Analytical and Bioanalytical Chemistry最新文献

{"title":"Encapsulating Pt NCs with an AIE effect into a metal-organic framework as a novel fluorescence nanoprobe for discriminatively detecting bisphosphonate drugs.","authors":"Juan Du, Jinwen Chen, Huixiao Tong, Shenghua Liao","doi":"10.1007/s00216-025-06075-9","DOIUrl":"https://doi.org/10.1007/s00216-025-06075-9","url":null,"abstract":"<p><p>Fluorescent metal organic frameworks (FMOFs) constructed by encapsulating aggregation-induced emission luminogens (AIEgens) into frameworks have garnered increasing attention due to their ability to overcome the aggregation-caused quenching (ACQ) effect of traditional fluorescent materials such as organic dyes, quantum dots, and up-conversion nanoparticles. However, conventional AIEgens usually contain aromatic groups that may pose potential health risks. Additionally, the quality of bisphosphonates (BPs), first-line treatment drugs in skeletal-related diseases, is critical to assure their effectiveness and safety. In this work, a new nanocomposite, Pt NCs@ZIF-8, was designed by encapsulating platinum nanoclusters (Pt NCs) with aggregation-induced emission (AIE) properties into zeolitic imidazolate framework-8 (ZIF-8) through self-assembly between Zn²⁺ and Pt NCs. Owing to the spatial confinement provided by ZIF-8 and the AIE effect of Pt NCs, the resulting Pt NCs@ZIF-8 nanocomposite presents excellent photostability and anti-photobleaching. Furthermore, the framework of Pt NCs@ZIF-8 collapses in the presence of BP drugs due to the strong coordination interaction between the P = O groups in BPs and the Zn²⁺ in ZIF-8, leading to the release of Pt NCs. This release of Pt NCs causes effective fluorescence quenching of the Pt NCs@ZIF-8 nanocomposite, resulting from the increasing intramolecular motion of surface ligands, glutathione (GSH), on Pt NCs. Therefore, a new Pt NCs@ZIF-8-based nanoprobe has been rationally designed for BP detection. More importantly, our designed Pt NCs@ZIF-8-based nanoprobe also presents excellent discrimination capability toward four structurally similar BPs-Etidronate, Clodronate, Alendronate, and Risedronate, as confirmed by statistical techniques such as principal component analysis (PCA) and hierarchical clustering analysis (HCA). Finally, the proposed Pt NCs@ZIF-8 nanoprobe was successfully applied to quantify the labeled amount percentage of commercially available Sodium Risedronate and Sodium Alendronate tablets, achieving satisfactory limits of detection (LOD) spanning from 1.2 to 8.8 µM and excellent recovery ranging from 97.50% to 102.6%. This work not only provides a novel and convenient strategy for detecting and distinguishing BPs but also demonstrates the potential application of metal-organic frameworks encapsulating metal nanoclusters with AIE properties in pharmaceutical quality control.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144937797","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Sustainable and renewable approaches in biomaterial-based integrated dried urine sampling/solid-phase extraction devices for drugs and metabolites: challenges, bioanalytical applications, and future directions.","authors":"Sohan G Jawarkar, Pinaki Sengupta","doi":"10.1007/s00216-025-06043-3","DOIUrl":"https://doi.org/10.1007/s00216-025-06043-3","url":null,"abstract":"<p><p>Urine samples are gaining increasing importance as preferred biological matrices in bioanalytical studies due to their non-invasive collection, abundance, and ability to reflect systemic metabolism. However, conventional urine sampling and storage methods face significant challenges, especially in resource-limited areas where transportation and preservation infrastructure are inadequate. Traditional methods of liquid urine collection require refrigeration to prevent analyte degradation, making long-term storage and global sample transport costly and impractical. Biomaterial-based integrated dried urine sampling (DUS) devices integrate both storage and extraction into a single-step process, significantly enhancing efficiency and reliability. The porous sorbent is designed with a high surface area and tailored molecular binding capacity, ensuring the selective adsorption of target metabolites while minimizing matrix effects. Its structure allows for rapid drying and ambient-temperature storage, eliminating the need for cold-chain logistics. This feature is particularly beneficial for field studies and resource-constrained settings where maintaining refrigeration is impractical. Biomaterial-based DUS devices could be cost-effective and customizable and enable long-term stabilization of urinary metabolites. By reducing drying times, enhancing analyte recovery, and improving storage conditions, this technology enhances the feasibility of large-scale bioanalytical studies, therapeutic drug monitoring, and clinical diagnostics. Furthermore, its ability to selectively extract metabolites streamlines laboratory workflows and improves overall analytical precision. Porous biomaterial-based DUS devices could present a transformative, sustainable, and renewable solution for bioanalysis. By addressing the critical issues of sample stability, extraction efficiency, and cost-effectiveness, this approach has the potential to revolutionize metabolomics research, personalized medicine, and biomarker discovery in both clinical and research settings.