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Keywords = high performance liquid chromatography

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23 pages, 1157 KB  
Review
Unifying Phytochemistry, Analytics, and Target Prediction to Advance Dendropanax morbifera Bioactive Discovery
by SuHyun Kim, Damhee Lee, Kyujeong Won, Jinseop Lee, Wooseop Lee, Woohyeon Roh and Youngjun Kim
Life 2026, 16(1), 100; https://doi.org/10.3390/life16010100 (registering DOI) - 11 Jan 2026
Abstract
Dendropanax morbifera (DM; “Hwangchil”) is an evergreen tree native to southern Korea and Jeju Island, traditionally used for detoxification, anti-inflammatory, immunomodulatory, and neuroprotective purposes. Recent studies indicate that DM extracts and their constituents exhibit a broad range of biological activities, including antioxidant, anti-inflammatory, [...] Read more.
Dendropanax morbifera (DM; “Hwangchil”) is an evergreen tree native to southern Korea and Jeju Island, traditionally used for detoxification, anti-inflammatory, immunomodulatory, and neuroprotective purposes. Recent studies indicate that DM extracts and their constituents exhibit a broad range of biological activities, including antioxidant, anti-inflammatory, antimicrobial, anticancer, antidiabetic, hepatoprotective, and neuroprotective effects. Phytochemical investigations have revealed a chemically diverse profile comprising phenolic acids, flavonoids, diterpenoids, triterpenoids—most notably dendropanoxide—and polyacetylenes, with marked variation in compound distribution across plant parts. Despite this progress, translational application remains constrained by the lack of standardized extraction protocols, substantial variability in high-performance liquid chromatography (HPLC) methodologies, and limited mechanistic validation of reported bioactivities. This review proposes an integrated framework that links extraction strategies tailored to compound class and plant part with standardized C18 reverse-phase HPLC conditions to enhance analytical reproducibility. In parallel, in silico target prediction using SwissTargetPrediction is applied as a hypothesis-generating approach to prioritize potential molecular targets for subsequent experimental validation. By emphasizing methodological harmonization, critical evaluation of evidence levels, and systems-level consideration of multi-compound interactions, this review aims to clarify structure–activity relationships, support pharmacokinetic and safety assessment, and facilitate the rational development of DM-derived materials for medical, nutritional, and cosmetic applications. Full article
(This article belongs to the Section Pharmaceutical Science)
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16 pages, 1104 KB  
Article
Inhibitory Effect of Trichoderma longibrachiatum on Growth of Fusarium Species and Accumulation of Fumonisins
by Ruiqing Zhu, Ying Li, María Viñas, Qing Kong, Manlin Xu, Xia Zhang, Xinying Song, Kang He and Zhiqing Guo
J. Fungi 2026, 12(1), 49; https://doi.org/10.3390/jof12010049 (registering DOI) - 10 Jan 2026
Abstract
Fusarium spp. cause devastating crop diseases and produce carcinogenic mycotoxins such as fumonisins, threatening global food safety and human health. In this study, Trichoderma longibrachiatum A25011, isolated from apples in Aksu, Xinjiang, exhibited significant antagonistic activity with mycelial growth inhibition rates of 54.52% [...] Read more.
