Pub Date : 2025-10-02DOI: 10.1016/j.jchromb.2025.124812
Asena Avci Akca Ph.D. , Sefa Akca Ph.D.
Retention time (RT) prediction can greatly improve the efficiency of chromatographic workflows in forensic toxicology, especially in high-throughput or non-targeted analytical workflows. In the present study, we compare the performance of four ensemble machine learning models—Random Forest (RF), Extra Trees, XGBoost, and LightGBM—in predicting RTs of 229 structurally diverse forensic compounds. Each compound was represented by a minimal set of RDKit-derived descriptors and an extended feature space that combines Mordred descriptors and Morgan circular fingerprints. All RTs were experimentally measured under standardized reversed-phase liquid chromatographic conditions. Model performance was evaluated using coefficient of determination (R2) and root-mean-square error (RMSE). Results show that models trained on extended descriptors (>2000 molecular features) outperformed those trained on basic descriptors, with XGBoost showing the highest predictive power (R2 = 0.718, RMSE = 1.23). Feature importance analysis showed that RTs are not only affected by global molecular properties like hydrophobicity and size but also by topological and electronic features. These results highlight the value of ensemble learning in RT prediction and demonstrate its practical utility in compound screening and chromatographic method development in forensic toxicology.
{"title":"Retention time prediction of forensic compounds using ensemble machine learning and molecular descriptors","authors":"Asena Avci Akca Ph.D. , Sefa Akca Ph.D.","doi":"10.1016/j.jchromb.2025.124812","DOIUrl":"10.1016/j.jchromb.2025.124812","url":null,"abstract":"<div><div>Retention time (RT) prediction can greatly improve the efficiency of chromatographic workflows in forensic toxicology, especially in high-throughput or non-targeted analytical workflows. In the present study, we compare the performance of four ensemble machine learning models—Random Forest (RF), Extra Trees, XGBoost, and LightGBM—in predicting RTs of 229 structurally diverse forensic compounds. Each compound was represented by a minimal set of RDKit-derived descriptors and an extended feature space that combines Mordred descriptors and Morgan circular fingerprints. All RTs were experimentally measured under standardized reversed-phase liquid chromatographic conditions. Model performance was evaluated using coefficient of determination (R<sup>2</sup>) and root-mean-square error (RMSE). Results show that models trained on extended descriptors (>2000 molecular features) outperformed those trained on basic descriptors, with XGBoost showing the highest predictive power (R<sup>2</sup> = 0.718, RMSE = 1.23). Feature importance analysis showed that RTs are not only affected by global molecular properties like hydrophobicity and size but also by topological and electronic features. These results highlight the value of ensemble learning in RT prediction and demonstrate its practical utility in compound screening and chromatographic method development in forensic toxicology.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1267 ","pages":"Article 124812"},"PeriodicalIF":2.8,"publicationDate":"2025-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145234244","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 : 2025-10-02DOI: 10.1016/j.jchromb.2025.124809
Yunfan Zhao , Yang Xie , Hulei Zhao , Jinyan Wu , Xuemei Xu , Liuyue Yin , Yanmin Shi , Jianya Yang , Peng Zhao , Qingzhou Guan , Yan Du , Suyun Li , Jiansheng Li , Xinguang Liu
Triple quadrupole (QQQ) MS based pseudotargeted lipidomics combines high coverage and quantitative accuracy, is commonly established by selecting the most responsive lipid ion pairs identified from high-resolution mass spectrometry (HRMS), then sending them to QQQ MS for quantification by multiple reaction monitoring. Due to the low resolution of QQQ MS, it may result in faulty peak identification in integration and method transition from HRMS to QQQ MS, thus, directly establish pseudotargeted lipidomics on HRMS is needed to improve the accuracy of present methods. We propose a method for rapidly screening high-resolution ion pairs for constructing multiplex-HRMS-based pseudotargeted lipidomics (MHPL). Firstly, high-resolution precursor and product ions for lipid quantification were obtained by HRMS. To solve the problem of lower acquisition speed in HRMS, we used the multiplex mode to simultaneously fragment co-eluting lipid precursor ions within one isolation window, scan the MS/MS mass spectra, and quantify the unique product ions (UPI) of co-eluting lipid (defined as Quan-PIs). Meanwhile, we provide a group of scripts for searching for lipid Quan-PIs in multiplex mode. The proposed MHPL strategy could ensure accurate quantification of 460 lipids within 5 injections, and was applied in serum differential lipid discovery for chronic obstructive pulmonary disease (COPD) patients. As a result, 47 differential lipids were found to comprise a potential biomarker panel for COPD diagnosis. The lipid identification and quantification in this work were conducted in the same instrument, and faulty peak identification was considerably reduced in integration and when transitioning methods from HRMS to QQQ MS.
