Journal of separation science最新文献

{"title":"Prediction of Elution Profiles in Countercurrent Chromatography With a Linear Loss of Stationary Phase","authors":"Felix Rüttler,&nbsp;Rosalie Ormos,&nbsp;Walter Vetter","doi":"10.1002/jssc.70526","DOIUrl":"10.1002/jssc.70526","url":null,"abstract":"<p>Countercurrent chromatography (CCC) is a liquid-liquid separation method that is predominantly used for the preparative isolation of natural products. Key indicators for the development of CCC methods are the partition coefficient (<i>K</i> value) between the two liquid phases and the properties of the chosen biphasic solvent system. Based on this information, it is possible to predict elution volumes and peak widths with the probabilistic model for immiscible separations and extractions (ProMISE). However, a common challenge in reliably predicting CCC elution profiles is a persistent loss of stationary phase (i.e., bleeding), which leads to a decrease in the percentile share of stationary phase (<i>S_f</i> value) in the column as the separation progresses. In this study, two carotenoids were isolated and used as test compounds to investigate the effect of bleeding with the <i>n</i>-hexane/benzotrifluoride/acetonitrile (20:7:13, <i>v</i>/<i>v</i>/<i>v</i>) solvent system. After showing that the stationary phase bleeding rate was linear (and not exponential), experiments were carried out both in head-to-tail and in tail-to-head mode. Next, a simple calibration method was developed to predict elution volumes and peak widths with the ProMISE2 software in CCC separations with linear bleeding. Finally, based on the collected data, a method was developed to accurately predict the elution volume and peak width during a (long) CCC run, particularly for analytes with comparatively high elution volumes.</p>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"49 9","pages":""},"PeriodicalIF":3.6,"publicationDate":"2026-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13572847/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148963398","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Determination of Enantiomers of Deschloroketamine and Its Metabolites in Wistar Rat Brains","authors":"Natalie Paškanová,&nbsp;Vendulka Hrubá,&nbsp;Peter Frühauf,&nbsp;Bronislav Jurásek,&nbsp;Martin Kuchař,&nbsp;Pavel Řezanka,&nbsp;Michal Kohout","doi":"10.1002/jssc.70503","DOIUrl":"10.1002/jssc.70503","url":null,"abstract":"<p>Deschloroketamine (DXE) is a dissociative new psychoactive substance whose enantiomers and metabolites may exhibit distinct pharmacological and toxicological profiles. Reliable analytical methods enabling the chiral determination of DXE and its metabolites in biological matrices are thus essential. In this study, we evaluated supercritical fluid chromatography coupled to tandem mass spectrometry (SFC‑MS/MS) and capillary zone electrophoresis with ultraviolet detection (CZE‑UV) for the enantioselective analysis of DXE and its major metabolites in rat brain tissue. SFC using polysaccharide‑based chiral stationary phases provided baseline separation of the hydroxylated and demethylated metabolites, but severe peak distortion of DXE enantiomers prevented their quantification. Therefore, quantitative analysis was performed by CZE employing carboxymethyl‑β‑cyclodextrin as a chiral selector and a high‑sensitivity Z‑shaped detection cell. Analysis of brain tissue samples obtained following high-dose administration revealed near equimolar concentrations of the DXE enantiomers and a higher concentration of (<i>R</i>)-norDXE relative to (<i>S</i>)-norDXE. In this particular case, the combined use of SFC‑MS/MS and CZE‑UV provided a practical workflow for the chiral profiling of DXE and its metabolites in complex biological matrices.</p>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"49 9","pages":""},"PeriodicalIF":3.6,"publicationDate":"2026-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13572844/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148963400","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Systematic Evaluation of Partition-Dominated Retention in Hydrophilic Interaction Liquid Chromatography (HILIC) Under Varying Aqueous Components of the Mobile Phase: Revisiting the HILIC Column Test Scheme","authors":"Tomoaki Shimpo,&nbsp;Tohru Ikegami","doi":"10.1002/jssc.70520","DOIUrl":"10.1002/jssc.70520","url":null,"abstract":"<div>\u0000 \u0000 <p>The robustness of a test for characterizing hydrophilic interaction liquid chromatography (HILIC) columns against variations in aqueous mobile-phase composition was evaluated using mobile phases of acetonitrile/aqueous mixture (9:1, v/v) in the pH range of 3.6–6.8 of