Search Results (228)

Background: This study aimed to identify distinct metabolic signatures associated with disease progression by integrating high-resolution computed tomography (HRCT) visual scoring with comprehensive metabolomic profiling. Materials and Methods: This single-center, cross-sectional study enrolled 60 idiopathic pulmonary fibrosis/interstitial lung disease (IPF/ILD) patients with usual interstitial pneumonia pattern. Participants underwent standardized pulmonary function testing, HRCT imaging, and peripheral blood collection for metabolomic analysis using one-dimensional hydrogen nuclear magnetic resonance spectroscopy and ultra-high-performance liquid chromatography coupled to tandem mass spectrometry. Linear regression analysis integrated radiographic scores with metabolomic profiles, adjusted for multiple covariates. Results: Stable IPF/ILD exhibited moderate negative correlations between the six most significant metabolites and HRCT scores (r = −0.27 to −0.51), along with a high abundance of specific phospholipids (triacylglycerol, monoacylglycerol, phosphatidylglycerol, phosphatidylethanolamine, diacylglycerol), sphingomyelin, ceramide, and acylcarnitine. In contrast, progressive disease showed weak positive correlations between the six most significant metabolites and HRCT scores (r = 0.19–0.26), and moderate negative correlation between specific triacylglycerol species and HRCT scores (r = −0.37–0.4). Furthermore, metabolomic analysis in individuals with progressive disease revealed both high and low abundances of specific phospholipid species (including high and low triacylglycerol species, as well as low levels of phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidylserine, and phosphatidylinositol), along with high levels of certain sphingomyelin, ceramide, taurine, and purine bases, and low levels of xanthine and lactic acid observed. Conclusions: Integration of systematic HRCT semi-quantitative scoring with metabolomic profiling successfully differentiated stable from progressive IPF/ILD through distinct molecular-radiographic signatures. Full article

Background/Objectives: Flavonoids are widely used as lead structures in drug discovery, and their pharmacological and metabolic properties are strongly influenced by structural features such as positional isomerism. This study aimed to compare the metabolic profiles and underlying mechanisms of two isoflavone-based positional isomers, ACF-02 (2-(4-hydroxy-3-methoxyphenyl)-6,7-dimethoxy-3-(4-methoxyphenyl)-4H-chromen-4-one) and ACF-03 (2-(3-hydroxy-4-methoxyphenyl)-6,7-dimethoxy-3-(4-methoxyphenyl)-4H-chromen-4-one). Methods: The metabolic pathways of synthetically prepared ACF-02 and ACF-03 were investigated using an in vitro incubation system with human liver microsomes (HLMs) supplemented with an NADPH-regenerating system, followed by liquid chromatography–high-resolution tandem mass spectrometry (LC–HRMS/MS) analysis. Metabolites were identified based on LC–HRMS/MS data and molecular networking-based node connectivity with the parent compounds. Major metabolites were further characterized by CYP phenotyping using recombinant CYP450 isoforms, and the potential for drug–drug interactions of ACF-03 was evaluated using a CYP probe substrate cocktail approach. Results: HLM incubation of ACF-02 and ACF-03 produced both hydroxylated and O-demethylated metabolites, with O-demethylation as the predominant pathway; notably, the most abundant O-demethylated metabolite differed in an isomer-dependent manner, occurring at the B₂ ring for ACF-02 and at the A ring for ACF-03, with distinct CYP isoform involvement. Molecular networking supported the relationships between the parent compounds and their metabolites, and both compounds exhibited relatively high metabolic stability with limited CYP inhibition. Conclusions: Despite differing only in the position of a single methyl substituent, ACF-02 and ACF-03 exhibited distinct isomer-dependent metabolic profiles. These findings demonstrate that even subtle positional isomerism can significantly influence metabolic behavior and should be carefully considered during lead optimization and drug design. Full article

