Journal of Chromatography B最新文献

{"title":"Anabasine analysis in human plasma using liquid chromatography coupled to tandem mass spectrometry to Verify tobacco use: empirical challenges and limitations","authors":"Christof Manuel Schönenberger ,&nbsp;Aurelie Berthet ,&nbsp;Matthias Briel ,&nbsp;Alain Amstutz ,&nbsp;Matthias Cavassini ,&nbsp;Loïc Sartori ,&nbsp;Camille Rime ,&nbsp;Davide Staedler ,&nbsp;Fiorella Lucarini","doi":"10.1016/j.jchromb.2026.124942","DOIUrl":"10.1016/j.jchromb.2026.124942","url":null,"abstract":"<div><div>Anabasine is an alkaloid frequently quantified by LC–MS/MS to differentiate tobacco use from nicotine replacement therapy. While urine provides reliable measurement, plasma remains a poorly validated and analytically challenging matrix. This study assessed widely used sample preparation strategies for anabasine determination in human plasma. Across all methods, recovery in undiluted plasma was highly variable and largely outside acceptable analytical ranges, with marked ion suppression and poor reproducibility. Matrix dilution increased apparent signal but substantially worsened variability. Experiments in albumin-enriched saline excluded protein binding as the main determinant of analyte loss, indicating broader plasma-related matrix effects. In human plasma, including time-course sampling during and after smoking, anabasine remained consistently below quantifiable levels. None of the tested workflows met the robustness criteria required for quantitative LC–MS/MS analysis, indicating that current preparation approaches do not enable reliable anabasine measurement in plasma, a critical limitation for studies using anabasine as a biomarker of tobacco exposure.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1272 ","pages":"Article 124942"},"PeriodicalIF":2.8,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146075778","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":"The pharmacokinetics and network pharmacology of blood absorbed components for clarifying the potential anti-inflammatory action mechanism of Nelumbinis stamen","authors":"Yang Liu ,&nbsp;Shujing Chen ,&nbsp;Xue Meng ,&nbsp;Rui Wang ,&nbsp;Kunze Du ,&nbsp;Yanxu Chang ,&nbsp;Jin Li","doi":"10.1016/j.jchromb.2026.124931","DOIUrl":"10.1016/j.jchromb.2026.124931","url":null,"abstract":"<div><div>The dried stamen of <em>Nelumbo nucifera</em> Gaertn., holds significant value and is widely used in health teas, functional foods and herbal medicine. <em>Nelumbinis</em> Stamen possesses anti-inflammatory, antioxidant and anti-melanin properties. However, the blood-absorbed components and their mechanisms of action remain unclear. This study aimed to identify the blood-absorbed components and elucidate the anti-inflammatory mechanisms of <em>Nelumbinis</em> Stamen by integrating pharmacokinetics with network pharmacology. A validated UHPLC-MS/MS method was developed to quantify eleven flavonoids in rat plasma following the administration of the aqueous and alcohol extracts for pharmacokinetic comparisons between the two extracts. Five flavonoids were detected in plasma. Kaempferol-3-<em>O</em>-<em>β</em>-D-glucuronide and kaempferol-3-<em>O</em>-<em>β</em>-galactoside present in both extracts while hyperoside, isorhamnetin and isorhamnetin-3-<em>O</em>-<em>β</em>-D-glucoside were exclusive to the alcohol extract. Significant differences in the pharmacokinetic characteristics of these components were observed between the two extracts. Network pharmacology analysis identified PI3K-AKT, TNF and IL-17 signaling pathways as key pathways mediating the anti-inflammatory effects. It was further validated that the alcohol extracts can regulate the PI3K-AKT pathway through <em>in vitro</em> experiments. These findings highlight the effectiveness of integrating pharmacokinetics with network pharmacology in clarifying the bioactive components and mechanisms of <em>Nelumbinis</em> Stamen, providing a scientific basis for its therapeutic potential in inflammatory diseases.