Journal of Chromatography B最新文献

{"title":"Integrative metabolomics and lipidomics reveal Jian-Pi-Yi-Shen formula improves adenine-induced CKD rats by regulating intrarenal glycolipid metabolism","authors":"Xinhui Liu ,&nbsp;Huicong Li ,&nbsp;Youcai Xu ,&nbsp;Yu Peng ,&nbsp;Shanshan Wu ,&nbsp;Xiaoqin Ye ,&nbsp;Yilong Yang ,&nbsp;Jiandong Lu","doi":"10.1016/j.jchromb.2026.124957","DOIUrl":"10.1016/j.jchromb.2026.124957","url":null,"abstract":"<div><div>Chronic kidney disease (CKD) is a global health problem with limited therapeutic options. Derangements in glucose and lipid metabolism represent a key feature of CKD that drives tubular injury and fibrosis. The Jian-Pi-Yi-Shen formula (JPYSF), a traditional Chinese medicine prescription, has been used for treating CKD for decades. To clarify metabolic mechanisms underlying the clinical efficacy of the JPYSF, we used an adenine-induced CKD model in male Sprague–Dawley rats, assigning animals to control, CKD, or CKD + JPYSF groups. After 6 weeks, renal function and serum lipids were quantified biochemically. Renal pathology and fibrosis were assessed histologically. Targeted renal metabolomics profiled metabolites associated with glycolysis, pentose phosphate pathway (PPP), tricarboxylic acid (TCA) cycle, and gluconeogenesis. Untargeted renal lipidomics characterized lipid profiles of kidney tissues. The results showed that JPYSF improved body weight and kidney index, lowered serum creatinine and urea, attenuated tubular injury and collagen deposition, and reduced α-smooth muscle actin and vimentin, without affecting alanine aminotransferase and aspartate aminotransferase. CKD kidneys displayed heightened glycolysis and PPP intermediates with reduced TCA intermediates and gluconeogenic precursors; JPYSF partially normalized these metabolites and reversed concordant transcriptional changes. Systemically, JPYSF reduced triglycerides and increased high-density lipoprotein cholesterol, with downward trends in total cholesterol and low-density lipoprotein cholesterol. Lipidomics showed extensive CKD-associated remodeling (749 differential lipids; glycerophospholipid and sphingolipid pathways). JPYSF reprogrammed the renal lipidome (64 differential lipids), predominantly restoring glycerophospholipids and dampening sphingolipids, directionally reversing subsets of CKD-perturbed species. In conclusion, JPYSF ameliorates adenine-induced CKD and fibrosis. These beneficial effects are accompanied by the partial restoration of renal glycolipid metabolic homeostasis, suggesting that metabolic regulation may contribute to the therapeutic mechanism of JPYSF.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1273 ","pages":"Article 124957"},"PeriodicalIF":2.8,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146172458","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":"Study on the improvement of cough variant asthma by the Tongfu Zhike formula via the TLR4/MyD88/NF-κB signaling pathway","authors":"Zhen Yang ,&nbsp;Huiying Li ,&nbsp;Wei Zhao ,&nbsp;Yingxia Yu ,&nbsp;Yanling Lin ,&nbsp;Jianxuan Liu ,&nbsp;Xinghu Fan ,&nbsp;Moyan Wang ,&nbsp;Xin Teng ,&nbsp;Zhikai Qiu","doi":"10.1016/j.jchromb.2026.124955","DOIUrl":"10.1016/j.jchromb.2026.124955","url":null,"abstract":"<div><h3>Objective</h3><div>Evaluation of the therapeutic effect of Tongfu Zhike Formula (TFZKF) on mice with cough variant asthma (CVA) and exploration of its mechanism of action in improving CVA.</div></div><div><h3>Methods</h3><div>The main chemical components of TFZKF were detected and analyzed using UHPLC-MS/MS. Network pharmacology was employed to investigate the potential targets of TFZKF in CVA. After grouping and drug administration, the number of coughs and cough latency were observed in CVA mice. White blood cell differential counts in BALF, serum levels of IgE, IL-4, IL-5, IL-13 and IFN-γ, as well as pathological changes in lung tissue were assessed. Western blot and RT-qPCR were used to analyze the protein expression levels and mRNA changes of TLR4, MyD88, and NF-κB in the lung tissue of CVA mice.</div></div><div><h3>Results</h3><div>A total of 495 chemical components were detected in TFZKF under both positive and negative ion modes. Network pharmacology analysis identified 199 potential targets related to TFZKF and CVA, and found close associations with the TLR4 and NF-κB signaling pathways. Animal experiments showed that, compared to the Model group, TFZKF significantly reduced the number of coughs, prolonged cough latency, decreased serum levels of IgE, IL-4, IL-5, and IL-13, reduced the total number of white blood cells, neutrophils, and eosinophils in BALF, alleviated inflammatory cell infiltration and fibrosis in lung tissue, and downregulated the protein and mRNA expression of TLR4, MyD88, and NF-κB.