the aqueous part. Ten columns (commercially available and home-made columns) were examined under five mobile phases differing in anion species (formate or acetate), NH₄⁺ concentration and salt concentration. The specific hydrophilicity of the columns was assessed via the retention factor of uridine (<i>k</i>(U)), and the relative retention characteristics (i.e., retention selectivity: <i>α</i>) were evaluated using structure-related probe pairs. The <i>k</i>(U)-values exhibited low variability under low-salt conditions (20 mM in the aqueous part; CVs = 0.01–0.08), but higher variability under high-salt conditions (100 mM in the aqueous part; CVs = 0.03–0.18). Under comparable pH conditions (ammonium formate at pH 4.5 vs. ammonium acetate at pH 4.7), <i>k</i>(U)-values were consistently higher in the former, indicating that differences in NH₄⁺ concentration affect hydration at the stationary-phase interface and thereby influence retention. This suggests that <i>k</i>(U), commonly used as an indicator of hydrophilic retention, can be affected significantly by partition interactions that are controlled by ionic concentration rather than the pH value of the mobile phase. In contrast, the characteristic <i>α</i>-values primarily associated with solute retention via partitioning exhibited high robustness (CVs ≤ 0.03). Notably, <i>α</i>(<i>k</i>(U)/<i>k</i>(2′-deoxyuridine)) values correlated with the estimated phase ratio, suggesting that it reflects the effective volume of the hydrated layer on the surface of the packing materials. Furthermore, the relationships between <i>α</i>(<i>k</i>(1-β-D-arabinofuranosyl uracil)/<i>k</i>(U)) and <i>α</i>(<i>k</i>(vidarabine)/<i>k</i>(adenosine)) indicated that nucleobase‑dependent interactions contribute significantly to stationary‑phase retention. Overall, these results demonstrate that each <i>k</i>(U) and <i>α</i>, particularly those associated with partition-dominated retention, exhibit generally high robustness within this test framework and provide a basis for more robust and transferable HILIC column characterization.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"49 9","pages":""},"PeriodicalIF":3.6,"publicationDate":"2026-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148962946","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Selective Screening of Dual-Target Bioactive Constituents From Shaoqitongfei Formula Against Chronic Obstructive Pulmonary Disease by Receptor Chromatography","authors":"Yunshan Wang,&nbsp;Yangyang Zhang,&nbsp;Kexin Li,&nbsp;Zihu Wang,&nbsp;Jia Wang,&nbsp;Qian Li","doi":"10.1002/jssc.70535","DOIUrl":"https://doi.org/10.1002/jssc.70535","url":null,"abstract":"<div>\u0000 \u0000 <p>Current mainstream treatment of chronic obstructive pulmonary disease (COPD) relies largely on long-acting β₂-adrenergic receptor (β₂AR) agonists and muscarinic M₃ receptor (M₃R) antagonists, yet residual inflammation and variable treatment responses persist. Developing therapeutics from multicomponent herbal medicines is an effective drug development approach but is constrained by the difficulty of linking individual constituents to pharmacologically relevant targets and in vivo efficacy. This study established an integrated discovery workflow combining whole-formula efficacy testing, receptor-affinity chromatography, mass spectrometry (MS), and in vivo validation of candidate combinations. We applied this workflow to the Shaoqitongfei formula (SQTF), an investigator-developed three-herb prescription evaluated for ameliorating COPD, and selected the β₂AR and M₃R as clinically relevant bronchodilator targets. In porcine pancreatic elastase/lipopolysaccharide (LPS)-induced COPD mice, SQTF improved respiratory parameters, attenuated lung injury, and reduced inflammatory cytokine levels in serum and bronchoalveolar lavage fluid. Affinity columns containing immobilized β₂AR and M₃R stationary phases were utilized to screen the receptor-targeting components in formula extract. High-performance liquid chromatography-tandem MS analysis tentatively annotated albiflorin and kaempferol in the extract retained by both columns. In rats exposed to cigarette smoke and LPS, co-administration of albiflorin and kaempferol reproduced several of the pulmonary and anti-inflammatory effects of SQTF and shifted protein-expression patterns associated with signaling proteins to those observed in controls. Taken together, these findings establish a receptor-guided workflow for selecting pharmacologically relevant constituent combinations from complex herbal medicines and provide an analytical basis for the further development of SQTF-derived therapies for COPD.