Search Results (418)

Grapes are frequently contaminated by Aspergillus section Nigri fungi and ochratoxin A (OTA), with A. niger also capable of producing substantial fumonisin B₂ (FB₂) levels. Emerging evidence suggests that aflatoxigenic fungi may eventually replace ochratoxigenic fungi in certain regions due to better adaptation to changes in climatic conditions. However, research on the toxigenic fungal community and mycotoxins in grapes from organic vineyards remains limited. Research on Spanish conventional grapes is also deficient, with most of the available literature being outdated. The present study investigates the diversity of toxigenic fungi and the presence of mycotoxins in organically cultivated grape berries in Spain, which are renowned for their significant oenological tradition. This study employed species-specific PCR protocols for fungal characterization and optimized methods for the analysis of OTA, FB₂, and aflatoxin B₁ (AFB₁) by UPLC–ESI–MS/MS. The most prevalent species present were Aspergillus flavus, A. niger, A. parasiticus, A. steynii, A. carbonarius, and A. westerdijkiae (67.1%, 43.5%, 20.0%, 14.1%, 14.1%, and 11.8% of the samples, respectively). OTA was detected only in 16 samples (19%), averaging 0.48 ng/g and peaking at 0.7 ng/g, which were lower than previously reported for conventional grapes. There was no FB₂ or AFB₁ detected. This study is pioneering in its exploration of the occurrence of toxigenic mycobiota, beyond Nigri fungi, and subsequent potential for other serious mycotoxins to contaminate Spain’s organic grapes. Full article

Pollen–stigma interactions have been studied extensively because they play an important role in sexual reproduction and crop yield. The vast majority of studies have focused on dry stigmas, which are typical of many model and agricultural plants; however, the data obtained are difficult to apply to plants with wet stigmas, such as tomato and tobacco. Pollen germination in this case occurs in a liquid, an exudate, which has a complex, species-specific composition. UPLC-ESI-MS-based hormone screening was carried out for six plant genera belonging to Solanaceae, Bromeliaceae, and Gesneriaceae families and revealed jasmonic acid (JA), abscisic acid (ABA) and/or jasmonoyl-L-isoleucine (IleJA) in stigma exudates of tobacco, tomato, and Streptocarpus sp. To assess the physiological significance of plant hormones in stigma exudate we tested their effect in vitro, finding that JA, IleJA, and MeJa significantly stimulated germination of tobacco pollen, with JA being most effective in accordance with its predominance in the stigma exudate; furthermore, ABA stimulated pollen germination in all tested species including bromeliads despite the lack of this hormone in their exudates. Both JA and ABA had an anti-oxidant effect on germinating pollen. Possible functions of hormones and ROS in exudate as well as ways of implementing the anti-oxidant effect of phytohormones are discussed. Full article

Background: Aldrovanda vesiculosa L. is a small aquatic plant that produces snap traps for capturing zooplankton prey. Aldrovanda belongs to the family Droseraceae, which is well known for the production of secondary metabolites (especially naphthoquinones). However, compared to other species in this family (Drosera and Dionaea), A. vesiculosa has been very poorly studied in terms of metabolites. The Aim: To fill this gap in knowledge, we investigated what secondary metabolites are present in the shoots of these plants. A hypothesis was tested stating that there are more metabolites in the younger (apical) parts of the shoots, which protect them from herbivores. Methods: Shoots of A. vesiculosa were collected, and the plant material was extracted with methanol, followed by 80% methanol or pure acetone using the accelerated solvent extraction method. The phytochemical profile was established using UPLC-DAD-(ESI)-MS. Results: A. vesiculosa shoots contained gallic acid and its derivatives, ellagic acid and its derivatives, flavonoids, and naphthoquinones (plumbagin and hydroplumbagin hexoside). A gradient (apical–basal) of gallic acid, ellagic acid, plumbagin, and hydroplumbagin hexoside was observed in the shoots. Meanwhile, the total flavonoid content did not differ between the middle and apical parts but was significantly lower in the basal part. In general, the lowest concentrations of metabolites were found in the basal part and the highest in the apical part, with the exception of total flavonoids. The number of free flavonoid aglycones was significantly higher in the middle part, whereas the apical part was dominated by glycoside derivatives. Full article

