Search Results (257)

The presence of Alternaria mycotoxins in hepatic tissue of both human and animal origin remains unexplored. This work describes the development of an analytical method based on salt-assisted liquid–liquid extraction (SALLE) and ultrahigh-performance liquid chromatography coupled to quadrupole time of flight mass spectrometry (UHPLC-QTOF-MS) for the determination of six main Alternaria mycotoxins and related metabolites. Sample treatment was fully optimized, including sample mass, extraction solvent, and volume and sodium chloride mass. The method was validated, achieving calibration curve R² values above 0.99 and limits of detection between 0.01 and 1.46 µg kg⁻¹. Moreover, satisfactory trueness (apparent recoveries between 84% to 111%) and precision (RSD values below 10%) were achieved, complying with EU requirements. Matrix effects in terms of signal suppression/enhancement varied between 53% for TeA and 78% for AME. Applied to real liver samples (20 human and 20 animal), alternariol monomethyl ether (AME) was found in pig liver, while alternariol (AOH) and tentoxin (TEN) were found in human forensic liver tissues. No other Alternaria mycotoxin metabolites were detected. This methodology is the first validated approach for determining Alternaria mycotoxins in liver tissue. Full article

Breast cancer remains one of the most prevalent and lethal malignancies affecting women worldwide, underscoring the need for safer and more effective therapeutic strategies. This study investigated the phytochemical composition, antioxidant activity, and antiproliferative potential of POW9™, a proprietary botanical blend formulated from nine medicinal plant extracts. Comprehensive phytochemical profiling was performed using ultrahigh-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-QTOF-MS) in both positive and negative ionization modes. A total of 34 compounds were identified in negative mode and 27 compounds in positive mode, comprising flavonoids, terpenoids, steroids, organic acids, peptides, glycosides, and lipids. POW9™ exhibited high total phenolic content (190.3 ± 3.5 mg gallic acid equivalents/g) and total flavonoid content (115.2 ± 1.5 mg quercetin equivalents/g), along with strong antioxidant activity, demonstrated by a 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging with a half-maximal inhibitory concentration (IC₅₀) of 1.66 mg/mL (33.73 mg Trolox equivalents/g). Cytotoxicity assessment revealed minimal toxicity toward normal Vero cells. In contrast, POW9™ significantly inhibited the proliferation of human breast cancer cell lines in a concentration- and time-dependent manner. The IC₅₀ values were 6.75 mg/mL for MCF-7 cells and 18.08 mg/mL for MDA-MB-231 cells after 72 h of treatment, while prolonged exposure (96 h) further enhanced antiproliferative efficacy, reducing the IC₅₀ to 2.34 mg/mL. These findings demonstrate that POW9™ is a chemically diverse herbal formulation with potent antioxidant and selective anti-breast cancer activities, supporting its potential development as a complementary therapeutic or nutraceutical agent for breast cancer management. Full article

The transformation of Andean grains and tubers through fermentation and bioencapsulation has emerged as a key strategy to enhance their nutritional, functional, and biotechnological value, driven by advances in proteomic and metabolomic techniques. This study aimed to systematize recent evidence on the biochemical and functional modifications induced by these processes and their potential application in the development of functional foods. The methodology integrated 67 studies analyzed using tools such as R 4.5.1 with the JupyterLab interface 4.5.2, SCImago Graphica Beta 1.0.53, and VOSviewer 1.6.20, incorporating data generated through LC-MS/MS, UHPLC-QTOF, Orbitrap platforms, transcriptomics, and combined omics approaches, considering original studies published between 2020 and 2025. The main findings indicate substantial increases in free amino acids (up to 64.8%), phenolic compounds (2.9–5.2%), and antioxidant activity (up to 45%), along with the identification of 430 polyphenols, 90 flavonoids, 14 novel oxindole acetates, and bioactive peptides with IC50 values ranging from 0.51 to 0.78 mg/mL. Bioencapsulation showed controlled release of bioactive compounds, highlighting nanocapsules of 133–165 nm with a maximum release of 9.86 mg GAE/g. In conclusion, the combination of fermentation and encapsulation enhances the stability, bioavailability, and functionality of Andean crops, supporting their industrial adoption for the development of sustainable nutraceutical foods that improve health and promote the valorization of traditional resources. Full article