Fusarium spp. cause devastating crop diseases and produce carcinogenic mycotoxins such as fumonisins, threatening global food safety and human health. In this study, Trichoderma longibrachiatum A25011, isolated from apples in Aksu, Xinjiang, exhibited significant antagonistic activity with mycelial growth inhibition rates of 54.52% against F. verticillioides 48.62% against F. proliferatum, and 58.22% against F. oxysporum in confrontation assays. Enzyme activity detection revealed high chitinase (583.21 U/mg protein) and moderate cellulase (43.92 U/mg protein) production, which may have the capacity to degrade fungal cell walls. High-Performance Liquid Chromatography–Mass Spectrometry (HPLC-MS/MS) analyses enabled the quantification of fungal hormones including gibberellin A3 (GA3, 2.44 mg/L), cytokinins (cis-zeatin riboside (CZR): 0.69 mg/L; trans-zeatin riboside (TZR) : 0.004 mg/L; kinetin: 0.006 mg/L), and auxins (indole-3-acetic acid (IAA) : 0.35 mg/L; abscisic acid: 0.06 mg/L). Application of a T. longibrachiatum A25011 spore suspension around the roots of peanut plants enhanced growth by 13.20% (height), 5.65% (stem and leaf biomass), and 39.13% (root biomass). Notably, A25011 reduced F. proliferatum-derived fumonisin accumulation in rice-based cultures by 93.58% (6 d) and 99.35% (10 d), suggesting biosynthetic suppression. The results demonstrated that T. longibrachiatum strain A25011 exhibited excellent biocontrol capability against Fusarium spp., proving its dual role in simultaneously suppressing fungal growth and fumonisin accumulation while promoting plant growth. T. longibrachiatum A25011 could be applied as a multifunctional biocontrol agent in sustainable agriculture in the future. Full article
(This article belongs to the Special Issue Advances in the Control of Plant Fungal Pathogens)
15 pages, 1418 KB  
Article
Characterization of Newly Synthesized Nanobiomaterials for the Treatment of White Spot Lesions
by Andra Clichici, Diana Dudea, Cristina Gasparik, Camelia Alexandra Coadă, Ioana Bâldea, Stanca Cuc and Mărioara Moldovan
Biomolecules 2026, 16(1), 112; https://doi.org/10.3390/biom16010112 - 8 Jan 2026
Viewed by 54
Abstract
Background: White spot lesions (WSLs) are characterized by enamel demineralization. Minimally invasive treatments using infiltrating resins, such as the commercially available Icon®, are recommended. The need for such treatments justifies ongoing research into developing materials that can address existing limitations regarding [...] Read more.
Background: White spot lesions (WSLs) are characterized by enamel demineralization. Minimally invasive treatments using infiltrating resins, such as the commercially available Icon®, are recommended. The need for such treatments justifies ongoing research into developing materials that can address existing limitations regarding strength, durability, and biocompatibility. Objectives: This study aimed to synthesize and characterize four novel nanobiomaterials by evaluating their physicochemical properties and biocompatibility compared to the commercial material Icon®. Materials and methods: The recipes for the experimental nanobiomaterials NB3, NB6, NB3F, and NB6F contain varying proportions of TEGDMA, UDMA, HEMA, Bis-GMA, and HAF-BaF2 glass. Mechanical and physicochemical characteristics were evaluated, such as flexural strength, measured using the three-point test; water absorption and solubility; fluoride release; polymerization conversion; and residual monomers, assessed using High-Performance Liquid Chromatography (HPLC). In vitro cell viability was assessed via colorimetry using human dysplastic oral keratinocytes (DOKs). Results: NB6 and NB6F demonstrated the greatest polymerization potential. NB3 exhibited the lowest water absorption and solubility due to its hydrophobic nature. Additionally, the inclusion of UDMA enhanced the strength and elasticity of NB3 when compared to NB6. Among the samples with fluoride additives (NB3F and NB6F), the highest fluoride release on day 7 occurred with the material lacking UDMA. In contrast, the NB3F sample containing UDMA released the least amount of fluoride on the same day. In quantitative terms, NB3 and NB6F exhibited the lowest levels of residual monomers, whereas NB6 showed the highest levels. Both NB3 and NB6 were significantly better tolerated by the cells, showing higher cell viability compared to the commercial material Icon®. Conclusions: The materials’ mechanical and physicochemical properties varied with component proportions, enabling identification of a suitable formulation for targeted clinical applications. Biocompatibility tests showed that the experimental NB3 and NB6 were better tolerated than Icon®. Furthermore, the incorporation of filler particles improved the mechanical strength of the experimental nanobiomaterials. Full article
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19 pages, 1836 KB  
Protocol
Decoding Cerebrospinal Fluid: Integrative Metabolomics Across Multiple Platforms
by Antoine Presset, Sylvie Bodard, Antoine Lefèvre, Edward Oujagir, Camille Dupuy, Jean-Michel Escoffre and Lydie Nadal-Desbarats
Methods Protoc. 2026, 9(1), 8; https://doi.org/10.3390/mps9010008 - 8 Jan 2026
Viewed by 138
Abstract
Cerebrospinal fluid (CSF) is a key biological matrix that reflects the physiological and pathological states of the central nervous system (CNS). It supports brain function by regulating ionic balance, facilitating molecular transport, and clearing metabolic waste. In this article, we present a standardized [...] Read more.