Rhododendron tomentosum is an aromatic plant belonging to the Ericaceae family, widely used for different applications, but still lacking in its molecular signature. This work provides a complete chemical and biological characterization of the hydroalcoholic extract of R. tomentosum tips of twigs. Combining untargeted metabolomic analysis with bioassays, a correlation between chemical composition and biological activity was defined. To this regard, liquid chromatography high-resolution tandem mass spectrometry (LC-MS/MS) revealed a heterogeneous chemical composition, including flavonoids, such as quercetin, catechin, and their derivatives, as well as a first tentative identification of novel aesculin derivatives. Cell-based model experiments on stressed immortalized human keratinocytes demonstrated the antioxidant activity of the extract. Moreover, it exhibited significant antifungal and antibacterial effects against Trichoderma atroviride AGR2, Botrytis cinerea, and Clavibacter michiganensis, while promoting the growth of the beneficial bacterium Bacillus amyloliquefaciens. These findings highlight the rich diversity of bioactive molecules present in R. tomentosum hydroalcoholic extract, bridging its chemical composition to its functional properties. Overall, these results suggest its promising potential for applications in improving plant health, as well as in pharmaceutical, cosmetic, and agricultural industries. Full article

Pharmaceuticals, classified as emerging pollutants, are extensively applied in both human and veterinary medicine to treat various diseases. Their widespread application has led to their occurrence in water, soil, sediments, and living organisms. Even at trace levels (ng·L⁻¹–μg·L⁻¹), pharmaceuticals can significantly impact ecosystems and present threats to the environment and public health. Reliable analytical methods, such as liquid chromatography–tandem mass spectrometry (LC-MS/MS) and high-resolution mass spectrometry (LC-HRMS), are crucial for detecting and monitoring these compounds, including new or unknown substances. Removing pharmaceuticals from water remains challenging, as no single technique can completely eliminate all compounds and their toxic transformation products. Therefore, combining different treatment approaches, including membrane technologies, advanced oxidation processes (AOPs), and adsorption, into hybrid systems is necessary to improve removal efficiency and mitigate impacts on ecosystems and public health. In this review, a comprehensive overview of the occurrence of pharmaceuticals, their monitoring in food and water, and wastewater treatment is provided, highlighting current challenges and the need for further research. Full article

The efficient generation of structurally diverse rare ginsenosides from abundant precursors remains a significant challenge. In this study, a heterogeneous catalyst, 12-tungstosilicic acid supported on mesoporous silica (HSiW@mSiO₂), was developed for the transformation of ginsenoside Re in aqueous ethanol solution. The reaction was conducted under mild conditions, and the products were systematically analyzed using high-performance liquid chromatography coupled with multistage tandem mass spectrometry and high-resolution mass spectrometry. A total of 24 transformation products were identified, arising from deglycosylation, epimerization, dehydration, cyclization, and nucleophilic addition reactions. Structural elucidation revealed the formation of deglycosylated, hydrated and dehydrated derivatives, C-20 epimers, and novel ethoxylated protopanaxatriol-type ginsenosides resulting from solvent incorporation at the C-24(25) or C-20 position. Product distribution varied with reaction parameters, including solvent composition, reaction time, temperature, and catalyst dosage. The synthesized HSiW@mSiO₂ catalyst could be readily recovered by centrifugation and reused for five consecutive cycles, with complete conversion of ginsenoside Re maintained in the first two runs and a gradual decline in conversion to approximately 50% by the fifth cycle. This work demonstrates the efficacy of solid acid catalysts in enabling the structural diversification of ginsenosides through solvent-involved pathways. Full article

Music festivals have emerged as venues for the consumption of recreational drugs and novel psychoactive substances. This systematic review provides the first critical evaluation and synthesis of published wastewater analyses for detecting recreational drug use at music festivals worldwide. Following Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) guidelines, we systematically searched PubMed, Embase, MEDLINE, and SCOPUS databases using terms combining music festivals, drugs, and wastewater analysis. Twenty-three studies were included, spanning festivals with 6200 to 600,000 attendees. Two primary sampling approaches emerged: wastewater sampling (16 studies) and pooled urine sampling (7 studies), using Liquid Chromatography–Tandem Mass Spectrometry or Liquid Chromatography–High-Resolution Mass Spectrometry for chemical analysis. 3,4-Methylenedioxymethamphetamine (MDMA; ecstasy) emerged as the most consistently detected substance. Regional variations included a dominance of methamphetamine in Eastern Europe, MDMA use in Western Europe, and a high prevalence of cocaine use in South America. Regarding the music genre, electronic dance music events showed markedly higher MDMA rates. Limitations include geographic underrepresentation of African and Asian countries and a gender bias in pooled urine sampling. Future research should work on enhancing sampling infrastructure, analytical capabilities, public health surveillance, and harm reduction strategies. Full article

