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Search Results (288)

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Keywords = UHPLC Q-ToF MS

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35 pages, 4379 KB  
Article
Microgreen-Based Juices: UHPLC Q-ToF MS Metabolomic Analysis, Antioxidant Properties, and Sensory Acceptability
by Spasoje D. Belošević, Danijel D. Milinčić, Jovana M. Marković, Uroš M. Gašić, Steva M. Lević, Verica B. Đorđević, Mirjana B. Pešić and Viktor A. Nedović
Beverages 2026, 12(6), 73; https://doi.org/10.3390/beverages12060073 - 15 Jun 2026
Viewed by 271
Abstract
The aim of this study was to develop novel and sensory-acceptable microgreens–apple juices and characterize them using UHPLC Q-ToF MS-based metabolomic analysis. The obtained juices showed reduced sugar content and mild acidity compared with the control apple juice. Untargeted analysis revealed 71 compounds, [...] Read more.
The aim of this study was to develop novel and sensory-acceptable microgreens–apple juices and characterize them using UHPLC Q-ToF MS-based metabolomic analysis. The obtained juices showed reduced sugar content and mild acidity compared with the control apple juice. Untargeted analysis revealed 71 compounds, including phenolics, betalains, and one glucosinolate. Semi-quantification confirmed a high content of sinapic acid and its derivatives and hydroxybenzoic acid in broccoli–apple juice, as well as a predominant amount of phenolic acid diglycosides and hydroxycinnamoyl–isocitric acid derivatives in amaranth–apple juice. Apigenin C-glycosides were the main phenolics in red beet–apple juice, with the highest content of cytisoside derivatives. All microgreen-based juices contained apple-derived flavan-3-ols, procyanidins, dihydrochalcones, and certain flavonols. The formulated juices exhibited promising antioxidant potential evaluated by several screening assays (TPC, DPPH, ABTS, and FRAP), consistent with their phytochemical profiles, while differences among them may be linked to the amounts of certain bioactive molecules, mainly derived from microgreens. Moreover, these juices showed high overall quality and consumer acceptability, successfully masking typical microgreens flavors. Overall, these cold-pressed microgreen–apple juices can be classified as novel, low-calorie, and highly sensory-acceptable beverages, containing diverse bioactive compounds from both apple and microgreens; however, further in vitro and in vivo evidence is needed to support claims regarding their functionalities. Full article
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14 pages, 717 KB  
Article
Larrea ameghinoi Speg. (Zygophyllaceae) “Jarilla Rastrera”: UHPLC-ESI-QTOF-MS Analysis, Antioxidant, Antimicrobial Properties, and Inhibition of Enzymes of Interest to Human Health
by Jessica Gómez, Silvana M. Sede, Belén Ariza Sampietro, Daniel Zaragoza-Puchol, María Elisa Bressan Merlo, Duilio Caballero, Beatriz Lima, Alejandro Tapia and Mario J. Simirgiotis
Antioxidants 2026, 15(6), 668; https://doi.org/10.3390/antiox15060668 - 26 May 2026
Viewed by 318
Abstract
Larrea ameghinoi Speg., an endemic species of Argentine Patagonia traditionally used in folk medicine to treat fever, stomach disorders, respiratory conditions, back pain, and as an emmenagogue, among others, still remains chemically and biologically underexplored compared to the other four members of the [...] Read more.
Larrea ameghinoi Speg., an endemic species of Argentine Patagonia traditionally used in folk medicine to treat fever, stomach disorders, respiratory conditions, back pain, and as an emmenagogue, among others, still remains chemically and biologically underexplored compared to the other four members of the genus. This study aimed to perform a comprehensive metabolomic characterization of methanolic extracts from two populations (EMLaSAO and EMLaMAQ) using ultra-high-resolution liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC–ESI–QTOF–MS) and to evaluate their antioxidant, antimicrobial, and enzyme-inhibitory activities of relevance to human health. Thirty-three compounds were tentatively identified by extensive UHPLC–MS analysis, including flavones, two major lignans, and oleanane-type triterpenes. Both extracts exhibited high phenolic content (215–239 mg of gallic acid equivalents (GAE)/g extract) and strong free radical scavenging activity, as evidenced by 2,2-diphenyl-1-picrylhydrazyl (DPPH, EC50 ≈ 10 μg/mL), ferric-reducing antioxidant power (FRAP), and Trolox equivalent antioxidant activity (TEAC) assays. In addition, significant inhibition of butyrylcholinesterase (IC50 ≈ 50 μg extract/mL) and α-glucosidase, together with selective antibacterial activity against methicillin-sensitive and resistant Staphylococcus aureus (MIC = 125 μg extract/mL), were recorded. These findings suggest that L. ameghinoi possesses a distinctive phytochemical composition conferring multitarget bioactivity, differing from other Larrea species dominated by lignans such as nordihydroguaiaretic acid (NDGA) and its derivatives. Overall, this work supports the potential of L. ameghinoi as a novel source of bioactive metabolites for managing oxidative stress-related disorders and opportunistic infections. This warrants future in vivo studies investigating biological activities associated with oxidative stress and their relevance to human health. Full article
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18 pages, 7081 KB  
Article
Sakuranetin, a Natural Flavonoid, Promising to Manage Grapevine Diseases
by Corentin Griffon, Brice Dautruche, Bilal Loumi, Abdouramane Dosso, David Lesur, Emilie Isidore, Morad Chadni, Florent Allais, Christian Magro, Charles Monteux and Florence Fontaine
Molecules 2026, 31(8), 1368; https://doi.org/10.3390/molecules31081368 - 21 Apr 2026
Viewed by 540
Abstract
Botrytis cinerea and Plasmopara viticola, the causal agents of grey mold and downy mildew, respectively, are two major grapevine pathogens whose control largely relies on synthetic fungicides, raising environmental and health concerns. Plant-derived secondary metabolites, particularly flavonoids involved in plant defense, represent [...] Read more.