</div></div><div><h3>Conclusion</h3><div>The interaction within the TLR4/MyD88/NF-κB signaling pathway may be a potential key target for the treatment of cough variant asthma.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1273 ","pages":"Article 124955"},"PeriodicalIF":2.8,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146172455","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":"Liquid chromatography-tandem mass spectrometry-based urinary steroid profiling applied to primary aldosteronism diagnosis","authors":"Jian Zhong ,&nbsp;Xiaoli Ma ,&nbsp;Danchen Wang ,&nbsp;Wei Luo ,&nbsp;Yicong Yin ,&nbsp;Yutong Zou ,&nbsp;Ying Zhu ,&nbsp;Ming Li ,&nbsp;Shaowei Xie ,&nbsp;Songlin Yu ,&nbsp;Ling Qiu","doi":"10.1016/j.jchromb.2026.124930","DOIUrl":"10.1016/j.jchromb.2026.124930","url":null,"abstract":"<div><div>Steroid hormones are essential regulators of physiological homeostasis, which can help in diagnosing primary aldosteronism (PA). However, current methodologies for urinary steroid analysis face critical limitations, including narrow analyte coverage and tedious sample preparation workflows. Since the liquid chromatography-tandem mass spectrometry (LC-MS/MS) is considered the preferred technique for steroid measurement, there is a great need to establish an LC-MS/MS method combined with simplified and efficient sample pretreatment for multi-steroid profiling. We establish a novel LC-MS/MS method for quantifying 35 steroid hormones within a 25-min runtime. 40 μL of urine samples can be efficiently hydrolyzed at room temperature in just 0.5 h using transgenic β-glucuronidase. Detection is performed on the Waters Acquity I-Class UPLC-tandem with a Waters TQ-S triple quadrupole MS/MS system, and the method performance is systematically evaluated. Linearity is excellent, and the recovery rates range from 80.0 to 120.6%. Acceptable intra-assay and inter-assay precisions are achieved, with the coefficients of variation ranging from 1.86 to 15.10% and 3.91 to 19.75%, respectively. For clinical application, patients with PA (<em>n</em> = 37) exhibit significantly higher levels of aldosterone, tetrahydroaldosterone, 18-hydroxycorticosterone, 18-oxocortisol, and 18-hydroxycortisol compared to non-PA patients (<em>n</em> = 104). Combined steroid profiling demonstrates strong diagnostic performance for PA, yielding an area under the curve of 0.914, with a sensitivity of 0.87 and a specificity of 0.88. In conclusion, this study establishes a technically advanced LC-MS/MS method that integrates efficient enzymatic hydrolysis to enable the accurate quantification of urinary steroids for the diagnosis of endocrine disorders.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1273 ","pages":"Article 124930"},"PeriodicalIF":2.8,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146116632","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 combined TLC–GC approach for the regiospecific analysis of triacylglycerols in Yarrowia lipolytica","authors":"Runze Miao ,&nbsp;Yu Liu ,&nbsp;Hui Wang ,&nbsp;Guangwei Yu ,&nbsp;Shi'’an Wang ,&nbsp;Juanjuan Su ,&nbsp;Feng Han","doi":"10.1016/j.jchromb.2026.124945","DOIUrl":"10.1016/j.jchromb.2026.124945","url":null,"abstract":"<div><div>This study developed a robust, cost-effective thin-layer chromatography (TLC) combined with gas chromatography (GC) protocol for the regiospecific analysis of triacylglycerols (TAGs), with application to microbial oils from engineered <em>Yarrowia lipolytica</em>. Key steps, such as <em>sn</em>-1,3-specific lipase hydrolysis, TLC separation, staining, and fatty acid methyl ester (FAME) derivatization, were systematically optimized. Porcine pancreatic lipase was identified as the most efficient and economical enzyme for hydrolysis. A petroleum ether/diethyl ether/acetic acid (32:8:0.8, v/v/v) TLC solvent system provided optimal resolution, with iodine vapor as the preferred non-destructive staining method. BF₃-methanol methylation prevented artifact formation observed with methanol-H₂SO₄. Applied to oils from strains producing nervonic acid (NA) and γ-linolenic acid (GLA), the method revealed distinct regiospecific patterns: NA was almost exclusively esterified at the <em>sn</em>-1,3 positions, whereas approximately 64.3% of GLA resided at the <em>sn</em>-2 position. This TLC-GC approach offers an accessible alternative for rapid screening of fatty acid positional distribution in TAGs.