Cerebrospinal fluid (CSF) is a key biological matrix that reflects the physiological and pathological states of the central nervous system (CNS). It supports brain function by regulating ionic balance, facilitating molecular transport, and clearing metabolic waste. In this article, we present a standardized protocol for CSF collection along with an integrative multiplatform metabolomic workflow that combines proton nuclear magnetic resonance spectroscopy (1H-NMRS) and high-performance liquid chromatography coupled to mass spectrometry (HPLC-MS). Integrating these complementary analytical modalities enhances metabolite coverage and improves analytical robustness, enabling a more comprehensive and reliable characterization of the CSF metabolome. This workflow supports the discovery of potential biomarkers and advances our understanding of neurochemical alterations within the CNS. Full article
(This article belongs to the Section Omics and High Throughput)
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23 pages, 1668 KB  
Article
Evaluation of In Vitro Cytoprotective Activity, Antioxidant Activity and Proteomic Profiles of Novel Sorghum-Based Fermented Beverages
by David R. Katerere, Abel Navarré Dopazo, Raffaele Sessa, Silvia Trombetti, Michela Grosso and Luana Izzo
Beverages 2026, 12(1), 9; https://doi.org/10.3390/beverages12010009 - 8 Jan 2026
Viewed by 221
Abstract
Fermentation, one of the oldest food processing techniques, is known to play a pivotal role in improving the nutritional and functional characteristics of cereals, with positive implications for gut health and overall well-being. The present study aims to examine the phenolic acids, peptides, [...] Read more.
Fermentation, one of the oldest food processing techniques, is known to play a pivotal role in improving the nutritional and functional characteristics of cereals, with positive implications for gut health and overall well-being. The present study aims to examine the phenolic acids, peptides, and potential bioactive properties of 2 novel sorghum-based fermented beverages, Niselo and Delishe. A total of 48 phenolic compounds were identified through targeted and untargeted Ultra-High Performance Liquid Chromatography coupled with a Quadrupole Orbitrap High-Resolution Mass Spectrometer (UHPLC–Q-Orbitrap HRMS) analyses, revealing a higher content of phenolic acids in Niselo and a prevalence of flavonoids in Delishe. Niselo exhibited enhanced in vitro cytoprotective and reactive oxygen species (ROS)-scavenging activity and displayed a clear cytoprotective effect against hydrogen peroxide-induced oxidative stress in Caco-2 cells. Proteomic profiling using tryptic digestion revealed that Niselo has a substantial abundance of fragments of peptides matching several stress-related and antioxidant proteins, indicating a superior stress-response and/or defense capability. Overall, these findings highlight the functional potential of sorghum-based fermented beverages, supporting their role as health-promoting products. Full article
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24 pages, 18899 KB  
Article
Network Pharmacology of the Phytochemical Content of Sunflower Seed (Helianthus annuus L.) Extract from LC-MS on Wound-Healing Activity and the In Vitro Wound Scratch Assay
by Juthamat Ratha, Tanit Padumanonda, Chawalit Yongram, Pimolwan Siriparu, Suthida Datham, Muhammad Subhan, Chatchavarn Chenboonthai and Ploenthip Puthongking
Plants 2026, 15(2), 187; https://doi.org/10.3390/plants15020187 - 7 Jan 2026
Viewed by 265
Abstract
Sunflower seeds have been reported to be a healthy natural source of polyphenols. This study aimed to explore the mechanisms of potential compounds in sunflower seed extract involved in wound healing; major compounds were investigated through network pharmacology and molecular docking. In an [...] Read more.