{"title":"Screening biomarkers for diagnosis of COPD by multiplex-high-resolution mass spectrometry based pseudotargeted lipidomics","authors":"Yunfan Zhao , Yang Xie , Hulei Zhao , Jinyan Wu , Xuemei Xu , Liuyue Yin , Yanmin Shi , Jianya Yang , Peng Zhao , Qingzhou Guan , Yan Du , Suyun Li , Jiansheng Li , Xinguang Liu","doi":"10.1016/j.jchromb.2025.124809","DOIUrl":"10.1016/j.jchromb.2025.124809","url":null,"abstract":"<div><div>Triple quadrupole (QQQ) MS based pseudotargeted lipidomics combines high coverage and quantitative accuracy, is commonly established by selecting the most responsive lipid ion pairs identified from high-resolution mass spectrometry (HRMS), then sending them to QQQ MS for quantification by multiple reaction monitoring. Due to the low resolution of QQQ MS, it may result in faulty peak identification in integration and method transition from HRMS to QQQ MS, thus, directly establish pseudotargeted lipidomics on HRMS is needed to improve the accuracy of present methods. We propose a method for rapidly screening high-resolution ion pairs for constructing multiplex-HRMS-based pseudotargeted lipidomics (MHPL). Firstly, high-resolution precursor and product ions for lipid quantification were obtained by HRMS. To solve the problem of lower acquisition speed in HRMS, we used the multiplex mode to simultaneously fragment co-eluting lipid precursor ions within one isolation window, scan the MS/MS mass spectra, and quantify the unique product ions (UPI) of co-eluting lipid (defined as Quan-PIs). Meanwhile, we provide a group of scripts for searching for lipid Quan-PIs in multiplex mode. The proposed MHPL strategy could ensure accurate quantification of 460 lipids within 5 injections, and was applied in serum differential lipid discovery for chronic obstructive pulmonary disease (COPD) patients. As a result, 47 differential lipids were found to comprise a potential biomarker panel for COPD diagnosis. The lipid identification and quantification in this work were conducted in the same instrument, and faulty peak identification was considerably reduced in integration and when transitioning methods from HRMS to QQQ MS.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1267 ","pages":"Article 124809"},"PeriodicalIF":2.8,"publicationDate":"2025-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145217305","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 : 2025-10-02DOI: 10.1016/j.jchromb.2025.124807
Gjulten Nedjip , Eyyup Karaogul
This study investigated the effects of phenolic composition in olive leaf extract (OLE) obtained using microwave-assisted extraction (MAE) and conventional extraction (CE). Liquid chromatography-tandem mass spectrometry (LC-MS/MS) and response surface methodology (RSM) were applied to mathematically model and optimize extraction parameters. The study aims to enhance bioactive yields, antioxidant activity, total phenolic content (TPC), and total flavonoid content (TFC) for potential applications in the food and pharmaceutical industries. Olive leaf extract (OLE) was obtained using MAE (250–350 watt, 30–90 min) and CE (30–90 min). Response surface methodology identified optimal conditions at 300 W for 60 min (MAE; desirability: 0.962) and 90 min for CE (desirability: 0.998). Linear models most accurately described thymoquinone, total phenolic compounds (TPc) (P < 0.01), hydroxycinnamic acid (P < 0.05), fumaric acid, caffeic acid, hydroxybenzoic acid, and total hydrolyzed tannins (THT) (P > 0.05) under CE. Oleuropein, quercetin, total condensed tannins (TCT) (P < 0.0001), and luteolin (P > 0.05) were best fitted to quadratic models, while caffeic acid, hydroxybenzoic acid, and thymoquinone under MAE required quadratic fits, with cubic models for remaining compounds (P > 0.05). Antioxidant assays (DPPH, ABTS) showed stronger radical scavenging for maceration (Mc) compared to CE and MAE. Optimizing extraction parameters enhances bioactive compound yield in OLE. MAE produces more phenols and flavonoids, while CE is more energy-efficient and can sometimes achieve antioxidant activity comparable to Mc. Systematic modeling improves phytotherapy research, ensuring more accurate prediction of extraction efficiency.
