Pub Date : 2026-03-17DOI: 10.1016/j.jpba.2026.117464
Qurat Ul Ain Haidery, Bo Han, Rashid Saeed, Chenyue Yan, Xingyuan Ma, Wenyun Zheng
Cannabidiol (CBD), a non-psychoactive terpenophenolic compound derived from Cannabis sativa, exhibits broad-spectrum pharmacological activities but is highly susceptible to oxidative, photolytic, and thermal degradation, making it challenging to preserve and use in therapeutics. Therefore, this study compares the preservation efficiency of a powder-based hydroxypropyl-β-cyclodextrin (HP-β-CD) inclusion-complex system for cannabinoid stability and solubility and medium-chain triglyceride (MCT) oil, a common lipid-based carrier in commercial formulations. Formulation characterizations, including CBD content (HPLC), color change, solvent residues, moisture, particle size, and zeta potential, were measured. DPPH experiments revealed that the HP-β-CD complex exhibited higher antioxidant activity (89.2%, EC₅₀ = 45.3 µg/mL) than the MCT oil system (83.6%, EC₅₀ = 58.7 µg/mL), with ascorbic acid serving as the control (EC₅₀ = 12.4 µg/mL). Additionally, photostability testing (8000 lux) demonstrated superior CBD retention in the HP-β-CD complex (>90%) compared with MCT oil (∼20%), indicating substantial light-induced degradation in the oil system. HP-β-CD complex retained 92-93% CBD (k = 0.0084 week⁻¹; t₁/₂ = 82.5 weeks) versus 88% in MCT oil. At 40 °C, HP-β-CD complex preserved 86-87% CBD. GC-FID/GC-MS showed that HP-β-CD powder preserved CBD with only trace degradation (<0.1-1.0%). In contrast, MCT oil produced higher cannabinoid by-products (0.1-6.21%), and greater antioxidant degradation (2.5-8.6% vs 0.5-2.1%). The results suggested that CBD degradation follows matrix-dependent kinetics, with cyclodextrin encapsulation significantly reducing the apparent degradation rate constants relative to oil-based solubilization under identical stress conditions.
{"title":"Matrix-dependent degradation kinetics and impurity fingerprinting of cannabidiol (CBD) in polymeric and lipid preservation systems.","authors":"Qurat Ul Ain Haidery, Bo Han, Rashid Saeed, Chenyue Yan, Xingyuan Ma, Wenyun Zheng","doi":"10.1016/j.jpba.2026.117464","DOIUrl":"https://doi.org/10.1016/j.jpba.2026.117464","url":null,"abstract":"<p><p>Cannabidiol (CBD), a non-psychoactive terpenophenolic compound derived from Cannabis sativa, exhibits broad-spectrum pharmacological activities but is highly susceptible to oxidative, photolytic, and thermal degradation, making it challenging to preserve and use in therapeutics. Therefore, this study compares the preservation efficiency of a powder-based hydroxypropyl-β-cyclodextrin (HP-β-CD) inclusion-complex system for cannabinoid stability and solubility and medium-chain triglyceride (MCT) oil, a common lipid-based carrier in commercial formulations. Formulation characterizations, including CBD content (HPLC), color change, solvent residues, moisture, particle size, and zeta potential, were measured. DPPH experiments revealed that the HP-β-CD complex exhibited higher antioxidant activity (89.2%, EC₅₀ = 45.3 µg/mL) than the MCT oil system (83.6%, EC₅₀ = 58.7 µg/mL), with ascorbic acid serving as the control (EC₅₀ = 12.4 µg/mL). Additionally, photostability testing (8000 lux) demonstrated superior CBD retention in the HP-β-CD complex (>90%) compared with MCT oil (∼20%), indicating substantial light-induced degradation in the oil system. HP-β-CD complex retained 92-93% CBD (k = 0.0084 week⁻¹; t₁/₂ = 82.5 weeks) versus 88% in MCT oil. At 40 °C, HP-β-CD complex preserved 86-87% CBD. GC-FID/GC-MS showed that HP-β-CD powder preserved CBD with only trace degradation (<0.1-1.0%). In contrast, MCT oil produced higher cannabinoid by-products (0.1-6.21%), and greater antioxidant degradation (2.5-8.6% vs 0.5-2.1%). The results suggested that CBD degradation follows matrix-dependent kinetics, with cyclodextrin encapsulation significantly reducing the apparent degradation rate constants relative to oil-based solubilization under identical stress conditions.</p>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"275 ","pages":"117464"},"PeriodicalIF":3.1,"publicationDate":"2026-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147494229","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-03-16DOI: 10.1016/j.jpba.2026.117463
Zhan Xu, Bin Miao, Zhiqi Gu, Yimin Hu, Ding Tang, Jiadong Li