Sunflower seeds have been reported to be a healthy natural source of polyphenols. This study aimed to explore the mechanisms of potential compounds in sunflower seed extract involved in wound healing; major compounds were investigated through network pharmacology and molecular docking. In an in vitro wound-healing assay applied using an immortalised human keratinocyte (HaCaT) cell model, 10 µg/mL of the sunflower seed extract promoted cell migration in HaCaT cells and led to complete wound closure after 24 h; at a 1 µg/mL concentration, it led to complete wound closure after 72 h. The sunflower seed extract presented moderate-to-strong antioxidant activity. Liquid chromatography–mass spectrometry and high-performance liquid chromatography were used to identify the major compounds present in the sunflower seed extract. Forty-seven compounds were identified, among which chlorogenic acid was the most abundant phenolic compound. Network pharmacology was used to identify wound-healing-related targets. In total, 252 proteins were linked to the 47 compounds. Cyto-Hubba analysis identified 10 hub proteins with a strong correlation with wound healing. Molecular docking was used to assess the ability of the major compounds in the sunflower seed extract to combat NF-κB1, EGFR, and MMP9. Chlorogenic acid showed higher binding affinity to all targets. Moreover, its pharmacokinetic properties were well distributed in the plasma (VDss = 0.377 log L/kg), and they were not a carcinogen and did not cause skin sensitisation. In conclusion, the findings suggest that the sunflower seed extract is a potential source of bioactive compounds that can enhance wound healing and can be developed to create a transdermal application. Full article
(This article belongs to the Section Phytochemistry)
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13 pages, 2202 KB  
Article
Differential Glycosylation Patterns in Parkinson’s Disease: Emphasis on Male-Specific Changes Identified via HILIC-LC-MS
by Béla Demeter, Adriána Kutás, Béla Viskolcz, Csaba Oláh, Edina Petercsák, Attila Garami and Csaba Váradi
Int. J. Mol. Sci. 2026, 27(1), 552; https://doi.org/10.3390/ijms27010552 - 5 Jan 2026
Viewed by 150
Abstract
Parkinson’s disease (PD) is a progressive neurodegenerative disorder primarily characterized by the degeneration of dopaminergic neurons, leading to significant motor and non-motor symptoms. This study investigates glycosylation patterns with a significant emphasis on male Parkinson’s Disease (PD) patients, revealing unique alterations distinguishing PD [...] Read more.
Parkinson’s disease (PD) is a progressive neurodegenerative disorder primarily characterized by the degeneration of dopaminergic neurons, leading to significant motor and non-motor symptoms. This study investigates glycosylation patterns with a significant emphasis on male Parkinson’s Disease (PD) patients, revealing unique alterations distinguishing PD from healthy states, utilizing high-performance liquid chromatography coupled with mass spectrometry (HILIC-LC-MS). Findings reveal significantly altered serum N-glycosylation profiles between male and female patients, with increased levels of high-mannose glycans and reduced mono-sialylated glycans in male patients. ROC curve analysis indicates that these glycan changes are the most important features for distinguishing PD from healthy states, with AUC values of 0.71 for M5 and 0.85 for M6. This study underscores the critical role of glycosylation in the pathophysiology of Parkinson’s disease and highlights its potential in early detection and monitoring of disease progression. Full article
(This article belongs to the Special Issue Mass Spectrometry Approaches as Clinical Diagnostic/Prognostic Tools)
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13 pages, 414 KB  
Article
Effects of High-Dose Vitamin D Supplementation and Physical Exercise on Vitamin D Metabolites in Professional Football Players: A Pilot Study
by Anna Książek, Aleksandra Zagrodna and Konrad Kowalski
Nutrients 2026, 18(1), 175; https://doi.org/10.3390/nu18010175 - 5 Jan 2026
Viewed by 270
Abstract
Background/Objectives: Vitamin D plays an important role in muscle metabolism and recovery, yet its kinetics during and after football-specific physical activity remain poorly understood. Therefore, this study aimed to determine whether physical effort during a football match influences the concentration of vitamin [...] Read more.
