Journal of Chromatography B最新文献

{"title":"A Roadmap guide on bioanalysis challenges and practical solutions for accurate quantification of oligonucleotide-based novel therapeutic modalities using LC-MS","authors":"Devendra Kumar ,&nbsp;Manish Sharma ,&nbsp;Neerja Trivedi","doi":"10.1016/j.jchromb.2025.124900","DOIUrl":"10.1016/j.jchromb.2025.124900","url":null,"abstract":"<div><div>Therapeutic oligonucleotides have emerged as a transformative drug class, yet their physicochemical complexity poses unique analytical challenges in bioanalysis. Liquid chromatography mass spectrometry (LC-MS) has become a powerful platform for their quantification, offering high specificity and structural insight. However, accurate measurement requires addressing challenges such as nonspecific binding, matrix effects, nuclease degradation, and ion-pairing interferences from sample preparation to LC-MS analysis. This review provides a practical roadmap for establishing robust LC-MS workflows for oligonucleotides bioanalysis, emphasizing optimized sample preparation, column and mobile phase selection, ionization control, and fragmentation tuning. Key strategies to minimize analytical artifacts, improve recovery, and ensure regulatory compliance are discussed in the context of current FDA or EMA bioanalytical validation guidelines. Collectively, this work outlines the critical considerations and systematic optimizations needed to achieve reliable, reproducible, and sensitive quantification of therapeutic oligonucleotides in complex biological matrices, supporting their successful clinical translation by informing pharmacokinetics, therapeutic potential, and safety profiles.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1270 ","pages":"Article 124900"},"PeriodicalIF":2.8,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145788163","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":"Hepatoprotection of Apocynum venetum flavonoids: Targeting oxidative stress via coordinated NRF2/BCHE induction and AR inhibition","authors":"Yao Liu ,&nbsp;Huanyi Ren ,&nbsp;Qing Bin ,&nbsp;Kai Hu ,&nbsp;Hongya Zhu ,&nbsp;Qing Cheng ,&nbsp;Mengqing Zhang ,&nbsp;Chao Jiang ,&nbsp;Jiahao Feng","doi":"10.1016/j.jchromb.2025.124899","DOIUrl":"10.1016/j.jchromb.2025.124899","url":null,"abstract":"&lt;div&gt;&lt;h3&gt;Background&lt;/h3&gt;&lt;div&gt;Acute liver injury (ALI) represents a serious clinical condition characterized by rapid progression toward hepatic functional decompensation, potentially culminating in liver failure. Despite proposed therapeutic interventions, effective management strategies remain limited. Total flavonoids from Apocynum venetum leaves (TFAP), recognized for their antioxidant and anti-inflammatory properties, have demonstrated multi-organ protective effects in prior studies. However, the precise mechanisms underlying TFAP's role in attenuating ALI pathogenesis remain poorly elucidated.&lt;/div&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Methods&lt;/h3&gt;&lt;div&gt;Utilizing the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database and analysis platform, in conjunction with the drug mapping server, the active constituents of TFAP and their therapeutic targets for acute liver injury were identified. Specific targets pertinent to the treatment of acute liver injury were screened and compiled via the GEO database. Molecular docking techniques were employed to elucidate the interactions between the active compounds and their respective targets. Furthermore, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted to provide a comprehensive analysis of the targets associated with acute liver injury. To validate the network pharmacology findings, CCl4-induced acute liver injury cell and animal models were established. Cell viability was assessed using the Cell Counting Kit-8 (CCK8) solution, while flow cytometry, RT-qPCR, and Western blot assays were conducted to evaluate indicators related to cell apoptosis and the Nrf2 signaling pathway. Biochemical assays were employed to quantify the enzymatic activities of aspartate transaminase (AST), alanine transaminase (ALT), and superoxide dismutase (SOD), as well as the concentration of malondialdehyde (MDA) in both cellular and plasma samples. Hematoxylin-Eosin (HE) staining was conducted to examine the pathological alterations in hepatic tissues of mice across different experimental groups. Additionally, immunohistochemical methods were utilized to assess the expression levels of the proteins Nrf2, Keap1, HO-1, and GCLC in liver tissues.