Botrytis cinerea and Plasmopara viticola, the causal agents of grey mold and downy mildew, respectively, are two major grapevine pathogens whose control largely relies on synthetic fungicides, raising environmental and health concerns. Plant-derived secondary metabolites, particularly flavonoids involved in plant defense, represent promising sustainable alternatives. Among them, sakuranetin, a flavanone aglycone known for its antifungal activity in rice, remains poorly explored for grapevine protection. In this study, sakuranetin was purified from cherry branches (48 mg) and structurally characterized using UHPLC-ESI-QTOF-MS and NMR analyses. Its antifungal activity against B. cinerea and P. viticola was evaluated through in vitro, in vivo and in planta assays. For B. cinerea, our results showed a significant in vitro inhibition of mycelium growth, with EC50 values of 16.43 mg·L−1, while no protection of detached berries was observed. Against P. viticola, sakuranetin has no effect on the release of zoospores, but there is a total inhibition of spore germination at 1 mg·L−1 in vitro, confirmed in vivo on a foliar disc. In planta, no significant protection is observed at 25 mg·L−1, even if some targeted defense genes are induced. Further studies are needed to determine the best concentration of sakuranetin to use to manage B. cinerea and P. viticola in planta. Full article
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25 pages, 8322 KB  
Article
Untargeted Metabolomics Reveals Metabolic Reprogramming During Viable but Non-Culturable State Formation in Aeromonas hydrophila Under Preservative Stress
by Gururaj Moorthy, Jatuphol Pholtaisong, Anusara Wongkotsila, Soottawat Benjakul, Awanwee Petchkongkaew and Jirakrit Saetang
Foods 2026, 15(8), 1289; https://doi.org/10.3390/foods15081289 - 9 Apr 2026
Viewed by 1081
Abstract
Aeromonas hydrophila is a major seafood-borne pathogen capable of persisting under preservative-associated stress by entering a viable but non-culturable (VBNC) state, thereby evading culture-based detection. Here, untargeted metabolomics was applied as the primary analytical approach to elucidate metabolic reprogramming during VBNC formation under [...] Read more.
Aeromonas hydrophila is a major seafood-borne pathogen capable of persisting under preservative-associated stress by entering a viable but non-culturable (VBNC) state, thereby evading culture-based detection. Here, untargeted metabolomics was applied as the primary analytical approach to elucidate metabolic reprogramming during VBNC formation under seafood-relevant preservation conditions. Cells were incubated at 4 °C for 30 days in sodium benzoate-supplemented saline, comparing 0.85% NaCl (culturable condition) and 4% NaCl (VBNC-inducing condition), with sampling every 6 days. Under 4% NaCl with sodium benzoate, culturability declined from 6.18 log CFU/mL at day 0 to undetectable levels by day 30, while cell viability was retained, confirming VBNC induction. UHPLC–ESI–QTOF–MS profiling detected over 893 intracellular metabolic features, of which 518 metabolites were significantly altered between VBNC and culturable states at day 30. Principal component analysis revealed clear, time-dependent metabolic divergence, with the VBNC trajectory explaining 34.4% (PC1) and 11.5% (PC2) of total variance. Pathway enrichment analysis demonstrated significant remodeling of alanine, aspartate and glutamate metabolism (8/28 hits, FDR = 5.7 × 10−4); arginine biosynthesis (5/14 hits, FDR = 5.44 × 10−3); purine metabolism (10/70 hits, FDR = 8.34 × 10−3); and pyrimidine metabolism (7/39 hits, FDR = 1.35 × 10−2), indicating nitrogen conservation and metabolic downshifting. A robust biomarker panel, including depleted cyclic AMP, aminoadipic acid, hypotaurine, O6-CM-dG, and betaine, and enriched urocanic acid, pipecolic acid, proline, azelaic acid, and orcinol perfectly discriminated VBNC from culturable cells. These findings demonstrate that sodium benzoate-based preservation can induce a metabolically reprogrammed VBNC state in A. hydrophila, highlighting a hidden food safety risk beyond culture-based assessment. Full article
(This article belongs to the Section Foodomics)
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25 pages, 5908 KB  
Article
Mapping the Polar Neuro-Interactome of Garcinia mangostana Against the AD-PD-ALS Nexus
by Rahni Hossain, Sirirat Surinkaew, Pradoldej Sompol, Nasmah K. Bastaki, Rifat Jafrin, Nazim Sekeroglu and Jitbanjong Tangpong
Life 2026, 16(4), 580; https://doi.org/10.3390/life16040580 - 1 Apr 2026
Viewed by 670
Abstract
Background/Objectives: Neurodegenerative diseases like Alzheimer’s, Parkinson’s, and Amyotrophic lateral sclerosis (ALS) share common molecular pathways, including neuroinflammation and oxidative stress, which complicate the effectiveness of single-target treatments. Garcinia mangostana L. (mangosteen) has shown neuroprotective properties, but previous studies focused on lipophilic xanthones, [...] Read more.