The epiphytic community of Laminaria hyperborea, dominated by red algae, is typically discarded during industrial processing despite its potential as a source of high-value natural products. This study aims to valorize this underutilized biomass by characterizing its secondary metabolites and evaluating the biological activities of its major bromophenols. A combined chromatographic workflow enabled the isolation and structural elucidation of five bromophenols (1–5), including one previously undescribed compound (5). Among these, compound 4 exhibited the strongest cytotoxicity against the acute myeloid leukemia (AML) cell line MOLM-13 (EC₅₀ = 6.23 μM) and induced pronounced apoptotic features. When tested on two normal cell lines (NRK and H9c2) and in zebrafish larvae, it showed moderate toxicity at higher concentrations, indicating a reasonable selectivity window. In contrast, compound 5 was more toxic to normal cells than to MOLM-13 in vitro, while showing no acute toxicity in zebrafish; however, interpretations are preliminary due to compound purity. Bromophenols 1–4 were also tested for antioxidant activity, with 4 being the most potent (ABTS EC₅₀ = 22.1 μM), although this did not translate into protection against doxorubicin-induced cardiotoxicity. Additionally, non-targeted UHPLC-QTOF MS/MS analysis tentatively identified nine additional bromophenols and provided an estimation of their origin species within the epiphytic assemblage. Full article

Background and Rationale: Tinospora crispa (L.) Hook.f. & Thomson (T. crispa) is a climbing medicinal plant with long-standing ethnopharmacological use, particularly in inflammatory and hepatic disorders and cancer-related conditions. There is a knowledge gap regarding how wild versus cultivated ecotypes differ in chemotype, bioactivity, and safety, and how this might support or refine traditional use. Study Objectives: This study aimed to compare wild and cultivated ecotypes of T. crispa from the Nile Delta (Egypt) in terms of quantitative and qualitative phytochemical profiles; selected in vitro biological activities (especially antioxidant and cytotoxic actions); genetic markers potentially associated with metabolic variation; and short-term oral safety in an animal model. Core Methodology: Standardized extraction of plant material from wild and cultivated ecotypes. Determination of total phenolics, total flavonoids, and major phytochemical classes (alkaloids, tannins, terpenoids). Metabolomic characterization using UHPLC-ESI-QTOF-MS, supported by NMR, to confirm key compounds such as berberine, palmatine, chlorogenic acid, rutin, and borapetoside C. In vitro bioassays including: Antioxidant activity (e.g., radical-scavenging assay with EC₅₀ determination). Cytotoxicity against human cancer cell lines, with emphasis on HepG2 hepatoma cells and calculation of IC₅₀ values. Targeted genetic analysis to detect single-nucleotide polymorphisms (SNPs) in the gen1 locus that differentiate ecotypes. A 14-day oral toxicity study in rats, assessing liver and kidney function markers and performing histopathology of liver and kidney tissues. Principal Results: The wild ecotype showed a 43–65% increase in total flavonoid and polyphenol content compared with the cultivated ecotype, as well as substantially higher levels of key alkaloids, particularly berberine (around 12.5 ± 0.8 mg/g), along with elevated chlorogenic acid and borapetoside C. UHPLC-MS and NMR analyses confirmed the identity of the main bioactive constituents and defined a distinct chemical fingerprint for the wild chemotype. Bioassays demonstrated stronger antioxidant activity of the wild extract than the cultivated one and selective cytotoxicity of the wild extract against HepG2 cells (IC₅₀ ≈ 85 µg/mL), being clearly more potent than extracts from cultivated plants. Genetic profiling detected a C → T SNP within the gen1 region that differentiates the wild ecotype and may be linked to altered biosynthetic regulation. The 14-day oral toxicity study (up to 600 mg/kg) revealed no evidence of hepatic or renal toxicity, with biochemical markers remaining within physiological limits and normal liver and kidney histology. Conclusions and Future Perspectives: The wild Nile-Delta ecotype of T. crispa appears to be a stress-adapted chemotype characterized by enriched levels of multiple bioactive metabolites, superior in vitro bioactivity, and an encouraging preliminary safety margin. These findings support further evaluation of wild T. crispa as a candidate source for standardized botanical preparations targeting oxidative stress-related and hepatic pathologies, while emphasizing the need for: More comprehensive in vivo efficacy studies. Cultivation strategies that deliberately maintain or mimic beneficial stress conditions to preserve phytochemical richness. Broader geographical and genetic sampling to assess how generalizable the present chemotypic and bioactivity patterns are across the species. Full article