{"title":"Role of model equations and energy dynamics in understanding bioactive compounds of olive leaf extract by LC-MS/MS; their evaluation using a cluster approach","authors":"Gjulten Nedjip , Eyyup Karaogul","doi":"10.1016/j.jchromb.2025.124807","DOIUrl":"10.1016/j.jchromb.2025.124807","url":null,"abstract":"<div><div>This study investigated the effects of phenolic composition in olive leaf extract (OLE) obtained using microwave-assisted extraction (MAE) and conventional extraction (CE). Liquid chromatography-tandem mass spectrometry (LC-MS/MS) and response surface methodology (RSM) were applied to mathematically model and optimize extraction parameters. The study aims to enhance bioactive yields, antioxidant activity, total phenolic content (TPC), and total flavonoid content (TFC) for potential applications in the food and pharmaceutical industries. Olive leaf extract (OLE) was obtained using MAE (250–350 watt, 30–90 min) and CE (30–90 min). Response surface methodology identified optimal conditions at 300 W for 60 min (MAE; desirability: 0.962) and 90 min for CE (desirability: 0.998). Linear models most accurately described thymoquinone, total phenolic compounds (TPc) (<em>P</em> < 0.01), hydroxycinnamic acid (<em>P</em> < 0.05), fumaric acid, caffeic acid, hydroxybenzoic acid, and total hydrolyzed tannins (THT) (<em>P</em> > 0.05) under CE. Oleuropein, quercetin, total condensed tannins (TCT) (<em>P</em> < 0.0001), and luteolin (P > 0.05) were best fitted to quadratic models, while caffeic acid, hydroxybenzoic acid, and thymoquinone under MAE required quadratic fits, with cubic models for remaining compounds (<em>P</em> > 0.05). Antioxidant assays (DPPH, ABTS) showed stronger radical scavenging for maceration (Mc) compared to CE and MAE. Optimizing extraction parameters enhances bioactive compound yield in OLE. MAE produces more phenols and flavonoids, while CE is more energy-efficient and can sometimes achieve antioxidant activity comparable to Mc. Systematic modeling improves phytotherapy research, ensuring more accurate prediction of extraction efficiency.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1267 ","pages":"Article 124807"},"PeriodicalIF":2.8,"publicationDate":"2025-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145294960","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}
Tripchlorolide is a promising therapeutic compound with potent pharmacological activity and an improved safety profile compared to triptolide. However, its pharmacokinetics and tissue distribution remain poorly characterized. In this study, we developed and validated a rapid and sensitive liquid chromatography–mass spectrometry method for the quantification of tripchlorolide in biological matrices, using triptolide as the internal standard. Quantification was performed in selective ion monitoring mode, following liquid–liquid extraction with ethyl acetate. Chromatographic separation was achieved on a 3.5 μm Agilent ZORBAX Eclipse Plus-C18 column under isocratic elution with a methanol–water mobile phase. Calibration curves were linear over the range of 0.16–200 ng/mL in rat plasma. The method was successfully applied to a pharmacokinetic study in rats and tissue distribution analysis in mice. Tripchlorolide exhibited an absolute bioavailability of 72.97 % after intraperitoneal administration and a half-life of approximately 45 min, with no significant sex-based differences in pharmacokinetic parameters. Tissue distribution following intravenous administration (400 μg/kg) in mice revealed the highest accumulation in the liver, followed by the kidney, spleen, testis, heart, intestine, and brain. These findings provide essential preclinical data for further development of tripchlorolide as a safe and effective therapeutic candidate.
{"title":"Pharmacokinetics and tissue distribution of tripchlorolide in rodents using liquid chromatography tandem mass spectrometry","authors":"Yanping Deng , Lele Zhou , Zhengyan Gu , Zhou Chen","doi":"10.1016/j.jchromb.2025.124805","DOIUrl":"10.1016/j.jchromb.2025.124805","url":null,"abstract":"<div><div>Tripchlorolide is a promising therapeutic compound with potent pharmacological activity and an improved safety profile compared to triptolide. However, its pharmacokinetics and tissue distribution remain poorly characterized. In this study, we developed and validated a rapid and sensitive liquid chromatography–mass spectrometry method for the quantification of tripchlorolide in biological matrices, using triptolide as the internal standard. Quantification was performed in selective ion monitoring mode, following liquid–liquid extraction with ethyl acetate. Chromatographic separation was achieved on a 3.5 μm Agilent ZORBAX Eclipse Plus-C18 column under isocratic elution with a methanol–water mobile phase. Calibration curves were linear over the range of 0.16–200 ng/mL in rat plasma. The method was successfully applied to a pharmacokinetic study in rats and tissue distribution analysis in mice. Tripchlorolide exhibited an absolute bioavailability of 72.97 % after intraperitoneal administration and a half-life of approximately 45 min, with no significant sex-based differences in pharmacokinetic parameters. Tissue distribution following intravenous administration (400 μg/kg) in mice revealed the highest accumulation in the liver, followed by the kidney, spleen, testis, heart, intestine, and brain. These findings provide essential preclinical data for further development of tripchlorolide as a safe and effective therapeutic candidate.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1267 ","pages":"Article 124805"},"PeriodicalIF":2.8,"publicationDate":"2025-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145246285","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}