Wumei (WM), a historical food and medicine homology fruit in China, is reported to have anti-allergic effect, yet its active components and mechanisms remain unclear. Here, a novel spleen tyrosine kinase-based piezoresistive cantilever array (Syk-PCA) biosensor was developed for screening anti-allergic components in WM. Once the biosensor composition was confirmed by X-ray photoelectron spectroscopy (XPS), its specificity was evaluated based on the differential electrochemical responses toward piceatannol and protocatechualdehyde. The biosensors were subsequently used to screen for Syk-binding compound of WM extract as a predictor of potential anti-allergic properties, including succinic acid, tartaric acid, chlorogenic acid, citric acid, and ursolic acid. The biosensor demonstrated high specificity and sensitivity, achieving a femtomolar-level interaction for ursolic acid with a surface equilibrium dissociation constant (KD, sur) of 2.37 fM and a linear detection range from 1 fM to 1 pM. Using this platform, ursolic acid was identified as the key active component targeting Syk. The activity was validated through in vitro and in silico approaches, including RBL-2H3 cell model, cellular thermal shift assay (CETSA), molecular docking and molecular dynamics simulation. The binding affinity between ursolic acid and Syk was demonstrated by the elevated thermostability of ursolic acid-Syk complex. Molecular docking and molecular dynamics simulation results indicated that the complex remained stable, with interactions mediated by hydrogen bonds and hydrophobic interactions. These findings reveal that ursolic acid exerts anti-allergic effects by targeting Syk, and the Syk-PCA biosensor provides a rapid, cost-effective tool for targeted drug discovery.
{"title":"Identification of ursolic acid from Wumei as a syk-targeting anti-allergic agent using a piezoresistive cantilever biosensor.","authors":"Zhan Xu, Bin Miao, Zhiqi Gu, Yimin Hu, Ding Tang, Jiadong Li","doi":"10.1016/j.jpba.2026.117463","DOIUrl":"https://doi.org/10.1016/j.jpba.2026.117463","url":null,"abstract":"<p><p>Wumei (WM), a historical food and medicine homology fruit in China, is reported to have anti-allergic effect, yet its active components and mechanisms remain unclear. Here, a novel spleen tyrosine kinase-based piezoresistive cantilever array (Syk-PCA) biosensor was developed for screening anti-allergic components in WM. Once the biosensor composition was confirmed by X-ray photoelectron spectroscopy (XPS), its specificity was evaluated based on the differential electrochemical responses toward piceatannol and protocatechualdehyde. The biosensors were subsequently used to screen for Syk-binding compound of WM extract as a predictor of potential anti-allergic properties, including succinic acid, tartaric acid, chlorogenic acid, citric acid, and ursolic acid. The biosensor demonstrated high specificity and sensitivity, achieving a femtomolar-level interaction for ursolic acid with a surface equilibrium dissociation constant (K<sub>D, sur</sub>) of 2.37 fM and a linear detection range from 1 fM to 1 pM. Using this platform, ursolic acid was identified as the key active component targeting Syk. The activity was validated through in vitro and in silico approaches, including RBL-2H3 cell model, cellular thermal shift assay (CETSA), molecular docking and molecular dynamics simulation. The binding affinity between ursolic acid and Syk was demonstrated by the elevated thermostability of ursolic acid-Syk complex. Molecular docking and molecular dynamics simulation results indicated that the complex remained stable, with interactions mediated by hydrogen bonds and hydrophobic interactions. These findings reveal that ursolic acid exerts anti-allergic effects by targeting Syk, and the Syk-PCA biosensor provides a rapid, cost-effective tool for targeted drug discovery.</p>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"276 ","pages":"117463"},"PeriodicalIF":3.1,"publicationDate":"2026-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147499123","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-03-15Epub Date: 2025-12-05DOI: 10.1016/j.jpba.2025.117306
Mintao Jian , Jiaqing Xiong , Kui Li
This study investigates the molecular mechanisms by which Xiangpi Shengji Ointment (XPSJO), combined with a heat-clearing and dampness-drying compound, accelerates postoperative wound healing in an anal fistula (AF) rat model, employing integrated proteomic and metabolomic approaches. An AF rat model was established and treated topically with the herbal combination. Wound healing progression was monitored postoperatively. Histopathological assessment (hematoxylin and eosin [H&E] staining), enzyme-linked immunosorbent assay [ELISA] quantification of inflammatory markers (e.g., tumor necrosis factor-alpha [TNF-α], interleukin [IL]-6) and vascular endothelial growth factor [VEGF], and proteomic (liquid chromatography-tandem mass spectrometry [LC-MS/MS]) and metabolomic (gas chromatography-mass spectrometry [GC-MS]) analyses identified critical pathways and biomolecules involved. In vitro, lipopolysaccharide (LPS)-stimulated THP-1 macrophages and human umbilical vein endothelial cells (HUVECs) were co-treated with XPSJO and Coptis chinensis-Phellodendron amurense Rupr. Extract (CGE). Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway modulation was examined via reverse transcription quantitative polymerase chain reaction (RT-qPCR), Western blot, and functional assays (cell counting kit [CCK]-8, flow cytometry, scratch wound, tube formation). Lentiviral NF-κB overexpression validated mechanistic specificity in vitro and in vivo. Results showed that XPSJO combined with the compound accelerated AF wound closuresuppressed NF-κB signaling pathway activation (evidenced by downregulation of pathway-associated proteins in proteomics and reduced p65 phosphorylation), decreased pro-inflammatory cytokine secretion, and enhanced VEGF expression. Critically, it promoted M2 macrophage polarization (increased Arg-1/CD206, decreased iNOS/CD80) and stimulated HUVEC migration/angiogenesis. Multi-omics analysis further confirmed NF-κB, TNF, and IL-17 pathways as central therapeutic targets. The study concludes that XPSJO combined with a heat-clearing and dampness-drying compound promotes AF healing through pharmacological inhibition of the NF-κB pathway, macrophage reprogramming, and tissue remodeling.
