Pub Date : 2025-11-04DOI: 10.1016/j.chroma.2025.466519
Jia-Hui Zhu , Sheng-Nan Lyu , Mo-Lin Li , Yu-Shen Liu , Lu-Liang Wang
Avermectins (AVMs) are widely used as broad-spectrum and potent antiparasitic activities. The AVMs residual in animal-derived products and poses potential risks to human health through the food chain. Existing detection methods struggle with ultra-trace residues and matrix interferences in complex samples, necessitating efficient sample pretreatment techniques. This study introduces a novel one-pot in situ synthesis of magnetic covalent organic framework (COF) nanoflowers (Fe3O4@TFB-TAPA) at room temperature, leveraging a green strategy that eliminates toxic solvents and high-pressure conditions. The unique Fe3O4@TFB-TAPA nanoflower enhanced surface area and adsorption efficiency, and were comprehensively characterization using XRD, FT-IR, TEM, VSM and TGA. Critical parameters for magnetic solid-phase extraction (MSPE) were optimized. The MSPE protocol was coupled with HPLC-FLD for sensitive detection. The Fe3O4@TFB-TAPA exhibited exceptional performance: high adsorption capacity due to π-π stacking and hydrogen bonding, superparamagnetic behavior and thermal stability. The method achieved rapid extraction within 5 min and low solvent consumption. Method validation showed wide linear ranges (0.2–600 ng/mL), low detection limits (0.06 ng/mL), and high precision (RSD < 5 %). Reusability exceeded 7 cycles with 96.5 % recovery.
{"title":"Facile room-temperature synthesis of magnetic covalent organic framework nanoflowers for efficient MSPE and detection of avermectins","authors":"Jia-Hui Zhu , Sheng-Nan Lyu , Mo-Lin Li , Yu-Shen Liu , Lu-Liang Wang","doi":"10.1016/j.chroma.2025.466519","DOIUrl":"10.1016/j.chroma.2025.466519","url":null,"abstract":"<div><div>Avermectins (AVMs) are widely used as broad-spectrum and potent antiparasitic activities. The AVMs residual in animal-derived products and poses potential risks to human health through the food chain. Existing detection methods struggle with ultra-trace residues and matrix interferences in complex samples, necessitating efficient sample pretreatment techniques. This study introduces a novel one-pot in situ synthesis of magnetic covalent organic framework (COF) nanoflowers (Fe<sub>3</sub>O<sub>4</sub>@TFB-TAPA) at room temperature, leveraging a green strategy that eliminates toxic solvents and high-pressure conditions. The unique Fe<sub>3</sub>O<sub>4</sub>@TFB-TAPA nanoflower enhanced surface area and adsorption efficiency, and were comprehensively characterization using XRD, FT-IR, TEM, VSM and TGA. Critical parameters for magnetic solid-phase extraction (MSPE) were optimized. The MSPE protocol was coupled with HPLC-FLD for sensitive detection. The Fe<sub>3</sub>O<sub>4</sub>@TFB-TAPA exhibited exceptional performance: high adsorption capacity due to π-π stacking and hydrogen bonding, superparamagnetic behavior and thermal stability. The method achieved rapid extraction within 5 min and low solvent consumption. Method validation showed wide linear ranges (0.2–600 ng/mL), low detection limits (0.06 ng/mL), and high precision (RSD < 5 %). Reusability exceeded 7 cycles with 96.5 % recovery.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1765 ","pages":"Article 466519"},"PeriodicalIF":4.0,"publicationDate":"2025-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145475390","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}
Pub Date : 2025-11-04DOI: 10.1016/j.chroma.2025.466516
Ahmed Elmekawy , Laura J Hayward , Masafumi Iwamoto , Jun Matsunami , Farzin Gharahdaghi
In the manufacturing of antisense oligonucleotides (ASOs), rapid analytical methods are crucial for efficient process control and quality assurance. However, current analytical techniques for ASO characterization are time-consuming, creating bottlenecks in the manufacturing process. To address this challenge, this study developed and implemented a fast chromatographic method for at-line analysis of ASO purified fractions, with a focus on integration with Multi-column Counter-current Solvent Gradient Purification (MCSGP). A fast 6.5-minute ion-pair liquid chromatography method with UV and MS detection was developed. Method parameters were optimised, including injection volume (2–4 µL) and sample concentration (0.1–0.2 mg/mL), to balance speed, sensitivity, and chromatographic performance. The method performance was comparable to a validated 40-minute LC-UV-MS method using various ASO samples, including conjugated crude and purified materials. The fast method demonstrated improved limits of detection and quantification compared to the conventional method, while maintaining linearity (R² > 0.99) and robust performance with high-salt MCSGP samples (up to 60 mM NaCl). Comparable accuracy was achieved for main components (UV purity, MS purity, % full length-n; %RSD < 2 %) across different sample types. The method's speed and compatibility with at-line and potential online analysis represent a significant advancement in ASO manufacturing process control.