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144938130","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative evaluation of phenyl isothiocyanate derivatization and \"dilute-and-shoot\" methods for HPLC-MS/MS-based targeted metabolomics analysis of amine-containing metabolites in plasma samples.","authors":"Kangkang Xu, Markus Aigensberger, Franz Berthiller, Heidi E Schwartz-Zimmermann","doi":"10.1007/s00216-025-06079-5","DOIUrl":"https://doi.org/10.1007/s00216-025-06079-5","url":null,"abstract":"<p><p>Metabolomics, the study of small molecule metabolites in biological systems, is essential for disease diagnosis and biomarker discovery. A key consideration in developing targeted metabolomics methods using HPLC-MS/MS for human or animal plasma is whether to employ derivatization of amino acids, amino acid-related compounds, and biogenic amines. Derivatization with phenyl isothiocyanate (PITC) enhances ionization and LC-separation, but complicates sample preparation and introduces potential errors. This study focuses on (1) the validation of a PITC derivatization method employing reversed-phase (RP) HPLC-MS/MS analysis and (2) a comparison of two analytical approaches for targeted metabolomics analysis of animal and human plasma: the PITC derivatization-based RP-LC-MS/MS method and a \"dilute-and-shoot\" approach using hydrophilic interaction chromatography (HILIC)-MS/MS and RP-LC-MS/MS analysis. The derivatization method was validated for porcine plasma, assessing limits of detection, lower limits of quantification (LLOQs), linearity, repeatability, recovery, and trueness. Derivatization reduced LLOQs for derivatized compounds in pure solvent solutions but, due to higher dilution factors, resulted in similar LLOQs for derivatized compounds and higher LLOQs for non-derivatized compounds in plasma compared to the \"dilute-and-shoot\" method. Derivatization improved chromatographic separation of isomers and reduced carryover but introduced challenges such as matrix effects, coelution with impurities, and calibration issues. The \"dilute-and-shoot\" method performed better for non-derivatized compounds and was less error-prone. Both methods were applied to plasma from various species, demonstrating comparable concentrations for most metabolites. The results also emphasize the importance of using different approaches for cross-validation. Above all, this study highlights the strengths and limitations of both the derivatization method and the \"dilute-and-shoot\" approach, providing guidance for their application in targeted metabolomics.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144938233","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Photonic resonator absorption microscopy: why consider metallic and magneto-plasmonic nano-assemblies over bare nanoparticles for digital biosensing?","authors":"Skye Shepherd, Weinan Liu, Seemesh Bhaskar, Brian T Cunningham","doi":"10.1007/s00216-025-06056-y","DOIUrl":"https://doi.org/10.1007/s00216-025-06056-y","url":null,"abstract":"<p><p>The unique optical interaction of species such as nanomaterials, proteins, viruses, antibodies, microRNA, and exosomes with the one-dimensional grating-based photonic crystals (PCs) has been leveraged in their detection using photonic crystal absorption microscopy (PRAM). While the principle and fundamental mechanism of such interfacial interactions are well delineated using wavelength and intensity modulations associated with the guided-mode resonance (GMR) of the PC, the effect of nano-assemblies in place of nanoparticles (NPs) has not been reported previously. In this work, the fundamental limitations observed with pristine NPs are overcome through the use of tunable AuNP assemblies synthesized via adiabatic cooling technology, where tunable nano-assemblies are obtained by subjecting the respective NPs to - 196 °C. Moreover, the higher contrast rendered by magneto-plasmonic, Fe₃O₄-Au hybrid nano-assemblies vis-à-vis metallic AuNP assemblies is corroborated with COMSOL Multiphysics simulations using electric and magnetic field hotspots. The high-contrast digital resolution enabled by magneto-plasmonic hybrid nano-assemblies, on account of synergistic coupling between the GMR of the underlying PC, delocalized Bragg, and localized Mie plasmons of dielectric-metal nano-assemblies, demonstrated excellent performance for microRNA-375-3p detection, opening a new window to explore hybrids of tunable \"permittivity + permeability\" as active probes in the design and development of microscopy-based biosensing modalities.