A highly sensitive and specific liquid chromatography-tandem mass spectrometry method was developed, fully validated, and successfully implemented for routine analysis to simultaneously quantify Bictegravir, Emtricitabine, Doravirine, Cabotegravir, Lenacapavir, Fostemsavir, Tenofovir alafenamide, and their metabolites, Temsavir and Tenofovir, in human plasma. The sample preparation employed a commercial liquid-liquid extraction kit optimized for low plasma volumes (50 μL), which also included the Internal Standard. The method demonstrated excellent precision, accuracy, and robustness, making it suitable for pharmacokinetic and therapeutic drug monitoring applications. Analyte separation was carried out using a gradient elution program over a total run time of seven minutes, with a flow rate of 0.35 mL/min. The mobile phase consisted of solvent A (water containing 0.1 % formic acid) and solvent B (acetonitrile containing 0.1 % formic acid). Detection was performed using a QTRAP® 5500 triple quadrupole mass spectrometer (SCIEX) equipped with an electrospray ionization source operating in positive ion mode. Ion monitoring was performed in multiple reaction monitoring (MRM) mode for all analytes. The method was validated in accordance with European Medicines Agency (EMA) guidelines across clinically relevant concentration ranges. The proposed method was successfully implemented in routine analysis. Following the initial months of application, biological samples from 165 patients were analyzed primarily to assess therapy adherence and confirm that drug blood concentrations reached the minimum threshold. Additionally, data on drug pharmacokinetics were obtained. Our findings indicate that the proposed method is a reliable and accurate tool for high-throughput screening that could be readily used by the clinicians to optimize therapeutic treatments, verify patients' adherence and reduce drug-related toxicities.
{"title":"Development, validation and clinical implementation of a HPLC-MS/MS method for the simultaneous quantification of bictegravir, emtricitabine, doravirine, cabotegravir, lenacapavir, fostemsavir, tenofovir alafenamide and the corresponding metabolites temsavir and tenofovir, in human plasma","authors":"Alessia Mattino , Davide Ferrari , Silvia Nozza , Camilla Muccini , Marco Ripa , Vincenzo Spagnuolo , Antonella Castagna , Massimo Locatelli , Eleonora Sabetta","doi":"10.1016/j.jchromb.2025.124803","DOIUrl":"10.1016/j.jchromb.2025.124803","url":null,"abstract":"<div><div>A highly sensitive and specific liquid chromatography-tandem mass spectrometry method was developed, fully validated, and successfully implemented for routine analysis to simultaneously quantify Bictegravir, Emtricitabine, Doravirine, Cabotegravir, Lenacapavir, Fostemsavir, Tenofovir alafenamide, and their metabolites, Temsavir and Tenofovir, in human plasma. The sample preparation employed a commercial liquid-liquid extraction kit optimized for low plasma volumes (50 μL), which also included the Internal Standard. The method demonstrated excellent precision, accuracy, and robustness, making it suitable for pharmacokinetic and therapeutic drug monitoring applications. Analyte separation was carried out using a gradient elution program over a total run time of seven minutes, with a flow rate of 0.35 mL/min. The mobile phase consisted of solvent A (water containing 0.1 % formic acid) and solvent B (acetonitrile containing 0.1 % formic acid). Detection was performed using a QTRAP® 5500 triple quadrupole mass spectrometer (SCIEX) equipped with an electrospray ionization source operating in positive ion mode. Ion monitoring was performed in multiple reaction monitoring (MRM) mode for all analytes. The method was validated in accordance with European Medicines Agency (EMA) guidelines across clinically relevant concentration ranges. The proposed method was successfully implemented in routine analysis. Following the initial months of application, biological samples from 165 patients were analyzed primarily to assess therapy adherence and confirm that drug blood concentrations reached the minimum threshold. Additionally, data on drug pharmacokinetics were obtained. Our findings indicate that the proposed method is a reliable and accurate tool for high-throughput screening that could be readily used by the clinicians to optimize therapeutic treatments, verify patients' adherence and reduce drug-related toxicities.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1267 ","pages":"Article 124803"},"PeriodicalIF":2.8,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145263140","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 : 2025-09-30DOI: 10.1016/j.jchromb.2025.124804
Yiting Zhang , Ruojia Li , Hangtian Song , Andrew McClain , Richard Ludwig , Li Tao , Jeff Beckman , Ming Zeng
High molecular weight species (HMW) is a critical quality attribute in biopharmaceuticals requiring comprehensive characterization and accurate quantification throughout drug development and manufacturing to ensure product efficacy and patient safety. In this study, the primary goal was to develop a robust size exclusion chromatography (SEC) method to quantify HMW varying in relative proportion to its parent molecule, an Fc-fusion protein therapeutic (FC1) for quality control release and stability testing. However, a unique “partially reversible” HMW kinetics behavior was observed, with dissociation towards equilibrium occurring slowly, up to several days, with rates dependent on time, temperature, concentration, and formulation components. This kinetic behavior posed challenges to process optimization and quality control testing, as the “true” quantitative value of HMW was highly variable based on these conditions, varying by up to an order of magnitude in relative proportion. To address this, mathematical models were built to describe HMW equilibrium kinetics which supported the development of a robust SEC test method that accurately quantifies this attribute under relevant conditions. The developed method was validated and implemented in quality control laboratory for drug release and stability testing. This work also provides new insights into the need to control protein aggregation dynamics as it relates to process optimization and drug product release requirements.