Neurodegenerative diseases, including Alzheimer’s disease (AD), represent one of the main global health challenges. Cannabis sativa synthesizes spermidine-type alkaloids, whose potential biological activities have been little studied. This study aimed to isolate bioactive alkaloids from an alkaloid-enriched extract (AEE) of C. sativa roots throughout a bioguided approach using conventional chromatographic techniques based on AChE and BuChE inhibitory activities. A qualitative and semiquantitative analysis by UPLC-ESI-MS/MS as well as molecular modeling simulations were performed. In addition, predictive in silico analyses were conducted to assess toxicity properties. The alkaloids cannabisativine (CS) and anhydrocannabisativine (ACS) were isolated, and showed highly selective BuChE inhibitory activity. The molecular modeling study revealed a conserved interaction profile across both alkaloids, indicating the amino acids TRP82, GLU197, TYR440, and HIS438 as the major contributors involved in the complex formation. Finally, CS and ACS exhibited low in silico predictive toxicity values. In conclusion, CS and ACS alkaloids emerge as new selective BuChE inhibitors with therapeutic potential that deserves the attention from the field of pharmacology in neurodegenerative disease research. Additionally, this approach promotes innovation and environmental sustainability through the use of C. sativa roots. Full article

Imperial chrysanthemum teas ‘Wuyuan Huangju’ (WYHJ) and ‘Jinsi Huangju’ (JSHJ), dried from the flowers of Chrysanthemum morifolium cv. Huangju, are traditional and popular herbal teas in China. However, their metabolite profiles and bioactivities remain unclear. In this study, we aimed to comprehensively elucidate the non-volatile and volatile metabolites of these two imperial chrysanthemum teas and assess their antioxidant activities and inhibitory effects on hyperglycemia and inflammation enzymes. Thus, we employed a widely targeted metabolomics approach based on UPLC-ESI-MS/MS and GC-MS/MS to characterize metabolite profiles of the two teas. In total, 1971 non-volatile and 1039 volatile metabolites were explored, and among these, 744 differential non-volatiles (classified into 11 categories) and 517 differential volatiles (classified into 12 categories) were identified. Further, 474 differential non-volatiles were upregulated in WYHJ, particularly flavonoids, terpenoids, and phenolic acids. In contrast, JSHJ exhibited a greater number of upregulated differential volatiles compared to WYHJ, contributing primarily to its sweet, fruity, and floral aroma. The results of scavenging activities towards DPPH·, ABTS·⁺, OH·⁻, and reducing power demonstrated that both imperial chrysanthemum teas, especially WYHJ, displayed high antioxidant capacity. We also noted that WYHJ exhibited stronger α-amylase, α-glucosidase, xanthine oxidase, and lipoxygenase inhibitory effects owing to its high active substance content. Therefore, this study provides insights into the metabolites of Chinese traditional medicinal herbal teas and highlights strategies for the comprehensive development and utilization of these traditional plant resources. Full article

Verifying the geographical origin of honey is crucial for its market value and for preventing fraudulent practices. This study aimed to characterize the chemical profiles of organic honeys from three distinct regions in Southeastern Türkiye—Şırnak Faraşin, Siirt Merkez, and Siirt Pervari—to establish a robust method for geographical authentication. A total of 51 multifloral honey samples were analyzed. The concentrations of 20 free amino acids (FAAs) and 16 phenolic compounds were quantified using (UPLC-ESI-MS/MS). The resulting data were subjected to both an unsupervised (PCA, CA) and supervised (PLS-DA, RF, SVM) chemometric analysis to identify biochemical markers for each region. The results revealed a distinct chemical fingerprint for each region. Based on the FAA profiles, the PLS-DA method provided the best overall classification, achieving an excellent discrimination with a total accuracy of 94.1% in the Şırnak Faraşin honeys. For the phenolic compound profiles, the RF method achieved the highest correct classification rate for Şırnak Faraşin honeys at 88.2%. This study demonstrates that an integrated approach, combining FAA and phenolic profiles with supervised chemometric methods, provides a successful and reliable model for determining the geographical origin of these multifloral honeys. Full article

