Central nervous system (CNS) infections require adequate drug exposure at the site of action, yet antibiotic meropenem (MER) shows limited cerebrospinal fluid (CSF) penetration and easily undergoes non-enzymatic degradation to an inactive open-ring metabolite (ORM). In this study, we developed a simple, sensitive, and fast liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of MER and ORM in human serum and CSF. Chromatographic separation was accomplished on an Agela Venusil MP C18 column, with MER-d6 and ORM-d6 as internal standards. Methanol was found to promote methanolysis, yielding a characteristic product (m/z 416.2). Therefore, acetonitrile was selected as both the organic phase and the protein-precipitation solvent. Method validation was conducted according to the ICH M10 guideline. Follow validation, the method was successfully applied to 57 serum and 16 CSF samples. ORM concentrations in human CSF were reported for the first time. This method provides a valuable tool to support MER monitoring in patients with CNS infections.
{"title":"Development and validation of an UPLC–MS/MS method for simultaneous determination of meropenem and its open-ring metabolite in human serum and cerebrospinal fluid with application to clinical samples","authors":"XiangLong Chen , Jinhui Xu , Chengliang Wang, Lijuan Yang, Jinwei Fan, Tongtong Li, Qian Zhang, Yanxia Yu, Lian Tang, Shenjia Huang","doi":"10.1016/j.jpba.2026.117389","DOIUrl":"10.1016/j.jpba.2026.117389","url":null,"abstract":"<div><div>Central nervous system (CNS) infections require adequate drug exposure at the site of action, yet antibiotic meropenem (MER) shows limited cerebrospinal fluid (CSF) penetration and easily undergoes non-enzymatic degradation to an inactive open-ring metabolite (ORM). In this study, we developed a simple, sensitive, and fast liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of MER and ORM in human serum and CSF. Chromatographic separation was accomplished on an Agela Venusil MP C18 column, with MER-d6 and ORM-d6 as internal standards. Methanol was found to promote methanolysis, yielding a characteristic product (<em>m/z</em> 416.2). Therefore, acetonitrile was selected as both the organic phase and the protein-precipitation solvent. Method validation was conducted according to the ICH M10 guideline. Follow validation, the method was successfully applied to 57 serum and 16 CSF samples. ORM concentrations in human CSF were reported for the first time. This method provides a valuable tool to support MER monitoring in patients with CNS infections.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"273 ","pages":"Article 117389"},"PeriodicalIF":3.1,"publicationDate":"2026-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146119315","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 : 2026-02-01DOI: 10.1016/j.jpba.2026.117387
Yinuo Wu , Xiaoli Li , Haixia Feng , Yifan Chen , Wenwen Wu , Siqin Wang
Insomnia is increasingly recognized as a disorder with complex metabolic underpinnings. We investigated the changes in fecal metabolite levels of 5-hydroxyindoleacetic acid (5-HIAA), octopamine (OA), oleoylethanolamide (OEA), and elaidic acid (EA) in patients with sleep disorders, as well as their correlations with insomnia severity. Sixty participants were divided into two groups, with thirty patients with sleep disorders hospitalized in the Department of Neurology, Zhongda Hospital, Southeast University (October 2024–March 2025) and 30 healthy controls recruited during the same period. Fecal samples were collected from all participants, and metabolite levels were analyzed via untargeted metabolomics analysis using liquid chromatography-mass spectrometry (LC-MS). The Chinese version of the Insomnia Severity Index (C-ISI) was employed to evaluate insomnia severity, and the correlations between insomnia severity and these four metabolites were subjected to statistical analysis. Both univariate and multivariate analyses revealed significant metabolic differences between groups. The experimental group showed significantly lower levels of 5-HIAA (FC = 0.947, P = 0.020) and OA (FC = 0.953, P < 0.001), but higher OEA (FC = 1.101, P < 0.001) and EA (FC = 1.026, P < 0.001). C-ISI scores correlated negatively with 5-HIAA (r = -0.380, P = 0.003) and OA (r = -0.448, P < 0.001), and positively with OEA (r = 0.500, P < 0.001) and EA (r = 0.408, P = 0.001). These fecal metabolites associate with insomnia severity and may serve as potential biomarkers for understanding its pathophysiology and developing interventions.