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144938055","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synthesis of a novel multifunctional stationary phase derived from diethanolamine, HILIC/RPLC/WAX mixed-mode HPLC applications, and investigation of the retention mechanism.","authors":"Tarık Aral, Meryem Dokdemir, Hayriye Aral, Kübra Tunç, Murat Sunkur, Mehmet Çolak","doi":"10.1007/s00216-025-06068-8","DOIUrl":"https://doi.org/10.1007/s00216-025-06068-8","url":null,"abstract":"<p><p>Mixed-mode chromatography is increasingly valued for retaining analytes with diverse polarity and charge by integrating hydrophilic interaction (HILIC), reversed-phase (RPLC), and ion-exchange mechanisms. However, designing stationary phases that are both easy to synthesize and chromatographically versatile remains challenging. This study presents DEA-Mix-SP, a novel silica-based stationary phase functionalized with diethanolamine via [2-(3,4-epoxycyclohexyl)ethyl]trimethoxysilane, offering a simple synthetic route for broad-spectrum separation. Three variants with different ligand densities were characterized by elemental analysis, FTIR, SEM, and BET. The columns showed excellent separation for various analytes: nine nucleobases/nucleosides were resolved within 13 min, achieving up to 35,000 theoretical plates per meter, and eight benzoic acid derivatives were fully separated in 9 min with efficiencies up to 50,000 N/m. Alkylbenzenes, PAHs, Sudan dyes, phenols, and anilines were also effectively separated. Retention mechanisms were studied using quantitative parameters such as logD, logS, and pK_a, confirming significant anion-exchange effects. Performance was compared with ACE C18 (RPLC) and aminopropyl (HILIC) columns: DEA-Mix-SP showed superior separation performance in HILIC over the aminopropyl column and better separation of phenols, benzoic acids, and anilines than ACE C18, while achieving comparable results for fully non-polar analytes like alkylbenzenes, Sudan dyes, and PAHs. DEA-Mix-SP thus combines cost-effectiveness and high mixed-mode selectivity, providing reliable separation for analytes with wide polarity and charge ranges, making it a strong option for pharmaceutical, environmental, and food analyses.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144938140","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comprehensive profiling of sulfated phenolic compounds in edible and infesting seaweeds by a dedicated software-assisted platform","authors":"Enrico Taglioni,&nbsp;Chiara Cavaliere,&nbsp;Andrea Cerrato,&nbsp;Aldo Laganà,&nbsp;Carmela Maria Montone,&nbsp;Anna Laura Capriotti","doi":"10.1007/s00216-025-06027-3","DOIUrl":"10.1007/s00216-025-06027-3","url":null,"abstract":"<div><p>Seaweeds, comprising green (Chlorophyta), brown (Phaeophyta), and red (Rhodophyta) algae, are recognized as valuable marine resources rich in bioactive phenolic compounds with nutraceutical potential. Despite growing interest in edible species, limited attention has been given to infesting seaweeds, which may represent an underexploited source of bioactives while addressing ecological challenges. In the present study, an innovative, structure-based data processing workflow was set up for the characterization of phenolic compounds in marine organisms and applied to eight edible and two infesting seaweed species from the major taxonomic groups by means of untargeted mass spectrometric data acquisition. The results reveal distinct phenolic profiles across seaweed taxa, with green algae exhibiting the highest abundances of phenol and phenolic acid sulfates, while brown algae were characterized by a predominance of phlorotannins. Sulfated phenolics outnumbered their non-sulfated counterparts, underscoring the importance of dedicated analytical workflows for marine-specific metabolites. Multivariate statistical analyses further distinguished seaweed classes based on phenolic content and compound classes, with infesting seaweeds demonstrating comparable phenolic profiles to their edible counterparts. This suggests the potential for valorizing infesting seaweeds as sustainable sources of bioactive compounds, aligning with Blue Economy principles. Our findings provide comprehensive insights into the chemical diversity of seaweed phenolics and highlight the unexplored potential of invasive species for nutraceutical applications, contributing to sustainable marine resource management and the development of functional foods.