Background/Objectives: Neurodegenerative diseases like Alzheimer’s, Parkinson’s, and Amyotrophic lateral sclerosis (ALS) share common molecular pathways, including neuroinflammation and oxidative stress, which complicate the effectiveness of single-target treatments. Garcinia mangostana L. (mangosteen) has shown neuroprotective properties, but previous studies focused on lipophilic xanthones, which have poor bioavailability and uncertain blood–brain barrier permeability. Methods: In the current study, polar metabolites from G. mangostana peel aqueous extract (GMPE) were assessed for potential multi-target interactions via UHPLC-QTOF-MS-based metabolomics, systems pharmacology, and molecular docking analysis. Further, in silico ADMET screening and network-based analyses assessed for overlap between GMPE compounds and genes associated with neurodegeneration (AD, PD, ALS). Results: Analysis of genes linked to AD, PD, and ALS revealed 121 common molecular targets influenced by GMPE metabolites. Network and enrichment analyses indicated that the compounds derived from GMPE may be involved in common pathways related to oxidative stress, neuroinflammation, and neuronal survival. Molecular docking analyses suggest that selected metabolites are likely to exhibit moderate binding affinities to their respective protein targets. Conclusions: The results presented in this study provide evidence that GMPE may possess potential multi-target interactions within common neurodegenerative pathways. However, since the data are based on computational and predictive approaches, these results should be considered hypothesis-generating and warrant further experimental validation. Full article
(This article belongs to the Special Issue Neurodegenerative Diseases: From Risk Factors to Treatments)
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17 pages, 3091 KB  
Article
Ultrasound-Assisted Extraction of Polyphenols from Hericium erinaceus: Optimization, Bioactivities and LC-MS-Based Chemical Profiling
by Hongfei Liu, Cong Zhao, Shuyue Pang, Yuting Shu, Lina Chen, Jing Wang and Helong Bai
Molecules 2026, 31(7), 1138; https://doi.org/10.3390/molecules31071138 - 30 Mar 2026
Viewed by 725
Abstract
In this study, the Box–Behnken Design (BBD) was adopted to optimize the ultrasound-assisted extraction (UAE) conditions of polyphenols from Hericium erinaceus (H. erinaceus) on the basis of single-factor experiments, with extraction time, solid–liquid ratio and ethanol concentration as the key investigation [...] Read more.