The aim of this study was to investigate the metabolites in Aspergillus subramanianii 1901NT-1.40.2 extract using UPLC-MS, isolate and elucidate the structure of individual compounds, and study the antimicrobial and cytotoxic activities of the isolated compounds. The structures of two previously unreported ergostane triterpenoid aspersubrin A (1) and pyrazine alkaloid ochramide E (2) were established using NMR and HR ESI-MS. The absolute configuration of 1 was determined using quantum chemical calculations. Moreover, the known polyketides sclerolide (3) and sclerin (4); the indolediterpene alkaloid 10,23-dihydro-24,25-dehydroaflavinine (5); the bis-indolyl benzenoid alkaloids kumbicin D (6), asterriquinol D dimethyl ether (7), petromurin C (8); and the cyclopentenedione asterredione (9) were isolated. The effects of compounds 3-9 on the growth and biofilm formation of the yeast-like fungus Candida albicans and the bacteria Staphylococcus aureus and Escherichia coli were investigated. Compounds 5 and 6 inhibited C. albicans growth and biofilm formation at an IC₅₀ of 7–10 µM. Moreover, the effects of compounds 3-9 on non-cancerous H9c2 cardiomyocytes, HaCaT keratinocytes, MCF-10A breast epithelial cells, and breast cancer MCF-7 and MDA-MB-231 cells were also investigated. Compound 8 (10 µM) significantly decreased the viability of MCF-7 cells, inhibited colony formation, and arrested cell cycle progression and proliferation in monolayer culture. Moreover, 8 significantly decreased the area of MCF-7 3D spheroids by approximately 30%. A competitive test with 4-hydroxytamoxyfen and molecular docking showed that estrogen receptors (ERβ more than ERα) were involved in the anticancer effect of petromurin C (8). Full article

Background: Nephrolepis exaltata (sword fern) possesses a considerable amount of phytochemicals and different biological activities. The current study investigates the anti-biofilm potential of greenly synthesized bimetallic nanoparticles of Nephrolepis exaltata leaf methanol extract (NEME-MnO₂-MgO BNPs). Methods: The NEME was subjected to UPLC/MS analysis, followed by characterization of its NPs by size, zeta potential, FTIR, entrapment efficiency, and release. Then, antioxidant, antimicrobial and antibiofilm assays were employed, followed by in silico studies. Results: The UPLC/MS analysis of NEME led to the tentative identification of 27 metabolites, mostly phenolics. The MnO₂-MgO BNPs presented a uniform size and distribution and exhibited IC₅₀ values of 350 and 215.6 μg/mL, in the DPPH and ABTS assays, respectively. Moreover, the NPs exhibited antimicrobial and anti-biofilm efficacies against Pseudomonas aeruginosa, Klebsiella pneumonia (ATCC-9633), Staphylococcus aureus (ATCC-6538), Escherichia coli, Bacillus cereus, and C. albicans, with MIC values of 250–500 μg/mL. The MnO₂-MgO BNPs inhibited Candida albicans biofilms with a % inhibition of 66.83 ± 2.45% at 1/2 MIC. The network pharmacology highlighted epigallocatechin and hyperoside to be the major compounds responsible for the anti-biofilm potential. The ASKCOS facilitated the prediction of the redox transformations that occurred in the green synthesis, while the docking analysis revealed enhanced binding affinities of the oxidized forms of both compounds towards the outer membrane porin OprD of P. aeruginosa, with binding scores of −4.6547 and −5.7701 kcal/mol., respectively. Conclusions: The greenly synthesized Nephrolepis exaltata bimetallic nanoparticles may provide a promising, eco-friendly, and sustainable source for antimicrobial agents of natural origin with potential biofilm inhibition. Full article

Veterinary drug residues in aquatic products are often overlooked, yet they pose significant environmental risks and potential threats to human health. In this study, a rapid and sensitive analytical method was developed for the simultaneous determination of nine commonly used macrolide antibiotics in largemouth bass (Micropterus salmoides) muscle using ultrahigh-performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS). Sample extraction was performed using 80% acetonitrile in water, followed by purification with Cleanert MAS-Q cartridges. Chromatographic separation was achieved on a Waters ACQUITY UPLC BEH C₁₈ column (50 mm × 2.1 mm; 1.7 μm), equipped with a Waters VanGuard^TM BEH C₁₈ guard column (1.7 μm), using a mobile phase consisting of 0.1% formic acid in water and 0.1% formic acid in acetonitrile. Mass spectrometric detection was conducted in positive electrospray ionization mode (ESI⁺) using multiple reaction monitoring (MRM). The method demonstrated excellent linearity in the concentration range of 0.2–30 ng/mL, with determination coefficients (R²) exceeding 0.9980 for all analytes. Average recoveries ranged from 89.3% to 108.4%, with intraday and interday relative standard deviations (RSDs) of 2.9–11.6% and 4.1–12.5%, respectively. The limits of detection (LOD) and quantification (LOQ) for largemouth bass muscle were determined to be 0.4 μg/kg and 2.0 μg/kg, respectively. The decision limits (CC_α) and detection capabilities (CC_β) ranged from 2.13 to 215.71 μg/kg and 2.22 to 231.42 μg/kg, respectively. The developed method was successfully applied to the quantitative analysis of macrolide residues in 20 largemouth bass samples collected from local markets. Full article