{"title":"Navigating atypical equilibrium dynamics of protein aggregates for biopharmaceutical release and stability monitoring","authors":"Yiting Zhang , Ruojia Li , Hangtian Song , Andrew McClain , Richard Ludwig , Li Tao , Jeff Beckman , Ming Zeng","doi":"10.1016/j.jchromb.2025.124804","DOIUrl":"10.1016/j.jchromb.2025.124804","url":null,"abstract":"<div><div>High molecular weight species (HMW) is a critical quality attribute in biopharmaceuticals requiring comprehensive characterization and accurate quantification throughout drug development and manufacturing to ensure product efficacy and patient safety. In this study, the primary goal was to develop a robust size exclusion chromatography (SEC) method to quantify HMW varying in relative proportion to its parent molecule, an Fc-fusion protein therapeutic (FC1) for quality control release and stability testing. However, a unique “partially reversible” HMW kinetics behavior was observed, with dissociation towards equilibrium occurring slowly, up to several days, with rates dependent on time, temperature, concentration, and formulation components. This kinetic behavior posed challenges to process optimization and quality control testing, as the “true” quantitative value of HMW was highly variable based on these conditions, varying by up to an order of magnitude in relative proportion. To address this, mathematical models were built to describe HMW equilibrium kinetics which supported the development of a robust SEC test method that accurately quantifies this attribute under relevant conditions. The developed method was validated and implemented in quality control laboratory for drug release and stability testing. This work also provides new insights into the need to control protein aggregation dynamics as it relates to process optimization and drug product release requirements.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1267 ","pages":"Article 124804"},"PeriodicalIF":2.8,"publicationDate":"2025-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145254479","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 : 2025-09-29DOI: 10.1016/j.jchromb.2025.124800
Chunhui Du, Zhenyu Zhao
Objective
To systematically characterise adverse drug reactions (ADRs) of four alpha (α)-glucosidase inhibitors – acarbose, voglibose, miglitol and mulberry twig alkaloids – using real-world data from the World Health Organisation VigiAccess® database.
Methods
A retrospective descriptive analysis was conducted on 9410 ADR reports (1983–2024). Disproportionality analyses (reporting odds ratio [ROR]/empirical Bayesian geometric mean [EBGM]) identified drug-adverse-event associations.
Results
Gastrointestinal disorders were the most frequently reported system organ class for all drugs (acarbose: 43.10 %; voglibose: 39.02 %; miglitol: 40.81 %; mulberry twig alkaloids: 23.68 %). For acarbose, diarrhoea (report rate 7.51 %, ROR = 6.53, 95 %CI 6.07–7.02; EBGM = 6.11, 95 %CI 5.68–6.57) and abdominal distension (report rate 7.45 %, ROR = 56.69, 95 %CI 52.68–61.00; EBGM = 52.30, 95 %CI 48.60–56.27) showed the highest reporting rates. Notably, mulberry twig alkaloids exhibited unique cardiac signals, including chest pain (report rate: 7.89 %, ROR = 14.80, 95 % CI: 4.55–48.13; EBGM = 13.71, 95 % CI: 4.22–44.59) and supraventricular tachycardia (report rate: 2.63 %, ROR = 198.83, 95 % CI: 27.28–1449.26; EBGM = 193.61, 95 % CI: 26.56–1411.23).