人们越来越认识到失眠是一种复杂代谢基础的疾病。我们研究了睡眠障碍患者粪便代谢物5-羟基吲哚乙酸（5-HIAA）、章鱼胺（OA）、油基乙醇酰胺（OEA）和elaidic酸（EA）水平的变化及其与失眠严重程度的相关性。60名受试者分为两组，选取东南大学中大医院神经内科住院的睡眠障碍患者30例（2024年10月- 2025年3月）和同期招募的健康对照30例。收集所有参与者的粪便样本，并通过液相色谱-质谱（LC-MS）非靶向代谢组学分析分析代谢物水平。采用中文版失眠症严重程度指数（C-ISI）评估失眠严重程度，并对失眠严重程度与上述四种代谢物的相关性进行统计分析。单因素和多因素分析均显示各组之间的代谢差异显著。实验组5-HIAA （FC = 0.947, P = 0.020）和OA （FC = 0.953, P . 0.05）水平显著低于对照组
{"title":"Fecal metabolic biomarkers associated with insomnia severity: A study on 5-hydroxyindoleacetic acid, octopamine, oleoylethanolamide, and elaidic acid","authors":"Yinuo Wu , Xiaoli Li , Haixia Feng , Yifan Chen , Wenwen Wu , Siqin Wang","doi":"10.1016/j.jpba.2026.117387","DOIUrl":"10.1016/j.jpba.2026.117387","url":null,"abstract":"<div><div>Insomnia is increasingly recognized as a disorder with complex metabolic underpinnings. We investigated the changes in fecal metabolite levels of 5-hydroxyindoleacetic acid (5-HIAA), octopamine (OA), oleoylethanolamide (OEA), and elaidic acid (EA) in patients with sleep disorders, as well as their correlations with insomnia severity. Sixty participants were divided into two groups, with thirty patients with sleep disorders hospitalized in the Department of Neurology, Zhongda Hospital, Southeast University (October 2024–March 2025) and 30 healthy controls recruited during the same period. Fecal samples were collected from all participants, and metabolite levels were analyzed via untargeted metabolomics analysis using liquid chromatography-mass spectrometry (LC-MS). The Chinese version of the Insomnia Severity Index (C-ISI) was employed to evaluate insomnia severity, and the correlations between insomnia severity and these four metabolites were subjected to statistical analysis. Both univariate and multivariate analyses revealed significant metabolic differences between groups. The experimental group showed significantly lower levels of 5-HIAA (FC = 0.947, P = 0.020) and OA (FC = 0.953, P < 0.001), but higher OEA (FC = 1.101, P < 0.001) and EA (FC = 1.026, P < 0.001). C-ISI scores correlated negatively with 5-HIAA (r = -0.380, P = 0.003) and OA (r = -0.448, P < 0.001), and positively with OEA (r = 0.500, P < 0.001) and EA (r = 0.408, P = 0.001). These fecal metabolites associate with insomnia severity and may serve as potential biomarkers for understanding its pathophysiology and developing interventions.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"273 ","pages":"Article 117387"},"PeriodicalIF":3.1,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146119341","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 : 2026-01-30DOI: 10.1016/j.jpba.2026.117386
Michalina McDermott , Zachary Sargeant , Christopher E. Karlsen , Feng Li , R. Andrew Shalliker , Jake A. Cravino
The demand for rapid and reliable analytical methods in the pharmaceutical industry continues to grow, with Ultra-/High-Performance Liquid Chromatography (U/HPLC) remaining the gold standard for impurity profiling, quantification of active pharmaceutical ingredients, and degradation product analysis. However, traditional HPLC methods are often constrained by pressure limitations at higher flow rates, which can hinder analytical throughput. While recent advancements in column technology have improved performance, they typically exacerbate pressure-related challenges. In this study, we evaluate a novel column technology designed to address these limitations by enabling high-resolution separations at reduced pressures and increased flow rates. Our findings demonstrate that the column, when operated in Radial Flow Stream Splitting (RFS) mode, maintains quantitative accuracy and repeatability while achieving up to a 120 % improvement in separation efficiency and a 30 % reduction in backpressure compared to conventional operation. By way of assaying over-the-counter medication, we have found no difference in the quantitative reliability of the assay when in RFS vs stock mode, despite reducing the analysis time by up to 40 %.
