Journal of pharmaceutical and biomedical analysis最新文献

{"title":"Pharmacokinetic investigation on eight primary active constituents of Weifuchun capsules in healthy and chronic atrophic gastritis model rats","authors":"Wentao Shao ,&nbsp;Xinxin Zhao ,&nbsp;Qiuya Zhou ,&nbsp;Lvli Ma ,&nbsp;Wei Zhang ,&nbsp;Yi Tao","doi":"10.1016/j.jpba.2026.117335","DOIUrl":"10.1016/j.jpba.2026.117335","url":null,"abstract":"<div><div>Weifuchun capsule (WC) is clinically employed in treating chronic atrophic gastritis (CAG), yet differences in its pharmacokinetic profile between normal and chronic atrophic gastritis model rats remain insufficiently characterized. In this study, a reliable and precise ultra-high performance liquid chromatography–tandem mass spectrometry method was developed and validated for the simultaneous quantification of eight major active constituents including <em>p</em>‑coumaric acid, ginsenoside Rg1, kaempferol, luteolin, hesperetin, apigenin, naringenin, and tangeretin in rat plasma. This method was subsequently applied to investigate the pharmacokinetics of Weifuchun capsule. A chronic atrophic gastritis model was established using a modified chemical stimulation method. Plasma samples were pretreated via protein precipitation with chloramphenicol as the internal standard and then analyzed. The intra- and inter-day precision for all eight analytes was below 6.32 %, with accuracies within ±5.19 %. Extraction recoveries ranged from 89.15 % to 112.50 %, and matrix effects were between 95.69 % and 104.83 %. All analytes demonstrated satisfactory stability under various storage conditions. The validated method was successfully applied to a comparative pharmacokinetic study. Compared with the normal group, the chronic atrophic gastritis model group exhibited significantly increased C_max and AUC_0→t values for ginsenoside Rg1, hesperetin, and naringenin (<em>p</em> &lt; 0.05), along with significantly elevated C_max for kaempferol and apigenin (<em>p</em> &lt; 0.05). Additionally, t_1/2 was significantly shortened for ginsenoside Rg1 and <em>p</em>-coumaric acid. These findings suggest enhanced absorption and accelerated elimination of certain bioactive components of Weifuchun capsules under chronic atrophic gastritis pathological conditions. The altered pharmacokinetic behavior of multiple active compounds of Weifuchun capsules in chronic atrophic gastritis model rats provides important insights into the pharmacological mechanisms of Weifuchun capsules in the treatment of chronic atrophic gastritis.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117335"},"PeriodicalIF":3.1,"publicationDate":"2026-01-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145917927","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}

{"title":"Importance in optimization of sample volume in the IgG-immunoaffinity UPLC-MS/MS assay for quantification of a therapeutic antibody in monkey serum","authors":"Kenji Kita ,&nbsp;Yuji Mano","doi":"10.1016/j.jpba.2026.117334","DOIUrl":"10.1016/j.jpba.2026.117334","url":null,"abstract":"<div><div>E6011 is a novel monoclonal antibody developed for the treatment of rheumatoid arthritis. Ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC–MS/MS) has become increasingly important in the assays of therapeutic antibodies, yet achieving high sensitivity remains a major challenge. For the E6011 assay, an immunoaffinity (IgG-IA) method employing protein G in combination with UPLC-MS/MS was utilized and qualified in monkey serum. Method development revealed that increasing the matrix volume did not always lead to improved sensitivity when using a fixed volume of protein G magnetic beads, due to their limited capture capacity. Using only 1.8 µL of monkey serum, E6011 was quantitatively analyzed at concentrations as low as 1 µg/mL. Accuracy and precision were within ±20 %, meeting the acceptance criteria, however, the selectivity result at 1 µg/mL highlighted the need for evaluation of selectivity across multiple individuals. E6011 concentrations in monkey serum were determined using the IgG-IA-UPLC-MS/MS method, and the resulting pharmacokinetic parameters were comparable to those previously reported using UPLC-MS/MS with ammonium sulfate precipitation and a ligand-binding assay. These findings highlight the importance of optimizing matrix volume in assay development and demonstrate the successful application of the established method in a monkey pharmacokinetic study.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117334"},"PeriodicalIF":3.1,"publicationDate":"2026-01-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145926822","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}

