Journal of Chromatography A最新文献

{"title":"Simultaneous quantification of cytidine, methylcytidine, and hydroxymethylcytidine by isotope-dilution LC–MS/MS with application to mouse liver samples","authors":"Pooja Mishra ,&nbsp;Jing Ma ,&nbsp;Huan Xie ,&nbsp;Fang Li ,&nbsp;Robert Y.L. Tsai ,&nbsp;Dong Liang","doi":"10.1016/j.chroma.2026.466721","DOIUrl":"10.1016/j.chroma.2026.466721","url":null,"abstract":"<div><div>DNA methylation and hydroxymethylation are important epigenetic modifications that play key roles in cancer development and aging processes by regulating gene expression and genome stability. Traditionally, bisulfite conversion-based or antibody-based enzyme-linked immunosorbent assays are used to find DNA methylation. These tests are non specific, tedious, and not able to differentiate the difference between methylation and hydroxymethylation. To address these issues, we developed a sensitive, reproducible, and specific LC–MS/MS method for simultaneous quantification of two major DNA methylation products, 5-methyl-2′-deoxycytidine (5-mdC) and 5-hydroxymethyl-2′-deoxycytidine (5-hmdC), as well as 2′-deoxycytidine (2-dC), using corresponding stable isotope-labeled internal standards: 5-methyl-2′-deoxycytidine-d₃, 5-(hydroxymethyl)-2′-deoxycytidine-d₃, and 2′-deoxycytidine-¹³C,¹⁵N₂. We purified DNA samples from mouse liver tissue, broke them down with enzymes, filtered them, added internal standards, and then run them through a SCIEX 6500+ Triple Quad LC–MS/MS system with an Atlantis T3 C18 column under a binary gradient. The method showed great chromatographic separation and specificity, with MRM transitions of m/z 228.154 to 112.1 for 2-dC, 242.143 to 126.2 for 5-mdC, and 258.135 to 142.1 for 5-hmdC.Peak area ratio of analyte to internal standard exhibited linearity across calibration ranges of 5–5000 ng /mL for 2-dC, 0.5–500 ng/ mL for 5-mdC, and 0.05–10 ng/mL for 5-hmdC (R² &gt; 0.999), using 2 µL injection and a total runtime of 9 min. The 5-hmdC level in female mouse liver significantly increased with aging from two to sixteen months old (0.0958 % to 0.1984 %; <em>P</em> <em>&lt;</em> <em>0.001</em>), whereas 5-mdC remained unchanged (3.47 % to 3.56 %; n.s.). These data confirm the accurate and reproducible quantification of DNA methylation and hydroxymethylation in tissue samples using the developed LC-MS/MS assay and indicate a broad application to cell culture and clinical biomarker studies.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1769 ","pages":"Article 466721"},"PeriodicalIF":4.0,"publicationDate":"2026-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146045931","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Preparation of two new anionic β-cyclodextrin-based hybrid monolithic chiral stationary phases for CEC enantioseparations of drugs and molecular modeling of enantiomer-chiral selector interactions.","authors":"Feixue Zhou, Qiufang Bai, Jingwen Ma, Wanru Wang, Yuran Shi, Yan Feng, Xiao Zhai, Feng Yang, Xin Wang, Yaxuan Song, Ye Sun, Linwei Li, Guangjun Fan, Linlin Fang","doi":"10.1016/j.chroma.2026.466724","DOIUrl":"https://doi.org/10.1016/j.chroma.2026.466724","url":null,"abstract":"<p><p>Two anionic β-cyclodextrin-based nanohybrid monolithic columns were synthesized and applied in CEC enantioseparation with UV detection. The first monolithic column was fabricated via a \"one-pot\" approach using glycidyl methacrylate-functionalized sulfated β-cyclodextrin (GMA-S-β-CD), sodium 3-mercaptopropanesulfonate, and alkoxysilanes. The second column was prepared similarly using glutamate-modified β-cyclodextrin (GMA-Glu-β-CD) as the chiral stationary phase. Compared to the Glu-β-CD monolith, the S-β-CD column exhibited significantly higher separation efficiency, achieving baseline separation for 19 out of 27 chiral drugs-including antibiotics, antihistamines, azoles, and other therapeutic agents-whereas the Glu-β-CD monolith resolved only 14. Repeatability was excellent, with relative standard deviations (RSDs) below 4.84% for run-to-run, day-to-day, and column-to-column analyses. Furthermore, quantum mechanics-molecular mechanics (QM-MM) simulations were employed to elucidate the differences in interaction strengths between enantiomers and cyclodextrins within the inclusion complexes.