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1273 ","pages":"Article 124945"},"PeriodicalIF":2.8,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146133960","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":"Applications of mass spectrometry in the routine diagnostic medical laboratory – a status report 2025","authors":"Michael Vogeser,&nbsp;Katharina Habler","doi":"10.1016/j.jchromb.2026.124958","DOIUrl":"10.1016/j.jchromb.2026.124958","url":null,"abstract":"<div><div>Mass spectrometry methods have become an essential part of the methodological portfolio of laboratory medicine over the past three decades. At present, however, their application is still largely limited to highly specialized laboratories in relatively few countries. Nevertheless, the technology provides important impulses for laboratory diagnostics overall—for example, in clinical pharmacology through innovative applications in therapeutic drug monitoring and precision dosing. After relatively slow progress in the area of automation, the first fully automated, closed MS-based analytical systems have recently been introduced for routine medical laboratories. In terms of usability, these systems are comparable to standard platforms based on photometry or ligand-binding techniques. The aim of this article is to describe the current medical, analytical, and organizational aspects of MS applications in diagnostics.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1273 ","pages":"Article 124958"},"PeriodicalIF":2.8,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146160076","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":"Anisodamine hydrobromide alleviates LPS-induced inflammation via the miR-1195/G3bp1/NF-κB axis in RAW264.7 cells","authors":"Yingli Cai ,&nbsp;Jun Liang ,&nbsp;Yiming Shao","doi":"10.1016/j.jchromb.2026.124956","DOIUrl":"10.1016/j.jchromb.2026.124956","url":null,"abstract":"<div><h3>Background</h3><div>Anisodamine Hydrobromide (Ani) and microRNAs (miRNAs) are essential regulators of immune responses, but the mechanisms of many Ani-related miRNAs remain insufficiently characterized. This study focused on miR-1195 as a potential regulator of Ani and its role in inflammation modulation in Lipopolysaccharide (LPS)-treated RAW264.7 cells.</div></div><div><h3>Methods</h3><div>Ani concentration was optimized using CCK-8. RAW264.7 cells were stimulated with LPS, with or without Ani pre-treatment. Inflammatory cytokines (TNFα and IL-1β) levels were assessed by RT-qPCR and ELISA. MiRNA sequencing was used to identify differentially expressed miRNAs, which were then validated by RT-qPCR. The interaction between miR-1195 and the 3’ UTR of G3bp1 was confirmed using luciferase assays in 293 T cells. For functional analysis, RAW264.7 cells were transfected with miR-1195 mimics, inhibitors, and G3bp1 inhibitors, followed by assessment of TNFα and IL-1β expression via RT-qPCR and ELISA. Western blot was performed to detect the protein levels of phosphorylated p65 (p-p65) and P65 to explore the downstream pathway of G3bp1.</div></div><div><h3>Results</h3><div>Ani reduced TNFα and IL-1β expression at both mRNA and protein levels, and reversed the LPS-induced downregulation of miR-1195. Overexpressing miR-1195 led to a marked reduction in TNFα and IL-1β levels in LPS-treated RAW264.7 cells, particularly at higher LPS concentrations. Besides, miR-1195 directly targeted and downregulated G3bp1 expression. Inhibition of miR-1195 enhanced cytokine release and promoted NF-κB activation, as indicated by increased p-p65 expression; these effects were reversed by G3bp1 knockdown.</div></div><div><h3>Conclusion</h3><div>Ani reduces LPS-induced inflammation by modulating miR-1195 and regulating G3bp1, partly through the NF-κB pathway, offering novel therapeutic insights for inflammatory control.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1273 ","pages":"Article 124956"},"PeriodicalIF":2.8,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146169186","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":"Ferrostatin-1 attenuates sepsis-induced lung injury by inhibiting ferroptosis via activation of the SLC7A11/GSH/GPX4 signaling pathway","authors":"Haidan Zhang,&nbsp;Hongyao Li,&nbsp;Shixian Liu,&nbsp;Jiahui Zheng,&nbsp;Peiwu Li","doi":"10.1016/j.jchromb.2026.124962","DOIUrl":"10.1016/j.jchromb.2026.124962","url":null,"abstract":"<div><h3>Background</h3><div>Ferroptosis is increasingly recognized as a pathological mechanism implicated in sepsis-induced lung injury. The study investigated the mechanism by which the ferroptosis inhibitor Ferrostatin-1 (Fer-1) attenuates lung injury through modulation of the solute carrier family 7 member 11 (SLC7A11)/glutathione (GSH)/glutathione peroxidase 4 (GPX4) signaling pathway.