&lt;/div&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Results&lt;/h3&gt;&lt;div&gt;A comprehensive transcriptome analysis combined with network pharmacology has indicated that TFAP may exert therapeutic effects on alcoholic hepatitis (ALD) by modulating the AR and BCHE signaling pathways. In vitro studies demonstrated that TFAP extended the survival of hepatocytes subjected to injury, significantly decreased the levels of AST, ALT, LDH, ROS, and MDA, and enhanced SOD activity in CCl₄-induced LO2 cells. Furthermore, treatment with these flavonoids resulted in a reduction of AR protein expression while increasing the expression of BCHE and Nrf2 proteins. In vivo experiments revealed that, in comparison to the blank control group, the model group exhibited significantly elevated levels of li","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1270 ","pages":"Article 124899"},"PeriodicalIF":2.8,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145837741","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":"Chemical composition and spatiotemporal distribution characteristics of Gu-ben-Qing-Hua formula in ulcerative colitis rats based on UPLC-Q-Exactive-MS and MALDI mass spectrometry imaging","authors":"Mengli Chang ,&nbsp;Xinghong Li ,&nbsp;Fengrong Zhang ,&nbsp;Ye Zhao ,&nbsp;Xianyu Li ,&nbsp;Liying Tang ,&nbsp;Hongwei Wu ,&nbsp;Hongjun Yang","doi":"10.1016/j.jchromb.2025.124898","DOIUrl":"10.1016/j.jchromb.2025.124898","url":null,"abstract":"<div><div>Gu-Ben-Qing-Hua formula (GBQHF) is a traditional Chinese medicine (TCM) formula that exhibits significant efficacy in the treatment of ulcerative colitis (UC) in a clinical setting. A combination of ultrahigh-performance liquid chromatography with Q Exactive mass spectrometry (UPLC-Q-Exactive-MS) in both positive and negative ionization modes and matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) in positive ionization mode was used to identify the components of GBQHF and determine its distribution after oral administration. A total of 240 chemical compounds, including 100 flavonoids, 27 saponins, 22 terpenoids, 20 phenolics, 18 alkaloids, 17 volatile compounds, and 12 organic acids, and 24 other compounds, were identified in GBQHF with high mass accuracy (within 5 ppm) and using MS/MS data based on online, local, and self-built databases for GBQHF using UPLC-Q-Exactive-MS. Moreover, 67 prototype compounds were identified in the intestinal tissues. The characteristics of the identified compounds, including the numbers as well as the structure and source in the duodenum, ileum, proximal colon, and distal colon, were elucidated. Among these 67 compounds, the distribution of licoricesaponin A3 and astragaloside IV known for their anti-inflammatory properties was analyzed using MALDI-MSI in 1.5-, 6-, and 24-h postdose sections of the ileum and the proximal and distal colon. Licoricesaponin A3 was significantly distributed in the basolateral region of the proximal colon, especially at 1.5 h. Astragaloside IV showed a significant distribution both in the mucosal side and basolateral region of the distal colon, especially at 6 after administration. This study systematically and comprehensively characterizes the chemical constituents of GBQHF, thereby establishing an effective strategy to characterize compounds in TCM formulae used to treat UC.