</p>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1770 ","pages":"466724"},"PeriodicalIF":4.0,"publicationDate":"2026-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146117259","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Measuring fluorotelomer alcohols by thermal desorption-gas chromatography-tandem mass spectrometry: Interlaboratory study results","authors":"Clara M.A. Eichler ,&nbsp;Hannah Calder ,&nbsp;Bharat Chandramouli ,&nbsp;Matthew Curtis ,&nbsp;Heidi Hayes ,&nbsp;Benjamin Kim ,&nbsp;Ruth Marfil-Vega ,&nbsp;Cristina Matos Mejías ,&nbsp;Laura Miles ,&nbsp;Alan Owens ,&nbsp;Jack Stuff ,&nbsp;Kurt Thaxton ,&nbsp;Jochen Vandenberg ,&nbsp;Nicola Watson ,&nbsp;David Wevill ,&nbsp;Jackie A. Whitecavage ,&nbsp;Xiaoyu Liu","doi":"10.1016/j.chroma.2026.466725","DOIUrl":"10.1016/j.chroma.2026.466725","url":null,"abstract":"<div><div>Fluorotelomer alcohols (FTOHs) are a group of volatile and semi-volatile per- and polyfluoroalkyl substances (PFAS) commonly found in indoor air and contribute to PFAS inhalation exposure. Improving our understanding of the contribution of FTOHs to human exposure to PFAS indoors is of high interest. Consensus standard test methods play an essential role in environmental risk assessment and management. We present the outcome of an international interlaboratory study (ILS) conducted with nine laboratories to evaluate the precision of ASTM International Standard Test Method D8591. The ILS was organized by the United States Environmental Protection Agency. The test method specifies the analysis of four FTOHs (4:2, 6:2, 8:2 and 10:2 FTOH) collected on PFAS-specific thermal desorption tubes by gas chromatography coupled with tandem mass spectrometry. During the ILS, participating laboratories were instructed to use the test method to analyze three samples (A, B, and C), each three times. Each sample contained the target FTOHs at defined concentrations unknown to the laboratories. The results from seven laboratories show that the relative reproducibility standard deviation (<em>RSD_R</em>) of the method ranges from 14% to 26% and the relative repeatability standard deviation (<em>RSD_r</em>) ranges from 4.6% to 11%, with RSDs decreasing with decreasing volatility of the FTOHs. Bias ranged from -13% to 6.0% and was generally larger and negative for less volatile FTOHs. The test method in conjunction with the precision statistics from this ILS will provide a reliable, defendable method that can be used in the context of studying PFAS sources, transport, and human exposure.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1769 ","pages":"Article 466725"},"PeriodicalIF":4.0,"publicationDate":"2026-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146023326","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Box-Behnken optimized salting-out assisted liquid-liquid extraction coupled with LC-MS/MS for sustainable amisulpride quantification in human plasma","authors":"Ahmed Serag ,&nbsp;Manal E. Alosaimi ,&nbsp;Maram H Abduljabbar ,&nbsp;Adnan Alharbi ,&nbsp;Faisal Alsenani ,&nbsp;Farooq M. Almutairi ,&nbsp;Muneef M. Aldhafeeri ,&nbsp;Atiah H. Almalki","doi":"10.1016/j.chroma.2026.466727","DOIUrl":"10.1016/j.chroma.2026.466727","url":null,"abstract":"<div><div>A sustainable salting-out assisted liquid-liquid extraction coupled with liquid chromatography-tandem mass spectrometry (SALLE-LC-MS/MS) method was developed and validated for amisulpride quantification in human plasma according to ICH M10 guidelines. Box-Behnken experimental design systematically optimized SALLE parameters through evaluation of sample pH (5.0–10.0), acetonitrile volume (500–1500 μL), salt concentration (2.0–6.0 mol/L), and centrifugation time (3.0–10.0 min). The developed polynomial model demonstrated strong predictive capability with subsequent numerical optimization identifying optimal conditions of pH 8.5, acetonitrile 1200 μL, ammonium acetate 4.5 mol/L, and 3-minute centrifugation. These conditions achieved 97.6% extraction recovery for amisulpride with minimal deviation from predicted values. Following optimization, chromatographic separation was achieved using a C18 column with isocratic elution (acetonitrile:water with 0.1% formic acid, 70:30 v/v) and 4-minute analysis time, with multiple reaction monitoring employing transitions <em>m/z</em>370.1→242.1 for amisulpride and <em>m/z</em>376.2→165.1 for haloperidol internal standard. Comprehensive validation subsequently demonstrated linearity across 2–1500 ng/mL with acceptable accuracy, precision, and stability under clinical storage conditions. Matrix effects ranged from 88–105% with low variability, while extraction recovery exceeded 98% across all quality control levels. Clinical application in healthy volunteers (<em>n</em> = 5) following 200 mg oral amisulpride administration successfully characterized key pharmacokinetic parameters including C_max (506 ng/mL), t_max (3.8 h), and t_1/2 (13.3 h) over 48 h. Finally, multi-metric sustainability assessment using CaFRI (75/100), BAGI (77.5/100), and RGB12 (83.1/100 whiteness) frameworks confirmed achievement of white analytical chemistry through balanced analytical performance, environmental friendliness, and practical implementation feasibility. The method offers significant advantages including minimal waste generation, reduced solvent consumption, and enhanced throughput while maintaining regulatory compliance for amisulpride therapeutic drug monitoring applications.