Conclusion
This study provides the first large-scale comparison of natural versus synthetic α-glucosidase inhibitors, highlighting gastrointestinal toxicity as a shared risk and cardiotoxicity as a novel concern for mulberry twig alkaloids. These findings underscore the need for cardiac monitoring in patients receiving natural inhibitors.
{"title":"Characteristic analysis of adverse reactions to natural and synthetic alpha-glucosidase inhibitors: a descriptive analysis using World Health Organisation-VigiAccess","authors":"Chunhui Du, Zhenyu Zhao","doi":"10.1016/j.jchromb.2025.124800","DOIUrl":"10.1016/j.jchromb.2025.124800","url":null,"abstract":"<div><h3>Objective</h3><div>To systematically characterise adverse drug reactions (ADRs) of four alpha (α)-glucosidase inhibitors – acarbose, voglibose, miglitol and mulberry twig alkaloids – using real-world data from the World Health Organisation VigiAccess® database.</div></div><div><h3>Methods</h3><div>A retrospective descriptive analysis was conducted on 9410 ADR reports (1983–2024). Disproportionality analyses (reporting odds ratio [ROR]/empirical Bayesian geometric mean [EBGM]) identified drug-adverse-event associations.</div></div><div><h3>Results</h3><div>Gastrointestinal disorders were the most frequently reported system organ class for all drugs (acarbose: 43.10 %; voglibose: 39.02 %; miglitol: 40.81 %; mulberry twig alkaloids: 23.68 %). For acarbose, diarrhoea (report rate 7.51 %, ROR = 6.53, 95 %CI 6.07–7.02; EBGM = 6.11, 95 %CI 5.68–6.57) and abdominal distension (report rate 7.45 %, ROR = 56.69, 95 %CI 52.68–61.00; EBGM = 52.30, 95 %CI 48.60–56.27) showed the highest reporting rates. Notably, mulberry twig alkaloids exhibited unique cardiac signals, including chest pain (report rate: 7.89 %, ROR = 14.80, 95 % CI: 4.55–48.13; EBGM = 13.71, 95 % CI: 4.22–44.59) and supraventricular tachycardia (report rate: 2.63 %, ROR = 198.83, 95 % CI: 27.28–1449.26; EBGM = 193.61, 95 % CI: 26.56–1411.23).</div></div><div><h3>Conclusion</h3><div>This study provides the first large-scale comparison of natural versus synthetic α-glucosidase inhibitors, highlighting gastrointestinal toxicity as a shared risk and cardiotoxicity as a novel concern for mulberry twig alkaloids. These findings underscore the need for cardiac monitoring in patients receiving natural inhibitors.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1267 ","pages":"Article 124800"},"PeriodicalIF":2.8,"publicationDate":"2025-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145351437","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 : 2025-09-24DOI: 10.1016/j.jchromb.2025.124802
Jiaxue Wang , Xuanxuan Zhou , Xiaoge Cheng , Renyi Li , Hongjin Wang , Lixin Sun
Huai Hua San (HHS), a traditional Chinese medicine (TCM) originated from the “Pu Ji Ben Shi Fang,” is primarily used to treat conditions such as colitis and hemorrhoids. Despite its demonstrated therapeutic effects on Ulcerative Colitis (UC), the metabolic characteristics in vivo and the mechanisms of action of bioactive components remain unclear. This study systematically investigated the metabolic differences of HHS in normal and UC model rats and explored the association between these variations and the pharmacological effects of key flavonoid components, providing a theoretical basis for further research into its chemical basis and mechanisms of action. The UHPLC-Orbitrap-MS/MS was performed to character the prototype and metabolites, and UPLC-MS/MS was used to elucidate the pharmacokinetic profiles of five main compounds of HHS in rats. Furthermore, we compared the differences of absorption and metabolism between normal and UC rats. Based on the comparison with reference substances, retention times, accurate masses, and characteristic MS/MS fragment patterns, a total of 181 and 167 HHS prototypes and metabolites were inferred in the normal and model groups. Sankey diagram visualization revealed a significant decrease in plasma flavonoid prototype compounds within the UC model group, accompanied by an increased abundance of flavonoid metabolites in urine. This suggests that intestinal absorption efficiency or enterohepatic circulation may be influenced by the inflammatory microenvironment. Further analysis indicated that metabolic pathways of flavonoid compounds, such as glucuronidation, sulfation, and methylation, were altered in UC rats. Furthermore, this investigation delved into the in vivo exposure profiles of five core flavonoid components in HHS (quercetin, naringenin, hesperetin, luteolin, and genistein). The results revealed that naringenin and hesperetin exhibited relatively higher exposure levels, while UC model rats displayed significantly enhanced quercetin absorption rates and markedly increased systemic exposure of genistein. Collectively, this study uncovers alterations in metabolic profiles and changes of pharmacokinetic behavior of HHS under UC pathological conditions, elucidating the dynamic metabolism-exposure-efficacy interrelationships of flavonoid components. These findings provide a theoretical basis for further research on the relationship between the chemical constituents of HHS and its pharmacological mechanisms of action.