In this study, the Box–Behnken Design (BBD) was adopted to optimize the ultrasound-assisted extraction (UAE) conditions of polyphenols from Hericium erinaceus (H. erinaceus) on the basis of single-factor experiments, with extraction time, solid–liquid ratio and ethanol concentration as the key investigation factors. The optimal extraction parameters were determined as follows: extraction time of 56.85 min, solid–liquid ratio of 1:56.71 g/mL and ethanol concentration of 44.64%, under which the actual yield of the total polyphenol crude extract (TPCE) reached 0.9985 ± 0.03%, which was highly consistent with the theoretical predicted value of 0.9960%, verifying the good fitting degree of the established model. Taking L-ascorbic acid as the positive control, the antioxidant activity of TPCE was evaluated by determining its scavenging capacity against ABTS·+, ·OH and DPPH· free radicals, and the half-maximal effective concentration (EC50) values were measured to be 0.8850, 0.9490 and 4.198 mg/mL, respectively. With acarbose as the reference drug, the inhibitory effects of TPCE on α-amylase and α-glucosidase related to carbohydrate metabolism were assayed, and the corresponding half-maximal inhibitory concentration (IC50) values were 0.0135 and 130.3 mg/mL, respectively. Furthermore, ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) was employed for the tentative identification of bioactive components in TPCE, and a total of 48 and 64 chemical constituents were characterized in negative and positive ion modes, respectively, providing a chemical basis for the biological activities of TPCE. This study confirmed that UAE is an efficient and feasible technology for extracting polyphenols from H. erinaceus, which lays a theoretical foundation for the development and utilization of its polyphenols, and also provides novel insights into the development of natural functional ingredients and potential therapeutic agents for the intervention of type 2 diabetes. Additionally, the findings further validate edible fungi as a valuable reservoir of natural bioactive substances, with promising application prospects in the research and development of functional foods and pharmaceuticals targeting metabolic diseases. Full article
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20 pages, 8089 KB  
Article
Therapeutic Effects of Akebia quinata Seeds Through Apoptosis and Immunogenic Cell Death in Non-Small Cell Lung Cancer
by Mibae Jeong, In Jin Ha, Chang-Seob Seo, Mi-Kyung Jeong, Kwang Seok Ahn and Jaemoo Chun
Int. J. Mol. Sci. 2026, 27(7), 3114; https://doi.org/10.3390/ijms27073114 - 30 Mar 2026
Viewed by 709
Abstract
Plant-derived saponins have attracted significant interest for their potential to promote apoptotic cell death and enhance antitumor immune responses through immunogenic cell death (ICD). Akebia quinata, a saponin-rich medicinal plant, exhibits diverse pharmacological properties; however, studies on its seeds are limited, and [...] Read more.
Plant-derived saponins have attracted significant interest for their potential to promote apoptotic cell death and enhance antitumor immune responses through immunogenic cell death (ICD). Akebia quinata, a saponin-rich medicinal plant, exhibits diverse pharmacological properties; however, studies on its seeds are limited, and their immunomodulatory activity in cancer remains largely unexplored. In this study, A. quinata seeds were extracted using 70% ethanol, and the phytochemical profile was characterized using UHPLC–QTOF MS/MS. We investigated the anticancer properties of A. quinata seed extract (AQSE), focusing on its role in inducing apoptosis and ICD in non-small cell lung cancer (NSCLC). In human NSCLC cell lines (A549 and H460), AQSE exhibited potent cytotoxic effects in a dose-dependent manner. Flow cytometric analysis confirmed the induction of apoptosis, evidenced by a significant increase in Annexin V-positive cells and an elevated sub-G1 population. Mechanistically, AQSE treatment induced cell death by simultaneously inhibiting the survival-promoting MEK/ERK/CREB axis and activating the stress-responsive JNK pathway. Furthermore, AQSE triggered hallmark features of ICD, characterized by surface exposure of calreticulin and the release of extracellular HMGB1 and ATP. Most importantly, an in vivo vaccination assay using a syngeneic mouse model demonstrated that immunization with AQSE-treated dying cells significantly suppressed tumor growth upon rechallenge, confirming the establishment of antitumor immunological memory. Additionally, bioassay-guided fractionation revealed that the anticancer activity was primarily concentrated in the ethyl acetate fraction. These findings suggest that AQSE exerts anticancer effects via the induction of apoptosis and ICD, highlighting its potential as a promising natural candidate for the development of novel therapeutic strategies against NSCLC. Full article
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18 pages, 2060 KB  
Article
BPA Disrupts Hepatic Lipid and Carbohydrate Metabolism in Female Zebrafish: Protective Effects of Probiotics Revealed by FTIRI and Lipidomics
by Christian Giommi, Chiara Santoni, Fabrizia Carli, Amalia Gastaldelli, Francesca Maradonna, Hamid R. Habibi, Elisabetta Giorgini and Oliana Carnevali
Int. J. Mol. Sci. 2026, 27(7), 2978; https://doi.org/10.3390/ijms27072978 - 25 Mar 2026
Viewed by 629
Abstract
Bisphenol A (BPA) is a widespread endocrine disruptor that interferes with metabolism in humans and animals by inducing oxidative stress, lipid peroxidation, and cell death. Probiotics, conversely, have shown potential in promoting host health and reducing the toxicity of endocrine-disrupting chemicals (EDCs). This [...] Read more.