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1770 ","pages":"Article 466727"},"PeriodicalIF":4.0,"publicationDate":"2026-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146090511","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of an LC–ESI-MS/MS method for the determination of contaminants of emerging concern – towards extending quality surveillance of water resources","authors":"Nataliia Leonova ,&nbsp;Olga Konovalova ,&nbsp;Peter Tarábek","doi":"10.1016/j.chroma.2026.466717","DOIUrl":"10.1016/j.chroma.2026.466717","url":null,"abstract":"<div><div>Inclusion of emerging contaminants into water quality monitoring programs can serve as an additional layer for protecting water resources against deterioration. Based on prior non-target screening using LC–QTOF, a direct injection liquid chromatography–electrospray ionization-tandem mass spectrometry with multiple reaction monitoring method was developed for the determination of 23 micropollutants in natural waters. The combination of methodological simplicity, flexibility, environmental compatibility, and demonstrated performance under real-world monitoring conditions represents a significant advancement over existing chromatographic strategies for the determination of relevant and emerging pollutants. The method provides high sensitivity and reliability, enabling the simultaneous quantification of target analytes (pesticides, pharmaceuticals and products of their transformation) spanning a wide polarity range (XLogP3 0.3–6.1) and extending the applicability of multianalytes chromatographic analysis to complex environmental matrices. The limits of detection and quantification and linearity were evaluated in deionized water, groundwater, and surface water, while accuracy in terms of recovery and precision were determined in surface water. Pesticides provided satisfactory recoveries ranging from 83 to 120%, with quantification limits between 0.02 µg L⁻¹ and 0.18 µg L⁻¹, whereas pharmaceuticals displayed recoveries of 83–119%, with quantification limits of 0.03–0.20 µg L⁻¹ in different types of water. Using the developed method, 599 water samples were analyzed between 2023 and 2025. The most frequently detected compounds at concentrations above the limit of quantification were terbumeton-desethyl, prometon and benzenesulfonamide in groundwater; fluconazole and sulfamethoxazole in surface water; and propazine-2‑hydroxy, terbuthylazine-2‑hydroxy and carbamazepine in both types of water samples.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1768 ","pages":"Article 466717"},"PeriodicalIF":4.0,"publicationDate":"2026-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146034352","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluation of portable atmospheric-pressure solids analysis probe–quadrupole mass spectrometry and UPLC−MS for the rapid screening of illicit substances in human urine","authors":"Doreen N.B Chandra Siri ,&nbsp;Si Ling Ng ,&nbsp;Ngee Sing Chong ,&nbsp;Philip J. Marriott ,&nbsp;Yong Foo Wong","doi":"10.1016/j.chroma.2026.466720","DOIUrl":"10.1016/j.chroma.2026.466720","url":null,"abstract":"<div><div>Atmospheric-pressure solids analysis probe (ASAP) is an ambient pressure ionisation mass spectrometry technique that allows the direct analysis of liquids and solids, with or without sample preparation, producing rapid, efficient, and real-time results. In this study, portable ASAP−quadrupole mass spectrometry (ASAP−QMS) method in positive ion mode was demonstrated for the rapid analysis of 9 drugs of abuse (amphetamine, methamphetamine, oxycodone, methadone, morphine, 3,4-methylenedioxymethamphetamine, ketamine, 6-monoacetylmorphine, and cocaine), fortified in water and urine samples, with analysis time of ∼30 s. Initially, a direct analysis approach without sample preparation was applied to fortified water and urine samples. However, significant urine matrix effects were observed, leading to ion suppression and detector saturation. A simple solid-phase extraction (SPE) procedure was then applied to reduce matrix effects arising from the urine samples. The influence of cone voltages on the fragmentation and abundance of molecular ions for the illicit compounds was evaluated. The analytes exhibited limits of detection in the range of 0.3 – 2 µg mL^-1, which were considerably higher than the UPLC−ESIMS (0.05 – 0.1 µg mL^-1) method. Drug compounds identification was performed via MS spectrum matching against an in-house database. Comparative assessment with UPLC−ESI linear ion trap MS using United Nations Office on Drugs and Crime International Collaborative Exercises urine samples showed detection accuracies of 43% for ASAP−QMS and 100% for the UPLC−MS method. The lower accuracy of ASAP−QMS was attributed to the poor spectral library matching (similarity scores &lt; 850) in complex urine matrices, particularly for low-concentration drug compounds. Nevertheless, portable ASAP−QMS (with SPE) achieved a higher greenness assessment score (0.54) compared to UPLC−MS (0.37), highlighting its potential as a rapid and environmentally friendly drug screening approach.