{"title":"Metabolic and pharmacokinetic differences of Huai Hua san in Sprague Dawley and ulcerative colitis rats","authors":"Jiaxue Wang , Xuanxuan Zhou , Xiaoge Cheng , Renyi Li , Hongjin Wang , Lixin Sun","doi":"10.1016/j.jchromb.2025.124802","DOIUrl":"10.1016/j.jchromb.2025.124802","url":null,"abstract":"<div><div>Huai Hua San (HHS), a traditional Chinese medicine (TCM) originated from the “Pu Ji Ben Shi Fang,” is primarily used to treat conditions such as colitis and hemorrhoids. Despite its demonstrated therapeutic effects on Ulcerative Colitis (UC), the metabolic characteristics in vivo and the mechanisms of action of bioactive components remain unclear. This study systematically investigated the metabolic differences of HHS in normal and UC model rats and explored the association between these variations and the pharmacological effects of key flavonoid components, providing a theoretical basis for further research into its chemical basis and mechanisms of action. The UHPLC-Orbitrap-MS/MS was performed to character the prototype and metabolites, and UPLC-MS/MS was used to elucidate the pharmacokinetic profiles of five main compounds of HHS in rats. Furthermore, we compared the differences of absorption and metabolism between normal and UC rats. Based on the comparison with reference substances, retention times, accurate masses, and characteristic MS/MS fragment patterns, a total of 181 and 167 HHS prototypes and metabolites were inferred in the normal and model groups. Sankey diagram visualization revealed a significant decrease in plasma flavonoid prototype compounds within the UC model group, accompanied by an increased abundance of flavonoid metabolites in urine. This suggests that intestinal absorption efficiency or enterohepatic circulation may be influenced by the inflammatory microenvironment. Further analysis indicated that metabolic pathways of flavonoid compounds, such as glucuronidation, sulfation, and methylation, were altered in UC rats. Furthermore, this investigation delved into the in vivo exposure profiles of five core flavonoid components in HHS (quercetin, naringenin, hesperetin, luteolin, and genistein). The results revealed that naringenin and hesperetin exhibited relatively higher exposure levels, while UC model rats displayed significantly enhanced quercetin absorption rates and markedly increased systemic exposure of genistein. Collectively, this study uncovers alterations in metabolic profiles and changes of pharmacokinetic behavior of HHS under UC pathological conditions, elucidating the dynamic metabolism-exposure-efficacy interrelationships of flavonoid components. These findings provide a theoretical basis for further research on the relationship between the chemical constituents of HHS and its pharmacological mechanisms of action.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1267 ","pages":"Article 124802"},"PeriodicalIF":2.8,"publicationDate":"2025-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145260498","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 : 2025-09-22DOI: 10.1016/j.jchromb.2025.124799
Aliz Széles , Károly Schöll , Gábor Hirka , Katalin Monostory , Tibor Renkecz
Protoporphyrin IX (PPIX) plays a pivotal role in the heme biosynthesis pathway and serves as both a valuable biomarker in clinical diagnostics and a photosensitizer in photodynamic applications. Despite its physiological importance, accurate quantification of endogenous PPIX in biological matrices remains challenging due to the lack of an analyte-free authentic control matrix and inherent baseline variability. This study describes the development and validation of a high-performance liquid chromatography method with fluorescence detection (HPLC-FLD) for PPIX quantification in rat plasma, applying a surrogate matrix strategy in full compliance with the International Council for Harmonisation M10 guideline.
Endogenous PPIX was removed from rat plasma by visible light-induced analyte stripping, enabling the in-house preparation of a surrogate matrix, for accurate calibration sample generation. The method showed appropriate selectivity, excellent linearity over the range of 10 and 700 ng/mL (r ≥ 0.995), satisfactory precision and accuracy across all validation levels. The lower limit of quantification was established at 10 ng/mL. The stability of both the analyte and internal standard was confirmed under various conditions, including 3 freeze–thaw cycles, short- and long-term storage, and autosampler residence. The method was successfully applied in an in vivo study in which male rats were treated with aminolevulinic acid to induce PPIX formation, thereby confirming its suitability for study sample analysis.
This fluorescence-based HPLC method offers a practical and cost-effective solution for monitoring PPIX in plasma samples. The bioanalytical method validation using a surrogate matrix approach for endogenous PPIX quantification fully aligned with current international regulatory standards may set a precedent for future method development in this field.
