Journal of Chromatography B最新文献

{"title":"Determination of glucose impurities in glucose-salt complex pharmaceutical products by HPLC-RID method: Analytical method development and validation study","authors":"Harun Ergen ,&nbsp;Yasin Gökekin ,&nbsp;Remziye Azra Kartop ,&nbsp;Büşra Çetin Ersen ,&nbsp;Müge Güleli ,&nbsp;Sevgi Kocaoba ,&nbsp;Cem Çalışkan","doi":"10.1016/j.jchromb.2025.124717","DOIUrl":"10.1016/j.jchromb.2025.124717","url":null,"abstract":"<div><div>Pharmaceutical products containing glucose and salt are widely used in the health sector and play a vital role. These pharmaceutical products are usually encountered as a part of drugs used in treatment and medical procedures. However, the analytical determination and quantification of active ingredients and impurities belonging to active ingredients in such products is a complicated process. This is because the retention time of impurities belonging to the glucose molecule in these pharmaceutical structures and the presence of salts that give peaks in similar places complicate the analysis process. Moreover, when the British Pharmacopoeia (BP), European Pharmacopoeia (EP) and US Pharmacopoeia (USP) are examined, it is seen that there are only monographs belonging to glucose raw material and there is no record regarding the determination of impurities belonging to glucose in pharmaceutical products containing glucose-salt complex structures. A new method has been developed within the scope of the study to prevent this complexity. As a result of the method development studies, the specific solvent method was optimized and a relatively simple pre-purification process with high recovery was applied to avoid salt interference. In addition, high-precision determination and validation studies of glucose and its impurities were carried out using the High Performance Liquid Chromatography-Refractive Index Detector (HPLC-RID) method. This new analytical approach is expected to contribute to the development of reliable and high-quality products in the healthcare field by raising industry standards.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1264 ","pages":"Article 124717"},"PeriodicalIF":2.8,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144572137","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 combination of multiple LC-MS approaches for the comprehensive characterization of recombinant herpes zoster vaccine","authors":"Zhen Long ,&nbsp;Chen Wei ,&nbsp;Xiang Zhu ,&nbsp;Xi Luo ,&nbsp;Xiangjun Li ,&nbsp;Suli Zhao","doi":"10.1016/j.jchromb.2025.124712","DOIUrl":"10.1016/j.jchromb.2025.124712","url":null,"abstract":"<div><div>Recombinant herpes zoster (HZ) vaccines were reported as the most effective strategy to reduce the burden of herpes zoster and related complications. To ensure the quality, safety, and efficacy of recombinant HZ vaccines, comprehensive characterization of its antigen, recombinant glycoprotein E (RgE), is critical. However, due to the extreme complexity and heterogeneity of RgE, a single analytical platform is insufficient for comprehensive characterization. This study developed an innovative LC-MS-based workflow that combines multiple-parallel-protease digestion, various separation techniques (nano-flow LC, high-flow LC, and native separation), and high-resolution mass spectrometry (HR-MS) with multiple ion activation methods to achieve a comprehensive analysis of RgE. As a result, 21 and 21 <em>O</em>-glycosylation and phosphorylation sites were identified by EThcD-induced LC-MS/MS. Two <em>N</em>-glycosylation sites (N207 and N406) were identified with various <em>N</em>-glycans. The most abundant <em>N</em>-glycan for N207 was A2S2F. The top five <em>N</em>-glycans detected for N406 were also identified. Additionally, five disulfide linkages were detected for RgE. They were Cys 195-Cys 205, Cys 401-Cys 411, Cys 177-Cys 189, Cys 365-Cys 374, and Cys 356-Cys 382. For host cell protein (HCP) analysis, a high-sensitivity nano LC-MS/MS approach was applied, identifying 50 host cell proteins (HCPs), including two high-risk proteins, clusterin and peroxiredoxin-1. Additionally, molecular weight measurements obtained under both native and denatured conditions agreed well with the amino acid sequence acquired by peptide mapping. Following the optimization of the LC-MS workflow using RgE, the method was applied to evaluate recombinant zoster vaccines produced through different manufacturing processes. The results demonstrated that this workflow is highly effective for characterizing new recombinant zoster vaccines and may also serve as a valuable tool for the development of other complex recombinant vaccines.