</p><h3>Graphical Abstract</h3><div><figure><div><div><picture><source><img></source></picture></div></div></figure></div></div>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":"417 23","pages":"5199 - 5209"},"PeriodicalIF":3.8,"publicationDate":"2025-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12432060/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144938207","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Structural characterization of intra- and intermolecular disulfide bonds in voltage-dependent anion channel 3 (VDAC3) protein from Rattus norvegicus by high-resolution mass spectrometry.","authors":"Maria Gaetana Giovanna Pittalà, Annamaria Cucina, Stefano Conti-Nibali, Vincenzo Cunsolo, Antonella Di Francesco, Giuseppe Battiato, Simona Reina, Salvatore Foti, Vito De Pinto, Rosaria Saletti","doi":"10.1007/s00216-025-06074-w","DOIUrl":"https://doi.org/10.1007/s00216-025-06074-w","url":null,"abstract":"<p><p>Voltage-dependent anion channels, the most abundant proteins of the mitochondrial outer membrane, are responsible for the exchange of ions and metabolites between cytosol and mitochondria. They participate in the control of glycolytic metabolism through interaction with numerous enzymes and play a key role in the regulation of mitochondria-mediated apoptosis, cancer, and neurodegenerative diseases. The enzymatic digestion procedure in solution, originally developed in our laboratory, followed by high-resolution mass spectrometry coupled with UHPLC, has proven to be a powerful tool for the structural characterization of these \"difficult\" proteins. In this work, we used this procedure for the localization of intramolecular disulfide bonds in rVDAC3 and also for the characterization of intermolecular disulfide bonds formed by this protein with other VDAC isoforms. As a result, three intramolecular and seven intermolecular disulfide bonds between rVDAC3 with rVDAC1 and rVDAC2 were uniquely characterized. Furthermore, evidence was obtained for the existence of two additional intramolecular disulfide bonds between Cys2/Cys8 with Cys36 and Cys122, although these identifications were not supported by MS/MS spectra. The formation of intermolecular disulfide bonds helps to explain the previously observed VDAC oligomerization and demonstrates that disulfide bridges are directly involved in their homo- or hetero-oligomerization. Data are available via ProteomeXchange with identifier PXD064110.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144938120","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"LC-HRMS analysis of ultrashort- and short-chain PFAS on porous graphitic carbon column and study of accumulation in plants.","authors":"Timur M Baygildiev, Kirill S Gutenev, Anastasia E Karnaeva, Mikhail F Vokuev, Grigory I Tsizin","doi":"10.1007/s00216-025-06080-y","DOIUrl":"https://doi.org/10.1007/s00216-025-06080-y","url":null,"abstract":"<p><p>In this study, a robust LC-HRMS (QTOF) method was developed for the simultaneous determination of ultrashort- and short-chain perfluoroalkyl substances (PFAS) in complex plant matrices using a porous graphitic carbon column. The method enabled sensitive and selective detection of seven PFAS (C2-C10) in 18 min, including trifluoroacetic acid (TFA), which is known to exhibit poor retention on conventional C18 phases. The retention behavior of the analytes was systematically investigated under isocratic and gradient conditions, revealing a non-linear dependence on methanol content. Based on these findings, an optimized gradient elution program was proposed, ensuring excellent separation and peak shape even when injecting methanol-rich plant extracts-one of the advantages of porous graphitic carbon columns. Calibration curves for all analytes demonstrated good linearity (R² > 0.99) over relevant concentration ranges with accuracy within ± 15%. The method showed good recovery (71-103%) and precision (RSD < 7%) in rapeseed and burr medic methanolic extracts. Application to real plant samples exposed to PFAS-contaminated media (10 µg/mL) revealed substantial accumulation of ultrashort-chain PFAS, especially TFA and perfluoropropionic acid (PFPrA). These results support the potential of plant-based remediation strategies and highlight the importance of monitoring short- and ultrashort-chain PFAS in environmental and agricultural systems.