Background/Objectives: Vitamin D plays an important role in muscle metabolism and recovery, yet its kinetics during and after football-specific physical activity remain poorly understood. Therefore, this study aimed to determine whether physical effort during a football match influences the concentration of vitamin D metabolites and to explore the effect of a single high-dose cholecalciferol supplementation combined with physical exercise on the levels of vitamin D metabolites in professional football players. Methods: Twenty professional football players participated in a three-phase, randomized placebo-controlled pilot study. Baseline fitness and blood samples were collected, followed by pre- and post-match measurements during two games. In the final phase, half of the players received a single 500,000 IU dose of vitamin D3 before a simulated match. Blood samples were collected before and after each session to analyze vitamin D metabolites using the isotope-dilution liquid chromatography–tandem mass spectrometry (ID-LC-MS/MS) method. Results: Physical exercise during the football match significantly increased serum concentrations of 25-(OH)D3, 24,25-(OH)2D3, and 3-epi-25-(OH)D3 (by up to 25%, p < 0.001). Following supplementation, these effects were further amplified, with 25-(OH)D3 rising by 98% and 3-epi-25-(OH)D3 by 424% (p < 0.001). Significant alterations in vitamin D metabolite ratios after exercise and supplementation suggest enhanced metabolic turnover and dynamic regulation of vitamin D pathways in response to physical effort. Conclusions: Football-specific physical activity appears to stimulate the release of vitamin D metabolites. High-dose cholecalciferol supplementation was well tolerated and may rapidly increase vitamin D status in professional athletes. These findings may have implications for optimizing recovery and performance, though larger trials are needed. Full article
(This article belongs to the Section Sports Nutrition)
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23 pages, 2464 KB  
Article
Biosynthesis of UV-Absorbing Mycosporine-like Amino Acids and Transcriptomic Profiling of Differential Gene Expression in Green Microalga Under Abiotic Stresses
by Georgia Tsintzou, Evmorfia Bataka, Georgia Tagkalaki, Sofoklis Keisaris, Nikolaos Tsiropoulos, Nikolaos Labrou and Panagiotis Madesis
Int. J. Mol. Sci. 2026, 27(1), 537; https://doi.org/10.3390/ijms27010537 - 5 Jan 2026
Viewed by 144
Abstract
Microalgae display remarkable resilience to harsh environments, partly through the biosynthesis of diverse secondary metabolites. Cyanobacteria and red algae are well known to produce mycosporine-like amino acids (MAAs)—low-molecular-weight, water-soluble UV-absorbing compounds with anti-inflammatory, anticancer, and antimicrobial activities. By contrast, green microalgae typically lack [...] Read more.
Microalgae display remarkable resilience to harsh environments, partly through the biosynthesis of diverse secondary metabolites. Cyanobacteria and red algae are well known to produce mycosporine-like amino acids (MAAs)—low-molecular-weight, water-soluble UV-absorbing compounds with anti-inflammatory, anticancer, and antimicrobial activities. By contrast, green microalgae typically lack detectable MAAs under standard conditions, and their responses under abiotic stress remain poorly characterized. Here, we investigated the freshwater green microalga Jaagichlorella luteoviridis grown under three stressors (salinity, heat, and UV) and assessed MAA induction. High-performance liquid chromatography (HPLC) revealed that stressed cultures accumulated multiple MAAs, whereas untreated controls showed no such accumulation. All stress treatments (UV, salinity, and heat) produced a substantial increase in peak intensity at 323–350 nm, whereas the control samples showed significantly lower absorption in this region. We also optimized an MAA extraction protocol suitable for “green” downstream applications in the pharmaceutical, nutraceutical, and cosmeceutical sectors and formulated an emulsion showing preliminary positive results and exhibiting an increased SPF index from 3.60 (control) to 3.78 when 0.2% MAA extract was added. Transcriptomic profiling against a reference genome revealed stress-specific differential gene expression and overexpression of specific genes of the MAA pathway, like ArioC and AroM/Aro1 SAM methyltransferases, thus identifying candidate targets for engineering enhanced MAA production. Given market demand for environmentally friendly and safe bioactives, microalgae represent a promising source of these valuable molecules. Full article
(This article belongs to the Special Issue Recent Research of Natural Products from Microalgae and Cyanobacteria)
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20 pages, 3007 KB  
Article
Plant-Derived Secondary Metabolites Tetrahydropalmatine and Rutaecarpine Alleviate Paclitaxel-Induced Neuropathic Pain via TRPV1 and TRPM8 Modulation
by Keun-Tae Park, Hyesang Yun, Juyeol Kang, Jae-Chul Lee and Woojin Kim
Metabolites 2026, 16(1), 46; https://doi.org/10.3390/metabo16010046 - 4 Jan 2026
Viewed by 193
Abstract
Background: Chemotherapy-induced peripheral neuropathy (CIPN) is a major dose-limiting adverse effect of paclitaxel and is characterized by cold and mechanical allodynia. Effective therapeutic strategies for CIPN remain limited. This study evaluated the analgesic potential of Corydalis yanhusuo (CY) and Evodia rutaecarpa (ER), as [...] Read more.