Background: The intestinal microflora is a population of microorganisms that resides in the human gastrointestinal tract and is important in maintaining metabolic and immune homeostasis in the body. Bacteria residing in the intestine produce short-chain fatty acids (SCFAs), which communicate with, among other things, the brain–gut axis—disorders of which are one of the causes of MS-like pathologies. A particular property of SCFAs is the induction of regulatory T cells, which are finding their way into pioneering therapies for MS patients. The aim of the study is to evaluate SCFA secretion in patients with multiple sclerosis from the West Pomeranian region depending on the genotypes of rs778986 and rs3894326 polymorphisms of the FUT3 gene. Methods: The study group included 47 patients clinically diagnosed with MS. Genotyping was performed by real-time PCR using TaqMan probes. Analysis of short-chain fatty acids in faeces was performed on a quadrupole mass spectrometer coupled to a time-of-flight (QTOF) analyser coupled to an AB Sciex high-performance liquid chromatograph (UHPLC). Results: Statistical analysis did not reveal any statistically significant differences in the prevalence of the studied polymorphisms in MS patients compared to the healthy control group. It was observed that the intestinal microflora and SCFA production in MS patients may be disturbed, while the studied FUT3 gene polymorphisms probably do not have a significant effect on their concentrations. A statistical tendency towards higher caproic acid content in heterozygotes of the rs778986 polymorphism and higher valeric acid secretion in homozygotes of rs3894326 was demonstrated. Conclusions: In summary, the studied FUT3 gene polymorphisms are not overrepresented in patients with MS. The rs778986 FUT3 polymorphism may affect the caproic acid content in the faeces of patients with MS, and the rs3894326 polymorphism may affect valeric acid secretion. Due to the small sample size and sparse genotype groups, the study has limited power and negative findings may reflect Type II error; replication in larger cohorts is warranted. Full article

Chamomile (Matricaria chamomilla L.) and feverfew (Tanacetum parthenium L. Sch. Bip.), both members of the Asteraceae family, are widely distributed in Serbia and traditionally used for their medicinal properties. Chamomile is primarily known for its gastrointestinal effects, while feverfew is noted for its antimigraine activity. Although the biological activity of each plant has been individually studied, there has been a lack of research related to their blends. So, the aim of this study was to prepare various chamomile/feverfew blends and their extracts with special focus on extraction temperature, to obtain superior herbal extract with the best functional characteristics. In order to characterize the obtained blend extracts this study included spectrophotometric and UHPLC Q-ToF MS analysis of prepared (selected) extracts, as well as evaluation of their antioxidant (ABTS, DPPH, FRAP, and CUPRAC) and antimicrobial properties. Antibacterial activity was evaluated against two Gram-positive and four Gram-negative bacterial strains using the broth microdilution method. Untargeted analysis showed the same phytochemical profile for both selected extracts (B3 and B9), as well as differences in distribution and abundance of identified compounds depending on applied extraction temperature (cold or heat-assisted). These differences in profile most probably contributed to variations in the antioxidant and antimicrobial properties of the extracts. The most potent antioxidant activity (123.04 µM TEAC/g) was observed for the 3:1 feverfew/chamomile blend (ABTS assay), while the highest metal-chelating capacity (1288.95 µM VCEAC/g) was recorded in extracts obtained by heat-assisted extraction (CUPRAC assay). Antibacterial activity of all blends ranged from 0.625 to 2.5 mg/mL, regardless of the extraction method. The findings indicate that combined extracts of chamomile and feverfew represent a promising source of bioactive compounds with potential applicability in both food science and pharmaceutical (biomedical) research. Full article