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1770 ","pages":"Article 466720"},"PeriodicalIF":4.0,"publicationDate":"2026-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146048916","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rapid detection of perfluorinated compounds in tap water using confined methanol","authors":"Xinyang Zhang,&nbsp;Huilan Piao,&nbsp;Mengqing Cui,&nbsp;Xiangzi Jin,&nbsp;Meihui Tong,&nbsp;Donghao Li,&nbsp;Yilin Zou","doi":"10.1016/j.chroma.2026.466718","DOIUrl":"10.1016/j.chroma.2026.466718","url":null,"abstract":"<div><div>Perfluorinated compounds (PFCs), characterized by their persistence and bioaccumulative properties, have been extensively detected across diverse environmental compartments, including aquatic systems, sediments, and organisms. These contaminants enter human bodies via food chain transfer, subsequently undergoing internal transport and metabolic transformation. Meanwhile, the detection of samples is also difficult to achieve sensitivity analysis due to the low content of the analytes and the complex matrix. Based on this, we propose a novel approach for analyzing the derivatives of 9 PFCs (derivatization reaction conditions: 1 min, room temperature) in tap water using carbon nanofibers/carbon fibers (CNFs/CFs) nanoconfined solvent extraction method, termed nanoconfined liquid phase nanoextraction (NLPNE), coupled with gas chromatography-mass spectrometry (GC–MS). The effect of various experimental parameters on extraction efficiency was investigated using a Box-Behnken design (BBD), such as volume of nanoconfined solvent, extraction time and desorption time, which method requires a less solvent-consumption (20 μL) and time ≤ 1 min, meeting the requirements for green environmental and sustainable development. Under optimal conditions, the developed method shown good linearity within the studied concentration range. Accuracy was evaluated by analyzing the extraction recovery of spiked tap water sample, the satisfactory recoveries (71.31%-126.15%) with RSD ≤ 17.45%. In addition, the limits of detection (LODs) for the analytes were in the range of 0.2–3 μg L⁻¹, which the sensitivity has been significantly improved. Considering the versatility of nanoconfined solvent extraction method, indicated that the NLPNE is a promising pretreatment technique for detecting PFCs in real samples.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1768 ","pages":"Article 466718"},"PeriodicalIF":4.0,"publicationDate":"2026-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146034306","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Advances in desorption electrospray ionization mass spectrometry imaging: Research progress and applications","authors":"Xujun Zhang ,&nbsp;Shan Wang ,&nbsp;Bo Sui ,&nbsp;Yan Wang ,&nbsp;Lingbo Ji ,&nbsp;Jianxun Zhang ,&nbsp;Ajuan Yu ,&nbsp;Wu Fan ,&nbsp;Wuduo Zhao","doi":"10.1016/j.chroma.2026.466719","DOIUrl":"10.1016/j.chroma.2026.466719","url":null,"abstract":"<div><div>Desorption electrospray ionization mass spectrometry imaging (DESI-MSI) is regarded as a novel molecular imaging approach that integrates DESI source with conventional mass spectrometry to provide spatial information and molecular composition on the sample’s surface. In this review, the principle of DESI-MSI was elucidated, including its desorption process and ionization mechanism. Through the applications in biological tissues and organs, the advantages of DESI-MSI technology were demonstrated in exploring the tissue functional roles and disease mechanisms, including brain, liver, kidney, and lung. Building on these findings, we review DESI-MSI’s unique capability to map both exogenous and endogenous biomolecules, thereby providing insights into spatial distribution of exogenous organisms and endogenous metabolic of transformation pathways and pharmacokinetics. Finally, the recent applications of DESI-MSI in biomedicine, environmental monitoring, and food safety are also reviewed. With the continuous improvement of instruments and analysis methods, DESI-MSI is expected to bring meaningful changes across a growing number of fields, opening up exciting opportunities to support innovative technologies and applied research.