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"49 9","pages":""},"PeriodicalIF":3.6,"publicationDate":"2026-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148943321","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Selective Screening of Dual-Target Bioactive Constituents From Shaoqitongfei Formula Against Chronic Obstructive Pulmonary Disease by Receptor Chromatography","authors":"Yunshan Wang,&nbsp;Yangyang Zhang,&nbsp;Kexin Li,&nbsp;Zihu Wang,&nbsp;Jia Wang,&nbsp;Qian Li","doi":"10.1002/jssc.70535","DOIUrl":"https://doi.org/10.1002/jssc.70535","url":null,"abstract":"<div>\u0000 \u0000 <p>Current mainstream treatment of chronic obstructive pulmonary disease (COPD) relies largely on long-acting β₂-adrenergic receptor (β₂AR) agonists and muscarinic M₃ receptor (M₃R) antagonists, yet residual inflammation and variable treatment responses persist. Developing therapeutics from multicomponent herbal medicines is an effective drug development approach but is constrained by the difficulty of linking individual constituents to pharmacologically relevant targets and in vivo efficacy. This study established an integrated discovery workflow combining whole-formula efficacy testing, receptor-affinity chromatography, mass spectrometry (MS), and in vivo validation of candidate combinations. We applied this workflow to the Shaoqitongfei formula (SQTF), an investigator-developed three-herb prescription evaluated for ameliorating COPD, and selected the β₂AR and M₃R as clinically relevant bronchodilator targets. In porcine pancreatic elastase/lipopolysaccharide (LPS)-induced COPD mice, SQTF improved respiratory parameters, attenuated lung injury, and reduced inflammatory cytokine levels in serum and bronchoalveolar lavage fluid. Affinity columns containing immobilized β₂AR and M₃R stationary phases were utilized to screen the receptor-targeting components in formula extract. High-performance liquid chromatography-tandem MS analysis tentatively annotated albiflorin and kaempferol in the extract retained by both columns. In rats exposed to cigarette smoke and LPS, co-administration of albiflorin and kaempferol reproduced several of the pulmonary and anti-inflammatory effects of SQTF and shifted protein-expression patterns associated with signaling proteins to those observed in controls. Taken together, these findings establish a receptor-guided workflow for selecting pharmacologically relevant constituent combinations from complex herbal medicines and provide an analytical basis for the further development of SQTF-derived therapies for COPD.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"49 9","pages":""},"PeriodicalIF":3.6,"publicationDate":"2026-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148943235","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bipyridine-functionalized Covalent Organic Framework as a Reusable Adsorbent for Trace BTEX Detection in Water Samples","authors":"Zhiqun Xu,&nbsp;Fangpei Ma,&nbsp;Tianju Chen,&nbsp;Bo Hou,&nbsp;Quan Cheng,&nbsp;Pingping Fan,&nbsp;Yueping Zheng,&nbsp;Lan Fang,&nbsp;Jianhua Wang","doi":"10.1002/jssc.70529","DOIUrl":"10.1002/jssc.70529","url":null,"abstract":"<div>\u0000 \u0000 <p>Benzene, toluene, ethylbenzene, and xylenes (BTEX) are hazardous volatile organic compounds frequently detected in environmental waters, posing serious risks to human health and ecosystems. Therefore, sensitive and reliable analytical methods for trace BTEX monitoring are urgently needed. In this work, a novel imine-linked covalent organic framework (TAPB-Bpy) containing bipyridine units was synthesized via Schiff-base reaction and applied as a dispersive solid-phase extraction (d-SPE) adsorbent for the preconcentration of BTEX coupled with gas chromatography-flame ionization detection (GC-FID). Benefiting from its highly ordered porous structure, good thermal stability and abundant π-electron-rich sites, TAPB-Bpy exerts strong π-π stacking and weak C-H···N hydrogen bonding interactions with BTEX molecules, as verified by XPS analysis. Under optimized conditions, the method achieved a linear range of 0.003–10 µg L⁻¹, limits of detection (LODs) of 0.001 µg L⁻¹ for all four analytes, and good reproducibility with relative standard deviations (RSDs) below 8.1%. The adsorbent retained stable extraction efficiency after at least 30 reuse cycles, showing favorable cost-effectiveness. Compared with recently reported SPE/SPME methods for BTEX determination, our approach provides 2-10-fold lower LODs, a broader linear range, and superior reusability. When applied to real water samples including tap water, river water, rainwater, and industrial wastewater, satisfactory recoveries ranging from 90.9% to 109.6% were obtained, with RSDs between 1.0% and 9.0%. These results indicate that TAPB-Bpy is a promising d-SPE adsorbent for trace BTEX analysis in complex environmental matrices.