Journal of pharmaceutical and biomedical analysis最新文献

{"title":"Simultaneous determination of four PDE5 inhibitors and metabolites in rat plasma by UPLC-MS/MS","authors":"Xiaonan Xia ,&nbsp;Zhe Song ,&nbsp;Weiwen He ,&nbsp;Zhou Qiao ,&nbsp;Fanglin Wang ,&nbsp;Shaoyuan Li","doi":"10.1016/j.jpba.2025.116883","DOIUrl":"10.1016/j.jpba.2025.116883","url":null,"abstract":"<div><div>Herein, we developed a sensitive UPLC-MS/MS method for the simultaneous determination of four PDE5 inhibitor and twelve metabolites in rat plasma. A total of 16 chemicals were separated on gradient elution with mobile phase A (0.1 % formate acetonitrile) and mobile phase B (0.1 % formate and 2 mmol/L ammonium formate aqueous solution) on an ACQUITY UPLC HSS T3 column (100 mm×2.1 mm, 1.8 μm) after protein precipitation, followed by the detection on the positive ion mode of electrospray ion source (ESI) via dynamic multi-reaction monitoring mode (MRM). The method provided good linearity over the range of 0.25–20 ng/mL for all compounds with correlation coefficients R greater than 0.99. The recoveries were higher than 75.5 %, the matrix factors were 2.1 %-20.4 %. The precision, accuracy and stability were also within acceptable criteria. A rat model was established to investigate the metabolic profile of Sildenafil, Vardenafil, Acetildenafil and Tadalafil. Blood samples from the experimental group were analyzed using ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS). Notably, desmethyltadalafil and methyl-desmethyltadalafil were not detected in the plasma. However, their glucuronide conjugates, desmethyltadalafil glucuronide and methyl-desmethyltadalafil glucuronide, were identified and characterized via tandem mass spectrometry (MS/MS). At present, cases of poisoning or even death due to overdose of phosphodiesterase type 5 (PDE5) inhibitors are often encountered in the practice of forensic science. Few studies on PDE5 inhibitors have been reported in forensic science toxicology identification, and the existing detection methods cannot meet the need for simultaneous and rapid analysis of multiple trace PDE5 inhibitors in vivo. Therefore, it is of great importance to establish and optimize the detection methods for PDE5 inhibitors drugs in biological samples. The significance of this method is to provide a reference for the determination of PDE5 inhibitors poisoning cases.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"262 ","pages":"Article 116883"},"PeriodicalIF":3.1,"publicationDate":"2025-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143828634","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":"Stability of the antibiotic ceftaroline fosamil with focus on degradation products and decomposition pathways – Integrative analysis by LC-DAD, ESI-QTOF and in silico prediction","authors":"Marianna Coelho Mendes ,&nbsp;Lilian Fanfa Machioli ,&nbsp;Graciela Carlos ,&nbsp;Tiago Franco de Oliveira ,&nbsp;Andreas Sebastian Loureiro Mendez","doi":"10.1016/j.jpba.2025.116876","DOIUrl":"10.1016/j.jpba.2025.116876","url":null,"abstract":"<div><div>Stability studies are important tools for pharmacovigilance, enabling the investigation of new compounds made available to the population under different hospital conditions. Indicated for the treatment of complicated skin and soft tissue infections and community-acquired pneumonia, ceftaroline fosamil is a fifth-generation cephalosporin marketed as Zinforo®, a novel antibiotic given the long development time, in the form of powder for infusion. Due to the presence of a phosphate group, ceftaroline fosamil is a prodrug that is converted by hepatic enzymes to active form ceftaroline. The aim of the present study was to investigate the stability of ceftaroline fosamil through forced degradation studies (thermal stress) under exposures to 40°C and 60°C, and clinical use conditions under exposures to 4°C and 25°C, in the forms of powder, reconstituted in purified water, and diluted in 5 % glucose solution and 0.9 % sodium chloride solution (2.4 mg/mL, 5.0 mg/mL, and 8.4 mg/mL), for time periods of 10 min, 30 min, 60 min, 2 h, 24 h, 48 h, and 72 h. The preliminary analysis performed by HPLC-DAD showed that ceftaroline fosamil diluted at 2.4 mg/mL in a 5 % glucose solution was the least stable under clinical use conditions, with a decay of approximately 65 % of ceftaroline fosamil. Through mass spectrometry analysis by ESI-QTOF, the degradation products <em>m/z</em> 303, <em>m/z</em> 337, <em>m/z</em> 442, <em>m/z</em> 483, and <em>m/z</em> 543 and their degradation pathways were proposed. It was possible to partially relate the results of mass spectrometry and <em>in silico</em> experimental model for predicting degradation products.