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1769 ","pages":"Article 466719"},"PeriodicalIF":4.0,"publicationDate":"2026-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146023324","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"L-Cys-functionalized Zr-based MOF NU-1000: Fabrication and application as chiral stationary phase in open tubular capillary electrochromatography for chiral separation","authors":"Gege Zhu ,&nbsp;Li Qi ,&nbsp;Bing Niu ,&nbsp;Qi Zhang ,&nbsp;Qin Chen ,&nbsp;Dan Zhong ,&nbsp;Xiaodong Sun","doi":"10.1016/j.chroma.2026.466716","DOIUrl":"10.1016/j.chroma.2026.466716","url":null,"abstract":"<div><div>Metal-organic frameworks (MOFs) are promising chiral separation materials owing to their well-defined pore structures and tunable functionality. In this work, mesoporous/microporous NU-1000 was functionalized with L-Cysteine (L-Cys) via solvent-assisted ligand exchange to produce L-Cys-NU-1000, which was applied as a stationary phase in capillary electrochromatography (CEC). The resulting L-Cys-NU-1000@capillary system achieved baseline separation of five chiral amino acid enantiomers with excellent intra-day, inter-day, and batch reproducibility (RSDs &lt; 5%). Notably, the system retained high enantioselectivity even after 100 consecutive injections. This study represents the first application of NU-1000 in chiral separation, opening new avenues for its use and laying a foundation for broader future applications. The system demonstrates distinct advantages for quality control of amino acids, offering high efficiency, robustness, and reproducible enantiomeric separation crucial for food and pharmaceutical analysis.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1768 ","pages":"Article 466716"},"PeriodicalIF":4.0,"publicationDate":"2026-01-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146028160","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Integrating hybrid modeling and high throughput screening: A modular process development platform for flowthrough chromatography","authors":"Llian Mabardi ,&nbsp;Janani Ram ,&nbsp;Chris Gerberich ,&nbsp;André Dumetz ,&nbsp;Lucas Kimerer ,&nbsp;Kerri Mendola ,&nbsp;Justin Godinho ,&nbsp;Nicholas Vecchiarello","doi":"10.1016/j.chroma.2026.466715","DOIUrl":"10.1016/j.chroma.2026.466715","url":null,"abstract":"<div><div>As the biopharmaceutical industry continues to advance towards intensified manufacturing and increasingly complex therapeutic pipelines, there is a growing demand for more efficient processes and process development strategies. Flowthrough and frontal loading operations are particularly attractive due to their high loading capacities, low buffer consumption, and operational simplicity. High throughput screening combined with mechanistic modeling provides a powerful framework for rapid development of these processes, but model calibration needed across the multitude of resins and conditions during screening remains resource intensive, and accurately capturing transport and kinetic effects is challenging. In this work, neural network and closed-form symbolic regression models were developed to predict protein loading and product yield, which can exclusively use high throughput plate-based data as inputs. To support training these models, a large database of general rate model (GRM) simulations spanning both high molecular weight (HMW) and low molecular weight (LMW) separations was created. This was accompanied by development of novel analytical expressions for loading and yield derived from fundamental principles under ideal chromatography conditions, enabling transport limitations to be treated as perturbations from the ideal case. The resulting machine learning models trained with these data rely solely on easy-to-measure, experimentally accessible inputs from standard plate-based high throughput experiments and therefore provide accurate predictions without the full input parameter set required by the GRM. Neural networks achieved the highest accuracy, while symbolic regression models performed nearly as well with the added advantages of simplicity, interpretability, and ease of implementation. Further, Monte Carlo analysis demonstrated that predictive errors of the machine learning models were comparable to or smaller than those of mechanistic simulations when typical experimental uncertainty in difficult-to-measure input parameters was considered. Building on this, closed-form expressions for productivity were developed that use the machine learning models as inputs to identify resins and operating conditions that maximize performance as a function of residence time. Finally, a platform workflow is proposed that applies these models to translate high throughput data into practical guidance for resin and condition selection, with predicted purities, yields, and productivities aligning well with experimentally-measured column performances. Overall, this proposed platform provides a foundation for seamlessly integrating high throughput screening and hybrid modeling into process development.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1768 ","pages":"Article 466715"},"PeriodicalIF":4.0,"publicationDate":"2026-01-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146034304","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}