{"title":"Validation and sample bioanalysis of protoporphyrin IX in rat plasma by the surrogate matrix approach","authors":"Aliz Széles , Károly Schöll , Gábor Hirka , Katalin Monostory , Tibor Renkecz","doi":"10.1016/j.jchromb.2025.124799","DOIUrl":"10.1016/j.jchromb.2025.124799","url":null,"abstract":"<div><div>Protoporphyrin IX (PPIX) plays a pivotal role in the heme biosynthesis pathway and serves as both a valuable biomarker in clinical diagnostics and a photosensitizer in photodynamic applications. Despite its physiological importance, accurate quantification of endogenous PPIX in biological matrices remains challenging due to the lack of an analyte-free authentic control matrix and inherent baseline variability. This study describes the development and validation of a high-performance liquid chromatography method with fluorescence detection (HPLC-FLD) for PPIX quantification in rat plasma, applying a surrogate matrix strategy in full compliance with the International Council for Harmonisation M10 guideline.</div><div>Endogenous PPIX was removed from rat plasma by visible light-induced analyte stripping, enabling the in-house preparation of a surrogate matrix, for accurate calibration sample generation. The method showed appropriate selectivity, excellent linearity over the range of 10 and 700 ng/mL (<em>r</em> ≥ 0.995), satisfactory precision and accuracy across all validation levels. The lower limit of quantification was established at 10 ng/mL. The stability of both the analyte and internal standard was confirmed under various conditions, including 3 freeze–thaw cycles, short- and long-term storage, and autosampler residence. The method was successfully applied in an <em>in vivo</em> study in which male rats were treated with aminolevulinic acid to induce PPIX formation, thereby confirming its suitability for study sample analysis.</div><div>This fluorescence-based HPLC method offers a practical and cost-effective solution for monitoring PPIX in plasma samples. The bioanalytical method validation using a surrogate matrix approach for endogenous PPIX quantification fully aligned with current international regulatory standards may set a precedent for future method development in this field.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1267 ","pages":"Article 124799"},"PeriodicalIF":2.8,"publicationDate":"2025-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145155300","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 : 2025-09-21DOI: 10.1016/j.jchromb.2025.124795
Cangming Yang , Zachary Z. Brown , Tao Meng , John R. Frost , Ashwin Rao , Xiaohong Zhu , Natalya Pissarnitski , Tianying Jian , Jianping Pan , Min Liu
Peptides are attracting broad interest across the pharmaceutical industry owing to their vast therapeutic potential for drugging high-value targets and flexible dosing and formulation options to improve patient access. As the industry increases focus on peptide-based discovery programs, fast, high-throughput, and green purification methods have become increasingly valuable. Due to the vast sequence and physicochemical diversity of peptides, expedited purification of large libraries has remained a challenge. Herein, we describe the development of a High-Throughput Purification (HTP) approach for peptides wherein each purification is achieved in 6 min or less, representing a substantial reduction in time and solvent consumption compared with conventional methods, all while maintaining high purity levels. This approach relies on Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC), leveraging a dual pump preparative system and short focused solvent gradients to reduce purification times and achieve highly efficient and accelerated high-throughput purification.
{"title":"Enabling faster and greener peptide purification in discovery chemistry","authors":"Cangming Yang , Zachary Z. Brown , Tao Meng , John R. Frost , Ashwin Rao , Xiaohong Zhu , Natalya Pissarnitski , Tianying Jian , Jianping Pan , Min Liu","doi":"10.1016/j.jchromb.2025.124795","DOIUrl":"10.1016/j.jchromb.2025.124795","url":null,"abstract":"<div><div>Peptides are attracting broad interest across the pharmaceutical industry owing to their vast therapeutic potential for drugging high-value targets and flexible dosing and formulation options to improve patient access. As the industry increases focus on peptide-based discovery programs, fast, high-throughput, and green purification methods have become increasingly valuable. Due to the vast sequence and physicochemical diversity of peptides, expedited purification of large libraries has remained a challenge. Herein, we describe the development of a High-Throughput Purification (HTP) approach for peptides wherein each purification is achieved in 6 min or less, representing a substantial reduction in time and solvent consumption compared with conventional methods, all while maintaining high purity levels. This approach relies on Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC), leveraging a dual pump preparative system and short focused solvent gradients to reduce purification times and achieve highly efficient and accelerated high-throughput purification.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1267 ","pages":"Article 124795"},"PeriodicalIF":2.8,"publicationDate":"2025-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145155301","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}
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