{"title":"Banxia Xiexin Decoction modulates short-chain fatty acid metabolism and mitigates ulcerative colitis by reshaping the intestinal microbiota","authors":"Ji Wanli ,&nbsp;Song Xingyue ,&nbsp;Liang Xinning ,&nbsp;Zheng Yashuo ,&nbsp;Qian Xiaojing ,&nbsp;Hu Cheng ,&nbsp;Huo Yan","doi":"10.1016/j.jpba.2025.117332","DOIUrl":"10.1016/j.jpba.2025.117332","url":null,"abstract":"<div><div>Ulcerative colitis (UC) is a chronic inflammatory disorder that significantly impacts the quality of life for patients. Dysbiosis of gut microbiota and changes in short-chain fatty acid (SCFA) metabolism play a role in both the initiation and progression of UC. Banxia Xiexin Decoction (BXD), a formula in traditional Chinese medicine, has shown therapeutic effects; however, its underlying mechanism remains unclear. A rat model of colitis induced by dextran sulfate sodium (DSS) was created, and various doses of BXD were administered. The principal components of BXD were analyzed through high-pressure liquid chromatography (HPLC). In order to clarify the mechanisms, 16S rRNA sequencing, serum metabolomics, targeted profiling of SCFAs via gas chromatography-mass spectrometry (GC-MS), and assessments of enzyme activity were conducted. BXD exerted protective effects against DSS-induced UC, as indicated by attenuated histological damage and reduced expression of pro-inflammatory cytokines. Untargeted metabolomics revealed that BXD modulated multiple metabolic pathways, enhancing SCFA-related processes, such as propanoate and butanoate metabolism. GC–MS revealed that BXD could increase the level of acetate, propionate, butyrate, isobutyrate, valerate, isovalerate and hexanoate. Microbiome sequencing indicated that BXD increased the abundance of beneficial taxa (e.g. Firmicutes, Bacteroidetes), while reducing pro-inflammatory genera. Altogether, BXD restored the microbial balance and promoted anti-inflammatory metabolite production. Our results demonstrated that BXD ameliorated UC by reprogramming gut microbial composition and enhancing SCFA biosynthesis, thereby suppressing intestinal inflammation. The potential of BXD as a therapy aimed at the microbiota for UC is emphasized in these studies, which also offer mechanistic insights using multi-omics approaches.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117332"},"PeriodicalIF":3.1,"publicationDate":"2026-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145917938","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}

{"title":"Elucidating the effects of ginger processing on Magnolia bark: A multi-platform strategy linking chemical composition to taste and bioactivity","authors":"Zhiyan Xie ,&nbsp;Haochen Wu ,&nbsp;Huixian Qing ,&nbsp;Yu Yin ,&nbsp;Zicheng Ma ,&nbsp;Guoqin Zhang ,&nbsp;Meiqi Liu ,&nbsp;Shiwei Chai ,&nbsp;Xiaoliang Ren","doi":"10.1016/j.jpba.2026.117333","DOIUrl":"10.1016/j.jpba.2026.117333","url":null,"abstract":"<div><div><em>Magnolia officinalis</em> (Houpo, HP) is a prominent Traditional Chinese Medicine(TCM). Despite its traditional processing with ginger to reduce irritation, enhance efficacy, and improve taste, the underlying mechanisms connecting this chemical transformation to its sensory and gastrointestinal effects are not fully understood. Existing studies lack systematic analysis of shared targets across gastrointestinal diseases and the identification of quality markers (Q-markers), limiting scientific justification for its processing and clinical use. This study therefore applied an integrated UPLC-Q-TOF/MS, chemometrics, and network pharmacology approach to elucidate the effects of ginger processing. In this study a total of 46 batches of samples (22 raw, 24 ginger-processed) were analyzed by UPLC-Q-TOF/MS to identify and quantify chemical constituents. Chemometric models were used to discriminate sample types and identify potential Q-markers. The correlation between taste and chemical profiles was assessed using an electronic tongue combined with Pearson correlation analysis. Network pharmacology was employed to predict targets related to functional dyspepsia (FD), postoperative nausea and vomiting (PONV), and chronic gastritis (CG), followed by GO/KEGG enrichment analysis. Molecular docking was performed to validate binding affinities between key bioactive components and core targets. The experimental results showed that Fifty-four compounds were characterized, and chemometric models clearly distinguished raw from ginger-processed HP (Jianghoupo, JHP). Electronic tongue analysis revealed that Magnoloside A plays a key role in reducing bitterness after ginger processing. Network pharmacology identified 51 overlapping targets across FD, PONV, and CG, with AKT1, TNF, CTNNB1, IL1B, and STAT3 as core nodes in the network. Molecular docking confirmed stable binding between principal components and these targets. An integrative “component transformation–taste modulation–gastrointestinal regulation” model was established, illustrating how ginger processing improves both palatability and pharmacological efficacy. In summary, ginger processing transforms the chemical and sensory profiles of HP into a state of improved palatability and strengthened gastrointestinal regulatory function. These findings provide a mechanistic rationale and scientific foundation for the traditional processing practice of combining ginger with HP in clinical use.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"272 ","pages":"Article 117333"},"PeriodicalIF":3.1,"publicationDate":"2026-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145928867","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}