With the advent of new analytical technologies and the urgent environmental problem, reopening investigations into polluting waste matrices becomes a priority. Olive mill wastewater is a pollutant and phytotoxic by-product of olive oil production. An untargeted UHPLC-QTOF analysis of three olive mill wastewaters from three different olive cultivars was performed, and modern informatic platforms were involved to characterize the chemical components in-depth. Data elaboration and statistical analysis confirmed the differences between samples and revealed a total of 364 annotated compounds, including iridoids, phenolic compounds, flavonoids, lignans, cinnamic acid derivatives, and pyrrolidine derivatives. Many of these metabolites, including compounds with known antioxidant and bioactive potential, are scarcely reported in olive products and by-products. The outcomes of this work could be useful for rethinking olive mill wastewater as a source of bioactive compounds to develop and optimize new detoxification strategies. Full article

Horseradish (Armoracia rusticana L.) root (HRP) and leaf (HLP) pomaces, by-products of juice production by cold-pressing, were analyzed as a novel potential source of natural antioxidants. Chromatography analysis (UHPLC Q-ToF MS) of the bioactive compounds of pomaces was performed along with spectrophotometric determination of total phenolic content (TPC), total flavonoid content (TFC), total phenolic acid (hydroxycinnamic) content (TPAC), and antioxidant capacity (via 1,1-diphenyl-2-picrylhydrazyl (DPPH^•) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic-acid) (ABTS^•+) radicals’ scavenging activity and ferric reducing antioxidant power (FRAP)). The concentrations of TPC, TFC, and TPAC differed among the pomaces, significantly favoring HLP. However, both horseradish pomaces (HRP and HLP) contained a considerable amount of various phenolics, with kaempferol and its glucosides dominating. In addition, they exhibit pronounced antioxidant activity, which is confirmed by all three methods used (DPPH, ABTS, and FRAP). These results highlight the potential of valorizing horseradish processing waste as a natural, reliable source of health-promoting bioactive compounds and functional ingredients in food products, thereby fortifying food, preventing oxidation, and prolonging shelf-life. In addition, this study supports endeavors to reduce food waste by providing new insights into the valorization of horseradish pomace, thus contributing to sustainable development and environmental protection. Full article

Banisteriopsis and Stigmaphyllon are among the most species-rich liana genera in the Neotropical Malpighiaceae family, known for their diverse chemical and biological activities. However, metabolomic information correlating these genera with their biome of occurrence, humidity levels, and plant habit (liana vs. shrub) remains limited. This study explored in detail the metabolomic profiles of 15 species (29 specimens) of Banisteriopsis and 26 species (35 specimens) of Stigmaphyllon using Partial Least Squares Discriminant Analysis (PLS-DA). Leaf extracts obtained with ethanol/water (4:1, v/v) were analyzed by Ultra Performance Liquid Chromatography coupled to Quadrupole Time-of-Flight Mass Spectrometry (UHPLC-QTOF-MS/MS). Comparative analyses were performed at the genus level and across biome, humidity, and plant habit variables. The UHPLC-MS/MS profiling enabled the annotation of compounds, including previously unreported metabolites identified in B. quadriglandula and S. saxicola. Shared metabolites between the two genera were also characterized. PLS-DA revealed discriminant metabolites associated with the different comparative parameters, notably glucose (plant habit), coumaroyl hexoside, myricetin-3-galactoside, quercetin (genus), and quercetin-3-O-robinobioside (environment). With 95% confidence, our results demonstrate that environmental and morphological factors significantly influence metabolite biosynthesis in Banisteriopsis and Stigmaphyllon, providing valuable insights for future phytochemical and ecological research in the Neotropics. Full article