Bisphenol A (BPA) is a widespread endocrine disruptor that interferes with metabolism in humans and animals by inducing oxidative stress, lipid peroxidation, and cell death. Probiotics, conversely, have shown potential in promoting host health and reducing the toxicity of endocrine-disrupting chemicals (EDCs). This study examined whether sub-chronic BPA exposure disrupts hepatic lipid metabolism in female zebrafish (Danio rerio), and whether co-administration of probiotics mitigates these effects. Adult females were exposed for 28 days to the following treatments: 10 µg/L BPA via water (BPA); 109 CFU/g body weight/day of probiotic formulation (P); and both treatments (BPA+P). An untreated group served as a control (CTRL). Hepatic lipid composition was analyzed using UHPLC-QTOF-MS, while liver sections were investigated by Fourier Transform Infrared Imaging (FTIRI) spectroscopy. BPA exposure decreased 14 unsaturated triacylglycerols and lysophosphatidylcholine 18:0, suggesting steatosis onset and inflammation, while in the group exposed to BPA+P, the decrease was limited to 8 triacylglycerols and the reduction in lysophosphatidylcholine 18:0 was prevented. Analyses of pooled liver samples precluded modeling tank-level effects; thus, the results are interpreted as semi-quantitative. Partial least square discriminant analysis built on the comparison of all groups together confirmed an intermediate phenotype for BPA+P fish between BPA and P groups. The observed beneficial role of probiotics in counteracting BPA-related metabolic disturbances was also supported by FTIRI, evidencing the ability to mitigate the effects of BPA on lipid and glycosylated compound metabolism. These findings highlight the potential of probiotic supplementation as a practical and accessible strategy to mitigate BPA-induced metabolic disturbances, contributing to the development of mitigating approaches against environmental contaminant-related liver dysfunction. Full article
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25 pages, 5026 KB  
Article
DNA Barcoding and Chemical Profile Using UHPLC, GC-MS and LC-MS/QTOF of Mitragyna speciosa Variation and Allied Species for Quality Control of Kratom Materials
by Phunsuk Anantaworasakul, Warunya Arunotayanun, Siripat Chaichit, Suthiwat Khamnuan, Chatchai Ngernsaengsaruay, Chuda Chittasupho, Nisa Leksungnoen, Mingkwan Na Takuathung, Ruedeemars Yubolphan, Apisada Jiso, Tachpon Techarang and Aekkhaluck Intharuksa
Plants 2026, 15(7), 1003; https://doi.org/10.3390/plants15071003 - 25 Mar 2026
Viewed by 1172
Abstract
Kratom (Mitragyna speciosa Korth.) has gained increasing global attention due to its traditional use, psychoactive properties, and emerging therapeutic potential; however, concerns regarding adulteration, substitution, and inconsistent quality of commercial products necessitate robust authentication strategies. This study aimed to integrate DNA barcoding [...] Read more.
Kratom (Mitragyna speciosa Korth.) has gained increasing global attention due to its traditional use, psychoactive properties, and emerging therapeutic potential; however, concerns regarding adulteration, substitution, and inconsistent quality of commercial products necessitate robust authentication strategies. This study aimed to integrate DNA barcoding and comprehensive chemical profiling to authenticate kratom variants and discriminate them from closely allied Mitragyna species for quality control and forensic applications. Nine DNA barcoding regions were analyzed, alongside chemical characterization using UHPLC, GC–MS, and LC–MS/QTOF. Among the tested loci, the internal transcribed spacer (ITS) and ITS2 regions exhibited the highest interspecific variation and effectively distinguished kratom from allied species. UHPLC and GC–MS analyses confirmed that mitragynine was exclusively detected in kratom variants, with Kan Khiao exhibiting the highest content (94.33 ± 0.14 mg/g) when quantified against the mitragynine standard using UHPLC analysis. LC–MS/QTOF profiling revealed an alkaloid-rich chemotype in kratom dominated by mitragynine and 7-hydroxymitragynine, whereas M. diversifolia, M. hirsuta, and M. rotundifolia showed distinct profiles enriched in phenolic acids and flavonoid glycosides. Multivariate analyses further identified procyanidin B1, datiscetin-3-O-rutinoside, mitragynine, and 7-hydroxymitragynine as key discriminatory markers. Overall, the combined molecular and chemical workflow provides a robust framework for kratom authentication, supporting regulatory monitoring, quality assurance, and forensic identification of kratom materials. Full article
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28 pages, 4809 KB  
Article
Exploring the Multifaceted Phytochemical Profile of Nigella sativa and the Therapeutic Potential of Thymoquinone
by Mohamed A. Fareid, Gamal M. El-Sherbiny, Nancy M. Elafandy, Nagat E. Eltoum, Mohamed S. Othman, Mohamed Shawky, Ahmad S. El-Hawary, Fatma A. Hamada and Amira Salah El-Din Youssef
Pharmaceuticals 2026, 19(3), 503; https://doi.org/10.3390/ph19030503 - 18 Mar 2026
Cited by 2 | Viewed by 1567
Abstract
Background: Nigella sativa (black cumin) seeds are renowned for their ethnomedicinal significance and are rich in bioactive phytochemicals, which contribute to food preservation and the prevention of various diseases through their antimicrobial and antioxidant properties. Accordingly, this study aimed to characterize the [...] Read more.