{"title":"Separation of mephentermine from other structural analogues of phenethylamine and amphetamine in biological matrices using Liquid Chromatography-Tandem Mass Spectrometry","authors":"Tusha Tripathi ,&nbsp;Kamna Sharma ,&nbsp;Awanish Upadhyay ,&nbsp;Ashok Singh ,&nbsp;Puran Lal Sahu ,&nbsp;Tarun Handa ,&nbsp;Meghna Choudhary ,&nbsp;Bikash Medhi","doi":"10.1016/j.jpba.2025.117328","DOIUrl":"10.1016/j.jpba.2025.117328","url":null,"abstract":"<div><div>Stimulants exert euphorigenic and relaxing effects which warrant intervention of regulatory authorities in non-medical usage of these doping agents. Thus, a semi-quantitative method utilizing Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry (UHPLC-MS/MS) was developed to monitor potential misuse of 10 structural analogues of amphetamine and phenylethylamine derivatives. The extraction protocol for urine was dilute and shoot (DS) with 1 mL aliquot size, while that for plasma/serum was solid-phase extraction (SPE) with a 100 µL aliquot size. Chromatographic separation was achieved over an 18-minute gradient on a Phenomenex Luna Omega UHPLC C18 column. The mobile phase consisted of water and acetonitrile, both containing formic acid as a modifier, specifically at 1 % (v/v) for urine and 0.1 % (v/v) for plasma/serum analyses. Positive electrospray ionization (ESI+) was employed for mass spectrometric detection with acquisition in multiple reaction monitoring (MRM) mode. Method validation was performed in accordance with World Anti-Doping Agency (WADA) requirements for analyzing prohibited substances with minimum reporting limits (MRLs). Limits of identification (LOI) determined for urine and plasma/serum matrices were as follows: 0.5 ng/mL and 1 ng/mL for Oxethazaine; 1 ng/mL and 2.5 ng/mL for BHPT and BHMPT; 12.5 ng/mL and 25 ng/mL for PT and 5 ng/mL and 12.5 ng/mL for MPT, NEAMP, DMAMP, 4MMAMP, NEBMPEA and NNBTPEA, respectively. An analytical method characterized by rapidity, simplicity and cost-effectiveness was implemented for the chromatographic separation of stimulant structural analogues, facilitating the determination of potential stimulant abuse within human performance sports.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117328"},"PeriodicalIF":3.1,"publicationDate":"2025-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145926823","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}

{"title":"A general strategy to screen and evaluate antitussive, anti-inflammatory, and expectorant active components from raw and honey-fired Farfarae Flos based on β2-AR/CysLT1R/M3R chromatography and mice models","authors":"Yaqi Wang,&nbsp;Ping Shu,&nbsp;Mei Lv,&nbsp;Yonghao Xing,&nbsp;Haoran Zhang,&nbsp;Xiaojing Yan,&nbsp;Bo Wang,&nbsp;Tingting Huang,&nbsp;Minfeng Fang","doi":"10.1016/j.jpba.2025.117331","DOIUrl":"10.1016/j.jpba.2025.117331","url":null,"abstract":"<div><div>Farfarae Flos and its honey-processed product are used to treat respiratory diseases; yet their active components and therapeutic targets remained unclear. Herein, we developed an integrated strategy combining receptor chromatography and mice models to screen and evaluate active components in both raw and honey-processed Farfarae Flos. We focused on three key receptors,β₂-AR, CysLT₁R, and M₃R, known to be involved in antitussive, anti-inflammatory, and expectorant activities. These receptors were immobilized onto silica gels to construct affinity columns. Using these columns, we identified the bioactive compounds in both forms of Farfarae Flos. The results revealed an identical set of target-specific compounds across raw and honey-processed samples: chlorogenic acid (CGA) and caffeic acid (CA) targeted β₂-AR; CGA and rutin bound to CysLT₁R; and CGA along with isochlorogenic acids A, B, and C interacted with M₃R. Notably, the interactions of rutin with CysLT₁R and isochlorogenic acids A, B, and C with M₃R are reported for the first time. These findings indicate that both raw and honey-fired Farfarae Flos share the same material basis for their antitussive, anti-inflammatory, and expectorant effects, with no significant alteration in pharmacological efficacy due to honey processing. Furthermore, the binding affinities of the bioactive compounds for the receptors correlated well with the molecular docking-predicted binding energies of the corresponding compound-receptor complexes. To our knowledge, this study represents the first target-centric comparative analysis of raw and processed Farfarae Flos. It establishes a generalizable framework for investigating processing-induced changes in herbal pharmacology and provides empirical evidence connecting metabolomic profiles to clinical efficacy.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117331"},"PeriodicalIF":3.1,"publicationDate":"2025-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145911637","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}