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1270 ","pages":"Article 124898"},"PeriodicalIF":2.8,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145829591","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 liquid chromatography-mass spectrometry method for profiling global phospholipid methylation via Headgroup analysis revealisng cell-type specificity","authors":"Hongjun Luo ,&nbsp;Hui Li ,&nbsp;Yongyin Zhou ,&nbsp;Wenhong Luo ,&nbsp;Zhexuan Lin","doi":"10.1016/j.jchromb.2025.124903","DOIUrl":"10.1016/j.jchromb.2025.124903","url":null,"abstract":"<div><div>Global analysis of phospholipid methylation is crucial for understanding the biological roles of phospholipids in mammalian membranes. This study developed and validated a robust liquid chromatography-mass spectrometry (LC-MS) method for the simultaneous determination of ethanolamine (EA) and choline (CL) released from the hydrolysis of phosphatidylethanolamine (PE) and phosphatidylcholine (PC), respectively. The extraction, hydrolysis, and detection procedures were optimized. The compounds were separated on an HILIC column with isocratic elution and detected using a mass spectrometer with electrospray ionization in positive ion mode. The selected ions monitored were <em>m</em>/<em>z</em> 104.1 for CL, m/z 113.1 for d9-CL (internal standard for CL), m/z 62.1 for EA, and m/z 66.1 for d4-EA (internal standard for EA). The method showed satisfactory linearity (r² &gt; 0.999), precision (intra- and inter-day RSDs ≤6.3 %), and accuracy (−2.1–6.8 %). The limits of detection (LOD) were 0.30 μM for EA and 0.02 μM for CL. This method was applied to human erythrocytes and several tumor cell lines (Hela, EC109, Saos-2), revealing significant differences in the EA/CL ratio (reflecting the PE/PC ratio) among different cell types. This study indicated that the global phospholipid methylation status is cell-type specific and established the approach as a valuable tool for investigating membrane lipid composition in health and disease.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1270 ","pages":"Article 124903"},"PeriodicalIF":2.8,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145822648","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":"Quantitative LC-MS/MS Assay for Evocalcet in Human Plasma: Comparative Pharmacokinetic Evaluation in Fasted and Fed Healthy Chinese Volunteers","authors":"Keran Chen ,&nbsp;Orwa Siddig ,&nbsp;Jucui Yang ,&nbsp;Mohammed Dhawelbate ,&nbsp;Alrazi Eisa Shogar ,&nbsp;Mohammed Abdelrahman ,&nbsp;Xinrui Wu ,&nbsp;Tai-jun Hang","doi":"10.1016/j.jchromb.2025.124890","DOIUrl":"10.1016/j.jchromb.2025.124890","url":null,"abstract":"<div><div>A validated LC-MS/MS analytical method was established for quantifying evocalcet in human plasma, using evocalcet-d4 as the internal standard. Chromatographic separation was performed on a Thermo ODS C8 column (150 × 4.6 mm, 5 μm) with gradient elution of water and methanol, both containing 0. formic acid, as the mobile phases. Detection was carried out in positive ion mode using an electrospray ionization source. The plasma sample was spiked with an internal standard solution, followed by protein precipitation, and the resulting supernatant was injected into the LC-MS/MS system. The method, validated under ICH guidelines for quantitative biological analysis, demonstrated linearity from 1 to 1000 ng/mL (R² &gt; 0.99) with an LLOQ of 1 ng/mL. Accuracy and precision were confirmed by recovery studies, and all other validation parameters were within acceptable limits. This validated method was successfully applied to characterize the pharmacokinetic profile of evocalcet in fasting and postprandial healthy Chinese volunteers after oral administration. Results showed significantly higher C_max and AUC values in the fasting group compared to the postprandial group, as well as delayed T_max (<em>p</em> &lt; 0.01). The pharmacokinetic parameters, including C_max, T_max, t_1/2, and AUC_0-∞, were consistent with those reported in earlier studies. Moreover, hyperthyroid patients exhibited longer T_max and t_1/2, along with increased AUC_0-t and AUC_0-∞ values, compared to healthy individuals. These findings offer valuable methodological insights and pharmacokinetic data to advance clinical research on evocalcet, thereby contributing to a deeper understanding of its safety and efficacy profile.