{"title":"Integrated proteomics and metabolomics reveal NF-κB pathway modulation by Xiangpi Shengji ointment and Qingre Zaoshi compound in anal fistula wound healing","authors":"Mintao Jian , Jiaqing Xiong , Kui Li","doi":"10.1016/j.jpba.2025.117306","DOIUrl":"10.1016/j.jpba.2025.117306","url":null,"abstract":"<div><div>This study investigates the molecular mechanisms by which Xiangpi Shengji Ointment (XPSJO), combined with a heat-clearing and dampness-drying compound, accelerates postoperative wound healing in an anal fistula (AF) rat model, employing integrated proteomic and metabolomic approaches. An AF rat model was established and treated topically with the herbal combination. Wound healing progression was monitored postoperatively. Histopathological assessment (hematoxylin and eosin [H&E] staining), enzyme-linked immunosorbent assay [ELISA] quantification of inflammatory markers (e.g., tumor necrosis factor-alpha [TNF-α], interleukin [IL]-6) and vascular endothelial growth factor [VEGF], and proteomic (liquid chromatography-tandem mass spectrometry [LC-MS/MS]) and metabolomic (gas chromatography-mass spectrometry [GC-MS]) analyses identified critical pathways and biomolecules involved. <em>In vitro</em>, lipopolysaccharide (LPS)-stimulated THP-1 macrophages and human umbilical vein endothelial cells (HUVECs) were co-treated with XPSJO and <em>Coptis chinensis</em>-<em>Phellodendron amurense</em> Rupr. Extract (CGE). Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway modulation was examined via reverse transcription quantitative polymerase chain reaction (RT-qPCR), Western blot, and functional assays (cell counting kit [CCK]-8, flow cytometry, scratch wound, tube formation). Lentiviral NF-κB overexpression validated mechanistic specificity <em>in vitro</em> and <em>in vivo</em>. Results showed that XPSJO combined with the compound accelerated AF wound closuresuppressed NF-κB signaling pathway activation (evidenced by downregulation of pathway-associated proteins in proteomics and reduced p65 phosphorylation), decreased pro-inflammatory cytokine secretion, and enhanced VEGF expression. Critically, it promoted M2 macrophage polarization (increased Arg-1/CD206, decreased iNOS/CD80) and stimulated HUVEC migration/angiogenesis. Multi-omics analysis further confirmed NF-κB, TNF, and IL-17 pathways as central therapeutic targets. The study concludes that XPSJO combined with a heat-clearing and dampness-drying compound promotes AF healing through pharmacological inhibition of the NF-κB pathway, macrophage reprogramming, and tissue remodeling.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117306"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145733403","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-03-15Epub Date: 2025-12-08DOI: 10.1016/j.jpba.2025.117311
Carlos Pernas-Fraguela , Rosario Rodil , José Benito Quintana , Antonio Saez , Ruth Olmos , Joan Mestre , Manuel Isorna , Rosa Montes
Chemsex is a practice mostly extended between men who have sex with men under drugs effects to enhance and/or prolong sexual activities. Within chemsex, slamming consists of intravenous consumption of substances such as cathinones and amphetamines. Syringe residue analysis is a relatively new alternative for knowing with accuracy the substances consumed by the users. Although several studies have been carried out applying the technique, none of them was focused on this population group. In this work, syringes were obtained from a drug addiction assistance centre from Madrid (Spain). Samples were processed by rinsing with methanol and injected in a liquid chromatography coupled to high resolution mass spectrometry system. Data (acquired in data dependent mode) were processed using a suspect screening approach. In total, 122 syringes from three sampling campaigns were analyzed. The compounds belonging to the cathinones’ group were the most frequently detected (99 %), followed by amphetamines and cocaine. The most consumed cathinone was chloromethcathinone, but also others as methylmethcathinone, pentedrone or N,N-dimethylpentylone were identified. Furthermore, temporal trends were detected during the different sampling campaigns. Thus, this technique allowed us to determine in an accurate way the substances consumed during slamming sessions, which may differ from the self-declared ones in surveys. Moreover, temporal trends were observed, that align with the market changes and regulation of cathinones.