Background: Nigella sativa (black cumin) seeds are renowned for their ethnomedicinal significance and are rich in bioactive phytochemicals, which contribute to food preservation and the prevention of various diseases through their antimicrobial and antioxidant properties. Accordingly, this study aimed to characterize the phytochemical composition of N. sativa seed extracts, isolate thymoquinone, and assess their antibacterial, antibiofilm, antioxidant, anti-inflammatory and antidiabetic activities. Methods: Nigella sativa seed extracts were prepared using solvents of increasing polarity and analyzed for phytochemical content. Metabolite profiling was performed using UHPLC/QTOF-MS. Thymoquinone, the major constituent, was isolated via thin-layer chromatography (TLC), further purified using semi-preparative reverse-phase high-performance liquid chromatography (RP-HPLC), and evaluated in vitro for antibacterial, antibiofilm, antioxidant, anti-inflammatory, and antidiabetic activities. Results: Extraction yields ranged from 5.5% to 8.4% (w/w), with methanol yielding the highest phenol (6.34 ± 0.31 mg GAE/mL) and flavonoid (5.12 ± 0.26 mg QE/mL) contents. UHPLC/QTOF-MS revealed a chemically diverse profile dominated by thymoquinone (58% relative abundance), alongside p-cymene, carvacrol, longifolene, and nigellidine. Thymoquinone (Rf = 0.56) was initially isolated from the methanolic extract with a yield of 270 mg/g and further purified from preparative TLC fractions using semi-preparative RP-HPLC, affording 82 mg of >95% pure compound with a 68.3% recovery, suitable for subsequent biological assays. It inhibited Gram-positive and Gram-negative bacteria, with MICs of 62.5 µg/mL against Staphylococcus aureus, Bacillus subtilis, and Listeria monocytogenes; 125–250 µg/mL against Escherichia coli and Salmonella typhimurium; and 500 µg/mL against Pseudomonas aeruginosa. Thymoquinone reduced biofilm formation (>80% at 25–50 µg/mL; MBIC50 ≈ 5.4–11.6 µg/mL), exhibited antioxidant activity (DPPH IC50 = 52.3 ± 2. 1 µg/mL; ABTS IC50 = 41.6 ± 1.9 µg/mL), stabilized erythrocyte membranes (IC50 ≈ 14.8 µg/mL), and inhibited carbohydrate-hydrolyzing enzymes, with stronger inhibition of α-glucosidase (~92%) than α-amylase (~84%) at 128 µg/mL. Conclusions: Thymoquinone is a major bioactive constituent of N. sativa seeds, exhibiting consistent multi-target in vitro activity. These findings highlight its functional relevance and in vivo investigations to establish therapeutic potential. Full article
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35 pages, 4909 KB  
Article
Metabolomics, Molecular Networking and Phytochemical Investigation of Psiadia dentata (Cass.) DC., Endemic to Reunion Island: Discovery of Novel Bioactive Molecules
by Lantomalala Elsa Razafindrabenja, Keshika Mahadeo, Gaëtan Herbette, Lúcia Mamede, Michel Frederich, Carole Di Giorgio, Béatrice Baghdikian, Patricia Clerc, Hippolyte Kodja, Isabelle Grondin and Anne Gauvin-Bialecki
Molecules 2026, 31(6), 973; https://doi.org/10.3390/molecules31060973 - 13 Mar 2026
Viewed by 793
Abstract
The genus Psiadia (Asteraceae), widely distributed in Madagascar and the Mascarene Islands (Mauritius, La Réunion, Rodrigues), is traditionally used to treat bronchitis, asthma, colds, abdominal pain, and other inflammatory disorders. However, few studies have scientifically validated these traditional medicinal uses. To assess P. [...] Read more.
The genus Psiadia (Asteraceae), widely distributed in Madagascar and the Mascarene Islands (Mauritius, La Réunion, Rodrigues), is traditionally used to treat bronchitis, asthma, colds, abdominal pain, and other inflammatory disorders. However, few studies have scientifically validated these traditional medicinal uses. To assess P. dentata as a valuable source of bioactive natural products, a combined 1H NMR-based metabolomic, molecular networking, and phytochemical study was conducted. Multivariate analysis (PLS-DA) of crude extracts from Psiadia species collected on Reunion Island enabled rapid discrimination of active extracts from P. dentata and revealed two methoxylated flavonoids and one coumarin as metabolites correlated with its antiplasmodial and anti-inflammatory activities. Additionally, UHPLC-DAD-ESI-QTOF-MS/MS molecular networking approach enabled detailed chemical profiling of this species, allowing the annotation of 25 compounds (125) in this species. Subsequent phytochemical investigation of P. dentata leaves led to the isolation and identification of 25 metabolites, including nine new diterpenes (2634), one new coumarin (35), and 15 known compounds (18, 11, 18, 19 and 3639) from the diterpenoid, flavonoid, and coumarin families. The structures of the new compounds were elucidated using spectroscopic methods, including extensive 1D and 2D NMR and HRESIMS analyses. Biological evaluation of the isolated compounds showed that compounds 1, 7, 26 and 27 showed antiplasmodial activity against Plasmodium falciparum (3D7 strain, IC50 = 7.25–13.46 μM). Compounds 7, 26, 27, 31 and 32 inhibited nitric oxide production (IC50 = 0.87–27.71 μM), indicating potential anti-inflammatory effects. Only compound 1 displayed moderate cytotoxicity against HepG2 and HT29 cancer cell lines (IC50 = 25.67 and 18.35 μM, respectively). Full article
(This article belongs to the Special Issue Chemical Constituents and Biological Activities of Natural Sources)
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14 pages, 2380 KB  
Article
Optimization of an Extraction Protocol for Untargeted Metabolomics of Vitis vinifera L. Leaves
by Laura Sánchez-Ochoa, Teresa Garde-Cerdán, Itziar Sáenz de Urturi, Miriam González-Lázaro and Eva P. Pérez-Álvarez
Plants 2026, 15(5), 696; https://doi.org/10.3390/plants15050696 - 26 Feb 2026
Cited by 1 | Viewed by 846
Abstract
Viticulture faces increasing challenges due to the susceptibility of Vitis vinifera L. to biotic and abiotic stresses, which trigger defense responses involving the synthesis of secondary metabolites. Untargeted metabolomics has become a powerful tool to explore these metabolic changes; however, the efficiency and [...] Read more.