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"262 ","pages":"Article 116876"},"PeriodicalIF":3.1,"publicationDate":"2025-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143823451","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":"Unique methotrexate polyglutamates distributions in peripheral blood mononuclear cells of rheumatoid arthritis patients: Development and validation of a UPLC-MS/MS method","authors":"Janani Sundaresan ,&nbsp;Marry Lin ,&nbsp;Gerrit Jansen ,&nbsp;Renske C.F. Hebing ,&nbsp;Maja Bulatović-Ćalasan ,&nbsp;Robert de Jonge ,&nbsp;Eduard A. Struys ,&nbsp;Maurits C.F.J. de Rotte","doi":"10.1016/j.jpba.2025.116882","DOIUrl":"10.1016/j.jpba.2025.116882","url":null,"abstract":"<div><div>Methotrexate is pivotal in treating immune-mediated inflammatory diseases. Intracellularly, methotrexate is metabolized to methotrexate-polyglutamates (MTX-PG_1–7), comprising up to six additional glutamate moieties, crucial for cellular retention and therapeutic efficacy. Hitherto, quantification of MTX-PG_1–6 in peripheral blood mononuclear cells (PBMCs) from methotrexate-treated patients was challenging due to their low abundance in blood and matrix effects. We present a robust validated UPLC-MS/MS method to quantify individual MTX-PG_1–6 in PBMCs. Stable-isotope labelled internal standard mixture of MTX-PG_1–6 was added to 5 million PBMCs, followed by deproteinization with perchloric acid, and additional sample clean-up using solid phase extraction columns. MTX-PG_1–6 were detected and quantified using UPLC-MS/MS. The method was validated for lower limit of quantification (LLOQ), linearity, carryover, recovery, matrix effects, precision and stability. We assessed MTX-PG_1–6 in PBMCs derived from five methotrexate-treated rheumatoid arthritis patients. For all MTX-PG_1–6, LLOQs were &lt; 1 fmol-MTX-PG_1–6/million cells with linearities R² &gt; 0.995. The recoveries, carryover and stability were acceptable and no matrix effects were observed. The intraday and interday precision %CVs of quality controls ranged from 2.7 % to 11.4 % and 3.5–14.9 % respectively. Interday precision using nine PBMCs aliquots from a single MTX-treated patient aligned similarly (%CV &lt;15 %). In patient-derived PBMC samples, MTX-PG₁ was the highest, with decreasing concentrations of MTX-PG₂ to MTX-PG₅. No signal for MTX-PG₆ was detected in the patient samples. We validated a new UPLC-MS/MS method to quantify MTX-PG_1–6 in PBMCs, thus facilitating PBMC-based therapeutic drug monitoring studies and understand associations between MTX-PG_1–6 concentration and therapy efficacy or adherence.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"262 ","pages":"Article 116882"},"PeriodicalIF":3.1,"publicationDate":"2025-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143820417","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":"Novel approaches in CRISPR/Cas12a-based sensing for HCC diagnosis - A review (2020–2025)","authors":"Cunhong Cen ,&nbsp;Xiyu Liu ,&nbsp;Wei He ,&nbsp;Xiaohong Tan ,&nbsp;Guiyin Li ,&nbsp;Na Jintong","doi":"10.1016/j.jpba.2025.116878","DOIUrl":"10.1016/j.jpba.2025.116878","url":null,"abstract":"<div><div>Early diagnosis of hepatocellular carcinoma (HCC) is crucial for improving patient survival and treatment outcomes and the early detection of biomarkers for HCC is key to achieving this goal. However, conventional detection methods often lack sufficient specificity and sensitivity. In recent years, CRISPR/Cas12a-based biosensing has gained significant attention due to its ease of use and high sensitivity, demonstrating its potential to address the limitations of conventional detection methods. This paper primarily reviews the research progress of CRISPR/Cas12a-based biosensors for HCC detection, introducing their fluorescence, electrochemical, colorimetric, and other detection principles, as well as practical applications in detail. Additionally, the differences in sensitivity, specificity, and detection speed among different types of CRISPR/Cas12a biosensors are comparatively analyzed. Finally, the potential future directions for the development and application of CRISPR/Cas12a technology in clinical settings are explored.