{"title":"Cost-effective routine pharmaceutical testing using radial flow stream splitting HPLC columns: Quantitative analysis and performance metrics in the analysis of over-the-counter drugs","authors":"Michalina McDermott , Zachary Sargeant , Christopher E. Karlsen , Feng Li , R. Andrew Shalliker , Jake A. Cravino","doi":"10.1016/j.jpba.2026.117386","DOIUrl":"10.1016/j.jpba.2026.117386","url":null,"abstract":"<div><div>The demand for rapid and reliable analytical methods in the pharmaceutical industry continues to grow, with Ultra-/High-Performance Liquid Chromatography (U/HPLC) remaining the gold standard for impurity profiling, quantification of active pharmaceutical ingredients, and degradation product analysis. However, traditional HPLC methods are often constrained by pressure limitations at higher flow rates, which can hinder analytical throughput. While recent advancements in column technology have improved performance, they typically exacerbate pressure-related challenges. In this study, we evaluate a novel column technology designed to address these limitations by enabling high-resolution separations at reduced pressures and increased flow rates. Our findings demonstrate that the column, when operated in Radial Flow Stream Splitting (RFS) mode, maintains quantitative accuracy and repeatability while achieving up to a 120 % improvement in separation efficiency and a 30 % reduction in backpressure compared to conventional operation. By way of assaying over-the-counter medication, we have found no difference in the quantitative reliability of the assay when in RFS vs stock mode, despite reducing the analysis time by up to 40 %.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"273 ","pages":"Article 117386"},"PeriodicalIF":3.1,"publicationDate":"2026-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146119296","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 : 2026-01-30DOI: 10.1016/j.jpba.2026.117379
Rachel Smith , Colin Guy , Rosie Upton , Sam Clawson , Henry Fisher , Mohammad Adam Nasar , David Firth , Allan Watkinson
A streamlined early phase formulation development workflow has been developed for monoclonal antibodies (mAbs) to provide a more efficient process, driving down costs and reducing timelines, without compromising the quality and hence patient safety. The proposed novel workflow combines liquid chromatography-mass spectrometry multi-attribute method (LC-MS MAM) and sensitive ligand binding using surface plasmon resonance (SPR). By linking the two methodologies it is possible to obtain a comprehensive understanding of a mAb’s critical quality attributes (CQAs) and provide a structure/function correlation. As LC-MS MAM cannot address all aspects of degradation, high throughput methods for the analysis of high molecular weight material (HMWM), and conformational and colloidal stability, were also evaluated. The workflow comprises an initial forced degradation study, to verify stability-indication and identify potential degradation routes. Secondly, optimal pH, based on conformational and colloidal stability, is determined. Finally, stabilising excipients are evaluated by design of experiment (DoE). We have verified this workflow using pembrolizumab. In an initial forced degradation study, LC-MS MAM and PD-1 ligand binding could identify the CQAs. Met105 oxidation, located in the CDR3 region, was identified as the major CQA. DoE demonstrated that 25 mM methionine inhibited Met105 oxidation and stabilised PD-1 binding. With this streamlined process, we were able to improve the stability of the protein by formulating in 20 mM histidine, 25 mM methionine, 0.02 % PS80 and 300 mM sucrose, at pH 5.5. The described workflow has the potential to decrease the demand for precious early development material as well as reduce costs and shorten timelines.
{"title":"A streamlined workflow for early phase formulation development of monoclonal antibodies comprising multi-attribute method and ligand binding assay","authors":"Rachel Smith , Colin Guy , Rosie Upton , Sam Clawson , Henry Fisher , Mohammad Adam Nasar , David Firth , Allan Watkinson","doi":"10.1016/j.jpba.2026.117379","DOIUrl":"10.1016/j.jpba.2026.117379","url":null,"abstract":"<div><div>A streamlined early phase formulation development workflow has been developed for monoclonal antibodies (mAbs) to provide a more efficient process, driving down costs and reducing timelines, without compromising the quality and hence patient safety. The proposed novel workflow combines liquid chromatography-mass spectrometry multi-attribute method (LC-MS MAM) and sensitive ligand binding using surface plasmon resonance (SPR). By linking the two methodologies it is possible to obtain a comprehensive understanding of a mAb’s critical quality attributes (CQAs) and provide a structure/function correlation. As LC-MS MAM cannot address all aspects of degradation, high throughput methods for the analysis of high molecular weight material (HMWM), and conformational and colloidal stability, were also evaluated. The workflow comprises an initial forced degradation study, to verify stability-indication and identify potential degradation routes. Secondly, optimal pH, based on conformational and colloidal stability, is determined. Finally, stabilising excipients are evaluated by design of experiment (DoE). We have verified this workflow using pembrolizumab. In an initial forced degradation study, LC-MS MAM and PD-1 ligand binding could identify the CQAs. Met105 oxidation, located in the CDR3 region, was identified as the major CQA. DoE demonstrated that 25 mM methionine inhibited Met105 oxidation and stabilised PD-1 binding. With this streamlined process, we were able to improve the stability of the protein by formulating in 20 mM histidine, 25 mM methionine, 0.02 % PS80 and 300 mM sucrose, at pH 5.5. The described workflow has the potential to decrease the demand for precious early development material as well as reduce costs and shorten timelines.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"273 ","pages":"Article 117379"},"PeriodicalIF":3.1,"publicationDate":"2026-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146132126","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 : 2026-01-30DOI: 10.1016/j.jpba.2026.117384