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1272 ","pages":"Article 124931"},"PeriodicalIF":2.8,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146006743","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":"High-sensitivity dual analysis of Baricitinib and remdesivir in serum and urine using HPLC-fluorescence and LC-MS approaches in COVID-19 therapy","authors":"Hazim M. Ali ,&nbsp;Lateefa A. Al-Khateeb ,&nbsp;Mohammed M. Ghoneim ,&nbsp;Maha M. Abdelrahman ,&nbsp;Ismail M. Ahmed ,&nbsp;Mohammed Gamal","doi":"10.1016/j.jchromb.2026.124927","DOIUrl":"10.1016/j.jchromb.2026.124927","url":null,"abstract":"<div><div>Sensitive and accurate analytical strategies for the simultaneous quantification of Baricitinib (BAR) and Remdesivir (REM) in biological fluids are essential for supporting their combined use in COVID-19 therapy. Currently, no analytical methods exist for the simultaneous quantification of both drugs in human biological fluids. To bridge this gap, we developed, optimized, and validated novel RP-HPLC-fluorescence and LC-MS methods for the concurrent assay of BAR and REM in serum and urine samples. A high-performance liquid chromatography method with fluorescence detection (HPLC-fluorescence) was optimized using an isocratic mobile phase of phosphoric acid (pH = 3) and ethanol (30:70, <em>v</em>/v) at a flow rate of 0.6 mL/min, with detection at excitation/emission wavelengths of 245/400 nm. Furthermore, an LC-MS approach was developed using a mobile phase of 0.1% formic acid with 0.05 M ammonium formate in water and methanol (80:20, v/v) at 0.5 mL/min in an isocratic approach. The two LC approaches were successfully validated and applied for the simultaneous analysis of spiked BAR and REM mixtures in human urine and serum samples within 5 and 6 min, respectively. In biological human samples, accuracy varied from 96.41% to 98.10% for the HPLC-fluorescence strategy and from 95.17% to 98.47% for the LC-MS approach. Additionally, the precision ranged from 0.83% to 1.23% for the HPLC-fluorescence approach and from 0.66% to 1.55% for the LC-MS strategy. The HPLC-FLD technique demonstrated higher sensitivity for quantifying both BAR and REM at trace levels, with a lower limit of quantification of 0.1 ng/mL and 0.5 ng/mL, respectively, whereas the LC-MS method offered a wider linear range, extending up to 2000 ng/mL for BAR and 1000 ng/mL for REM.</div><div>The assessment of green features using the Analytical GREEnness Metric Approach (AGREE) revealed that the HPLC-fluorescence approach, particularly due to its use of ethanol, is more sustainable (AGREE score: 0.68) than the LC-MS approach (AGREE score: 0.56). The practical functionality of both novel approaches was confirmed with high Blue Applicability Grade Index (BAGI) scores (77.5 and 75, respectively). Without bio-matrices interference, the optimized LC approaches showed very high levels of sensitivity and accuracy. Therefore, these methods are highly suitable for therapeutic drug monitoring and pharmacokinetic studies of the BAR-REM combination.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1272 ","pages":"Article 124927"},"PeriodicalIF":2.8,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146037396","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":"Identification of antithrombotic active components in Xuefu Zhuyu decoctionby thrombin targeted affinity ultrafiltration mass spectrometry","authors":"Ting Zou ,&nbsp;Xiangchang He ,&nbsp;Zhiying Yuan,&nbsp;Weihe He,&nbsp;Jie Dai,&nbsp;Siyu Li,&nbsp;Guangming Xu","doi":"10.1016/j.jchromb.2026.124944","DOIUrl":"10.1016/j.jchromb.2026.124944","url":null,"abstract":"<div><h3>Objective</h3><div>Since screening compound active ingredients is more complex than screening single ingredients, we developed a one-step high-throughput screening method using Affinity Ultrafiltration-Mass Spectrometry (AUF-UPLC-Q-TOF-MS) to rapidly identify the active anti-thrombotic components in Xuefu Zhuyu Decoction.</div></div><div><h3>Method</h3><div>In this study, 50 g of Xuefu Zhuyu Decoction was subjected to reflux extraction with 90% ethanol, followed by concentration and sequential extraction using ethyl acetate, n-butanol and water. The n-butanol fraction was selected as the key research target. Using thrombin as the target, the fraction was incubated with active/inactivated enzyme at 37 °C for 40 min, and the complexes were separated via a 10 kDa ultrafiltration tube. Potential ligands were preliminarily screened by UPLC-Q-TOF-MS combined with blank control experiments. Subsequent in vitro and in vivo validation was performed using molecular docking, chromogenic substrate assay, three coagulation indicators (APTT/PT/TT) and a zebrafish thrombosis model. By omitting the tedious steps of repeated column chromatography and activity tracking, this method exhibits advantages such as minimal sample consumption, high screening efficiency, and reduced experimental cost and time.