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1263 ","pages":"Article 124712"},"PeriodicalIF":2.8,"publicationDate":"2025-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144518899","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":"Optimization of preparative HPLC method for the purification of thymol from the essential oil of Trachyspermum ammi (L.)","authors":"Smija Kakkuth Puthenvitil,&nbsp;Palani Perumal","doi":"10.1016/j.jchromb.2025.124707","DOIUrl":"10.1016/j.jchromb.2025.124707","url":null,"abstract":"<div><div>The escalating demand for high-purity bioactive compounds in pharmaceutical research and quality control underscores the critical role of preparative High-Performance Liquid Chromatography. This study introduces a novel and optimized preparative HPLC protocol for the large-scale purification of thymol from <em>Trachyspermum ammi</em> (L.) essential oil. Addressing a notable knowledge gap, this is the first reported application of Prep-HPLC for thymol isolation from this specific botanical source, establishing a new, scalable protocol. An initial analytical HPLC method, optimized with Methanol:Water (60:40, <em>v</em>/v), was successfully scaled to a semi-preparati<em>v</em>e level. The optimized preparative method, utilizing Methanol:Water (90:10, <em>v</em>/v) as the mobile phase, achieved thymol purification with an impressive 98.87 % purity. The structural identity of the purified compound was unequivocally confirmed through comprehensive spectroscopic analyses, including GC–MS, FTIR, XRD, and NMR. Crucially, the purified thymol exhibited potent and broad-spectrum antifungal activity against both susceptible and multidrug-resistant strains of <em>Candida albicans</em> and <em>Candida auris</em>. Minimum inhibitory concentrations were determined to be as low as 0.125 μg/mL against multidrug-resistant <em>C. albicans</em> and 0.25 μg/mL against multidrug-resistant <em>C. auris</em>. This robust efficacy, particularly against challenging drug-resistant isolates, strongly positions the purified thymol as a promising candidate for further therapeutic development against candidiasis. This research significantly advances separation science and highlights the industrial potential of Prep-HPLC for high-value natural product isolation.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1263 ","pages":"Article 124707"},"PeriodicalIF":2.8,"publicationDate":"2025-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144501077","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 chemical-pharmacological continuum of Yucan formula: Integrated component-tissue exposure correlation informs diabetic kidney disease target network","authors":"Haowen Wang ,&nbsp;Huihui Shi ,&nbsp;Guangyu Sheng ,&nbsp;Ming Xue ,&nbsp;Wenqi Huang ,&nbsp;Xuejun Yang ,&nbsp;Tao Yang","doi":"10.1016/j.jchromb.2025.124715","DOIUrl":"10.1016/j.jchromb.2025.124715","url":null,"abstract":"<div><div>This study focuses on uncovering the material basis and therapeutic mechanisms of the Yucan Formula (YCF) in the treatment of diabetic kidney disease (DKD). Utilizing a combination of UHPLC-Q-Exactive Orbitrap HRMS and AB Sciex Triple Quad™ 4500 LC-MS, a comprehensive qualitative and quantitative analysis of YCF's chemical constituents was performed. A total of 166 compounds were identified, primarily flavonoids (57), terpenoids (32), organic acids (25), and phenylpropanoids (15), with fragmentation patterns established for the major classes and tissue distribution characteristics investigated. In vivo distribution studies identified prototype compounds present in both blood and kidney, and through peak area screening and methodological validation, eight key compounds (Senkyunolide H, Senkyunolide I, Nobiletin, 3-n-Butylphthalide, Senkyunolide A, Methylnissolin, Ferulic Acid and p-Coumaric acid) were selected for pharmacokinetic analysis. These compounds demonstrated rapid absorption, prolonged retention time, and strong pharmacological potential. By integrating data from TCMSP, SwissTargetPrediction, and other databases, a compound–target–disease network was constructed, revealing through GO and KEGG enrichment analyses that YCF primarily exerts its effects via pathways such as AGE-RAGE, PI3K-AKT, and MAPK, thereby regulating oxidative stress, inflammation, and fibrosis. These findings highlight the multi-component, multi-target, and multi-pathway synergistic nature of YCF in the treatment of DKD. Collectively, this study elucidates the in vivo behavior and active constituents of YCF while constructing a scientific mechanism-based network, offering a systematic pharmacological model for exploring traditional Chinese medicine formulas in the management of complex diseases.