Mohamed A. Gab-Allah , Hyojin Hwang , Mingyu Kim , Ngoc-Trinh Tran , Bong Jik Kim , Minyoung Kim , Jin Hee Han , Yehree Kim , Byung Yoon Choi , Jeongkwon Kim
Accurate detection of point mutations in mitochondrial DNA (mtDNA) is crucial for diagnosing various mitochondrial disorders. In this study, we developed an ultra-high-performance liquid chromatography coupled with high-resolution tandem mass spectrometry (UHPLC–HRMS/MS) method for the direct, label-free identification and localization of single-nucleotide mutations using synthetic 20- and 49-mer oligonucleotides as model fragments representing the pathogenic mtDNA point mutation (mt.3243 A>G). Three mobile phase systems, including ammonium bicarbonate (ABC), triethylamine/hexafluoroisopropanol (TEA/HFIP), and tributylamine/HFIP (TBA/HFIP), were systematically evaluated to assess their effects on oligonucleotide retention behavior and duplex stability under denaturing and non-denaturing conditions. The ABC buffer provided optimal performance for maintaining partial duplex integrity, while TEA/HFIP offered superior ionization efficiency for single-stranded analysis. Deconvoluted mass spectra revealed accurate monoisotopic mass differences between wild-type and mutant oligonucleotides, including ∼ + 16 Da for the sense strand (A>G), ∼ –15 Da for the antisense strand (T > C), and ∼ + 1 Da for the duplex, enabling confident mutation discrimination at the intact molecular level. High-resolution MS achieved excellent mass accuracy within ±3 ppm, and high-energy collision dissociation (HCD) MS/MS enabled sequence-specific fragmentation that localized the mutation site with high confidence when compared with theoretical fragments. Overall, this study establishes a reliable analytical framework for mutation detection in oligonucleotide models and highlights the potential of UHPLC–HRMS/MS as a complementary tool for targeted mtDNA fragment analysis.
{"title":"Detection and localization of single-nucleotide mutations in synthetic oligonucleotides by ultra-high-performance liquid chromatography coupled with high-resolution tandem mass spectrometry","authors":"Mohamed A. Gab-Allah , Hyojin Hwang , Mingyu Kim , Ngoc-Trinh Tran , Bong Jik Kim , Minyoung Kim , Jin Hee Han , Yehree Kim , Byung Yoon Choi , Jeongkwon Kim","doi":"10.1016/j.jpba.2026.117384","DOIUrl":"10.1016/j.jpba.2026.117384","url":null,"abstract":"<div><div>Accurate detection of point mutations in mitochondrial DNA (mtDNA) is crucial for diagnosing various mitochondrial disorders. In this study, we developed an ultra-high-performance liquid chromatography coupled with high-resolution tandem mass spectrometry (UHPLC–HRMS/MS) method for the direct, label-free identification and localization of single-nucleotide mutations using synthetic 20- and 49-mer oligonucleotides as model fragments representing the pathogenic mtDNA point mutation (mt.3243 A>G). Three mobile phase systems, including ammonium bicarbonate (ABC), triethylamine/hexafluoroisopropanol (TEA/HFIP), and tributylamine/HFIP (TBA/HFIP), were systematically evaluated to assess their effects on oligonucleotide retention behavior and duplex stability under denaturing and non-denaturing conditions. The ABC buffer provided optimal performance for maintaining partial duplex integrity, while TEA/HFIP offered superior ionization efficiency for single-stranded analysis. Deconvoluted mass spectra revealed accurate monoisotopic mass differences between wild-type and mutant oligonucleotides, including ∼ + 16 Da for the sense strand (A>G), ∼ –15 Da for the antisense strand (T > C), and ∼ + 1 Da for the duplex, enabling confident mutation discrimination at the intact molecular level. High-resolution MS achieved excellent mass accuracy within ±3 ppm, and high-energy collision dissociation (HCD) MS/MS enabled sequence-specific fragmentation that localized the mutation site with high confidence when compared with theoretical fragments. Overall, this study establishes a reliable analytical framework for mutation detection in oligonucleotide models and highlights the potential of UHPLC–HRMS/MS as a complementary tool for targeted mtDNA fragment analysis.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"273 ","pages":"Article 117384"},"PeriodicalIF":3.1,"publicationDate":"2026-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146119301","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 : 2026-01-28DOI: 10.1016/j.jpba.2026.117383
Jérôme Jonveaux, Marc Faudon, Pauline Heymes, Valentina Lucchini, Maria Fernanda Zuluaga Estrada, Michael Jahn, Mostafa Zarei
Isomerization of aspartic acid (Asp) to isoaspartic acid (isoAsp) within the complementarity-determining regions (CDRs) of monoclonal antibodies (mAbs) can lead to conformational changes that decrease antigen-binding affinity. Although isomerization can significantly alter the chromatographic and electrophoretic profiles, precise localization of this modification requires a mass spectrometry-based approach, such as peptide mapping. In this work, we present a case study that investigates various analytical strategies to identify the root cause of significant changes observed in the chromatographic and electrophoretic profiles of an mAb during formulation development. LC-MS analysis of reduced mAb using high-resolution mass spectrometry, peptide mapping using trypsin digestion, and fraction collection of the newly identified peak followed by trypsin digestion suggested that isomerization occurs within the CDR of the mAb. However, due to the presence of three Asp residues within a single tryptic peptide, this modification could not be precisely localized. To overcome this limitation, we developed a sequential enzymatic digestion strategy, utilizing trypsin followed by Asp-N digestion, which enabled accurate localization and quantification of the isomerization site. The resulting data indicated that the main isoAsp signal originated from isomerization at the DS motif that increased substantially over time in the liquid formulation, while no significant change was observed in the lyophilized formulation. The level of isomerization determined through this sequential digestion method correlated well with the LC-UV quantitation data of the reduced mAb.