</div></div><div><h3>Methods</h3><div>In vivo and in vitro models were established by performing cecal ligation and puncture (CLP) in rats and stimulating MLE-12 cells with lipopolysaccharide (LPS), respectively. These models were subsequently intervened with Fer-1 and Erastin. The 7-day survival rate of septic rats was monitored, and serum levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were measured. Histopathological examination of lung tissue was performed using hematoxylin and eosin (H&amp;E) staining, while the lung wet/dry weight (W/D) ratio and protein concentration in bronchoalveolar lavage fluid (BALF) were assessed to evaluate pulmonary edema. Levels of ferrous ions (Fe²⁺), prostaglandin-endoperoxide synthase 2 (PTGS2), and malondialdehyde (MDA)—were quantified. Lipid peroxidation was evaluated using the BODIPY™ 581/591 C11 fluorescent probe, and mitochondrial ultrastructural changes were examined via transmission electron microscopy. Additionally, the expression levels and activities of GPX4, SLC7A11 and GSH were determined.</div></div><div><h3>Results</h3><div>(1) Fer-1 administration effectively mitigates systemic inflammation in septic rats by reducing serum levels of IL-6 and TNF-α, while significantly improving the 7-day survival rate. Furthermore, Fer-1 attenuates pathological lung injury, decreases the lung W/D ratio, and reduces protein concentration in BALF. In vitro, Fer-1 treatment enhanced the viability of LPS-stimulated MLE-12 cells. (2) Ferroptosis was activated in septic rats and LPS-stimulated MLE-12 cells. Fer-1 treatment reduced ferroptosis markers such as PTGS2, malondialdehyde, and lipid peroxidation, alleviated mitochondrial damage—marked by decreased volume, lower membrane density, fewer cristae, and outer membrane rupture—and restored mitochondrial function. Additionally, Fer-1 enhanced expression of key components in the SLC7A11/GSH/GPX4 signaling pathway.</div></div><div><h3>Conclusion</h3><div>In sepsis-induced lung injury, the ferroptosis-related SLC7A11/GSH/GPX4 signaling pathway is downregulated. Ferrostatin-1 activates this pathway and protects the lungs.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1273 ","pages":"Article 124962"},"PeriodicalIF":2.8,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146172457","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 multi-residue analysis of 87 pesticides in Egyptian food commodities: Method validation and dietary risk assessment","authors":"Ahmed Ezzat ,&nbsp;Omar Khaled ,&nbsp;Hassan A. El-Gammal ,&nbsp;Mahmoud S. Elshabrawy ,&nbsp;Rasha M. El Nashar","doi":"10.1016/j.jchromb.2026.124960","DOIUrl":"10.1016/j.jchromb.2026.124960","url":null,"abstract":"<div><div>A comprehensive multi-residue analytical method was developed and validated for the simultaneous determination of 87 pesticides in diverse food matrices using a modified QuEChERS extraction protocol coupled with gas chromatography-tandem mass spectrometry (GC–MS/MS). Method validation according to SANTE/11312/2021 guidelines demonstrated excellent analytical performance, achieving a recovery range of 70–120% at three fortification levels (0.01, 0.05, and 0.1 mg/kg). Relative standard deviations for intraday and interday precision remained below 21% for all validated compounds. The method achieved limits of quantification ranging from 0.0005 to 0.01 mg/kg. The practical applicability was demonstrated through an extensive survey of 591 real food samples comprising eight commodities (apple, chamomile, caraway, dried mint, fennel, green beans, hibiscus, and rice), which revealed widespread pesticide contamination, with 325 samples (54.9%) containing detectable residues. Thirty-eight different pesticide compounds were identified, with chlorpyrifos, cypermethrin, and profenofos being the most frequently detected. Dried herbs showed the highest contamination rates (63.1% for mint, 64.2% for hibiscus), with some samples containing up to eight different pesticides simultaneously. Human health risk assessment through Target Hazard Quotient (THQ) calculations identified several pesticide-commodity combinations within the moderate concern range (THQ &gt; 0.1), particularly chlorpyrifos in rice (THQ = 0.286) and profenofos in dried mint (THQ = 0.098). These values, while below the action threshold of unity, indicate meaningful contributions to chronic pesticide exposure warranting enhanced regulatory monitoring and targeted intervention strategies. The environmental sustainability assessment yielded an AGREE score of 0.60, reflecting reasonable green analytical performance despite the inherent challenges of trace-level multi-residue analysis.