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"262 ","pages":"Article 116878"},"PeriodicalIF":3.1,"publicationDate":"2025-04-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143800273","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":"Pseudotargeted metabolomics profiles potential damage-associated molecular patterns as machine learning predictors for acute pancreatitis","authors":"Wen-Lu Cai ,&nbsp;Can Fang ,&nbsp;Hong-Xu Leng ,&nbsp;Jia-Yi Zheng ,&nbsp;Li-Fang Liu ,&nbsp;Guan-Wen Gong ,&nbsp;Gui-Zhong Xin","doi":"10.1016/j.jpba.2025.116874","DOIUrl":"10.1016/j.jpba.2025.116874","url":null,"abstract":"<div><div>Acute pancreatitis (AP) is a common gastrointestinal disease characterized by pancreatic cell damage and inflammation. Given the early clinical diagnosis and management challenges, exploring novel analytical frameworks from new orientations for interrogating AP is urgent. The release of damage-associated molecular patterns (DAMPs) and their receptor recognition initiate sterile inflammation, serving as key drivers in the development and progression of AP. Thus, this study aimed to delineate the underlying correlations between alterations in the DAMP profile and the AP state. We have developed a new framework combining potential DAMPs profiles obtained from pseudotargeted metabolomics method with machine learning (ML) models for AP prediction. 2-(1-Piperazinyl) pyrimidine chemical labeling was utilized to provide characteristic fragment ions and improve the quantitative sensitivity of targeted metabolites. A total of 49 potential DAMPs were identified and semi-quantified from collected serum samples (n = 84), positive or negative for APs. For modeling obtained datasets with five different ML algorithms, the support vector machine model was chosen as the optimal model to differentiate with high accuracy, achieving an area under the receiver-operating characteristic curve (AUROC) of 0.944. It also showed a strong performance in an external independent validation set (AUROC: 0.907). Moreover, the model was interpreted using the Shapley Additive exPlanations analysis to specify the important features and identify specific free fatty acids as key contributors. Overall, the novel framework enables high accuracy in predicting the presence of AP status. Meanwhile, it underlines the utility of DAMPs in inflammatory diseases and provides reference values for diagnosing in first-line clinics.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"262 ","pages":"Article 116874"},"PeriodicalIF":3.1,"publicationDate":"2025-04-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143815277","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":"Alcohol-based solvents as mobile phases for LC-MS characterization of therapeutic proteins","authors":"Mostafa Zarei ,&nbsp;Jérôme Jonveaux,&nbsp;Michael Jahn","doi":"10.1016/j.jpba.2025.116879","DOIUrl":"10.1016/j.jpba.2025.116879","url":null,"abstract":"<div><div>Acetonitrile (ACN) is currently the preferred solvent in reversed phase (RP) chromatography for protein characterization through peptide mapping. Despite its effective performance, ACN poses toxicity risks to humans and has adverse effects on environmental sustainability. In the current work, we developed a novel alcohol-based solvent system for peptide mapping by systematic evaluation of parameters such as organic eluent composition, solvent gradient, flow rate, and column temperature. We compared the chromatographic performance and MS response of peptides between the standard (ACN based) and the new developed solvent systems (EtOH/IPA based). The results of our study show that the EtOH/IPA based solvent system improves selectivity factor (α) and resolution (R), while the standard ACN based solvent system provides a lower peak width and hence a higher peak height. The majority of the analysed peptides exhibited shorter retention times, whereas hydrophobic peptides eluted later when using the EtOH/IPA solvent system. Several critical quality attributes (CQA) of a monoclonal antibody (mAb) were successfully characterized by this method without compromising the chromatographic separations and MS response of the peptides. Furthermore, the suitability of the new approach for LC-UV assessment of a mAb as part of an identity test of therapeutic proteins was demonstrated. Our proposed approach, which prioritizes safety, non-toxicity, and compatibility with all LC-MS instruments, offers significant support to a broad community of academic and biopharmaceutical scientists in their pursuit of a bottom-up green strategy.