{"title":"Sequential digestions enable identification and quantification of rapid aspartic acid isomerization in the CDR of a monoclonal antibody light chain","authors":"Jérôme Jonveaux, Marc Faudon, Pauline Heymes, Valentina Lucchini, Maria Fernanda Zuluaga Estrada, Michael Jahn, Mostafa Zarei","doi":"10.1016/j.jpba.2026.117383","DOIUrl":"10.1016/j.jpba.2026.117383","url":null,"abstract":"<div><div>Isomerization of aspartic acid (Asp) to isoaspartic acid (isoAsp) within the complementarity-determining regions (CDRs) of monoclonal antibodies (mAbs) can lead to conformational changes that decrease antigen-binding affinity. Although isomerization can significantly alter the chromatographic and electrophoretic profiles, precise localization of this modification requires a mass spectrometry-based approach, such as peptide mapping. In this work, we present a case study that investigates various analytical strategies to identify the root cause of significant changes observed in the chromatographic and electrophoretic profiles of an mAb during formulation development. LC-MS analysis of reduced mAb using high-resolution mass spectrometry, peptide mapping using trypsin digestion, and fraction collection of the newly identified peak followed by trypsin digestion suggested that isomerization occurs within the CDR of the mAb. However, due to the presence of three Asp residues within a single tryptic peptide, this modification could not be precisely localized. To overcome this limitation, we developed a sequential enzymatic digestion strategy, utilizing trypsin followed by Asp-N digestion, which enabled accurate localization and quantification of the isomerization site. The resulting data indicated that the main isoAsp signal originated from isomerization at the DS motif that increased substantially over time in the liquid formulation, while no significant change was observed in the lyophilized formulation. The level of isomerization determined through this sequential digestion method correlated well with the LC-UV quantitation data of the reduced mAb.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"273 ","pages":"Article 117383"},"PeriodicalIF":3.1,"publicationDate":"2026-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146119332","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 : 2026-01-28DOI: 10.1016/j.jpba.2026.117382
Meichen Liu , Yalin Yu , Yifei Jing , Yingxia Guo , Chuya Wang , Hongyu Xue , Hong Liu , Lei Yin , Meiyun Shi
While polyethylene glycol (PEG) is an extensively utilized pharmaceutical polymer with established biocompatibility and regulatory acceptance, the in vivo pharmacokinetics of its bifunctional derivatives, such as α,ω-dipropionic acid polyethylene glycol (PA-PEG-PA), remain largely unexplored. This study presents the development and validation of a highly sensitive and selective UPLC-MS/MS method for the accurate quantification of PA-PEG12-PA in various biological matrices, utilizing a straightforward protein precipitation protocol. After intravenous administration in rats (10 mg/kg), PA-PEG12-PA demonstrated rapid clearance (t1/2 = 3.99 ± 1.06 h). Tissue distribution analysis revealed a pronounced affinity for renal accumulation, with kidney concentrations (16473 ± 881 ng/g at 2 h) substantially surpassing those in the liver and lungs, followed by rapid depletion within 24 h. Excretion studies indicated renal clearance as the dominant pathway, with 55.93 % of the administered dose recovered unchanged in urine over 72 h, while fecal excretion was minimal (1.76 %). This work provides the first comprehensive pharmacokinetic and biodistribution profile of PA-PEG12-PA, underscoring its renal-driven clearance and tissue disposition. The findings offer crucial insights for the rational design of PEGylated drug delivery systems, and the robust UPLC-MS/MS assay established herein serves as a valuable tool for characterizing polymeric excipients in biological environments.