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"49 9","pages":""},"PeriodicalIF":3.6,"publicationDate":"2026-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148930198","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Selective Extraction of Genomic DNA From Animal Tissues Using a Hydrophobic Magnetic Ionic Liquid","authors":"Yu Hou,&nbsp;Xingchen He,&nbsp;Xin Tang,&nbsp;Xueqin Ding","doi":"10.1002/jssc.70533","DOIUrl":"10.1002/jssc.70533","url":null,"abstract":"<div>\u0000 \u0000 <p>The development of green and efficient methods for genomic DNA extraction from animal tissues is crucial for molecular diagnostics, food traceability, and genetic research. Conventional methods often involve toxic reagents, multiple centrifugation steps, and are time-consuming. In this study, a hydrophobic magnetic ionic liquid (MIL), <i>N</i>-octyl-4-dimethylaminopyridinium hexafluorophosphate MIL ([C₈DMAP][PF₆]‑Ni MIL), was synthesized and applied for the selective extraction of genomic DNA from various animal tissues. The material exhibited strong paramagnetic behavior, high thermal stability, and excellent hydrophobicity, enabling rapid phase separation under an external magnetic field. A mechanical shaking-assisted extraction method was developed, and key parameters including temperature, time, shaking speed, and [C₈DMAP][PF₆]-Ni MIL dosage were systematically optimized. The method demonstrated high selectivity for DNA over proteins, RNA, and amino acids, with a maximum recovery rate of 78.06 ± 1.91%. Compared to a commercial DNA extraction kit, the [C₈DMAP][PF₆]-Ni MIL-based approach provided higher yields from several tissues, including mouse liver, brain, and rabbit lung. Furthermore, the [C₈DMAP][PF₆]-Ni MIL could be reused for at least six cycles while maintaining extraction efficiency. This work not only provides a high-performance material for DNA extraction, but also demonstrates a sustainable and easily retrievable liquid-phase separation strategy, offering a generalizable platform for complex sample pretreatment.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"49 9","pages":""},"PeriodicalIF":3.6,"publicationDate":"2026-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148930258","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"UHPLC-Q/TOF-MS Characterization of Danshen–Chuanxiong and Mechanistic Insights Into Its Protective Effects Against Aβ1-42-Induced Brain Endothelial Injury Through Metabolomics","authors":"Hui Wang,&nbsp;Ling Li,&nbsp;Xiaoli He,&nbsp;Shuqi Shen,&nbsp;Liang Chao,&nbsp;Piaoxue You,&nbsp;Kaihong Ji,&nbsp;Yue Liu,&nbsp;Zhanying Hong","doi":"10.1002/jssc.70527","DOIUrl":"10.1002/jssc.70527","url":null,"abstract":"<div>\u0000 \u0000 <p>Danshen–Chuanxiong is a traditional herb pair used for cardiovascular and cerebrovascular disorders, but its chemical basis and potential effects in Alzheimer's disease (AD)-related endothelial injury remain incompletely defined. Here, an integrated analytical workflow combining reversed-phase liquid chromatography (RPLC) and hydrophilic interaction liquid chromatography (HILIC) with quadrupole time-of-flight mass spectrometry (Q-TOF-MS), cellular assays, metabolomics, and network pharmacology was applied in an amyloid-β_1-42 (Aβ_1-42) oligomer-induced human brain microvascular endothelial cell (hBMEC) injury model. The complementary RPLC/HILIC and positive/negative electrospray-ionization strategy expanded coverage across a broad polarity range. UHPLC-Q/TOF-MS detected 90 compounds, mainly phenolic acids, phthalides, tanshinones, diterpenoids, and other aromatic or terpenoid constituents. The reductions in cell viability and zonula occludens-1 expression following Aβ_1-42 exposure confirmed the successful establishment of the cellular injury model, whereas pretreatment with Danshen, Chuanxiong, or their combination increased cell viability and alleviated Aβ_1-42-induced cellular injury. Untargeted metabolomics annotated 38 differential metabolites, of which 30 showed a recovery-associated shift toward the control state after Danshen–Chuanxiong treatment; the associated pathways included amino-acid, purine, glutathione, and glycerophospholipid metabolism. Integrative analysis prioritized seven candidate targets of Danshen–Chuanxiong, namely ACHE, VEGFA, NOS2, NOS3, AKR1B1, MDM2, and XDH. These findings provide a separation-MS-based chemical characterization and mechanistic framework for further validation of Danshen–Chuanxiong in AD-related endothelial injury models.