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1270 ","pages":"Article 124890"},"PeriodicalIF":2.8,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145716836","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":"Glycan profiling of therapeutic monoclonal antibodies: Rapid, non-denaturing single-step digestion via pH-tuned enzymatic cleavage","authors":"Doudou Lou ,&nbsp;Jingke Fan ,&nbsp;Lin Fan ,&nbsp;Yang Song ,&nbsp;Jinlin Zhang","doi":"10.1016/j.jchromb.2025.124892","DOIUrl":"10.1016/j.jchromb.2025.124892","url":null,"abstract":"<div><div>N-glycosylation of monoclonal antibodies (mAbs) critically affects their efficacy, stability, and immunogenicity, necessitating efficient and sensitive glycan analysis. Hydrophilic Interaction Liquid Chromatography with Fluorescence Detection (HILIC-FLD) is widely used for antibody N-glycan profiling and glycosylation heterogeneity control due to its high sensitivity and strong applicability, primarily by cleaving N-glycans from antibodies with PNGase F followed by liquid chromatography or coupled mass spectrometry (MS) analysis. Conventional PNGase F digestion protocols often require protein denaturation before digestion and overnight incubation, which may cause the preparation process cumbersome, time-consuming and toxic. To address these limitations, we developed a rapid, non-denaturing enzymatic cleavage method by selecting an appropriate buffer system and regulating the pH of the reaction system for mAbs. The method requires only a single-step digestion and enhances enzyme access to N-glycans by modulating antibody conformation and surface charge through pH adjustment, enabling efficient cleavage.</div><div>This method achieves N-glycan release within 20 min—48 times faster than conventional overnight digestion—while eliminating toxic denaturants, delivering superior performance in time efficiency, operational simplicity, safety, and environmental friendliness. Validation with trastuzumab (IgG1) and sintilimab (IgG4) showed nearly identical N-glycan profiles to conventional methods, retaining chromatographic properties and yielding comparable FA2 peak areas and glycoform abundances. MS revealed consistent glycoform profiles between methods (27 vs. 25 glycoforms in optimized vs. conventional), with strong abundance correlation. These results demonstrate that the method is not only applicable to multi-sample scenarios but also fully MS compatible, offering a robust tool for biopharmaceutical development, batch consistency assessments, and biomarker discovery in glycobiology research.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1270 ","pages":"Article 124892"},"PeriodicalIF":2.8,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145735275","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":"Simultaneous chromatographic quantification of upadacitinib, nilotinib, and tryptophan decoding myeloid leukemia and liver cancer integrated with ELISA and Flow cytometry","authors":"Ahmed M. Saleh ,&nbsp;Ola A. Saleh ,&nbsp;Rabeay Y.A. Hassan ,&nbsp;Ahmed A. Abd-Rabou ,&nbsp;Amr M. Badawey ,&nbsp;Hoda M. Marzouk","doi":"10.1016/j.jchromb.2025.124891","DOIUrl":"10.1016/j.jchromb.2025.124891","url":null,"abstract":"<div><div>For the first time, an integrated dual analytical–therapeutic platform is presented for concurrent investigation of hematological (chronic myeloid leukemia, CML) and solid (hepatocellular carcinoma, HCC) malignancies. A six-dimensional analytical framework combining RP-HPLC–DAD, UV–Vis spectrophotometry, ELISA, MTT assay, single-cell fluorescence imaging, and flow cytometry enabled comprehensive biochemical profiling. A novel QbD-driven RP-HPLC–DAD method was developed for the simultaneous quantification of Nilotinib (NIL, tyrosine kinase inhibitor, TKI), Upadacitinib (UPA, Janus kinase inhibitor, JAKI) and tryptophan (TRP) as a critical metabolic biomarker, successfully applied in serum matrices. Because CML and immune-mediated inflammatory diseases (IMIDs) share aberrant JAK–STAT signaling, and both NIL and UPA are metabolized via CYP3A4, continuous quantification is essential for safe co-therapy. Chromatographic separation was achieved on a carefully selected fluorinated Pursuit PFP column (150 × 4.6 mm, 3 μm), a key factor enabling efficient separation of the