{"title":"The role of syringe residue analysis in identifying substances consumed in sexualized drug use practices","authors":"Carlos Pernas-Fraguela , Rosario Rodil , José Benito Quintana , Antonio Saez , Ruth Olmos , Joan Mestre , Manuel Isorna , Rosa Montes","doi":"10.1016/j.jpba.2025.117311","DOIUrl":"10.1016/j.jpba.2025.117311","url":null,"abstract":"<div><div><em>Chemsex</em> is a practice mostly extended between men who have sex with men under drugs effects to enhance and/or prolong sexual activities. Within <em>chemsex</em>, <em>slamming</em> consists of intravenous consumption of substances such as cathinones and amphetamines. Syringe residue analysis is a relatively new alternative for knowing with accuracy the substances consumed by the users. Although several studies have been carried out applying the technique, none of them was focused on this population group. In this work, syringes were obtained from a drug addiction assistance centre from Madrid (Spain). Samples were processed by rinsing with methanol and injected in a liquid chromatography coupled to high resolution mass spectrometry system. Data (acquired in data dependent mode) were processed using a suspect screening approach. In total, 122 syringes from three sampling campaigns were analyzed. The compounds belonging to the cathinones’ group were the most frequently detected (99 %), followed by amphetamines and cocaine. The most consumed cathinone was chloromethcathinone, but also others as methylmethcathinone, pentedrone or N,N-dimethylpentylone were identified. Furthermore, temporal trends were detected during the different sampling campaigns. Thus, this technique allowed us to determine in an accurate way the substances consumed during <em>slamming</em> sessions, which may differ from the self-declared ones in surveys. Moreover, temporal trends were observed, that align with the market changes and regulation of cathinones.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117311"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145733401","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-03-15Epub Date: 2025-11-29DOI: 10.1016/j.jpba.2025.117296
Abulimiti Yasen , Yuxia Zhou , Wenhan Gao , Zhongxin Zhang , Reyihan Tudi , Mei Xiang , Bumaliya Abulimiti
Laser-Induced Breakdown Spectroscopy (LIBS) holds significant value for rapid elemental detection; however, strong spectral interference, matrix effects, and high-dimensional data characteristics pose considerable challenges to accurate quantitative analysis. To enhance the performance of LIBS quantitative analysis, this study proposes a novel machine learning framework that integrates XGBoost and Multilayer Perceptron (MLP), optimized by a Multi-dimensional Adaptive Sparrow Search Algorithm (MA-SSA). The framework employs XGBoost for automated feature selection, eliminating redundant spectral variables while retaining critical information, and utilizes MA-SSA to optimize the hyperparameters of the MLP in regression tasks, significantly improving model stability and prediction accuracy. Experimental results demonstrate that the proposed method achieves 100 % accuracy in multi-class classification, outperforming traditional classifiers such as Random Forest, XGBoost, and standalone MLP. In terms of quantitative detection, the MA-SSA-optimized model achieves an RMSE of 4.43 µg/g, surpassing other hybrid optimization models including XGBoost-SSA-MLP (RMSE=4.62 µg/g), XGBoost-PSO-MLP (RMSE=5.225 µg/g), and XGBoost-GA-MLP (RMSE=5.584 µg/g). XGBoost-based feature selection effectively reduces spectral dimensionality while maintaining predictive performance. The proposed MA-SSA algorithm further enhances convergence efficiency and generalization capability. This study provides a robust, efficient, and scalable solution for LIBS analysis, with broad application potential in the field of real-time quantitative detection.