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1263 ","pages":"Article 124715"},"PeriodicalIF":2.8,"publicationDate":"2025-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144524023","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 ADME evaluation of Akuamma alkaloids","authors":"Abhishek Gour ,&nbsp;Simone M. Creed ,&nbsp;Andrew P. Riley ,&nbsp;Abhisheak Sharma","doi":"10.1016/j.jchromb.2025.124713","DOIUrl":"10.1016/j.jchromb.2025.124713","url":null,"abstract":"<div><div>The seeds of the akuamma tree (<em>Picralima nitida</em>) have been traditionally used to treat fever and pain. The seed contains at least five major alkaloids: akuammine, akuammicine, akuammiline, picraline, and pseudo-akuammigine, which possess diverse pharmacological effects, including analgesic, anti-inflammatory, and mood-enhancing properties. The present study aimed to assess <em>in vitro</em> and <em>in vivo</em> pharmacokinetics of akuamma alkaloids using a sensitive ultra-performance liquid chromatography−tandem mass spectrometry (UPLC-MS/MS)-based bioanalytical method. The method was found to be simple, sensitive, and rapid, effectively evaluating absorption, distribution, metabolism, and oral/intravenous pharmacokinetics in male Sprague-Dawley rats. These alkaloids demonstrated high permeability using human colorectal adenocarcinoma cell monolayers. Akuammine and akuammiline showed half-lives of 13.5 and 30.3 min in reduced nicotinamide adenine dinucleotide phosphate-supplemented rat liver microsomes. Pseudo-akuammigine showed the highest plasma protein binding, followed by the other alkaloids. Oral dosing of the ground seed suspension revealed significant systemic exposure of only akuammine, with low oral bioavailability. This research is the first comprehensive report on the ADME properties and pharmacokinetics of akuamma alkaloids, providing critical insights for developing these compounds as potential phytotherapeutic agents for pain management or other medicinal uses.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1263 ","pages":"Article 124713"},"PeriodicalIF":2.8,"publicationDate":"2025-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144501078","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":"Development and validation of an LC-MS/MS method for quantifying a new G protein selective μ-opioid receptor agonist YZJ-4729 and its major metabolite M10 in human plasma to support a phase I study in healthy subjects","authors":"Yufeng Ni ,&nbsp;Chenxin Ding ,&nbsp;Minlu Cheng ,&nbsp;Li Ding","doi":"10.1016/j.jchromb.2025.124714","DOIUrl":"10.1016/j.jchromb.2025.124714","url":null,"abstract":"<div><div>The use of conventional opioids that act on μ-receptors is a recognized care agent for moderate-to-severe acute treatment. However, recent attention has shifted to a new class of μ-receptor agonists which can selectively activate the G-protein pathway and thus have better efficacy and fewer side effects. Recently, YZJ-4729 was developed as a new G protein selective μ-opioid receptor agonist. And for its clinical trial investigation usage, a rapid LC-MS/MS method was established for the concurrent measurement of YZJ-4729 and its major metabolite M10 in human plasma. A step involving the precipitation of proteins was used for plasma sample preparation. The chromatography separation was done on a Poroshell 120 EC-C18 analytical column (2.1 × 50 mm, 2.7-μm, Agilent). Gradient elution was performed with 5.0 mM ammonium acetate (NH₄Ac) and 0.1 % formic acid (FA) water solution as the mobile phase A and pure acetonitrile (ACN) as the mobile phase B. Detection occurred in the mode of positive ion electrospray ionization through multiple reaction monitoring using deuterium YZJ-4729 (d6-YZJ-4729) as the internal standard. The ionic transitions used were YZJ-4729: <em>m/z</em> 409.3 → 244.2; d6-YZJ-4729: <em>m/z</em> 415.3 → 244.2; M10: <em>m/z</em> 425.3 → 260.2;). The method showed excellent linearity across the ranges of 0.500 to 500 ng/mL for YZJ-4729 and 0.0500 to 50.0 ng/mL for M10. The method was utilized to assess the plasma concentrations of YZJ-4729 and M10 in healthy volunteers after a 30-min intravenous infusion in the phase I clinical study, and the clinical pharmacokinetic profiles of YZJ-4729 and M10 were described.