{"title":"A critical review of molecularly imprinted polymer technology for aflatoxin sensing: Current status and future directions","authors":"Ahmet Cetinkaya ,&nbsp;S. Irem Kaya ,&nbsp;Burcak Demirbakan ,&nbsp;Mehmet Altay Unal ,&nbsp;Mustafa Kemal Sezginturk ,&nbsp;Sibel A. Ozkan ,&nbsp;Evrim Gunes Altuntas","doi":"10.1016/j.jpba.2025.117330","DOIUrl":"10.1016/j.jpba.2025.117330","url":null,"abstract":"<div><div>Since the first discovery and identification of aflatoxins almost 70 years ago, their occurrence, potential risks, and health hazards remain of great importance. Therefore, several chromatography-, colorimetry-, and immunoassay-based analytical applications for aflatoxin detection are reported in the literature. Molecularly imprinted polymers (MIPs) have attracted considerable interest in sensor design due to their unique potential as synthetic recognition elements, providing high selectivity and excellent stability. These features help overcome the disadvantages of conventional sensing systems. The ongoing advancement of MIP-based sensors presents new opportunities for future applications and offers a reliable and versatile approach to meet the growing demands of current analytical research. This review focuses on studies published since 2023 concerning the detection of aflatoxins using MIP-based sensors, which hold a prominent and critical position in the literature. Additionally, this review addresses several limitations of MIP-based sensors, particularly in the context of aflatoxin detection, and discusses proposed solutions to overcome these challenges. Lastly, current advancements and future expectations in the topic are also highlighted.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117330"},"PeriodicalIF":3.1,"publicationDate":"2025-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145881328","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}

{"title":"Rapid one-tube sputum processing for tuberculosis diagnosis via azide-functionalized magnetic nanoplatforms with selective bacterial capture","authors":"Bilsen Tural ,&nbsp;Alican Bilden ,&nbsp;Erdal Ertaş ,&nbsp;Emre Tural ,&nbsp;Hakan Temiz ,&nbsp;Erdal Özbek ,&nbsp;Servet Tural","doi":"10.1016/j.jpba.2025.117329","DOIUrl":"10.1016/j.jpba.2025.117329","url":null,"abstract":"<div><div>Tuberculosis (TB) remains a global health challenge requiring rapid and reliable diagnostic tools. Here, azide-functionalized magnetic nanoparticles (MNPs-N₃) were synthesized, characterized, and applied for one-tube detection of <em>Mycobacterium tuberculosis</em> (Mtb) in sputum. Structural analyses including Fourier-transform infrared (FT-IR), transmission electron microscopy (TEM), scanning electron microscopy (SEM), dynamic light scattering (DLS), and vibrating sample magnetometry (VSM) confirmed successful functionalization, uniform morphology, and preserved superparamagnetism. MNPs-N₃ were integrated into modified Ehrlich-Ziehl-Neelsen (MNPs-N₃-assisted EZN staining) and auramine-rhodamine (MNPs-N₃-assisted AR staining) staining protocols to enhance bacterial capture and visualization without decontamination or centrifugation. Control experiments using non-functionalized magnetic nanoparticles showed no bacterial co-localization, supporting the specificity of the azide-mediated interaction. The entire process was completed within one hour, offering a rapid alternative to conventional culture requiring ≥ 41 days. Using Mycobacterial Growth Indicator Tube (MGIT) culture as the reference, MNPs-N₃-assisted AR staining achieved 99 % sensitivity and 97 % specificity, outperforming MNPs-N₃-assisted EZN staining (95 % and 96 %, respectively). Diagnostic indices, including Youden index (0.96) and F1-score (0.98), demonstrated excellent agreement with culture results. These findings establish MNPs-N₃ as a fast, efficient, and cost-effective tool for Mtb diagnosis. The single-tube workflow minimizes contamination risk and simplifies laboratory handling, supporting potential application in resource-limited settings. Further optimization and large-scale clinical validation are still warranted.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117329"},"PeriodicalIF":3.1,"publicationDate":"2025-12-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145878462","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}