</div></div><div><h3>Conclusion</h3><div>AUF-UPLC-Q-TOF-MS identified seven components with binding rates exceeding 10% from 11 differential peaks, among which Rhoifolin, Ginsenoside Ro, and Licoricesaponin G2 were confirmed for the first time to exhibit significant thrombin inhibitory efficacy. Compared to the traditional “separation-purification-verification” approach, this method reduces sample consumption by 90% and cuts both time and cost by 80%. It provides a replicable template for rapid, eco-friendly, and cost-effective screening of active components in traditional Chinese medicine formulations, while also establishing a solid experimental foundation for the efficient discovery of antithrombotic lead compounds.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1272 ","pages":"Article 124944"},"PeriodicalIF":2.8,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146121740","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":"Unravelling hepatic metabolism patterns of Gegen Qinlian decoction through a comparative in vivo and in vitro study","authors":"Mengyuan Wang,&nbsp;Yunuo Fan,&nbsp;Ye Zhang,&nbsp;Xingyu Li,&nbsp;Bin Li,&nbsp;Ping Li","doi":"10.1016/j.jchromb.2026.124941","DOIUrl":"10.1016/j.jchromb.2026.124941","url":null,"abstract":"<div><div>The liver plays a central role in drug metabolism, and understanding its metabolic processes is crucial for effective drug development. Gegen Qinlian Decoction (GQD) is widely used in clinical therapy, but its systematic liver metabolic characteristics are still unclear. By integrating UPLC-QTOF-MS with mass defect filtering (MDF), in-house database screening, and feature-based molecular network (FBMN), this study established a multi-dimensional data mining strategy to systematically characterize the <em>in vivo</em> and <em>in vitro</em> liver metabolism characteristics of GQD and clarify its biotransformation pathways. A total of 220 GQD-related components were annotated, of which 179 and 140 were identified in rat liver tissue and human liver microsomes, respectively, and 99 components overlapped between these two biological matrices. Comprehensive analysis shows that alkaloids mainly undergo demethylation and hydroxylation, flavonoids are mainly metabolized by glucuronidation and sulfation, and triterpenoids mainly undergo glucuronidation, hydroxylation, and hydrolysis. The interspecies comparison further demonstrates that alkaloids show more diverse metabolic reactions in rat liver microsomes (RLMs), and the metabolite characteristics largely include those in HLMs and show comparable metabolic rates. In contrast, flavonoids are metabolized faster and extensively in HLMs, resulting in greater product diversity. <em>In vitro</em> and <em>in vivo</em> identification data further support these observations. These findings show that the rat model provides a reasonable representation for the study of alkaloid metabolism, while flavonoid metabolism needs to be verified using HLMs or humanized liver models to improve the clinical translatability of predictions.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1272 ","pages":"Article 124941"},"PeriodicalIF":2.8,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146075780","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":"An optimized HPLC-MS/MS assay for uracil and dihydrouracil in plasma: Improving dihydropyrimidine dehydrogenase phenotyping for individualized fluoropyrimidine treatment","authors":"Yahia Bennani ,&nbsp;Carla Chehade ,&nbsp;Caroline Samer ,&nbsp;Thibaud Kössler ,&nbsp;Aurélien Thomas ,&nbsp;Youssef Daali","doi":"10.1016/j.jchromb.2026.124926","DOIUrl":"10.1016/j.jchromb.2026.124926","url":null,"abstract":"<div><div>Dihydropyrimidine dehydrogenase (DPD) is a key phase I drug-metabolizing enzyme responsible for catabolizing fluoropyrimidine chemotherapeutics such as 5-fluorouracil, capecitabine and tegafur. Its activity shows marked interindividual variability due to both genetic polymorphisms and environmental factors. Reduced DPD activity results in excessive fluoropyrimidine exposure, potentially causing severe and life-threatening toxicities. Although DPD testing is recommended before initiating fluoropyrimidine therapy, no single analytical strategy has been universally adopted, leading to substantial variability in clinical practice. While genotyping offers high specificity, phenotyping provides greater sensitivity by capturing both genetic and non-genetic influences on enzyme activity. In this study, we developed and validated an HPLC-MS/MS assay for the simultaneous quantification of endogenous uracil and dihydrouracil in human plasma, widely used endogenous biomarkers for DPD phenotyping. Sample preparation involved extraction with lipid removal, followed by chromatographic separation on porous graphitic carbon (100 × 2.1 mm, 5 μm; Hypercarb™, Thermo