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1273 ","pages":"Article 124960"},"PeriodicalIF":2.8,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146172456","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 pharmacokinetics and metabolite profiling of the novel anti-tumor polypeptide CPP-4-2 in rats using UHPLC-MS/MS","authors":"Anci Zhu ,&nbsp;Shuo Ma ,&nbsp;Juan Cao ,&nbsp;Peng Li ,&nbsp;Ran Liu ,&nbsp;Bosai He","doi":"10.1016/j.jchromb.2026.124954","DOIUrl":"10.1016/j.jchromb.2026.124954","url":null,"abstract":"<div><div>CPP-4-2, a novel anti-tumor peptide, has been shown to disrupt tumor growth by competitively binding to Staphylococcal nuclease domain-containing protein 1 (SND1), inducing its degradation and inhibiting the SND1-Metadherin (MTDH) interaction. Despite the established efficacy of this approach, comprehensive in vivo analytical methodologies remain limited. Therefore, based on the physicochemical characteristics of CPP-4-2, its anti-hepatocellular carcinoma activity was confirmed using CCK-8, colony formation, and cell migration assays. A comprehensive quality standard for CPP-4-2, encompassing description, identification, tests, assay, and related substances, was established, alongside a validated HPLC method for in vitro quantification. Additionally, a highly sensitive, specific, reproducible, and rapid UHPLC-QTRAP-MS/MS method for in vivo CPP-4-2 quantification was developed and fully validated, meeting the requirements for biological sample analysis in plasma, heart, liver, spleen, lung, and kidney tissues. Application of this method to SD rats following intravenous injection (10 mg/kg) revealed a rapid peak plasma concentration, followed by a swift decline, indicating a primary efficacy window of 0–2 h. 16 metabolites, including the parent compound, were identified using UHPLC-Q-TOF-MS/MS. This study highlights the potential of CPP-4-2, establishes quality control, provides key in vivo data, and lays the foundation for further structural modifications, development of novel dosage forms, and investigations into safe medication practices.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1273 ","pages":"Article 124954"},"PeriodicalIF":2.8,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146160071","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 two-dimensional hyphenated ion-exchange chromatographic platform for simultaneous assessment of AAV8 capsids and impurity fingerprinting in upstream processes","authors":"Timotej Žvanut ,&nbsp;Tjaša Kašček ,&nbsp;Lara Vogrinčič ,&nbsp;Petra Dekleva ,&nbsp;Sandra Potušek ,&nbsp;Anže Martinčič Celjar ,&nbsp;Ivana Petrović Koshmak ,&nbsp;Blaž Goričar ,&nbsp;Mitja Martelanc ,&nbsp;Aleš Štrancar ,&nbsp;Andreja Gramc Livk","doi":"10.1016/j.jchromb.2026.124959","DOIUrl":"10.1016/j.jchromb.2026.124959","url":null,"abstract":"<div><div>Reliable monitoring of full (F) versus empty (E) adeno-associated virus (AAV) capsids, along with impurity fingerprinting, is essential for optimising upstream processes (USPs). Conventional analytical workflows for USP monitoring, including PCR-ELISA and one-dimensional (1D) liquid chromatography (LC), often lack robustness, selectivity, reliability, and throughput, especially when analysing complex and impurity-rich samples. To overcome these challenges, we introduce a two-dimensional (2D) LC workflow that combines cation-exchange (CEX) capture in the first dimension with subsequent anion-exchange (AEX) separation in the second. The system incorporates multi-wavelength UV–Vis, fluorescence, and light scattering (LS), offering greater control over analysis and deeper insights into AAV8 sample heterogeneity while simplifying production analysis.</div><div>The platform facilitates direct analysis of complex AAV8 USP samples—without the need for offline purification—achieving high sensitivity, high throughput, and robust separation. The two-dimensional system consistently resolved E and F capsids, distinguished partially filled intermediates, and enabled concomitant impurity monitoring. Validation studies demonstrated high recovery rates, negligible carryover, exceptional linearity, and reproducibility across matrices relevant to USP standards. Furthermore, the 2D method optimised for AAV8 was demonstrated to be applicable to other AAV serotypes. Compared to conventional one-dimensional liquid chromatography, orthogonal techniques confirm the superior selectivity and practicality of this method by obviating the need for costly, time-consuming, multi-step offline procedures. The integrated multi-detector two-dimensional workflow addresses a critical analytical gap in AAV manufacturing and provides a compelling tool for process monitoring, method standardisation, and quality assurance within biopharmaceutical analytics.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1273 ","pages":"Article 124959"},"PeriodicalIF":2.8,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146172459","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}