Phenolic molecules represent one of the most prevalent and biologically important categories of secondary metabolites. Within this diverse group, phenolic acids and flavonoids are the most extensively studied categories, primarily due to their structural diversity and broad spectrum of reported bioactivities. We first provide an overview of the physicochemical characteristics of flavonoids and phenolic acids and discuss how these properties relate to mass spectrometry (MS) fragmentation patterns and chromatographic behavior, including retention characteristics and isomer resolution. Next, we systematically examine the utilization of MS-based procedures for the characterization of flavonoids and phenolic acids in complex natural matrices. We initially examine targeted liquid chromatography–tandem mass spectrometry (LC–MS/MS) utilizing triple-quadrupole (QQQ) platforms, focusing on selected/multiple reaction monitoring (SRM/MRM) and associated scanning techniques (product-ion and precursor-ion scans). We summarize validated methodologies and strategies for both absolute and relative quantification, including stable-isotope dilution, matrix-matched calibration or standard addition, and internal-standard normalization. We subsequently analyze untargeted high-resolution mass spectrometry methodologies (direct injection and coupled to liquid chromatography), highlighting recent progress in data acquisition while addressing ongoing challenges in computational processing. Finally, we present a brief evaluation of commonly used extraction and clean-up processes, highlighting their practical impact on phenolic recoveries. Full article

Per- and polyfluoroalkyl substances (PFAS) are persistent environmental contaminants that tend to accumulate in solid matrices such as sewage sludge, raising concerns regarding their fate and potential ecological risks. This study aimed to develop and validate a robust analytical method for the accurate determination of PFAS in dehydrated sludge. A liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was optimized for 28 PFAS, including perfluoroalkyl carboxylic acids (PFCAs) and sulfonic acids (PFSAs). Solid–liquid extraction with basic methanol was followed by cleanup using a cartridge packed with ferrite and sodium sulfate to remove moisture and particulate interferences. Chromatographic separation was performed with an Avantor^® ACE^® PFAS Delay column coupled to an Agilent triple quadrupole MS operating in negative electrospray ionization mode. The method achieved excellent sensitivity (MDL < 0.02 µg/g dry weight for most compounds), satisfactory precision (RSD < 15%), and recoveries between 80–118%. Optimization of mobile phase additives, gradient conditions, and MS parameters enhanced chromatographic resolution and signal-to-noise ratio. The validated method demonstrates high reliability for PFAS determination in complex solid matrices and can be applied as a valuable tool for environmental monitoring and risk assessment of sludge management practices. Full article

Background/Objectives: The aim of this study was to reveal the influence of the natural nanoparticles (Nnps) isolated from Gegen–Qinlian Decoction (GQD), i.e., GQD-Nnps, on the intestinal absorption and pharmacokinetic properties of several representative active GQD constituents. Methods: The morphology of GQD-Nnps was examined using scanning electron microscopy (SEM). Protein and polysaccharide contents were measured using the bicinchoninic acid (BCA) assay and phenol–sulfuric acid method, respectively. Major GQD constituents were quantified by liquid chromatography–tandem mass spectrometry (LC-MS/MS). Formation mechanisms were explored using dynamic light scattering (DLS), Fourier transform infrared spectroscopy (FTIR), and high-resolution mass spectrometry (HRMS). Pharmacokinetic studies were conducted in mice with dextran sulfate sodium (DSS)-induced UC. Results: GQD-Nnps were spherical, with a size of 110.9 ± 8.1 nm and a zeta potential of −13.7 ± 1.5 mV. GQD-Nnps were primarily composed of proteins and polysaccharides. FTIR analysis revealed significant hydrogen bonding interactions between the small molecular and macromolecular constituents of GQD. HRMS analyses indicated complex formation among small molecules, particularly berberine, baicalin, and glycyrrhizic acid. DLS demonstrated good stability of GQD-Nnps in artificial gastric and intestinal fluids. Pharmacokinetic studies showed that, except for puerarin, blood and liver exposure levels of several constituents in the GQD-Nnps group were significantly higher than those in the GQD extract group, suggesting enhanced colonic absorption and hepatic distribution. Conclusions: GQD-Nnps create an oral drug delivery system through complex interactions, significantly enhancing the colonic absorption and hepatic distribution of several active GQD constituents. Full article

