1016/j.toxicon.2026.109195
  49. Rapid Commun Mass Spectrom. 2026 Sep 15. 40(17): e70127
    Metabolic Stability and Metabolic Profile of Schisandrol B in Rat and Human Liver Microsomes Employing HPLC-MS/MS Analysis.
    Tao Hu, Yicong Bian, Sheng Ma, Linsheng Liu, Qingqing Deng, Liyan Miao.
       RATIONALE: Schisandrol B (Sol B) is a major bioactive lignan compound extracted from the dried fruits of Schisandra chinensis (Turcz.) Baill. with comprehensive pharmacological activities. The study aims to investigate the metabolic stability and metabolic characteristics of Sol B in rat liver microsomes (RLMs) and human liver microsomes (HLMs) and further clarify the metabolic differences among different species.
    METHOD: The in vitro incubation systems included 5 μM Sol B and 0.5 mg/mL HLMs or RLMs in the presence of NADPH (5 mM) at 37°C in 200 μL phosphate-buffered saline. The residual amount of Sol B in the incubation system at different time points was determined to calculate the metabolic stability. The main metabolites of Sol B were identified and further elucidated the main metabolic pathways of Sol B.
    RESULTS: After incubation in RLMs and HLMs for 60 min, the remaining percentage of parent drug was 26.42% and 55.04%, and the t1/2 was calculated as 33.6 min and 75.21 min, respectively. A total of seven metabolites were detected in RLMs and HLMs systems. Metabolites M402-1 and M384-1 were detected only in the RLMs incubation system, while all other metabolites were monitored in both RLMs and HLMs incubation systems. Main metabolic pathways of Sol B in RLMs and HLMs mainly include dehydration, O-demethylation, and oxidative dehydrogenation.
    CONCLUSIONS: The metabolic behavior of Sol B in RLMs and HLMs exhibited certain species differences. Overall, Sol B was more stable in HLMs, with relatively simpler metabolic pathways.
    Keywords:  HPLC‐MS/MS; metabolic profile; metabolic stability; schisandrol B
    DOI:  https://doi.org/10.1002/rcm.70127
  50. Biomed Chromatogr. 2026 Jul;40(7): e70513
    Development and Validation of a Stability-Indicating RS HPLC Method for Leuprolide Acetate and Its Related Substances: Characterization of Degradation Products With LC-MS.
    Pydi Simhachalam, Thatituri Sabithakala.
      A new stability-indicating related substance method was developed and validated for leuprolide acetate (LPA) and its related impurities and established the degradation profile for the LPA drug that was exposed to various stress conditions. During the stress study, the drug showed substantial degradation in acidic (14.34%), basic (7.47%), and thermal (5.70%) conditions. The high-performance liquid chromatography method was developed as an isocratic method, mobile phase A (triethylamine pH 3.0), and mobile phase B (acetonitrile/n-propanol [300:200]). The chromatographic separation was attained by using an analytical HPLC column Inertsil ODS 3; 100 × 4.6 mm 3 μ. The degradation products (DPs) were characterized by using the LC-MS with an electron spray ionization technique. The method was validated according to ICH guidelines. The method showed a good limit of detection (0.092-0.274 μg/mL) and quantification limits (0.244-0.829 μg/mL) for LPA and its known impurities. During the linearity test, the method showed correlation coefficient (R2) values between 0.9999 and 0.9990. The method showed acceptable precision % of RSD values (0.01-5.70) and accuracy values in terms of % of recovery (95.10-109.60) for LPA and its related impurities.
    DOI:  https://doi.org/10.1002/bmc.70513
  51. Analyst. 2026 Jun 16.
    An ultra-performance convergence chromatography tandem mass spectrometry strategy for quantification of monodisperse polyethylene glycol polymers coupled with multiple fragmentation to enhance sensitivity.
    Jiarui Zhang, Hui Miao, Jiale Liu, Xinyue Zhou, Yue Deng, Shuang Feng, Yalin Xi, Meiyun Shi, Lei Yin.
      Accurate quantification of hydrophilic monodisperse polyethylene glycol (PEG) derivatives in complex biological matrices remains challenging due to substantial matrix interference and poor retention in conventional chromatographic systems. This study presents an environmentally sustainable analytical strategy based on ultra-performance convergence chromatography coupled with triple-stage mass spectrometry (UPC2-MS3) for the highly selective determination of HO-PEG8-OH in MCF-7 cell lysates. By harnessing the orthogonal separation mechanism of supercritical CO2 and the exceptional noise reduction capability of MS3 technology, the developed method achieves a rapid analysis time of 1.7 minutes while eliminating the need for sample pretreatment. The sustainability and practical applicability of the platform were systematically evaluated using a triple-metric framework, yielding favorable scores for GEMAM (7.166), AGSA (72.22), and CACI (75). Application of this sensitive approach to intracellular pharmacokinetic studies revealed an extremely low internalization rate of 0.0355% following 48 h exposure. Compared to existing methodologies, the UPC2-MS3 strategy offers a more robust, selective, and eco-friendly tool for elucidating the biological fate of monodisperse polymers in cellular systems.
    DOI:  https://doi.org/10.1039/d6an00305b
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