{"title":"A MA-SSA-optimized XGBoost-MLP framework using LIBS for rapid classification and quantitative analysis of heavy metals in traditional chinese medicines","authors":"Abulimiti Yasen , Yuxia Zhou , Wenhan Gao , Zhongxin Zhang , Reyihan Tudi , Mei Xiang , Bumaliya Abulimiti","doi":"10.1016/j.jpba.2025.117296","DOIUrl":"10.1016/j.jpba.2025.117296","url":null,"abstract":"<div><div>Laser-Induced Breakdown Spectroscopy (LIBS) holds significant value for rapid elemental detection; however, strong spectral interference, matrix effects, and high-dimensional data characteristics pose considerable challenges to accurate quantitative analysis. To enhance the performance of LIBS quantitative analysis, this study proposes a novel machine learning framework that integrates XGBoost and Multilayer Perceptron (MLP), optimized by a Multi-dimensional Adaptive Sparrow Search Algorithm (MA-SSA). The framework employs XGBoost for automated feature selection, eliminating redundant spectral variables while retaining critical information, and utilizes MA-SSA to optimize the hyperparameters of the MLP in regression tasks, significantly improving model stability and prediction accuracy. Experimental results demonstrate that the proposed method achieves 100 % accuracy in multi-class classification, outperforming traditional classifiers such as Random Forest, XGBoost, and standalone MLP. In terms of quantitative detection, the MA-SSA-optimized model achieves an RMSE of 4.43 µg/g, surpassing other hybrid optimization models including XGBoost-SSA-MLP (RMSE=4.62 µg/g), XGBoost-PSO-MLP (RMSE=5.225 µg/g), and XGBoost-GA-MLP (RMSE=5.584 µg/g). XGBoost-based feature selection effectively reduces spectral dimensionality while maintaining predictive performance. The proposed MA-SSA algorithm further enhances convergence efficiency and generalization capability. This study provides a robust, efficient, and scalable solution for LIBS analysis, with broad application potential in the field of real-time quantitative detection.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117296"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145691109","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-03-15Epub Date: 2025-11-16DOI: 10.1016/j.jpba.2025.117269
Yaqin Sun , Junyin Zhou , Xiaoli Hou , Tianqi Ma , Ye Qiao , Luxiang Sun , Hong Cheng
Crepidatin, a bioactive bibenzyl derivative isolated from multiple plants of Dendrobium, has recently gathered significant interest due to its activity against lung cancer. The aim of this study was to elucidate the metabolic fates of crepidatin using human liver microsomes and hepatocytes. To achieve this goal, a specific HPLC-MS/MS method for the determination of crepidatin in human liver microsomes and hepatocytes was developed and validated. Separation was achieved within 2 min on an ACQUITY BEH C18 column (1.7 μm, 2.1 × 50 mm) with a mobile phase consisting of 0.1 % formic acid in water and methanol/acetonitrile (30:70, v/v), running at 0.4 mL/min. Detection employed positive electrospray ionization in multiple reaction monitoring (MRM) mode, with the transition m/z 319.1→151.1 for both crepidatin and internal standard (erianin). The developed method displayed excellent linearity over the concentration range of 1.0–1000 nM (r > 0.999). Crepidatin showed rapid metabolism both in human liver microsomes (t1/2: 17.2 min) and hepatocytes (t1/2: 10.8 min). Metabolite identification was achieved using HPLC-Q-Orbitrap-HRMS operated in full MS/dd-MS2 scan mode. A total of 15 metabolites across five metabolic pathways were structurally characterized through accurate mass measurement and fragmentation analysis. Hydroxylation (M5, M7) and glucuronidation (M14) emerged as the predominant metabolic pathways. Furthermore, incubation with glutathione (GSH)-supplemented human liver microsomes led to the detection of three GSH conjugates, providing direct evidence for crepidatin bioactivation via the formation of ortho-quinone and quinone-methide species. Crepidatin was primarily metabolized by CYP3A4, 2C8, 2C19, UGT1A1, UGT1A8, and UGT1A9. This study describes the first integrated HPLC-MS/MS and HPLC-Q-Orbitrap-HRMS method for its in vitro metabolic profiling, and the results facilitate prediction of in vivo pharmacokinetics.