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144937994","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"LC-QTOF-MS/MS for tentative identification of antioxidant peptides in defatted Spirulina platensis cakes obtained by supercritical CO₂ extraction.","authors":"Lucía Cuesta-Ramos, Nerea Sánchez-Moreno, Manuel Salgado-Ramos, Juan Manuel Castagnini, Francisco J Martí-Quijal, Emilia Ferrer, Francisco J Barba, Noelia Pallarés","doi":"10.1007/s00216-025-06073-x","DOIUrl":"https://doi.org/10.1007/s00216-025-06073-x","url":null,"abstract":"<p><p>This work tackles a supercritical fluid extraction (SFE) of microalga spirulina (Arthrospira platensis) to recover potential products from a nutritional and bioactive point of view. The execution of a response-surface model (RSO) through Box-Behnken design yielded 86% defatting, which significantly contrasts traditional Soxhlet extraction (9%). Contrarily to most of the reported works in the literature that focused on the analysis of the recovered oil, this study points out the added value of the remaining cakes after processing (spirulina residues). Namely, protein content determined by the DUMAS method concluded that a noted increase in the SFE cake was produced with respect to the raw material (from 69.2 to 74.4 g/100 g). In this regard, a tentative identification of antioxidant peptides through LC-QTOF-MS/MS was carried out. More deeply, 14 peptide sequences containing specific amino acids with potential antioxidant capacity were found in the SFE remaining cakes, of which the peptide sequences \"SIVNADAEARYLSPGELDRIK\" and \"RYLSPGELDRIKSFVT\" were only detected in these cakes and not in the Soxhlet solids. In addition, 4 short amino acid groups associated with potential antioxidant capacity were also identified in both cases (\"EL\", \"RY\", \"RYL\", and \"YL\"). ICP-MS served to determine the mineral profile in these materials, with most of them remaining in the cakes after SFE and Soxhlet extractions. With respect to oils, they were rich in Mg, P, K, and Fe minerals. The SFE oil extract had a much higher antioxidant capacity, phenolic content, chlorophyll A, and carotenoid content than the Soxhlet extract. Overall, this study presents an advanced extraction technique that efficiently recovers bioactive compounds and nutrients with potential food applications.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144938071","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization and metabolic profiling of oxymetholone and methasterone metabolites studied with human liver S9 model using GC-Orbitrap-HRMS for anti-doping purposes.","authors":"Siying Zheng, Yuqi Ge, Xian Fang, Mengpan Liu, Haoyi Sun, Xiaojun Deng, Lei Liao","doi":"10.1007/s00216-025-06066-w","DOIUrl":"https://doi.org/10.1007/s00216-025-06066-w","url":null,"abstract":"<p><p>In vitro metabolic models provide a means to circumvent the ethical concerns associated with human administration research, allowing for preliminary predictions of human metabolism while generating high concentrations of metabolites for characterization. As S1.1-class anabolic androgenic steroids on the World Anti-Doping Agency (WADA) Prohibited List, oxymetholone and methasterone have consistently appeared in the top 20 substances identified in adverse analytical findings (AAFs) in recent years, reflecting their persistent abuse patterns in sports. Given their exogenous nature, the metabolites of these steroid hormones fall within the scope of doping control, making metabolic studies a crucial aspect of anti-doping research. In this study, human liver S9 fractions were employed as a model for the characterization and metabolic profiling of oxymetholone and methasterone via gas chromatography-orbitrap-high-resolution mass spectrometry (GC-Orbitrap-HRMS). The full scan mode of GC-Orbitrap-HRMS was utilized to detect free and two conjugated fractions of metabolites, comparing these with control groups to confirm the metabolites during in vitro incubation. Possible metabolite structures were inferred from EI mass spectra, and the metabolic pathways for both drugs were discussed. In vitro, three oxymetholone and five methasterone metabolites were identified, and among them, two metabolites, OMT-M3 (2α,17α-methyl-5ξ-androstan-3α,6β,17β-triol) and MTS-M3 (2α,17α-dimethyl-5ξ-androstane-3α,12ξ,16ξ,17β-tetrol), were characterized as novel metabolites based on recent human in vivo metabolic studies. These metabolites exhibited diverse metabolic pathways, and their structures were corroborated through complementary in vitro and in vivo metabolic analyses. This study provides a comprehensive evaluation of the applicability of the human liver S9 model in the metabolic studies of anabolic steroids in vitro, verifying novel human metabolites and providing valuable insights for future research in this field.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144938222","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}