{"title":"Determination of N-Lactoyl-phenylalanine, phenylacetylglutamine and trimethylamine-N-oxide related metabolites in hypertensive patients by liquid chromatography-mass spectrometry and the application to blood pressure regulation","authors":"Xin Tan ,&nbsp;Xianyun Jiang ,&nbsp;Lu Ren ,&nbsp;Ziyu Meng ,&nbsp;Jingyi Yang ,&nbsp;Yuan Wang","doi":"10.1016/j.jpba.2025.117327","DOIUrl":"10.1016/j.jpba.2025.117327","url":null,"abstract":"<div><div>Metabolites, such as N-lactoyl-phenylalanine (Lac-Phe), phenylacetylglutamine, and trimethylamine-N-oxide related precursors, may be associated with hypertension. A rapid, sensitive and accurate method, with high throughput, low sample volume, and easy operation to analyze these metabolites would facilitate their measurement and provide important insights into their role in hypertension. We developed an ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for the simultaneous measurement of eight metabolites in 5 µL human plasma samples with a 4.2-min chromatographic run and easy operation. The method achieved good linearity (R² &gt; 0.99), sensitivity (limit of quantification: 0.1–1 ng/mL), accuracy (within-day: −5.33 % to 11.38 %; between-day: −3.35 % to 11.50 %), precision (within-day: 1.35–13.25 %; between-day: 1.74–12.73 %), matrix effect (−5.93 % to 14.81 %), recovery rates (91.49 % to 103.31 %), bench-top stability (coefficient of variations [CVs]: 1.02–6.16 %), long-term stability (CVs: 3.63–12.94 %), on-instrument stability (CVs: 0.53–5.09 %) and freeze-thaw stability (biases: −12.60 % to −0.13 %). We successfully utilized the UHPLC–MS/MS method to measure plasma samples obtained from 204 healthy individuals and 204 patients with hypertension. Plasma Lac-Phe was reduced in patients with hypertension, and the potential effects of blood pressure regulation by Lac-Phe were explored. In angiotensin II-induced hypertensive mice, Lac-Phe alleviated the increases in systolic and diastolic blood pressure. These findings highlight the therapeutic potential of Lac-Phe in hypertension.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117327"},"PeriodicalIF":3.1,"publicationDate":"2025-12-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145881327","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}

{"title":"In-depth O-glycosylation characterization and comparison of commercially available etanercept biosimilars by using Electron Activated Dissociation","authors":"Wenhong Fan ,&nbsp;Xiang Li ,&nbsp;Wentao Wang ,&nbsp;Tie Gao ,&nbsp;Ji Luo ,&nbsp;Zoe Zhang ,&nbsp;Bingjie Liu ,&nbsp;Chenggang Liang ,&nbsp;Hongxu Chen","doi":"10.1016/j.jpba.2025.117326","DOIUrl":"10.1016/j.jpba.2025.117326","url":null,"abstract":"<div><div>Glycosylation is one of the important post-translational modifications in protein drugs expressed by eukaryotic cells. It is closely related to the conformation, stability, and solubility of therapeutic proteins and holds vital biological significance. Etanercept contains 26 O-glycosylation sites with high glycosylation heterogeneity, which makes the analysis of O-glycosylation highly challenging. In this paper, using liquid chromatography-mass spectrometry (LC-MS), 11 O-glycosylation on the single chain of etanercept were first identified at the intact protein level, with Core1 and Core2 O-glycans. Second, 11 O-glycosylation sites were identified with the electron activation dissociation (EAD) fragmentation mode at the glycopeptide level, and their glycans were comprehensively characterized. The major O-glycan structures were Core1 and Core2, which were consistent with those observed at the intact protein level. Third, the etanercept innovator (Y1) and six biosimilars or follow-on products (Y2-Y7) from different manufacturers were also compared. Compared with the etanercept innovator, the results showed that etanercept biosimilars or follow-on products from different manufacturers exhibited pronounced differences in O-glycosylation modification due to variation in production processes. Based on a comprehensive assessment of consistency in deconvoluted spectra and the glycosylation ratios, etanercept biosimilar Y2 was the most similar to the innovator Y1. In-depth studies of etanercept’s O-glycosylation can effectively be applied to the etanercept innovator's comparative analysis and biosimilars. Furthermore, the LC-MS method combined with the EAD fragmentation technique can offer detailed and precise data support for the comprehensive quality control of complex glycoprotein drugs and the optimization of production processes.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117326"},"PeriodicalIF":3.1,"publicationDate":"2025-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145881326","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}