{"title":"Characterization of In Vitro metabolic profiles of crepidatin in human liver microsomes and hepatocytes using HPLC-MS/MS and HPLC-Q-Orbitrap-HRMS","authors":"Yaqin Sun , Junyin Zhou , Xiaoli Hou , Tianqi Ma , Ye Qiao , Luxiang Sun , Hong Cheng","doi":"10.1016/j.jpba.2025.117269","DOIUrl":"10.1016/j.jpba.2025.117269","url":null,"abstract":"<div><div>Crepidatin, a bioactive bibenzyl derivative isolated from multiple plants of Dendrobium, has recently gathered significant interest due to its activity against lung cancer. The aim of this study was to elucidate the metabolic fates of crepidatin using human liver microsomes and hepatocytes. To achieve this goal, a specific HPLC-MS/MS method for the determination of crepidatin in human liver microsomes and hepatocytes was developed and validated. Separation was achieved within 2 min on an ACQUITY BEH C<sub>18</sub> column (1.7 μm, 2.1 × 50 mm) with a mobile phase consisting of 0.1 % formic acid in water and methanol/acetonitrile (30:70, <em>v/v</em>), running at 0.4 mL/min. Detection employed positive electrospray ionization in multiple reaction monitoring (MRM) mode, with the transition <em>m/z</em> 319.1→151.1 for both crepidatin and internal standard (erianin). The developed method displayed excellent linearity over the concentration range of 1.0–1000 nM (<em>r</em> > 0.999). Crepidatin showed rapid metabolism both in human liver microsomes (<em>t</em><sub>1/2</sub>: 17.2 min) and hepatocytes (<em>t</em><sub>1/2</sub>: 10.8 min). Metabolite identification was achieved using HPLC-Q-Orbitrap-HRMS operated in full MS/dd-MS<sup>2</sup> scan mode. A total of 15 metabolites across five metabolic pathways were structurally characterized through accurate mass measurement and fragmentation analysis. Hydroxylation (M5, M7) and glucuronidation (M14) emerged as the predominant metabolic pathways. Furthermore, incubation with glutathione (GSH)-supplemented human liver microsomes led to the detection of three GSH conjugates, providing direct evidence for crepidatin bioactivation via the formation of ortho-quinone and quinone-methide species. Crepidatin was primarily metabolized by CYP3A4, 2C8, 2C19, UGT1A1, UGT1A8, and UGT1A9. This study describes the first integrated HPLC-MS/MS and HPLC-Q-Orbitrap-HRMS method for its in vitro metabolic profiling, and the results facilitate prediction of <em>in vivo</em> pharmacokinetics.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117269"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145555412","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Health hazards associated with the ingestion of dietary supplements containing illegal adulterants have been reported. High-performance liquid chromatographysingle quadrupole mass spectrometry (LC-MS) is useful for identification of illegal adulterants in dietary supplements because of its moderate sensitivity and wide accessibility. Previously, we reported a method for the simultaneous identification of 18 illegal adulterants in dietary supplements using LC-MS. Considering numerous analogs of pharmaceutical ingredients recently identified, developing a more convenient screening method to identify a broader range of such compounds is essential. In this study, we introduced another LC-MS-based method to simultaneously identify 50 illegal adulterants in dietary supplements, including newly identified compounds such as descarbonsildenafil and N-phenylpropoxyphenylcarbodenafil. The proposed method achieved clear separation of the targeted peaks in their extracted ion chromatograms. All coefficients of determination (r2) exceeded 0.9794, with most compounds showing good linearity above 0.99. Recovery tests confirmed the specificity of detection for each compound, with recovery rates ranging from 74.9 % to 127.7 % and relative standard deviations below 21.3 %. Validation results demonstrated the method’s suitability for identifying all 50 targeted compounds and quantifying 32 compounds with acceptable accuracy and precision. The method also successfully detected illegal adulterants in positive samples. Thus, the proposed method may be useful for identifying—and partially quantifying—illegal adulterants present in dietary supplements.
{"title":"Simultaneous identification of 50 illegal adulterants in dietary supplements using high-performance liquid chromatography–single quadrupole mass spectrometry","authors":"Takayuki Sakai , Misa Tanaka , Yuki Azuma, Yusuke Sakamoto, Takaomi Tagami, Akiko Asada, Takahiro Doi","doi":"10.1016/j.jpba.2025.117300","DOIUrl":"10.1016/j.jpba.2025.117300","url":null,"abstract":"<div><div>Health hazards associated with the ingestion of dietary supplements containing illegal adulterants have been reported. High-performance liquid chromatography<img>single quadrupole mass spectrometry (LC-MS) is useful for identification of illegal adulterants in dietary supplements because of its moderate sensitivity and wide accessibility. Previously, we reported a method for the simultaneous identification of 18 illegal adulterants in dietary supplements using LC-MS. Considering numerous analogs of pharmaceutical ingredients recently identified, developing a more convenient screening method to identify a broader range of such compounds is essential. In this study, we introduced another LC-MS-based method to simultaneously identify 50 illegal adulterants in dietary supplements, including newly identified compounds such as descarbonsildenafil and <em>N</em>-phenylpropoxyphenylcarbodenafil. The proposed method achieved clear separation of the targeted peaks in their extracted ion chromatograms. All coefficients of determination (r<sup>2</sup>) exceeded 0.9794, with most compounds showing good linearity above 0.99. Recovery tests confirmed the specificity of detection for each compound, with recovery rates ranging from 74.9 % to 127.7 % and relative standard deviations below 21.3 %. Validation results demonstrated the method’s suitability for identifying all 50 targeted compounds and quantifying 32 compounds with acceptable accuracy and precision. The method also successfully detected illegal adulterants in positive samples. Thus, the proposed method may be useful for identifying—and partially quantifying—illegal adulterants present in dietary supplements.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117300"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145714540","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-03-15Epub Date: 2025-11-25DOI: 10.1016/j.jpba.2025.117283