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"262 ","pages":"Article 116879"},"PeriodicalIF":3.1,"publicationDate":"2025-04-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143815274","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":"HPLC method for novel cannabinoid separation with electrochemical detection on boron-doped diamond electrode","authors":"Zeynab Belbasi ,&nbsp;Jan Petr ,&nbsp;Jan Hrbac ,&nbsp;David Jirovsky","doi":"10.1016/j.jpba.2025.116875","DOIUrl":"10.1016/j.jpba.2025.116875","url":null,"abstract":"<div><div>In this study, we developed a novel and environmentally friendly method for the determination of selected cannabinoids — cannabidiol, cannabinol, tetrahydrocannabinol, JWH 073 synthetic cannabinoid, hexahydrocannabinol (HHC), and hexahydrocannabinol-O-acetate — using high-performance liquid chromatography with electrochemical detection. The method employs a boron-doped diamond working electrode, which offers significant advantages in comparison with commonly used materials. Its main strengths include low background currents, reduced fouling, and an extended potential window, making it suitable for detecting compounds that are not readily oxidizable. The chromatographic system utilized a reversed-phase Gemini C18 column with a mobile phase consisting of acetonitrile and 25 mM phosphate buffer (pH 4.4) in step-gradient mode from 68 % to 88 % acetonitrile. The separation and determination of cannabinoids were optimized to be completed in less than 25 min. Hydrodynamic voltammetry was performed to identify the optimal potential for the amperometric detection, with a working potential of + 1400 mV (vs. Pd/H₂) providing the best analytical response based on S/N ratio. Under these optimal separation and detection conditions, the limits of detection were within the range of 6.1–666 nM (for cannabidiol and 9(R)-hexahydrocannabinol-O-acetate), while the response was linear within the concentration range measured up to 10 μM. The method demonstrated good linearity, precision, and sensitivity, with limits of detection and quantitation reaching nanomolar levels. The developed method was successfully applied to commercial samples of HHC jelly bears and HHC distillate.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"262 ","pages":"Article 116875"},"PeriodicalIF":3.1,"publicationDate":"2025-04-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143815278","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":"Intervention study of Yanghe decoction on plasma cell mastitis based on mammary microecology and metabolomics investigation","authors":"Feixia Ma ,&nbsp;Yi Xiao ,&nbsp;Luyao Qian ,&nbsp;Shuo Zhang","doi":"10.1016/j.jpba.2025.116870","DOIUrl":"10.1016/j.jpba.2025.116870","url":null,"abstract":"<div><div>Plasma cell mastitis (PCM), a chronic breast inflammatory disease, is characterized by ductal dilatation and plasma cell infiltration. Traditional Chinese Medicine (TCM) Yanghe decoction is used to relieve mastitis symptoms clinically. However, its efficacy and mechanism on PCM remain undefined. The PCM patients receiving dexamethasone and metronidazole tablets, served as control group, with the other group receiving combination of Yanghe decoction, 15 cases each. Main symptom indicators included mass size, skin colour, skin temperature, breast pain, blood cell count (WBC), C reactive protein (CRP), interleukin (IL)-6, IL-4, serum prolactin (PRL), erythrocyte sedimentation rate (ESR), immunoglobulin A (IgA), IgM, and IgG levels. Secondary indicators included TCM syndrome and anxiety/depression scores. Breast differential flora (DF) and metabolites (DAMs) post-treatment between two groups were detected by 16S rRNA sequencing and metabolomics, with further correlation analysis. Post-treatment symptom scores exhibited lower than pre-treatment in both groups, with greater decline on WBC, CRP, IL-6, IL-4, PRL, and ESR, and enhanced IgA, IgM, and IgG levels in Yanghe decoction group than in control group. Notably, Yanghe decoction group demonstrated no significant DF, with decreased <em>Corynebacterium</em> and <em>Rhodococcus,</em> and elevated <em>Staphylococcus</em>, which correlated significantly with the above indicators. Moreover, 43 DAMs were detected between these two groups, with Glycerophosphocholine, 9,10-Epoxyoctadecenoic acid, Arachidic acid, and 4,5-Dihydroorotic acid showing strong correlations with the above flora. Based on the clarification of PCM improvement with Yanghe decoction, we preliminarily explored potential roles of breast tissue microorganisms and DAMs, providing scientific basis for its clinical application and potential clinical biomarkers in PCM.