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"49 9","pages":""},"PeriodicalIF":3.6,"publicationDate":"2026-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148912666","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Machine Learning-Assisted Headspace Gas Chromatography Ion Mobility Spectrometry for Characterization and Geographical Authentication of Perillae Folium","authors":"Chenyu Wang,&nbsp;Xinyu Yue,&nbsp;Ruizi Shan,&nbsp;Hongjing Dong,&nbsp;Shuang Liu","doi":"10.1002/jssc.70525","DOIUrl":"10.1002/jssc.70525","url":null,"abstract":"<div>\u0000 \u0000 <p>Perillae Folium, derived from the leaves of <i>Perilla frutescens</i> (L.) Britt., is recognized as both a medicinal substance and an edible ingredient in China, with its market demand steadily rising. The nutritional value of Perillae Folium varies markedly across different geographical origins, posing a constraint on its further industrial development. To date, there remains a paucity of robust and reliable approaches for tracing the geographical origin of Perillae Folium samples. This study employed headspace gas chromatography ion mobility spectrometry to profile VOCs in Perillae Folium, revealing that those originating from Hunan province contained comparatively higher volatile organic component levels. Subsequently, a total of 54 volatile organic components were putatively identified in Perillae Folium, and six featured VOCs including 2-decanone, 3-octanol, 2,6-dimethyl-5-heptenal, 3-methyl-3-buten-1-ol, ethyl 3-hydroxybutyrate, and 1-(1-methylethyl)-4-methylbenzene were identified. Eight machine learning classifiers were built based on the selected featured VOCs. Among the eight models, the linear and polynomial kernel support vector machines achieved the highest predictive accuracies, both exceeding 90%, indicating their strong potential for practical application. This study provides a reference for the study of volatile organic components in Perillae Folium from different geographical origins, and offers a simple, accurate, and reliable method for distinguishing Perillae Folium from these regions.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"49 9","pages":""},"PeriodicalIF":3.6,"publicationDate":"2026-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148912677","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enantioselective Determination of Upadacitinib in Pharmaceutical Formulations Using Reversed-Phase Chiral Liquid Chromatography","authors":"Sakine Atila Karaca,&nbsp;Sona Aliyeva,&nbsp;Duygu Yeniceli","doi":"10.1002/jssc.70531","DOIUrl":"10.1002/jssc.70531","url":null,"abstract":"<p>Stereochemical impurities can significantly alter a drug's pharmacological profile and compromise patient safety even in small levels. Therefore, the development of selective chiral analytical methods is critical for quality control assurance. In this study, a novel chiral HPLC method was developed for the enantiomeric separation and quantification of upadacitinib, a selective JAK-1 inhibitor used in the treatment of rheumatoid arthritis, psoriatic arthritis, ulcerative colitis, and Crohn's disease. The Box–Behnken design, a design of experiments approach, was applied to evaluate the effects of mobile phase pH, acetonitrile content, and column temperature on resolution and peak width responses. Based on the design of experiment results, the chromatographic conditions were optimized to achieve optimal separation performance. Chiral separation was achieved under reversed-phase conditions using a Chiralcel OJ–RH column (4.6 × 150 mm, 5 µm) at 40°C. The mobile phase consisted of 20 mM phosphate buffer (pH 6) and acetonitrile (84:16, v/v), delivered isocratically at a flow rate of 0.7 mL/min, with detection performed at 340 nm. Baseline enantiomeric separation was achieved with a resolution value of 2.51. The method was validated according to ICH Q2(R2) guidelines in terms of specificity, linearity, accuracy, precision, and robustness. Good linearity was obtained over the concentration range of 0.1–12.8 µg/mL for upadacitinib and its 3<i>R</i>,4<i>S</i> enantiomer. The validated method was successfully applied to the quantification of upadacitinib in a commercial tablet formulation. Its environmental impact and practicality were evaluated using various tools. This study presents the first validated reversed-phase chiral HPLC method reported for the enantiomeric analysis of upadacitinib.</p>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"49 9","pages":""},"PeriodicalIF":3.6,"publicationDate":"2026-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jssc.70531","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148912683","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}