Scientific). Validation was performed according to international guidelines, demonstrating appropriate selectivity, sensitivity, linearity, accuracy, precision, carry-over, and stability. The method was applied to 28 human plasma samples, with uracil concentrations compared against an external laboratory using an independent LC–MS/MS platform. Deming regression showed no significant constant bias at the 95% confidence level, although underestimation occurred at higher concentrations (y = 0.63× + 4.86). Bland–Altman analysis indicated a small mean difference of −0.02 ng/mL but wide limits of agreement (−9.61 to +9.57 ng/mL), highlighting the impact of analytical variability near clinical decision thresholds. Further studies are warranted to evaluate the clinical utility of this approach in larger cohorts and to assess its correlation and complementarity with DPD genotyping and clinical outcomes. This method represents a valuable analytical tool to support personalized fluoropyrimidine-based chemotherapy in patients with solid tumors.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1272 ","pages":"Article 124926"},"PeriodicalIF":2.8,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146047065","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 metabolomics, network pharmacology, experimental validation and quantitative analysis to reveal and evaluate the pharmacological substances of Paris polyphylla var. yunnanensis on anti-inflammatory","authors":"Pengjuan Li ,&nbsp;Qiwei Zhang ,&nbsp;Hongbo Xu ,&nbsp;Yuangui Yang","doi":"10.1016/j.jchromb.2026.124932","DOIUrl":"10.1016/j.jchromb.2026.124932","url":null,"abstract":"<div><div>This study employed an integrated approach combining <em>in vivo</em> metabolism analysis, network pharmacology, experimental validation and quantitative analysis to investigate the anti-inflammatory material basis and mechanism of action of <em>Paris polyphylla</em> var. <em>yunnanensis</em> (PPY). Firstly, rats were orally administered extracts from the roots and leaves of PPY, and plasma, urine, and feces samples were collected at different time points post-administration. Using UHPLC-Q/TOF-HRMS technology, a total of 26 xenobiotics (18 prototypes and 8 metabolites) were identified from the rat samples. Subsequently, network pharmacology analysis predicted 19 core targets and preliminarily screened 10 key bioavailable components with potential anti-inflammatory activity. Further <em>in vitro</em> cell experiments evaluated the bioactivity of these key components, revealing that polyphyllin II exhibited the most significant anti-inflammatory effect, while pennogenin and diosgenin also demonstrated anti-inflammatory activity within certain concentration ranges. Finally, HPLC-DAD analysis was used to compare the content differences of major active components in PPY from different geographical origins. The results showed that samples from Hanzhong (Shaanxi) and Chengdu (Sichuan) contained higher levels of pennogenin-type saponins, whereas those from Honghe (Yunnan) and Yuxi (Yunnan) were richer in diosgenin-type saponins. In summary, this comprehensive strategy significantly contributes to elucidating the anti-inflammatory material basis and mechanism of PPY, while also providing a reference for the systematic discovery of active components in the quality evaluation of traditional Chinese medicines.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1272 ","pages":"Article 124932"},"PeriodicalIF":2.8,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146075777","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":"Sustainable fabric-phase sorptive extraction membrane characterization and comprehensive greenness assessment for detecting pharmaceutical adulterants in herbal slimming products","authors":"Aslıhan Gürbüzer ,&nbsp;Halil İbrahim Ulusoy ,&nbsp;İbrahim Narin ,&nbsp;Abuzar Kabir ,&nbsp;Marcello Locatelli","doi":"10.1016/j.jchromb.2026.124928","DOIUrl":"10.1016/j.jchromb.2026.124928","url":null,"abstract":"<div><div>This study presents a sustainable analytical approach centered on the characterization and environmental evaluation of a sol-gel Carbowax 20M-coated fabric-phase sorptive extraction (FPSE) membrane designed for detecting pharmaceutical adulterants—specifically fluoxetine and sibutramine—in herbal slimming products. The membrane was synthesized via in situ sol-gel immobilization and characterized using FTIR and SEM, confirming the formation of a uniform and porous sorbent layer that maintained the structural integrity and permeability of the cellulose fabric substrate. Under optimized