Background: Chemotherapy-induced peripheral neuropathy (CIPN) is a major dose-limiting adverse effect of paclitaxel and is characterized by cold and mechanical allodynia. Effective therapeutic strategies for CIPN remain limited. This study evaluated the analgesic potential of Corydalis yanhusuo (CY) and Evodia rutaecarpa (ER), as well as their major alkaloids tetrahydropalmatine (THP) and rutaecarpine, in a mouse model of paclitaxel-induced neuropathic pain. Methods: Neuropathic pain was induced by paclitaxel administration (2 mg/kg, i.p., four injections). CY and ER extracts were orally administered at doses of 100 or 300 mg/kg, either alone or in combination, and cold and mechanical allodynia were assessed from days 0 to 8. The analgesic effects of THP and rutaecarpine were also examined. Gene and protein expression analyses were performed to evaluate the involvement of TRPV1 and TRPM8 signaling pathways, and high-performance liquid chromatography (HPLC) was used to confirm the presence of THP in CY and rutaecarpine in ER. Results: Paclitaxel reliably induced robust cold and mechanical hypersensitivity. Oral administration of CY or ER significantly alleviated allodynia in a dose-dependent manner, with greater efficacy at 300 mg/kg. Combined CY–ER treatment produced stronger anti-allodynic effects than either extract alone. THP and rutaecarpine also exhibited dose-dependent analgesic effects, and their co-administration yielded the most pronounced inhibition of paclitaxel-evoked hypersensitivity. Molecular analyses confirmed the involvement of TRPV1- and TRPM8-related pathways in these analgesic effects. Collectively, these findings indicate that CY, ER, and their representative alkaloids effectively attenuate paclitaxel-induced neuropathic pain and highlight CY–ER-based natural products as promising candidates for managing CIPN through modulation of TRPV1/TRPM8 signaling. Full article
(This article belongs to the Special Issue Plant Metabolites for Managing Chemotherapy-Induced Side Effects)
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25 pages, 2002 KB  
Article
Potential Nutraceutical Properties of Vicia faba L: LC-ESI-HR-MS/MS-Based Profiling of Ancient Faba Bean Varieties and Their Biological Activity
by Francesca Fantasma, Gilda D’Urso, Alessandra Capuano, Ester Colarusso, Michela Aliberti, Francesca Grassi, Maria Chiara Brunese, Gabriella Saviano, Vincenzo De Felice, Gianluigi Lauro, Alfonso Reginelli, Maria Giovanna Chini, Agostino Casapullo, Giuseppe Bifulco and Maria Iorizzi
Molecules 2026, 31(1), 184; https://doi.org/10.3390/molecules31010184 - 4 Jan 2026
Viewed by 172
Abstract
Vicia faba L. is a widely cultivated legume known to contain numerous specialised metabolites. In this study, the seed coats and cotyledons of two ancient V. faba L. varieties, historically consumed in southern Italy and distinguished by black and purple seed coats, were [...] Read more.
Vicia faba L. is a widely cultivated legume known to contain numerous specialised metabolites. In this study, the seed coats and cotyledons of two ancient V. faba L. varieties, historically consumed in southern Italy and distinguished by black and purple seed coats, were extracted using 80% methanol and 80% ethanol. Extracts were analysed for total polyphenol, flavonoid and proanthocyanidin contents, and antioxidant activity using DPPH, ABTS, and FRAP assays. The purple seed coats exhibited the highest levels of phenolics and antioxidant capacity, exceeding those of black seed coats. Next, liquid chromatography coupled with high-resolution mass spectrometry (LC-ESI-HR-MS) was used to characterise the bioactive metabolites in both seed coats and cotyledons. The purple variety showed a higher phytochemical content, with a greater level of flavonoids and proanthocyanidins in methanolic extract. Furthermore, the purple seed coat exhibited in vitro anti-inflammatory activity by inhibiting soluble epoxide hydrolase (sEH), a key enzyme in the arachidonic acid cascade, with an IC50 of 31.51 ± 1.16 µg/µL. Elemental analysis was performed for both varieties to assess their nutritional value. Specifically, the purple seed coats were found to represent a valuable source of bioactive compounds and micronutrients, highlighting their potential applications in nutraceutical, cosmetic, and food supplement sectors. Full article
(This article belongs to the Special Issue Bioactive Food Compounds and Their Health Benefits)
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16 pages, 4803 KB  
Article
The Effect of Acute Supplementation of Branched Chain Amino Acids on Serum Metabolites During Endurance Exercise in Healthy Young Males: An Integrative Metabolomics and Correlation Analysis Based on a Randomized Crossover Study
by Xinxin Zhang, Xintang Wang, Chenglin Luan, Yizhang Wang, Junxi Li, Wei Shan, Zhen Ni, Chunyan Xu and Lijing Gong
Metabolites 2026, 16(1), 41; https://doi.org/10.3390/metabo16010041 - 2 Jan 2026
Viewed by 247
Abstract
Background: Branched-chain amino acids (BCAAs) are popular as sports supplements due to their ability to enhance performance and recovery. However, the full spectrum of metabolic alterations triggered by acute supplementation with BCAAs in conjunction with exercise remains incompletely understood. Methods: A randomized crossover [...] Read more.