Background/Objectives: New synthetic opioids (NSOs) like nitazenes pose significant public health risks due to their high potency and increasing prevalence. Ethyleneoxynitazene, a benzofuran-containing nitazene, recently emerged on the illicit market and was identified in seizures in Europe. Although no intoxications have been reported to date, its µ-opioid receptor activity raises concern. This study investigated the metabolism of ethyleneoxynitazene to better understand its pharmacological profile, toxicity, and detectability in clinical and forensic contexts. Methods: Ethyleneoxynitazene was incubated with cryopreserved human hepatocytes pooled from 10 donors. Metabolites were detected by liquid chromatography coupled with high-resolution tandem mass spectrometry (LC-HRMS/MS) and identified using Compound Discoverer (Thermo Scientific; Waltham, MA, USA); detection and identification were assisted by in silico metabolite predictions with BioTransformer. Results: Sixteen metabolites were identified, with major biotransformations including N-deethylation at the N,N-diethylethanamine chain, hydroxylation at the dihydrofuran ring, and dihydrofuran ring opening via oxidative cleavage, leading to the formation of the corresponding ethanoic acid. Conclusions: This study provides the first characterization of the metabolism of a nitazene without an alkoxyphenyl moiety; the absence of this particular group reflects significant differences in the pharmacokinetic and pharmacodynamic profile compared to other nitazenes. We propose N-deethyl-3′-ethanoic acid-4′-hydroxy ethyleneoxynitazene, N-deethyl-hydroxy ethyleneoxynitazene, 3′-ethanoic acid-4′-hydroxy ethyleneoxynitazene, hydroxy ethyleneoxynitazene, and N-deethyl ethyleneoxynitazene as metabolite biomarkers of ethyleneoxynitazene consumption in clinical and forensic toxicology. Given the potential activity of some metabolites and interindividual variability in metabolic pathways, further studies are warranted to refine these findings through the analysis of biological samples from multiple ethyleneoxynitazene-positive cases. Full article

Background: Broccoli contains diverse phytochemicals, including glucosinolates and their hydrolysis products, with potential nutritional and bioactive properties. Accurate metabolite profiling requires optimized sample preparation and comprehensive databases. Methods: A rapid enzymatic deactivation method with 70% methanol, implemented prior to cryogrinding, was evaluated for processing freeze-dried and fresh broccoli florets, which were compared as plant materials. A widely targeted, organ-resolved metabolite database was constructed by integrating over 612 reported phytochemicals with glucosinolate degradation products. LC-HRMS combined with MS-DIAL and GNPS was employed for metabolite detection and annotation. Results: Freeze-dried samples yielded nearly twice the number of glucosinolates, isothiocyanates, and nitriles compared with standard-processed fresh tissue. Methanol pre-treatment preserved metabolite integrity in fresh samples, achieving comparable sensitivity to freeze-dried material. Using the integrated database, 998 metabolites were identified or tentatively characterized, including amino acids, carboxylic acids, phenolics, alkaloids, terpenoids, and glucosinolate derivatives. Cross-platform reproducibility was improved and false positives reduced. Conclusions: Optimized sample preparation combined with a curated metabolite database enables high-confidence, comprehensive profiling of broccoli florets phytochemicals. The resulting dataset provides a valuable reference for studies on genotype–environment interactions, nutritional quality, and functional bioactivity of cruciferous vegetables. Full article

Vegetarians must rely on supplements to meet the recommended daily intake of vitamin B12. Therefore, it is essential to establish a rapid, inexpensive, and reliable method of determining B12 levels in order to accurately characterize and assess the quality of supplements. This study describes a methodology for quantifying vitamin B12 in the form of methylcobalamin and cyanocobalamin following 2 min of ultrasound-assisted extraction performed at pH 4. Vitamin B12 was determined using UV-Vis spectrophotometry and liquid chromatography-tandem mass spectrometry. Thus, LC-MS/MS validated the cost-effective UV-Vis method. The content and form of vitamin B12 in the tested supplements were investigated, and serious discrepancies were found in the content or form of vitamin B12 in three out of ten supplements. To verify the quality of the analyzed supplements, the presence of metal impurities (as Cd, Hg, and Pb) was also assessed using high-resolution continuum source electrothermal atomic absorption spectrometry. No risk associated with the presence of these metals has been noted. Nevertheless, our findings underscore the need for stricter quality control in supplement manufacturing. Full article