fluorinated drugs, using methanol–water (75,25, <em>v</em>/v) at 1.0 m/min and 230 nm detection, within a QbD 2³ full factorial framework, exhibiting excellent linearity (1.0–50.0 μg/mL). Fluorescence-based determination using flow cytometry with ELISA revealed a pioneer significant pro-apoptotic effect of Upadacitinib in HCC cells via modulation of the Bax/Bcl-2 axis with combination therapy showing superior anticancer effect compared to standard chemotherapy of Doxorubicin (DOX). Sustainability evaluation using AGREE (greenness) and BAGI (blueness) with RGB12 algorithm (whiteness) and spider-diagram visualization, in addition to the recently launched Multi-Color Assessment (MA) tool to confirm the method's multidimensional eco-efficiency in strong alignment with the United Nations Sustainable Development Goals (SDGs). This study further harnesses the Need–Quality–Sustainability (NQS) index and Koel's pyramid principles for holistic evaluation and benchmarking against reported approaches toward sustainable analytical oncology with personalized medicine.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1270 ","pages":"Article 124891"},"PeriodicalIF":2.8,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145735274","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":"Comprehensive urinary proteomics using DIA and PRM for low-abundance protein profiling of Wilson disease","authors":"Huiling Zhou ,&nbsp;Simin Dong ,&nbsp;Ao Pan ,&nbsp;Huan Xu ,&nbsp;Xin Tang ,&nbsp;Xixi Wang ,&nbsp;Linshen Xie ,&nbsp;Yongxin Li","doi":"10.1016/j.jchromb.2025.124906","DOIUrl":"10.1016/j.jchromb.2025.124906","url":null,"abstract":"<div><div>Wilson disease (WD) is an inherited disorder of copper metabolism with early diagnostic challenges. Urinary proteomics shows promise for identifying WD-related biomarkers, but current strategies neglect the detection of low-abundance proteins critical for preclinical diagnosis. Urine samples from 53 newly diagnosed WD patients and 47 matched controls were analyzed using an optimized proteomics strategy integrating data-independent acquisition (DIA) with parallel reaction monitoring (PRM). Functional enrichment analysis, hierarchical clustering and other multidimensional analyses were employed to delve into low-abundance protein biomarkers. Recursive feature elimination (RFE) and support vector machine (SVM) were applied to identify the candidate biomarkers, and the performance of the diagnostic model was measured by the receiver operating characteristic (ROC) curve. Furthermore, the potential biomarkers were validated by ELISA in an independent validation cohort. The optimized DIA-based untargeted proteomics identified 2263 urine proteins, including 447 differentially expressed proteins (68 upregulated and 379 downregulated). After LC-PRM-MS verification, 46 new candidate biomarkers for WD were identified and 11 were included in the final model. The SVM model performed the best in classifying WD and healthy control, and the areas under the ROC curve in the training set and the test set were 0.95 and 0.94 respectively. Four proteins were validated by ELISA in an independent cohort, with expression levels consistent with the proteomics data. The proposed DIA-PRM proteomics analysis detect more low-abundance urinary proteins, and a urinary protein biomarker panel with high accuracy for non-invasive diagnosis in WD patients was identified.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1270 ","pages":"Article 124906"},"PeriodicalIF":2.8,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145852138","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":"Chiral separation of fluoroquinolones by capillary electrophoresis using glucosamine-functionalized carbon quantum dots as a Pseudostationary phase","authors":"Guoli Wu ,&nbsp;Chen Li ,&nbsp;Yue Zhang ,&nbsp;Tingli Qu","doi":"10.1016/j.jchromb.2025.124909","DOIUrl":"10.1016/j.jchromb.2025.124909","url":null,"abstract":"<div><div>Fluoroquinolones are a class of widely used antibiotics, most of which contain chiral centers. Significant differences exist between their enantiomers in terms of antimicrobial