This study characterized leaf extracts of Cymbopogon flexuosus (Ryukyu Lemongrass Corporation, Okinawa, Japan) and evaluated the bioaccessibility and bioactivities of phenolic compounds following a simulated in vitro gastrointestinal model of digestion (in vitro GID) of plant material. Undigested (controls, AqC, EtC) and digested aqueous (AqD) and ethanolic (EtD) extracts were analyzed. Control extracts contained higher total phenolics and flavonoids than digested ones, with EtC showing the highest values. UHPLC-QToF-MS (ultra-high-performance liquid chromatography system coupled to a quadrupole time-of-flight mass spectrometer) identified 32 compounds, including phenolic acids, flavone aglycones, C-glycosides, and derivatives. Hydroxybenzoic acids, coumaric acid, caffeic esters, flavones, tricin derivatives, vitexin, and isoorientin exhibited reduced recovery, while coumaric acid hexoside, ferulic acid hexoside, and isoschaftoside/schaftoside exceeded 100% recovery, suggesting release from the matrix. Some compounds were absent from AqD, and many were found in the pellet, indicating potential colonic metabolism. Antioxidant activity (DPPH, reducing power, β-carotene/linoleic acid) was stronger in controls but always weaker than BHT/ascorbic acid. Extracts mildly inhibited α-amylase but more strongly inhibited α-glucosidase as shown with applied enzyme inhibition assays, especially EtD (76.93% at a concentration of 10 mg/mL), which showed stronger activity than controls but remained below acarbose (87.74% at 1 mg/mL). All extracts promoted HaCaT keratinocyte growth and reduced HCT-116 colon cancer cell viability at 250 µg/mL, with the strongest effects in AqC and AqD. Overall, GID decreased antioxidant activity but enhanced antidiabetic potential, confirming the safety and selective anticancer effects of C. flexuosus extracts. Full article

Hepatocellular carcinoma (HCC), the most common primary liver malignancy, usually develops in patients with cirrhosis, yet the metabolic mechanisms that distinguish the two conditions remain poorly understood. This study aimed to explore metabolic dysregulations in HCC compared with cirrhosis and to identify potential biomarkers, especially lipids, with diagnostic and prognostic value. We prospectively studied 81 patients—41 with HCC and 40 with cirrhosis—using high-resolution UHPLC-QTOF-ESI⁺-MS to characterize their serum lipidome. Across both groups, 322 metabolites were identified, but their distribution was strikingly different. Patients with HCC showed higher levels of sphingolipids, glycerophospholipids, diglycerides, sterols, and certain fatty acids, reflecting tumor-related metabolic rewiring. In contrast, cirrhotic patients had increased D-glucose, 5-hydroxymethyluracil, lysophospholipids, acylcarnitines, and specific fatty acid derivatives. Several lipids, such as CerPE(d16:2/24:1(2OH)), SM(d18:0/14:0), PA(36:6), and GlcCer(d18:1/12:0), displayed excellent discriminative accuracy, highlighting their role as putative biomarkers. These findings underscore the importance of lipid metabolic reprogramming in HCC, characterized by membrane remodeling, energy adaptation, and oxidative stress resistance. Integrating lipidomic profiling into clinical practice could improve early detection and risk stratification in cirrhotic patients. Larger, multicenter studies are needed to validate these biomarkers and assess their therapeutic implications. Full article