Ilona Szabłowska-Gadomska, Marta Bochyńska-Czyż, Agnieszka Mroczko
Ensuring sterility is a key quality requirement in the manufacturing of cell-based medicinal products. This study evaluated the capability of the BACTEC automated microbial detection system to identify viable microorganisms in adipose-derived stem cell-based products suspended in Ringer’s Lactate. The validation process comprised four stages: i) sterility testing of uninoculated media to assess reagent quality; ii) growth promotion test to confirm medium performance; iii) suitability testing for compatibility with the sample matrix; iv) confirmation using pharmacopeial methods. All inocula met the European Pharmacopoeia recommendations. Validation was defined as the demonstration that the BACTEC system provides results equivalent to the pharmacopeial sterility test in terms of sensitivity, reliability, and detection time. Evaluated parameters included accuracy, repeatability, and detection limit, following European Pharmacopoeia requirements for alternative microbiological methods. The system successfully detected microbial contamination for all reference strains with a detection limit of 5 CFU and repeatability > 95 %. All microorganisms showed growth within 24–72 h, well inside the 7-day acceptance criterion. Quantitative comparison with the pharmacopeial method demonstrated full concordance (Cohen’s κ = 1.0) and a strong correlation of detection times (r = 0.92, p < 0.001). These results confirm that the BACTEC system is a validated and reliable alternative to the pharmacopeial sterility test, providing faster detection without loss of analytical accuracy. The validation supports its applicability for sterility assurance of cell-based materials and integration into quality management during the manufacture of advanced therapy medicinal products.
{"title":"Validation of the Bactec system for sterility testing of advanced therapy medicinal product suspensions in Ringer’s Lactate","authors":"Ilona Szabłowska-Gadomska, Marta Bochyńska-Czyż, Agnieszka Mroczko","doi":"10.1016/j.jpba.2025.117283","DOIUrl":"10.1016/j.jpba.2025.117283","url":null,"abstract":"<div><div>Ensuring sterility is a key quality requirement in the manufacturing of cell-based medicinal products. This study evaluated the capability of the BACTEC automated microbial detection system to identify viable microorganisms in adipose-derived stem cell-based products suspended in Ringer’s Lactate. The validation process comprised four stages: i) sterility testing of uninoculated media to assess reagent quality; ii) growth promotion test to confirm medium performance; iii) suitability testing for compatibility with the sample matrix; iv) confirmation using pharmacopeial methods. All inocula met the European Pharmacopoeia recommendations. Validation was defined as the demonstration that the BACTEC system provides results equivalent to the pharmacopeial sterility test in terms of sensitivity, reliability, and detection time. Evaluated parameters included accuracy, repeatability, and detection limit, following European Pharmacopoeia requirements for alternative microbiological methods. The system successfully detected microbial contamination for all reference strains with a detection limit of 5 CFU and repeatability > 95 %. All microorganisms showed growth within 24–72 h, well inside the 7-day acceptance criterion. Quantitative comparison with the pharmacopeial method demonstrated full concordance (Cohen’s κ = 1.0) and a strong correlation of detection times (r = 0.92, p < 0.001). These results confirm that the BACTEC system is a validated and reliable alternative to the pharmacopeial sterility test, providing faster detection without loss of analytical accuracy. The validation supports its applicability for sterility assurance of cell-based materials and integration into quality management during the manufacture of advanced therapy medicinal products.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117283"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145622391","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The present study was undertaken to compare the bioactive potential and presence of metabolites in Agaricus bisporus fruiting body (ABF) and Agaricus bisporus stipe (ABS) using Ultrasound-assisted extraction (UAE), which enhanced the extraction yield by 1.3-fold and 1.7-fold in ABF and ABS, respectively. ABF expressed a higher amount of total phenol content, which correlates with the antioxidant potential determined by DPPH, FRAP, and ABTS assays. Liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) determined 1225 and 1249 compounds in ABS and ABF, respectively. Among the untargeted compounds, gamma-sitosterol and phenylalanine were found abundant in ABS and ABF, respectively. Principal Component Analysis (PCA) further validated the correlation among the obtained metabolites from ABS and ABF. Furthermore, KEGG analysis was performed to understand the dominance of metaboloic pathways. To the best of our knowledge, herein we report the first comprehensive LC/MS-QTOF metabolite profiling of ABS, revealing its nutraceutical potential.