activity, pharmacokinetics, and toxicity. In this study, glucosamine chiral-functionalized carbon quantum dots (GA-CQDs) were successfully synthesized and employed as a pseudostationary phase. By optimizing the chromatographic conditions, an efficient and novel chiral separation platform based on capillary electrophoresis (CE) was established for the enantioseparation of six fluoroquinolones. Compared to the CE system using free glucosamine solely as the chiral selector, the CE system employing GA-CQDs as a pseudostationary phase demonstrated a substantial improvement in chiral resolution (Rs) for all six model analytes (Ofloxacin: 1.56 → 9.21; Pazufloxacin: 0 → 8.53; Prulifloxacin: 1.38 → 7.58; Lomefloxacin: 1.01 → 2.84; Gemifloxacin: 0 → 1.25; Balofloxacin: 0 → 0.88). The incorporation of the pseudostationary phase significantly enhanced the chiral separation capability. This proposed strategy is characterized by its operational simplicity and low cost, thereby offering a valuable conceptual framework and a highly promising approach for developing chiral separation methods for fluoroquinolone antibiotics. This study represents the first report on employing GA-CQDs as a chiral selective medium to construct a capillary electrophoresis system, which achieved satisfactory enantioselectivity. Furthermore, the chiral recognition mechanism of glucosamine was preliminarily investigated through molecular modeling simulations, with the binding energy providing insights into the separation process.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1270 ","pages":"Article 124909"},"PeriodicalIF":2.8,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145867098","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":"From seed to bioactive: unveiling sprouting-driven changes in chickpeas via targeted metabolomics and in silico modelling","authors":"Merna M. Sakr ,&nbsp;Reham S. Ibrahim ,&nbsp;Amira M. Beltagy ,&nbsp;Reham S. Darwish","doi":"10.1016/j.jchromb.2025.124904","DOIUrl":"10.1016/j.jchromb.2025.124904","url":null,"abstract":"<div><div>Sprouting is a widely practiced food processing method, particularly in Western countries. Chickpeas (<em>Cicer arietinum</em>) have been traditionally used in various cuisines worldwide for centuries. Their versatility and nutritional value make them a staple ingredient in many traditional dishes. This study investigated the impact of sprouting chickpea seeds on their phytochemical profile and anticancer properties. Chickpea seeds were germinated under light and dark conditions for eight days, with samples collected on intervals. Extracts were analyzed using UPLC-MS/MS and tested for cytotoxic activity against two different breast cancer cell lines, namely; MDA-MB-231 (triple-negative breast cancer) and HeLa (uterine cervical cancer). UPLC-MS/MS detected 117 metabolites, primarily flavonoids, phenolic acids, and amino acids. Germination affected phytochemical composition and sprout length, with longer sprouts in darkness. Metabolite abundance peaked on day eight, while cytotoxicity was highest on day six. An OPLS model linked cytotoxic effects on HeLa and MDA-MB-231 cells to key metabolites, identified <em>via</em> coefficient plots. Caffeoylquinic acid, naringenin, and biochanin B were major contributors against MDA-MB-231, while hydroxybenzoic acid hexoside, orobol, and biochanin A hexoside were prominent in HeLa cells. Molecular docking of the top three metabolites with IL2 and ABCB1 genes, revealed as common genes between bioactive compounds' genes and cancer genes, showed hydroxybenzoic acid hexoside and naringenin as top binders, aligning with OPLS findings. Although the results demonstrate promising <em>in vitro</em> anticancer potential, further <em>in vivo</em> and mechanistic studies are required to confirm efficacy, safety, and bioavailability before establishing chickpea sprouts as a functional food or a chemo-preventive agent.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1270 ","pages":"Article 124904"},"PeriodicalIF":2.8,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145880597","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}