The plant Synsepalum dulcificum is notable for its considerable edible and medicinal value, with a longstanding history as a folk remedy for diabetes. Its chemical constituents are rich and structurally diverse. However, there is limited information regarding the metabolic basis of these characteristics, and the biological activities and mechanisms underlying its blood glucose-lowering effects remain incompletely understood. In this study, we conducted a widely targeted metabolomics analysis of the stems, leaves, and fruits of S. dulcificum using UPLC-ESI-MS/MS to compare the differences in metabolite profiles among these three tissue types. Our analysis identified a total of 2544 secondary metabolites, primarily consisting of flavonoids and triterpenes, categorized into thirteen distinct compound classes. We selected differential metabolites through multivariate statistical analysis, revealing significant differences among the metabolite profiles of the three tissue types, with flavonoids being the most abundant compounds. Furthermore, we investigated the anti-diabetic mechanisms and potential pharmacological components of S. dulcificum utilizing network pharmacology and molecular docking techniques. Finally, the α-glucosidase inhibitory activity of the potential active components was evaluated using in vitro experiments. These findings establish a foundation for the future application of S. dulcificum in the prevention and treatment of diabetes. Full article

In this study, we integrated HSQC-based DeepSAT with UPLC-MS/MS to guide the isolation of omega-3 polyunsaturated fatty acid derivatives (PUFAs) from marine resources. Through this approach, four new (1–4) and nine known (5–13) PUFA analogues were obtained from large-scale cultures of the marine dinoflagellate Prorocentrum lima, with lipidomic profiling identifying FA18:5 (5), FA18:4 (7), FA22:6 (8), and FA22:6 methyl ester (11) as major constituents of the algal oil extract. Structural elucidation was achieved through integrated spectroscopic analyses of IR, 1D and 2D NMR, and HR-ESI-MS data. Given the pivotal role of microglia in Alzheimer’s disease (AD) pathogenesis, we further evaluated the neuroprotective potential of these PUFAs by assessing their regulatory effects on critical microglial functions in human microglia clone 3 (HMC3) cells, including chemotactic migration and amyloid-β42 (Aβ42) phagocytic clearance. Pharmacological evaluation demonstrated that FA20:5 butanediol ester (1), FA18:5 (5), FA18:4 (7), FA22:6 (8), and (Z)-10-nonadecenoic acid (13) significantly enhanced HMC3 migration in a wound-healing assay. Notably, FA18:4 (7) also significantly promoted Aβ42 phagocytosis by HMC3 microglia while maintaining cellular viability and avoiding pro-inflammatory activation at 20 μM. Collectively, our study suggests that FA18:4 (7) modulates microglial function in vitro, indicating its potential to exert neuroprotective effects. Full article

A sensitive method was developed for detecting diazepam residues in aquatic products using magnetic dispersive solid-phase extraction (MDSPE) coupled with ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS). Samples extracted with 1% ammonia–acetonitrile were purified using synthesized Fe₃O₄@SiO₂-PSA nanoparticles via MDSPE before UPLC-MS/MS analysis. Separation was performed on a C₁₈ column with gradient elution using 0.1% formic acid–2 mM ammonium acetate/methanol. Detection employed positive electrospray ionization (ESI⁺) in multiple reaction monitoring (MRM) mode. Characterization confirmed Fe₃O₄@SiO₂-PSA’s mesoporous structure with excellent adsorption capacity and magnetic properties. The method showed good linearity (0.1–10 μg/L, r > 0.99) with an LOD and LOQ of 0.20 μg/kg and 0.50 μg/kg, respectively. Recoveries at 0.5–15.0 µg/kg spiking levels were 74.9–109% (RSDs 1.24–11.6%). This approach provides rapid, accurate, and high-precision analysis of diazepam in aquatic products, meeting regulatory requirements. Full article