Viticulture faces increasing challenges due to the susceptibility of Vitis vinifera L. to biotic and abiotic stresses, which trigger defense responses involving the synthesis of secondary metabolites. Untargeted metabolomics has become a powerful tool to explore these metabolic changes; however, the efficiency and reproducibility of metabolomic studies strongly depend on the extraction protocol used. Current literature shows variability in sample handling, solvent composition, and extraction conditions. This study aimed to optimize an extraction protocol for secondary metabolites in grapevine leaves to ensure high recovery of compounds relevant to untargeted metabolomics. Leaves of Vitis vinifera L. cv. Tempranillo were collected, pooled, frozen, and ground under liquid nitrogen. A factorial design was used to evaluate the effects of sample mass, sample-to-solvent ratio, and solvent type on extraction efficiency. Extracts were analyzed using UHPLC-QTOF-MS in both positive and negative ionization modes, and multivariate statistical tools (PCA and OPLS-DA) were used to identify discriminant metabolites. Optimal extraction was achieved using 750 mg of leaf powder, a sample-to-solvent ratio of 100 mg/Ml, and methanol 80% acidified with 0.1% of formic acid. This protocol maximizes the recovery of metabolites and provides a robust basis for future untargeted metabolomics studies of grapevine responses. Full article
(This article belongs to the Section Phytochemistry)
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14 pages, 1115 KB  
Article
Renoprotective Potential of Beetroot Spent Extract Under Hyperglycemic Conditions
by Wachiraporn Tipsuwan, Onsaya Kerdto, Phronpawee Srichomphoo, Wittaya Chaiwangyen, Pongsak Angkasith, Yanping Zhong and Somdet Srichairatanakool
Foods 2026, 15(4), 769; https://doi.org/10.3390/foods15040769 - 20 Feb 2026
Viewed by 719
Abstract
Diabetic nephropathy is a major complication of diabetes mellitus, primarily driven by hyperglycemia-induced oxidative stress and renal tubular cell injury. Beetroot (Beta vulgaris L.) is rich in antioxidant phytochemicals, and its industrial processing generates large amounts of spent material that may retain [...] Read more.
Diabetic nephropathy is a major complication of diabetes mellitus, primarily driven by hyperglycemia-induced oxidative stress and renal tubular cell injury. Beetroot (Beta vulgaris L.) is rich in antioxidant phytochemicals, and its industrial processing generates large amounts of spent material that may retain significant bioactive compounds. This study evaluated the phytochemical profile, antioxidant capacity, and renoprotective potential of beetroot spent extracts under hyperglycemic conditions. Beetroot spent material was extracted using hot water and 70% ethanol. Total phenolic, flavonoid, and betalain contents were quantified, and antioxidant activity was assessed using the 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging assay. Phytochemical characterization was performed by ultra-high-pressure liquid chromatography–electrospray ionization–quadrupole time-of-flight mass spectrometry (UHPLC-ESI-QTOF-MS). Cytotoxicity was evaluated in peripheral blood mononuclear cells, SH-SY5Y, HEK-293, and MDA-MB-231 cells using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Renoprotective effects were investigated in HEK-293 renal tubular cells cultured under normal (5.5 mM) and high-glucose (200 mM) conditions. UHPLC-ESI-QTOF-MS was used to identify over 80 phenolic and flavonoid compounds including quercetin, epicatechin, and epigallocatechin gallate. The hot water extract exhibited superior antioxidant activity, achieving approximately 90% ABTS radical inhibition. Beetroot spent extract showed no cytotoxicity at concentrations below 1 mg/mL and significantly restored HEK-293 cell viability (>90%) under high-glucose conditions at concentrations ≥31.25 µg/mL. In conclusion, beetroot spent water extract possesses strong antioxidant and renoprotective activities against hyperglycemia-induced renal cell damage, supporting its valorization as a sustainable functional food ingredient for diabetes-related health applications. Full article
(This article belongs to the Special Issue Health Benefits of Bioactive Compounds from Vegetable Sources)
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16 pages, 1592 KB  
Article
Development and Validation of an UHPLC-ESI-QTOF-MS Method According to the ICH M10 Guideline for Quantification of the Clinical Drug Candidate RD2 in the Mouse Brain
by Alissa Jonas, Ian Gering, Elena Schartmann, Sarah Schemmert, Dieter Willbold, Beatrix Santiago-Schübel and Janine Kutzsche
Analytica 2026, 7(1), 15; https://doi.org/10.3390/analytica7010015 - 7 Feb 2026
Viewed by 726
Abstract
The all-d-enantiomeric-peptide RD2 was developed for the treatment of Alzheimer’s disease. This study aimed to develop a specific and highly sensitive liquid chromatography-mass-spectrometric (UHPLC-ESI-QTOF) method for quantifying RD2 in the mouse brain and to validate it according to the ICH M10 [...] Read more.