Long non-coding RNAs (lncRNAs) are increasingly recognized as key regulators of gene expression and cellular signaling in cancer. Their functions are primarily mediated through interactions with specific protein partners that modulate chromatin structure, epigenetic remodeling, transcription, and signal transduction. In this review, we explore reports and strategies for the proteomic characterization of lncRNA-associated proteins, particularly emphasizing high-throughput liquid chromatography–mass spectrometry (LC-MS)-based techniques. Affinity-based methods such as RNA pull-down, ChIRP MS, RAP-MS, BioID-MS, and SILAC-MS enable sensitive and specific mapping of lncRNA and protein complexes. These approaches reveal cancer-specific proteomic signatures, post-translational modifications, and mechanistic insights into tumor biology. The use of label-free quantification, bituminization, and crosslinking strategies further enhances the resolution of dynamic RNA–protein networks. Validation tools following bioinformatic analyses, such as Western blotting, immunohistochemistry, immunofluorescence, and ELISA, are used to prioritize and confirm findings. Candidate biomarkers from hepatocellular carcinoma to colorectal and prostate cancers, profiling lncRNA-associated proteins, hold promise for identifying clinically actionable biomarkers and therapeutic targets. This review highlights the translational relevance of lncRNA protein studies and advocates for their broader adoption in oncological research. In LC-MS workflows, proteins bound to lncRNAs are enzymatically digested into peptides, separated via nano-LC, and analyzed using high-resolution tandem MS. Label-free or isotope-labeled methods quantify differential enrichment, followed by bioinformatics-driven pathway annotation. Full article

A comprehensive metabolic profiling of Catharanthus roseus (L.) G. Don was performed using tandem mass spectrometry, along with an evaluation of the biological activities of its various solvent extracts. Among these, the methanolic leaf extract exhibited mild radical scavenging activity, low to moderate antimicrobial activity, and limited cytotoxicity in both the brine shrimp lethality assay and MTT assay against HeLa and A549 cell lines. High-performance liquid chromatography–electrospray ionization–high-resolution tandem mass spectrometry (HPLC-ESI-HRMS/MS) analysis led to the annotation of 34 metabolites, primarily alkaloids. These included 23 indole alkaloids, two fatty acids, two pentacyclic triterpenoids, one amino acid, four porphyrin derivatives, one glyceride, and one chlorin derivative. Notably, two metabolites—2,3-dihydroxypropyl 9,12,15-octadecatrienoate and (10S)-hydroxypheophorbide A—were identified for the first time in C. roseus. Furthermore, Global Natural Products Social Molecular Networking (GNPS) analysis revealed 18 additional metabolites, including epoxypheophorbide A, 11,12-dehydroursolic acid lactone, and 20-isocatharanthine. These findings highlight the diverse secondary metabolite profile of C. roseus and support its potential as a source of bioactive compounds for therapeutic development. Full article

Background/Objectives: The emergence of cathinone-based psychostimulants necessitates ongoing research and analysis of the characteristics and properties of novel derivatives. The metabolic fate of five novel cathinone-derived substances (ASProp, MASProp, MASPent, PySProp, and PySPent) containing a selenophene moiety was investigated in vitro and in vivo. Methods: All compounds were incubated individually with pooled human liver S9 fraction. A monooxygenase activity screening investigating the metabolic contribution of eleven recombinant phase I isoenzymes was conducted. Rat urine after oral administration was prepared by urine precipitation. Liquid chromatography–high-resolution tandem mass spectrometry was used for the analysis of all samples. Reversed-phase liquid chromatography (RPLC) and zwitterionic hydrophilic interaction liquid chromatography (HILIC) were used to evaluate and compare the metabolites’ chromatographic resolution. Results: Phase I reactions of ASProp, MASProp, MASPent, PySProp, and PySPent included N-dealkylation, hydroxylation, reduction, and combinations thereof. The monooxygenase activity screening revealed the contribution of various isozymes. Phase II reactions detected in vivo included N-acetylation and glucuronidation. Both chromatographic columns complemented each other. Conclusions: All substances revealed metabolic reactions comparable to those observed for other synthetic cathinones. Contributions from isozymes to their metabolism minimized the risk of drug–drug interactions. The identified metabolites should be considered as targets in human biosamples, especially in urine screening procedures. RPLC and HILIC can both be recommended for this purpose. Full article