Background: Betulins (betulin, betulinic acid and lupeol) demonstrated antitumor and chemopreventive activity but showed low bioavailability due to their self-aggregation in hydrophilic environments. To overcome these disadvantages, their incorporation into lipid nanoformulations (PEGylated liposomes and Lipid Nanostructured Carriers (NLCs)) has proven to represent a viable solution. Objectives: The purpose of this study is to evaluate the size and incorporation rate of these molecules in nanoformulations, as well as their delivery and metabolic fate (pure betulinic acid versus a standardized extract, TT) relative to Doxorubicin using an in vivo protocol. The investigation extended our previous in vitro investigations towards an in vivo evaluation of antitumor activity, metabolic fate and toxicity in Wistar rats bearing Walker 256 carcinoma tumors over 21 days. Since previous studies used oral or intratumor administration, this exploratory study applied intravenous administration via microbubble-assisted sonoporation, considering its higher relevance for translational studies. Methods: The delivery and metabolic fate of the parent molecules, the identification of their fragments and metabolites using UHPLC-QTOF-ESI⁺MS were investigated, along with the identification of some toxicity biomarkers in rat plasma, tumor tissues and urine. Results: Preferential accumulation of Doxorubicin in tumors was observed compared to betulinic acid and TT components, as well as their persistence in plasma or elimination in urine. Compared to PEGylated liposomes, NLC formulations (especially NLC Doxo) induced a lower survival rate, a decreased bioavailability and increased toxicity by around 20%. Conclusions: These data are a starting point and complement the contrast-enhanced imaging and histology evaluations, which may contribute to the actual knowledge about the in vivo fate of betulins. Full article

Blueberry mosaic virus (BlMaV) is a persistent pathogen that alters host physiology; however, its impact on secondary metabolism in blueberry fruits remains poorly characterized. In this study, the phenolic profile of the cultivar ‘Duke’ was systematically examined in healthy and BlMaV-infected plants over two successive years. Using UHPLC Q-ToF MS, a total of 46 phenolic compounds were detected, spanning flavonols, phenolic acids, and anthocyanins. Comparative analyses revealed consistent shifts in metabolite abundance between healthy and infected samples. Several flavonol aglycones and phenolic acid derivatives accumulated in infected fruits, whereas multiple anthocyanins and glycosides were reduced. To further explore metabolic relationships, color correlation analysis highlighted distinct co-variation patterns among compound classes. Principal component analysis clearly separated infected and healthy fruits, confirming that viral infection was the dominant source of variation, surpassing the influence of harvest year or environmental factors. Nevertheless, the antioxidant capacity remained unchanged, regardless of the presence of the virus or the variation in environmental conditions. These results provide novel biochemical evidence that BlMaV infection reshapes the phenolic composition of blueberries and lays the groundwork for future studies on the metabolic consequences of viral stress in fruit crops. Full article

Bacterial outer-membrane vesicles (OMVs) mediate stress tolerance, biofilm formation, and interkingdom communication, but their role in beneficial endophytes remains underexplored. We isolated 11 non-redundant isolates associated with Bacillus, Enterococcus, Kosakonia and Kocuria from Agave tequilana seeds, identified by MALDI-TOF MS and 16S rRNA gene sequencing. We focused on the catalase-negative Enterobacter cloacae SEA01, which exhibits plant-promoting traits and support agave growth under nutrient-poor microcosms. In addition, this endophyte produces OMVs. Time-resolved SEM documented OMV release and cell aggregation within 9 h, followed by mature biofilms at 24 h with continued vesiculation. Purified OMVs (≈80–300 nm) contained extracellular DNA and were characterized by dynamic light scattering and UHPLC–ESI–QTOF-MS lipidomics. The OMV lipidome was dominated by phosphatidylethanolamine (~80%) and was enriched in monounsaturated fatty acids (16:1, 18:1), while the stress-associated cyclopropane fatty acids (17:1, 19:1) were comparatively retained in the whole-cell membranes; OMVs also exhibited reduced ubiquinone-8. SEA01 is catalase-negative, uncommon among plant-associated Enterobacter, suggesting a testable model in which oxidative factors modulate OMV output and biofilm assembly. These may have implications for recognition and redox signaling at the root interface. Future works should combine targeted proteomics/genomics with genetic or chemical disruption of catalase/OMV pathways. Full article