The all-d-enantiomeric-peptide RD2 was developed for the treatment of Alzheimer’s disease. This study aimed to develop a specific and highly sensitive liquid chromatography-mass-spectrometric (UHPLC-ESI-QTOF) method for quantifying RD2 in the mouse brain and to validate it according to the ICH M10 guideline to investigate the pharmacokinetic profile of RD2 in its target organ. Sample preparation, chromatographic separation and quantification were very challenging due to RD2’s highly hydrophilic properties, the complex matrix and the required lower limit of quantification (LLOQ). Chromatographic separation was performed on an Acquity UPLC BEH C18 column (2.1 × 100 mm, 1.7 μm particle size) within 5 min at 50 °C with a flow rate of 0.5 mL·min−1. Mobile phases consisted of water and acetonitrile with 0.2% formic acid and 0.015% heptafluorobutyric acid. Ions were generated by electrospray ionization in the positive mode, and RD2 was quantified by QTOF-MS. The developed extraction method revealed complete recovery. The linearity of the calibration curve was in the range of 2 ng·mL−1 to 500 ng·mL−1 (R2 > 0.99) with a LLOQ of 5 ng·mL−1. The intraday and interday accuracy and precision ranged from 0.4% to 12.2% and from 1.0% to 12.0%. RD2 remained stable in the freshly homogenized brain even after several freeze–thaw cycles, but stability decreased over time during long-term storage at −80 °C. Using this validated method, RD2-spiked brain homogenate samples and samples of a pharmacokinetic study with RD2 in mice were analyzed. Full article
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18 pages, 3964 KB  
Article
Dynamic Metabolic Changes During Postmortem Aging of Yili Horsemeat Revealed by Untargeted Metabolomics
by Xixi Yang, Tongliang Wang, Jingtao Gan, Chen Meng, Jingxuan Shen, Zexu Li, Xueyan Li, Yaqi Zeng, Wanlu Ren, Xinkui Yao and Jun Meng
Animals 2026, 16(3), 508; https://doi.org/10.3390/ani16030508 - 5 Feb 2026
Viewed by 1140
Abstract
Postmortem aging is a crucial process for improving meat quality; however, the metabolic mechanisms underlying the postmortem maturation of Yili horsemeat remain unclear. In this study, untargeted metabolomics was employed to investigate the dynamic changes in metabolites during postmortem aging of Yili horse [...] Read more.
Postmortem aging is a crucial process for improving meat quality; however, the metabolic mechanisms underlying the postmortem maturation of Yili horsemeat remain unclear. In this study, untargeted metabolomics was employed to investigate the dynamic changes in metabolites during postmortem aging of Yili horse longissimus dorsi muscle. Samples from six Yili horses were subjected to aging at 4 °C for 0, 14, and 28 days. Intramuscular Environment and metabolic profiles were analyzed using UHPLC-QTOF-MS. Results showed that meat pH initially decreased and then increased over the aging period (p < 0.05), whereas muscle fiber diameter and cross-sectional area gradually declined (p < 0.05). Metabolomic analysis revealed significant differences in metabolic profiles among different aging stages. KEGG pathway enrichment analysis indicated that the differentially expressed metabolites (DEMs) were predominantly involved in purine metabolism, nucleotide metabolism, and amino acid biosynthesis pathways. Notably, the degradation of AMP and IMP into inosine and hypoxanthine and the accumulation of free amino acids such as tryptophan and phenylalanine were closely associated with the development of umami taste and overall flavor of horsemeat. These findings elucidate the key metabolic pathways regulating postmortem meat quality in Yili horses and provide a theoretical basis for its production and application. Full article
(This article belongs to the Section Equids)
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