Walnut (Juglans regia L.) shells are valuable agro-industrial by-products rich in polyphenols. This study investigated traditional (maceration) and advanced extraction techniques—ultrasound-assisted extraction (UAE), enzyme-assisted extraction (EAE), pressurized liquid extraction (PLE), and combined ultrasound–enzyme extraction (US-EAE)—to recover bioactive compounds from walnut shells. Extraction efficiency, total phenolic content (TPC), antioxidant capacity (ABTS^•+, DPPH•), and polyphenol composition were evaluated. UPLC-ESI-MS/MS identified key polyphenols including ellagic acid, 4-hydroxybenzoic acid, vanillin, taxifolin, and quercitrin. The highest TPC (5625 mg GAE/100 g dw) was found in extracts subjected to US-EAE, in which ultrasound pretreatment (200 W, 10 min) was followed by enzymatic extraction using 0.06 mL/g Viscozyme^® L at pH 3.5 and 45 °C. Under the same extraction conditions, UAE alone yielded the second highest TPC (4129 mg GAE/100 g dw). The highest ABTS^•+ scavenging activity (14,478 mg TE/100 g dw) and enhanced DPPH• activity (45.38 mg TE/100 g dw) were also observed in US-EAE extracts. Chemometric techniques (PCA and HCA) revealed meaningful clustering and variation patterns among methods. These findings highlight the potential of walnut shells as a sustainable source of polyphenols and demonstrate the effectiveness of innovative extraction technologies in maximizing bioactive compound recovery for potential functional applications. Full article

This study examined the phytochemical compositions of ten Sapindus mukorossi samples from two regions in Korea: Suwon and Daegu. The Folin–Ciocalteu method was used to calculate the total polyphenol content (TPC). Among all extracts tested, leaf samples from Suwon and Daegu (SLE and DLE) exhibited the highest TPC at 2.70 and 2.90 mg tannic acid equivalent/g extract. Similarly, a modified aluminum chloride colorimetric test was used to determine the total flavonoid content (TFC). Similar results were obtained, with SLE and DLE having TFC values of 40.71 and 41.07 mg quercetin equivalent/g extract, respectively. Liquid chromatography with tandem mass spectrometry was used to detect 13 compounds, whereas high-performance liquid chromatography was used to quantify the prominent compounds: rutin, nicotiflorin, and narcissin. Among these, rutin was the most abundant, especially in SLE and DLE (54.37 and 70.21 mg/g, respectively). Furthermore, rutin significantly contributed to the total content of these samples at 78.31 and 85.44 mg/g, respectively. There were significant variations in the distribution of these compounds across different parts of the plant. These findings highlight the importance of S. mukorossi as a source of natural bioactive chemicals and pave the way for further research into its potential applications in healthcare products. Full article

Structural characterization of natural products in complex herbal extracts remains a major challenge in phytochemical analysis. In this study, we present a novel post-acquisition data-processing strategy—key ion diagnostics–neutral loss filtering (KID-NLF)—combined with ultra-high-performance liquid chromatography–quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS) for systematic profiling of the medicinal plant Terminalia chebula. The strategy consists of four main steps. First, untargeted data are acquired in negative electrospray ionization (ESI⁻) mode. Second, a genus-specific diagnostic ion database is constructed by leveraging characteristic fragment ions (e.g., gallic acid, chebuloyl, and HHDP groups) and conserved substructures. Third, MS/MS data are high-resolution filtered using key ion diagnostics and neutral loss patterns (302 Da for HHDP; 320 Da for chebuloyl). Finally, structures are elucidated via detailed spectral analysis. The methanol extract of T. chebula was separated on a C18 column using a gradient of acetonitrile and 0.1% aqueous formic acid within 33 min. This separation enabled detection of 164 compounds, of which 47 were reported for the first time. Based on fragmentation pathways and diagnostic ions (e.g., m/z 169 for gallic acid, m/z 301 for ellagic acid, and neutral losses of 152, 302, and 320 Da), the compounds were classified into three major groups: gallic acid derivatives, ellagitannins (containing HHDP, chebuloyl, or neochebuloyl moieties), and triterpenoid glycosides. KID-NLF overcomes key limitations of conventional workflows—namely, isomer discrimination and detection of low-abundance compounds—by exploiting genus-specific structural signatures. This strategy demonstrates high efficiency in resolving complex polyphenolic and triterpenoid profiles and enables rapid annotation of both known and novel metabolites. This study highlights KID-NLF as a robust framework for phytochemical analysis in species with high chemical complexity. It also paves the way for applications in quality control, drug discovery, and mechanistic studies of medicinal plants. Full article