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"262 ","pages":"Article 116870"},"PeriodicalIF":3.1,"publicationDate":"2025-04-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143800274","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":"Assessment of the hepatoprotective effects of Bicyclol's forced degradation products using a zebrafish model","authors":"Mengqi Jia ,&nbsp;Runjuan Yang ,&nbsp;Cui Ma ,&nbsp;Chao Gu ,&nbsp;Dongying Wu ,&nbsp;Xucong Huang ,&nbsp;Lili Jing ,&nbsp;Guorong Fan","doi":"10.1016/j.jpba.2025.116871","DOIUrl":"10.1016/j.jpba.2025.116871","url":null,"abstract":"<div><div>Bicyclol (BIC), a synthetic hepatoprotective agent widely prescribed in China, lacks comprehensive safety and activity profiles for its degradation products (DPs). Here, we systematically investigated BIC’s forced degradation behavior using ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS). Three hydrolytic degradation products (DP1-DP3) were isolated via HPLC and structurally characterized, revealing methylenedioxy group hydrolysis as the primary degradation pathway. Molecular docking simulations demonstrated enhanced target binding affinities of DPs compared to BIC, supported by absorption, distribution, metabolism, and excretion (ADME) predictions showing improved drug-likeness. In an alcohol-induced fatty liver zebrafish model, both BIC and its DPs attenuated hepatic macrovesicular steatosis and inflammatory responses. Mechanistically, treatment normalized lipid metabolism by downregulating alcohol-induced expression of <em>FASN</em>, <em>SREBP1</em>, <em>PPARα</em>, and <em>PPARγ</em>, while reduced IL-6 and TNF-α levels confirmed anti-inflammatory efficacy. These findings demonstrate that BIC DPs exhibit dual pharmacological activity through lipid homeostasis modulation and inflammation suppression, providing critical insights for quality control and therapeutic applications.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"262 ","pages":"Article 116871"},"PeriodicalIF":3.1,"publicationDate":"2025-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143844988","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":"Integrated cartilage metabolomics and proteomics analysis reveals the therapeutic effect of Wenjing Tongluo Decoction on Knee osteoarthritis rats","authors":"Wei Wei ,&nbsp;Chenjian Peng ,&nbsp;Renjun Gu ,&nbsp;Xiwu Yan ,&nbsp;Jiapeng Ye ,&nbsp;An Kang ,&nbsp;Luning Sun","doi":"10.1016/j.jpba.2025.116869","DOIUrl":"10.1016/j.jpba.2025.116869","url":null,"abstract":"<div><div>Knee osteoarthritis (KOA)is an age-related degenerative whole-joint disease characterized by poor outcomes. Wenjing Tongluo Decoction (WJTLD), a Chinese herbal remedy, has demonstrated favorable clinical effects on KOA. However, the precise mechanisms therein remain poorly defined. In this study, we employed the method of anterior cruciate ligament transection (ACLT) method to establish a rat model of KOA. Following 8 weeks of oral administration of WJTLD, the morphology of knee joint cartilage was evaluated using Safranin-O/Fast green staining, H&amp;E staining, and micro-CT imaging. Utilizing GC-MS based untargeted metabolomics and nano-LC-QE-MS based proteomics, we identified altered metabolites and proteins associated with knee cartilage in different rat groups, which were further validated through western blotting and real-time PCR. Our findings indicate that WJTLD alleviates damage to knee joint cartilage and inhibits cartilage degradation. Proteomics data revealed that the altered proteins in OA and WJTLD treated group were related to the biological process including amoebiasis, platelet activation, ECM-receptor interaction, protein digestion and absorption, and ribosome function. Western blotting results confirmed that the expression levels of MMP8 and LDHA were significantly upregulated in the KOA group but were rescued by WJTLD treatment. According to untargeted metabolomics, the intensities of lactic acid, isoleucine, lysine, glutamate, myo-inositol, adenosine, and β-alanine were significantly elevated in the KOA group, however, these metabolites experienced a dramatic following WJTLD treatment. These results suggest that WJTLD exerts a therapeutic effect on KOA by suppressing inflammation and cartilage degradation, as well as regulating multiple pathways related to ECM degradation, amino acid metabolism, and energy metabolism, including glycolysis.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"262 ","pages":"Article 116869"},"PeriodicalIF":3.1,"publicationDate":"2025-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143800272","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}