{"title":"Ultrasonication-assisted extraction and exploration of untargeted metabolomics in Agaricus bisporus fruiting body and stipe using LC/MS Q-TOF","authors":"Charu Agnihotri , Bhim Pratap Singh , Shwet Kamal , Vijay Singh Sharanagat","doi":"10.1016/j.jpba.2025.117257","DOIUrl":"10.1016/j.jpba.2025.117257","url":null,"abstract":"<div><div>The present study was undertaken to compare the bioactive potential and presence of metabolites in <em>Agaricus bisporus</em> fruiting body (ABF) and <em>Agaricus bisporus</em> stipe (ABS) using Ultrasound-assisted extraction (UAE), which enhanced the extraction yield by 1.3-fold and 1.7-fold in ABF and ABS, respectively. ABF expressed a higher amount of total phenol content, which correlates with the antioxidant potential determined by DPPH, FRAP, and ABTS assays. Liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) determined 1225 and 1249 compounds in ABS and ABF, respectively. Among the untargeted compounds, gamma-sitosterol and phenylalanine were found abundant in ABS and ABF, respectively. Principal Component Analysis (PCA) further validated the correlation among the obtained metabolites from ABS and ABF. Furthermore, KEGG analysis was performed to understand the dominance of metaboloic pathways. To the best of our knowledge, herein we report the first comprehensive LC/MS-QTOF metabolite profiling of ABS, revealing its nutraceutical potential.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117257"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145622392","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-03-15Epub Date: 2025-11-22DOI: 10.1016/j.jpba.2025.117278
Sevda Hasanova , Eda Gumus , Erhan Zor
The incidence and mortality rates of cancer are rapidly increasing worldwide. As the global population grows and ages, cancer has become a leading cause of death, partly due to significant reductions in mortality rates from stroke and coronary heart disease in many countries. Despite significant advances in cancer therapy, there is a substantial interest in developing new anticancer agents with different mechanisms of action, as cancer cells have developed resistance to current treatments. Noteworthy anticancer agents in this category include doxorubicin, 5-fluorouracil, and methotrexate among others. There has been a concerted effort to develop rapid, sensitive, and non-destructive methods for detecting the effects and mechanisms of anticancer drugs both in vitro and in vivo. Currently, methods such as mass spectrometry-based liquid chromatography (LC-MS), capillary electrophoresis (CE), and mass spectrometry-based gas chromatography (GC-MS) are still widely used to study anticancer drugs. However, electroanalytical techniques have recently gained popularity due to their higher sensitivity, greater selectivity, eco-friendliness, shorter analysis time, and lower cost. These techniques are now frequently used to detect anticancer drugs. Identifying anticancer drugs at low concentrations and with high sensitivity is crucial for tracking these medications. In this study, we discuss the recent advances in electrochemical sensors for the detection of anticancer drugs between 2019 and 2024.
{"title":"Recent advances in electrochemical sensors for the detection of anticancer drugs","authors":"Sevda Hasanova , Eda Gumus , Erhan Zor","doi":"10.1016/j.jpba.2025.117278","DOIUrl":"10.1016/j.jpba.2025.117278","url":null,"abstract":"<div><div>The incidence and mortality rates of cancer are rapidly increasing worldwide. As the global population grows and ages, cancer has become a leading cause of death, partly due to significant reductions in mortality rates from stroke and coronary heart disease in many countries. Despite significant advances in cancer therapy, there is a substantial interest in developing new anticancer agents with different mechanisms of action, as cancer cells have developed resistance to current treatments. Noteworthy anticancer agents in this category include doxorubicin, 5-fluorouracil, and methotrexate among others. There has been a concerted effort to develop rapid, sensitive, and non-destructive methods for detecting the effects and mechanisms of anticancer drugs both in vitro and in vivo. Currently, methods such as mass spectrometry-based liquid chromatography (LC-MS), capillary electrophoresis (CE), and mass spectrometry-based gas chromatography (GC-MS) are still widely used to study anticancer drugs. However, electroanalytical techniques have recently gained popularity due to their higher sensitivity, greater selectivity, eco-friendliness, shorter analysis time, and lower cost. These techniques are now frequently used to detect anticancer drugs. Identifying anticancer drugs at low concentrations and with high sensitivity is crucial for tracking these medications. In this study, we discuss the recent advances in electrochemical sensors for the detection of anticancer drugs between 2019 and 2024.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117278"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145622394","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
扫码关注我们
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号