Background: Branched-chain amino acids (BCAAs) are popular as sports supplements due to their ability to enhance performance and recovery. However, the full spectrum of metabolic alterations triggered by acute supplementation with BCAAs in conjunction with exercise remains incompletely understood. Methods: A randomized crossover trial was conducted in 8 healthy active young males, who received either BCAA or placebo supplementation for three consecutive days prior to a high-intensity cycling test. Plasma samples were collected pre- and post-exercise and analyzed by ultra-high-performance liquid chromatography–quadrupole time-of-flight mass spectrometry, followed by correlation and enrichment analyses. Results: Acute BCAA supplementation was significantly associated with enhanced fat oxidation and attenuated post-exercise increases in plasma ammonia, creatine kinase, and lactate dehydrogenase, suggesting the potential improvements in energy supply and membrane stability. Metabolomics analysis identified differential metabolites primarily involved in lipid, amino acid, and glucose metabolism. Pathway enrichment revealed coordinated regulation of fatty acid oxidation (FAO) and tryptophan-related pathways. Correlation analysis further showed that changes in metabolite profiles were strongly associated with biochemical outcomes, particularly linking enhanced fat oxidation and ammonia clearance with BCAA intake. Conclusions: Short-term BCAA supplementation could enhance FAO and membrane stability via coordinated regulation of lipid and amino acid metabolism post exercise, supporting its potential role as a precision nutrition strategy. Full article
(This article belongs to the Special Issue The Role of Diet and Nutrition in Relation to Metabolic Health)
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15 pages, 1361 KB  
Article
Detecting and Grouping In-Source Fragments with Low-Energy Stepped HCD, Together with MS3, Increases Identification Confidence in Untargeted LC–Orbitrap Metabolomics of Plantago lanceolata Leaves and P. ovata Husk
by Vilmantas Pedišius, Tim Stratton, Lukas Taujenis, Valdas Jakštas and Vytautas Tamošiūnas
Metabolites 2026, 16(1), 42; https://doi.org/10.3390/metabo16010042 - 2 Jan 2026
Viewed by 288
Abstract
Background: Comprehensive and accurate compound composition characterization in natural sources has high relevance in food and nutrition, health and medicine, environmental and agriculture research areas, though profiling of plant metabolites is a challenging task due to the structural complexity of natural products. This [...] Read more.