{"title":"Comprehensive pharmacokinetic and tissue distribution study of α,ω-dipropionic acid polyethylene glycol (PA-PEG₁₂-PA) in rats using a validated UPLC-MS/MS method","authors":"Meichen Liu , Yalin Yu , Yifei Jing , Yingxia Guo , Chuya Wang , Hongyu Xue , Hong Liu , Lei Yin , Meiyun Shi","doi":"10.1016/j.jpba.2026.117382","DOIUrl":"10.1016/j.jpba.2026.117382","url":null,"abstract":"<div><div>While polyethylene glycol (PEG) is an extensively utilized pharmaceutical polymer with established biocompatibility and regulatory acceptance, the <em>in vivo</em> pharmacokinetics of its bifunctional derivatives, such as α,ω-dipropionic acid polyethylene glycol (PA-PEG-PA), remain largely unexplored. This study presents the development and validation of a highly sensitive and selective UPLC-MS/MS method for the accurate quantification of PA-PEG<sub>12</sub>-PA in various biological matrices, utilizing a straightforward protein precipitation protocol. After intravenous administration in rats (10 mg/kg), PA-PEG<sub>12</sub>-PA demonstrated rapid clearance (t<sub>1/2</sub> = 3.99 ± 1.06 h). Tissue distribution analysis revealed a pronounced affinity for renal accumulation, with kidney concentrations (16473 ± 881 ng/g at 2 h) substantially surpassing those in the liver and lungs, followed by rapid depletion within 24 h. Excretion studies indicated renal clearance as the dominant pathway, with 55.93 % of the administered dose recovered unchanged in urine over 72 h, while fecal excretion was minimal (1.76 %). This work provides the first comprehensive pharmacokinetic and biodistribution profile of PA-PEG<sub>12</sub>-PA, underscoring its renal-driven clearance and tissue disposition. The findings offer crucial insights for the rational design of PEGylated drug delivery systems, and the robust UPLC-MS/MS assay established herein serves as a valuable tool for characterizing polymeric excipients in biological environments.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"272 ","pages":"Article 117382"},"PeriodicalIF":3.1,"publicationDate":"2026-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146078995","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 : 2026-01-27DOI: 10.1016/j.jpba.2026.117381
Chiara Collevecchio, Serena Fiorito, Francesco Epifano, Salvatore Genovese
The present study explores the glutathione peroxidase (GPx)-like mimicking activity of selected natural phenylpropanoids, their oxyprenylated derivatives, and natural and semisynthetic cannabinoids using a revised version of Iwaoka’s assay based on the application of GC-MS and HPLC-DAD. Thus, naturally occurring phenylpropanoids, including ferulic acid, p-coumaric acid, and umbelliferone, together with their oxyprenylated derivatives, were synthesized and tested for catalytic effects on 1,4-dithiotrhreitol (DTTred) oxidation. While ferulic acid and p-coumaric acid displayed an appreciable GPx-like activity, their oxyprenylated counterparts exhibited a reduction in catalytic efficiency, suggesting that the carboxylic acid and free phenolic hydroxyl groups play a key role in the observed activity. Coumarin derivatives showed minimal activity, likely due to their rigid ring system. Among cannabinoids, cannabidiol (CBD) and cannabigerol (CBG) enhanced DTT oxidation, with CBG displaying a nearly comparable effect to the reference catalyst Ebselen®. Structure–activity relationship analysis of semisynthetic CBG ethers and esters revealed that substitution patterns strongly influence catalytic performance, with diethyl and di-n-propyl derivatives demonstrating the highest GPx-like behavior. Overall, this work identifies cinnamic acids, CBG, and some of their etherified analogues as promising GPx mimetics and provides mechanistic insight into the redox properties of phenylpropanoids and cannabinoids, which may support future antioxidant drug design.