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1263 ","pages":"Article 124714"},"PeriodicalIF":2.8,"publicationDate":"2025-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144490282","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":"Decoding serum N-Glycome signatures across COVID-19 symptomatic stages","authors":"Mehmet Emrah Yaman ,&nbsp;Haci Mehmet Kayili ,&nbsp;İzzet Avci ,&nbsp;Alperen Aksakal ,&nbsp;Omer Faruk Kocak ,&nbsp;Mehmet Atakay ,&nbsp;Bekir Salih","doi":"10.1016/j.jchromb.2025.124710","DOIUrl":"10.1016/j.jchromb.2025.124710","url":null,"abstract":"<div><div>SARS-CoV-2 infection causes a wide spectrum of clinical manifestations, ranging from asymptomatic cases to fatal outcomes. Although significant progress has been made in understanding SARS-CoV-2, the identification of reliable biomarkers for early risk assessment and disease severity prediction remains an unmet clinical need. In this study, we characterized the serum N-glycome profiles of a cohort comprising COVID-19 patients with mild to severe symptoms and healthy controls. After enzymatic deglycosylation, serum samples were analyzed using HILIC-FLD-QTOF-MS. In case-control comparisons, COVID-19 patients exhibited a significant decrease in oligomannose and hybrid-type glycans, along with an increase in tetra-antennary and tetra-galactosylated structures. In addition, four N-glycan structures were identified as having diagnostic potential to distinguish COVID-19 cases from healthy controls. When comparing severely symptomatic COVID-19 patients to those with mild symptoms and healthy controls, a significant increase was observed in antennary fucosylated N-glycans (SLex). Moreover, ROC analysis demonstrated that an isomer of Hex5HexNAc4Neu5Ac had strong diagnostic potential (AUC &gt; 0.8) in distinguishing severely symptomatic COVID-19 patients from healthy controls. Our study reveals a novel association between antennary fucosylation and an isomer of Hex5HexNAc4Neu5Ac in severe COVID-19, highlighting its potential relevance for biomarker discovery.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1263 ","pages":"Article 124710"},"PeriodicalIF":2.8,"publicationDate":"2025-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144518898","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":"Bioanalytical method development of nevirapine, fosamprenavir calcium and its metabolite amprenavir by RP-HPLC in rat plasma","authors":"Neha Srivastava ,&nbsp;Vijay Mishra ,&nbsp;Bimlesh Kumar ,&nbsp;Yachana Mishra ,&nbsp;Vasanth Raj Palanimuthu","doi":"10.1016/j.jchromb.2025.124702","DOIUrl":"10.1016/j.jchromb.2025.124702","url":null,"abstract":"<div><div>The current analytical work focuses on the development of a bioanalytical method for anti-HIV drugs, nevirapine, fosamprenavir calcium, and its metabolite amprenavir. Fosamprenavir is a prodrug of amprenavir, digested by cellular phosphatases <em>in vivo</em>. Rat plasma was isolated from the collected rat blood by centrifugation and used for preparing stock solutions. A reverse-phase column (ODS C-18) was used for the development of the method and validation. The bioanalytical method was developed using the isocratic mode. The solvent system used is Methanol: Water: ACN (800 mL: 200 mL: 50 mL). The method was run at a flow rate of 0.5 mL/min, and 258 nm was the detection wavelength. The developed method showed retention at 8.705 min for nevirapine, 11.923 min for fosamprenavir calcium, and 14.391 min for amprenavir. The calibration curve was linear with a correlation coefficient (r²) of 0.9916 for nevirapine, 0.9909 for fosamprenavir calcium, and 0.9879 min for amprenavir. The RSD of the accuracy, precision, and stability study of the method was found to be less than 2 % and was found to be acceptable. The method is reliable and does not show any kind of interference due to the plasma sample. Thus, the results support that a reliable, reproducible, and efficient method was developed, and validation was carried out for the estimation of the drug nevirapine, fosamprenavir, and its metabolite amprenavir in rat plasma samples.