Background: Comprehensive and accurate compound composition characterization in natural sources has high relevance in food and nutrition, health and medicine, environmental and agriculture research areas, though profiling of plant metabolites is a challenging task due to the structural complexity of natural products. This study delves into the identification and characterization of compounds within the Plantago genus, leveraging state-of-the-art analytical techniques. Methods: Utilizing an ultra-high-performance liquid chromatography (UHPLC) system in conjunction with Orbitrap™ IQ-X™ Tribrid™ mass spectrometer (MS), we employed a Phenyl-Hexyl HPLC column alongside optimized extraction protocols to analyze both husk and leaf samples. To maximize compound identification, we implemented data-dependent acquisition (DDA) methods including MS2 (ddMS2), MS3 (ddMS3), AcquireX™ deep scan, and real-time library search (RTLS). Results: Our results demonstrate a significant increase in the number of putatively yet confidently assigned compounds, with 472 matches in P. lanceolata leaves and 233 in P. ovata husk identified through combined acquisition methods. The inclusion of an additional fragmentation level (MS3) noticeably enhanced the confidence in compound annotation, facilitating the differentiation of isomeric compounds. Furthermore, the application of low-energy fragmentation (10 normalized collision energy (NCE) for higher-energy collisional dissociation (HCD)) improved the detection and grouping of MS1 fragments by 55% in positive mode and by 16% in negative mode, contributing to a more comprehensive analysis with minimal loss in compound identification. Conclusions: These advancements underscore the potential of our methodologies in expanding the chemical profile of plant materials, offering valuable insights into natural product analysis and dereplication of untargeted data. Full article
(This article belongs to the Section Advances in Metabolomics)
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10 pages, 1798 KB  
Article
Is the Reindeer Lichen Cladonia arbuscula Really Producing Isousnic Acid? A Chemotaxonomy Query
by Dagmar Ísleifsdóttir, Maonian Xu, Maia Biwersi, Marie-Jeanne Leblanc, Starri Heiðmarsson, Snæbjörn Pálsson, John L. Sorensen, Elvar Örn Viktorsson and Elín Soffía Ólafsdóttir
Molecules 2026, 31(1), 143; https://doi.org/10.3390/molecules31010143 - 1 Jan 2026
Viewed by 171
Abstract
Isousnic acid (isoUA) has been detected in a few usnic acid (UA)-producing lichens with chemotaxonomic values. IsoUA was first isolated from a specimen belonging to Cladonia arbuscula s.l. (referred to as C. mitis in the publication). However, the isolation and detection of isoUA [...] Read more.
Isousnic acid (isoUA) has been detected in a few usnic acid (UA)-producing lichens with chemotaxonomic values. IsoUA was first isolated from a specimen belonging to Cladonia arbuscula s.l. (referred to as C. mitis in the publication). However, the isolation and detection of isoUA in this Cladonia species have not been reproduced and confirmed with clear evidence. This study focused on C. arbuscula s.l. collected in Iceland and aimed to (1) identify the lichen specimen using DNA barcoding and (2) investigate whether isoUA is produced using a series of chromatographic methods. The fungal nuclear ribosomal internal transcribed spacer (nrITS) barcode was sequenced, and the specimen was identified as C. arbuscula, following recent circumscription recommendations. Routine metabolite profiling did not detect isoUA, and it could only be identified after vigorous chromatographic purification and concentration steps using flash chromatography and preparative high-performance liquid chromatography. IsoUA was found in trace quantities (~24 µg/g dry weight), which likely explains its absence in routine metabolite profiling. A rapid ultra-high-performance liquid chromatography (UHPLC) method using a pentafluorophenyl column was developed to separate UA and isoUA. Our study highlights the importance of an integrative approach combining DNA barcoding and detailed chromatographic analyses for lichen chemistry research. Full article
(This article belongs to the Section Chemical Biology)
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Article
Magnetic Deep Eutectic Solvent-Based Single-Drop Microextraction for the Determination of Triazine Herbicides in Environmental Waters
by Xinyuan Bi, Wenying Wu, Xiaorong Xue, Xu Jing and Qiang Zhang
Agronomy 2026, 16(1), 107; https://doi.org/10.3390/agronomy16010107 - 1 Jan 2026
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Abstract
A magnetic deep eutectic solvent-based single-drop microextraction technique coupled with high-performance liquid chromatography (MDES-SDME-HPLC) was established for the determination of five triazine herbicides in environmental water samples. MDES, used as the extraction solvent, was composed of heptanoic acid, methyltrioctylammonium chloride, and iron chloride. [...] Read more.
A magnetic deep eutectic solvent-based single-drop microextraction technique coupled with high-performance liquid chromatography (MDES-SDME-HPLC) was established for the determination of five triazine herbicides in environmental water samples. MDES, used as the extraction solvent, was composed of heptanoic acid, methyltrioctylammonium chloride, and iron chloride. This pretreatment method requires only 50 μL of MDES, thereby avoiding the use of large volumes of toxic organic solvents. The MDES containing the target triazine herbicides was rapidly separated from the aqueous matrix by applying an external magnetic force, thus eliminating the need for centrifugation or additional reagents to achieve phase separation. The method demonstrated a linear range of 0.2–20 μg L−1, with a limit of detection of 0.06 μg L−1. Recoveries obtained from different environmental water matrices ranged from 75.5% to 102.4%. The greenness of the method was confirmed using five independent green analytical assessment tools. This approach represents a green and efficient analytical technique for detecting triazine herbicides in environmental water samples. Full article
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