{"title":"Glutathione peroxidase-like activity of natural and semisynthetic phenylpropanoids and cannabinoids: An analytical investigation by a GC-MS- and HPLC-DAD-based Iwaoka’s assay","authors":"Chiara Collevecchio, Serena Fiorito, Francesco Epifano, Salvatore Genovese","doi":"10.1016/j.jpba.2026.117381","DOIUrl":"10.1016/j.jpba.2026.117381","url":null,"abstract":"<div><div>The present study explores the glutathione peroxidase (GPx)-like mimicking activity of selected natural phenylpropanoids, their oxyprenylated derivatives, and natural and semisynthetic cannabinoids using a revised version of Iwaoka’s assay based on the application of GC-MS and HPLC-DAD. Thus, naturally occurring phenylpropanoids, including ferulic acid, <em>p</em>-coumaric acid, and umbelliferone, together with their oxyprenylated derivatives, were synthesized and tested for catalytic effects on 1,4-dithiotrhreitol (DTT<sup>red</sup>) oxidation. While ferulic acid and <em>p</em>-coumaric acid displayed an appreciable GPx-like activity, their oxyprenylated counterparts exhibited a reduction in catalytic efficiency, suggesting that the carboxylic acid and free phenolic hydroxyl groups play a key role in the observed activity. Coumarin derivatives showed minimal activity, likely due to their rigid ring system. Among cannabinoids, cannabidiol (CBD) and cannabigerol (CBG) enhanced DTT oxidation, with CBG displaying a nearly comparable effect to the reference catalyst Ebselen®. Structure–activity relationship analysis of semisynthetic CBG ethers and esters revealed that substitution patterns strongly influence catalytic performance, with diethyl and di-<em>n</em>-propyl derivatives demonstrating the highest GPx-like behavior. Overall, this work identifies cinnamic acids, CBG, and some of their etherified analogues as promising GPx mimetics and provides mechanistic insight into the redox properties of phenylpropanoids and cannabinoids, which may support future antioxidant drug design.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"272 ","pages":"Article 117381"},"PeriodicalIF":3.1,"publicationDate":"2026-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146078996","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 : 2026-01-26DOI: 10.1016/j.jpba.2026.117380
Yanmei He , Xiao Lin , Wanwan Zhong , Xingyan Liu , Xiuyan Yang , Youlian Yao , Meimei Li , Jianyong Zhang
Shuganning injection (SGNI) is derived from Yinchenhao decoction and has the functions of promoting diuresis, reducing jaundice and protecting the liver. It is primarily utilized in the treatment of liver diseases, particularly damp-heat jaundice. However, the pharmacodynamic material basis and specific mechanisms remain unclear. Therefore, this study aims to elucidate the pharmacodynamic material basis and mechanism of SGNI in the treatment of damp-heat jaundice syndrome (DHJS). High-throughput metabolomics was employed to identify biomarkers associated with DHJS. The migratory prototype components and metabolites present in the blood of SGNI were characterized through serum pharmacochemistry. Correlation analysis was conducted to associate the biomarkers of DHJS with the migrating components in the blood of SGNI, clarifying the pharmacodynamic basis of SGNI in treating DHJS. Furthermore, metabolic pathway enrichment analysis was performed to explore the endogenous core metabolites and key metabolic pathways, providing insights into the mechanism of SGNI in treating DHJS from the metabolic perspective. Finally, a total of 25 metabolites were identified as biomarkers for DHJS, along with 24 prototype migratory components and 38 migratory metabolites of SGNI. It was established that the pharmacodynamic substance basis of SGNI in the treatment of DHJS comprises 15 components. These components primarily exert their effects through key metabolic pathways, including pentose and glucuronate interconversions, arachidonic acid metabolism, and primary bile acid biosynthesis. In conclusion, SGNI demonstrates a significant therapeutic effect on DHJS. It has preliminarily elucidated the pharmacodynamic substance basis and mechanism of action at the metabolic level, offering a scientific foundation for the clinical application and further development of SGNI.