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1263 ","pages":"Article 124702"},"PeriodicalIF":2.8,"publicationDate":"2025-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144481134","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":"Application of preparative high-speed countercurrent chromatography for isolation of a bicyclo-[3.2.1]-octanoid neolignan from Piper multinodum C. DC. and determination of its absolute stereochemistry","authors":"André Mesquita Marques ,&nbsp;Lavínia de Carvalho Brito ,&nbsp;Ana Carolina Ferreira de Albuquerque ,&nbsp;Arthur Ribeiro de Souza ,&nbsp;Fernando Martins dos Santos Junior ,&nbsp;Adélia Viviane de Luna ,&nbsp;Maria Raquel Figueiredo ,&nbsp;Ygor Jessé Ramos ,&nbsp;Davyson de Lima Moreira","doi":"10.1016/j.jchromb.2025.124711","DOIUrl":"10.1016/j.jchromb.2025.124711","url":null,"abstract":"<div><div>This study aimed to propose a straightforward approach for isolating a novel neolignan compound using high-speed countercurrent chromatography (HSCCC) in combination with spectrometric techniques. The bicycleneolignan compound was detected in the dichloromethane partition obtained from methanolic leaf extract of <em>Piper multinodum</em>. After evaluating several two-phase solvent systems, the <em>n</em>-hexane/ethyl acetate/methanol/water in ratio 3:1:1:1 (<em>v</em>/v/<em>v</em>/v) demonstrated the highest efficiency for HSCCC separation of the target compound, as evidenced by the optimal distribution coefficient K_<em>D</em> value of 1.55, using the tail-to-head elution mode, yielding 87.0 mg in 96.7 %. Its relative structure and absolute stereochemistry were unequivocally determined through 1D and 2D-NMR spectroscopy and HRMS data, complemented by NMR quantum-mechanical calculations, and application of the DP4+ method, as well as electronic circular dichroism data. Our findings confirmed the identification of the new compound multinodine -(7<em>R</em>,8<em>S</em>,1′<em>R</em>,3′<em>R</em>) (E)7-(benzo[3,4]-dioxol-1-yl)-3′,6′-dimethoxy-8-methyl-1′-(prop-7′-en-7′-yl) bicyclo-[3.2.1]-oct-5′-en-4′-one).</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1264 ","pages":"Article 124711"},"PeriodicalIF":2.8,"publicationDate":"2025-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144562830","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":"Automated DBS-online SPE–LC–MS/MS platform with dynamic solvent dilution: Robust quantification of antipsychotic drugs for decentralized therapeutic drug monitoring","authors":"Yanxiao Zhou ,&nbsp;Huirong Li ,&nbsp;Mengjia Lin ,&nbsp;Jiwei Yang ,&nbsp;Xiaofeng Yu","doi":"10.1016/j.jchromb.2025.124704","DOIUrl":"10.1016/j.jchromb.2025.124704","url":null,"abstract":"<div><div>This study introduces a novel fully automated dried blood spot (DBS)-online solid-phase extraction (SPE)-LC-MS/MS system for the quantification of 19 antipsychotic drugs in dried blood spots, addressing critical challenges in therapeutic drug monitoring (TDM). By integrating dynamic online dilution technology, the system overcomes solvent incompatibility between DBS eluents (high organic content) and hydrophilic SPE sorbents, minimizing analyte loss during trapping. Moreover, the automated workflow eliminates manual intervention, achieving precision (CV ≤12 %), accuracy (RE ≤ ±10 %) and matrix-agnostic performance (ME%: −11.4 %–10.8 %, CV &lt;10 %). When validated across serum and whole blood matrices, the method demonstrated exceptional linearity (R² &gt; 0.99) and sensitivity (LOD: 0.01–0.71 ng/mL). Stability studies confirmed DBS integrity at extreme temperatures (40 °C to 2–8 °C), enabling cost-effective sample transport via standard postal services. Comparative analysis with offline LC–MS–MS revealed strong agreement, underscoring the method's equivalence to gold-standard techniques. The platform's automation, minimal matrix dependency, and robustness position it as a transformative solution for decentralized TDM, particularly in resource-limited settings</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1263 ","pages":"Article 124704"},"PeriodicalIF":2.8,"publicationDate":"2025-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144365933","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}