conditions, the developed FPSE-HPLC-DAD method achieved limits of detection (LOD) of 4.28 ng mL⁻¹ for fluoxetine and 5.71 ng mL⁻¹ for sibutramine, with recoveries ranging between 94.2% and 112.5% and relative standard deviations (RSD) below 5.2%. The linear ranges extended from 15 to 900 ng mL⁻¹ for fluoxetine and 20–1200 ng mL⁻¹ for sibutramine, demonstrating strong linearity (R² &gt; 0.99). Beyond analytical validation, the environmental performance of the method was systematically assessed using six recognized greenness and applicability metrics: AGREE (score = 0.63), MoGAPI, AGSA, BAGI (&gt; 60), CACI (62%), and AGREEprep, all confirming high sustainability and practical applicability. These findings highlight that the sol-gel Carbowax 20M coated FPSE membrane provides a low-cost, solvent-efficient, and environmentally responsible platform for the routine screening of undeclared pharmaceutical compounds in complex herbal matrices.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1272 ","pages":"Article 124928"},"PeriodicalIF":2.8,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146006748","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 identification of chemical components in Polygonatum kingianum Collett & Hemsl. based on UHPLC-Q-Exactive orbitrap MS coupled with molecular networking","authors":"Qing Li ,&nbsp;Kailin Li ,&nbsp;Jiatong Yang ,&nbsp;Wen Cai ,&nbsp;Huiqing Yang ,&nbsp;Bowen Gao ,&nbsp;Wei Cai","doi":"10.1016/j.jchromb.2026.124943","DOIUrl":"10.1016/j.jchromb.2026.124943","url":null,"abstract":"<div><div><em>Polygonatum kingianum</em> Collett &amp; Hemsl. (PK) is a plant dually used as both medicine and food, renowned in traditional medicine for its abilities to replenish qi, nourish yin, moisten the lungs, and benefit the kidneys. Previous investigations have indicated that its chemical composition encompasses steroidal saponins, polysaccharides, and flavonoids. However, the PK's chemical components is still scarce. Therefore, it is essential to establish a systematic method for the identifying and characterizing its chemical components. In this study, a total of 153 chemical components, including 37 peptides, 26 organic acids, 20 saponins, 19 flavonoids, 7 amino acids, 7 nucleotides, 7 lipids, 6 terpenoids and other classes, was identified by UHPLC-Q-Exactive Orbitrap MS with molecular networking. This analysis revealed 118 new compounds from PK, which distinctly lays the foundation for the scientific exploration of the pharmacodynamic substance basis of PK as well as the development and application of the resources derived from it.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1272 ","pages":"Article 124943"},"PeriodicalIF":2.8,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146115083","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":"Impacts of methionine oxidation on product quality and anti-tumor activity of a therapeutic monoclonal antibody","authors":"Qiangzu Sun ,&nbsp;Zhenhua Miao ,&nbsp;Liqing Bian ,&nbsp;Fei Teng ,&nbsp;Huanhuan Guo ,&nbsp;Xu-Alan Lin ,&nbsp;Xichen Zhang ,&nbsp;Fan Zhang ,&nbsp;Dong-Qiang Lin","doi":"10.1016/j.jchromb.2026.124920","DOIUrl":"10.1016/j.jchromb.2026.124920","url":null,"abstract":"<div><div>Methionine oxidation in monoclonal antibodies (mAbs) could critically compromise drug stability and efficacy. This investigation comprehensively evaluated the structural and functional impacts of oxidation on a mAb An under long-term storage (1.5 years at 5 °C) and forced oxidation conditions. Orthogonal analytical approaches including high-performance liquid chromatography and mass spectrometry revealed preferential oxidation at M253 and M429 residues, concomitant with increased charge heterogeneity (basic peaks) and hydrophobicity. Notably, while the oxidation attenuated Complement-Dependent Cytotoxicity (CDC) activity by ∼70%, it did not alter Antibody-Dependent Cellular Cytotoxicity (ADCC), binding affinity (FcγRIIIa), or <em>in-vivo</em> anti-tumor efficacy in murine models. The pharmacokinetics was similarly unaffected. These findings demonstrate that methionine oxidation, despite altering physicochemical properties, does not impact mAb A's essential biological functions. Crucially, forced oxidation samples effectively simulated the oxidation progression of mAb An under subsequent storage conditions, supporting its stability during long-term storage.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1271 ","pages":"Article 124920"},"PeriodicalIF":2.8,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145974242","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}