{"title":"The discovery of pharmacodynamic material basis and mechanism of Shuganning injection in treating damp-heat jaundice syndrome: Combining metabolomics and serum pharmacochemistry","authors":"Yanmei He , Xiao Lin , Wanwan Zhong , Xingyan Liu , Xiuyan Yang , Youlian Yao , Meimei Li , Jianyong Zhang","doi":"10.1016/j.jpba.2026.117380","DOIUrl":"10.1016/j.jpba.2026.117380","url":null,"abstract":"<div><div>Shuganning injection (SGNI) is derived from Yinchenhao decoction and has the functions of promoting diuresis, reducing jaundice and protecting the liver. It is primarily utilized in the treatment of liver diseases, particularly damp-heat jaundice. However, the pharmacodynamic material basis and specific mechanisms remain unclear. Therefore, this study aims to elucidate the pharmacodynamic material basis and mechanism of SGNI in the treatment of damp-heat jaundice syndrome (DHJS). High-throughput metabolomics was employed to identify biomarkers associated with DHJS. The migratory prototype components and metabolites present in the blood of SGNI were characterized through serum pharmacochemistry. Correlation analysis was conducted to associate the biomarkers of DHJS with the migrating components in the blood of SGNI, clarifying the pharmacodynamic basis of SGNI in treating DHJS. Furthermore, metabolic pathway enrichment analysis was performed to explore the endogenous core metabolites and key metabolic pathways, providing insights into the mechanism of SGNI in treating DHJS from the metabolic perspective. Finally, a total of 25 metabolites were identified as biomarkers for DHJS, along with 24 prototype migratory components and 38 migratory metabolites of SGNI. It was established that the pharmacodynamic substance basis of SGNI in the treatment of DHJS comprises 15 components. These components primarily exert their effects through key metabolic pathways, including pentose and glucuronate interconversions, arachidonic acid metabolism, and primary bile acid biosynthesis. In conclusion, SGNI demonstrates a significant therapeutic effect on DHJS. It has preliminarily elucidated the pharmacodynamic substance basis and mechanism of action at the metabolic level, offering a scientific foundation for the clinical application and further development of SGNI.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"272 ","pages":"Article 117380"},"PeriodicalIF":3.1,"publicationDate":"2026-01-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146079092","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 : 2026-01-26DOI: 10.1016/j.jpba.2026.117378
Lifeng Zhao , Siyang Wu , Xin Yu , Zhishan Huang , Luyang Liu , Xuehao Cheng , Zheng Yuan , Yingfei Li
Shengmaiyin oral liquid (SMY), formulated with schisandra chinensis, red ginseng, and Ophiopogon japonicus, is widely used for treating cardiac dysfunction (CD). However, its functional chemical basis and pharmacological profiles remain insufficiently explored. This study employed advanced analytical strategies to characterize the bioactive components of SMY and investigate their in silico pharmacodynamics and in vivo pharmacokinetics, providing mechanistic insights into their roles in CD treatment. Using UPLC-Q-TOF-MS/MS, 132 compounds were identified in SMY, with 31 detected in the plasma of SMY-treated mice. Among these, 21 chemicals (12 lignans and 9 saponins) were identified as key bioactives against CD. Network pharmacology and molecular docking revealed their multi-target interactions and varied binding affinities with CD-related proteins. Pharmacokinetic (PK) analysis showed that 5 compounds had high plasma exposure and rapid elimination in CD model mice. All 21 chemicals exhibited significant tissue distribution following prolonged SMY administration. The globally pharmacokinetic seeking (GPS) box analysis revealed two distinct fast and slow PK patterns among the chemicals. Notably, a highly tissue-specific and time-dependent alteration of lignan and saponin clusters was observed in the hearts of CD mice within 8 h post-administration. This study highlights the GPS box as an innovative platform for multi-dimensional PK analysis. These findings advance the integration of traditional herb medicine with modern analytical methodologies, offering new perspectives for developing precision medicine approaches in ethnopharmacology.
{"title":"Chemical profiling and multi-dimensional pharmacokinetic analysis of shengmaiyin oral liquid for cardiac dysfunction","authors":"Lifeng Zhao , Siyang Wu , Xin Yu , Zhishan Huang , Luyang Liu , Xuehao Cheng , Zheng Yuan , Yingfei Li","doi":"10.1016/j.jpba.2026.117378","DOIUrl":"10.1016/j.jpba.2026.117378","url":null,"abstract":"<div><div>Shengmaiyin oral liquid (SMY), formulated with schisandra chinensis, red ginseng, and Ophiopogon japonicus, is widely used for treating cardiac dysfunction (CD). However, its functional chemical basis and pharmacological profiles remain insufficiently explored. This study employed advanced analytical strategies to characterize the bioactive components of SMY and investigate their in silico pharmacodynamics and in vivo pharmacokinetics, providing mechanistic insights into their roles in CD treatment. Using UPLC-Q-TOF-MS/MS, 132 compounds were identified in SMY, with 31 detected in the plasma of SMY-treated mice. Among these, 21 chemicals (12 lignans and 9 saponins) were identified as key bioactives against CD. Network pharmacology and molecular docking revealed their multi-target interactions and varied binding affinities with CD-related proteins. Pharmacokinetic (PK) analysis showed that 5 compounds had high plasma exposure and rapid elimination in CD model mice. All 21 chemicals exhibited significant tissue distribution following prolonged SMY administration. The globally pharmacokinetic seeking (GPS) box analysis revealed two distinct fast and slow PK patterns among the chemicals. Notably, a highly tissue-specific and time-dependent alteration of lignan and saponin clusters was observed in the hearts of CD mice within 8 h post-administration. This study highlights the GPS box as an innovative platform for multi-dimensional PK analysis. These findings advance the integration of traditional herb medicine with modern analytical methodologies, offering new perspectives for developing precision medicine approaches in ethnopharmacology.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"272 ","pages":"Article 117378"},"PeriodicalIF":3.1,"publicationDate":"2026-01-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146078997","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