在反义寡核苷酸（ASOs）的生产中，快速分析方法对于有效的过程控制和质量保证至关重要。然而，目前用于ASO表征的分析技术非常耗时，在制造过程中造成了瓶颈。为了解决这一挑战，本研究开发并实施了一种用于ASO纯化馏分的在线分析的快速色谱方法，重点是与多柱逆流溶剂梯度纯化（MCSGP）相结合。建立了一种6.5 min快速离子对液相色谱法，并结合紫外和质谱检测。优化进样量（2-4µL）和样品浓度（0.1-0.2 mg/mL），以平衡速度、灵敏度和色谱性能。该方法的性能与经过验证的40分钟LC-UV-MS方法相当，该方法使用各种ASO样品，包括共轭原油和纯化物质。与传统方法相比，该方法的检测限和定量范围有所提高，同时在高盐MCSGP样品（高达60 mM NaCl）中保持线性（R²> 0.99）和稳健的性能。不同样品类型的主要成分（紫外纯度，质谱纯度，%全长n； %RSD < 2%）的准确度相当。该方法的速度和与在线和潜在在线分析的兼容性代表了ASO制造过程控制的重大进步。
{"title":"At-line analysis of antisense oligonucleotide purified fractions using fast chromatographic methods","authors":"Ahmed Elmekawy , Laura J Hayward , Masafumi Iwamoto , Jun Matsunami , Farzin Gharahdaghi","doi":"10.1016/j.chroma.2025.466516","DOIUrl":"10.1016/j.chroma.2025.466516","url":null,"abstract":"<div><div>In the manufacturing of antisense oligonucleotides (ASOs), rapid analytical methods are crucial for efficient process control and quality assurance. However, current analytical techniques for ASO characterization are time-consuming, creating bottlenecks in the manufacturing process. To address this challenge, this study developed and implemented a fast chromatographic method for at-line analysis of ASO purified fractions, with a focus on integration with Multi-column Counter-current Solvent Gradient Purification (MCSGP). A fast 6.5-minute ion-pair liquid chromatography method with UV and MS detection was developed. Method parameters were optimised, including injection volume (2–4 µL) and sample concentration (0.1–0.2 mg/mL), to balance speed, sensitivity, and chromatographic performance. The method performance was comparable to a validated 40-minute LC-UV-MS method using various ASO samples, including conjugated crude and purified materials. The fast method demonstrated improved limits of detection and quantification compared to the conventional method, while maintaining linearity (R² > 0.99) and robust performance with high-salt MCSGP samples (up to 60 mM NaCl). Comparable accuracy was achieved for main components (UV purity, MS purity, % full length-n; %RSD < 2 %) across different sample types. The method's speed and compatibility with at-line and potential online analysis represent a significant advancement in ASO manufacturing process control.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1764 ","pages":"Article 466516"},"PeriodicalIF":4.0,"publicationDate":"2025-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145463605","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}
Pub Date : 2025-11-04DOI: 10.1016/j.chroma.2025.466517
Kathrin Stavenhagen , Manasses Jora , Carina Leandersson , Rebecca Rae , Julien Bourquin , Vahid Golghalyani , Werngard Czechtizky , Tomas Leek
Antisense oligonucleotides and small interfering RNA are effective RNA-targeting technologies to modulate gene expression. Early drug discovery programs involve extensive oligonucleotide synthesis and testing in in vitro and in vivo assays, demanding fast and flexible analytical characterization for assessing the oligonucleotide purity prior to testing. Rapid methods and data analysis represent a bottleneck in delivering high-throughput analytical solutions. Here, we present two liquid chromatography-mass spectrometry methods for oligonucleotide molecular weight confirmation, purity assessment and impurity profiling applicable across early drug discovery. A novel high-throughput screening method enables rapid purity checks of hundreds of oligonucleotide samples a day and can be utilized for fraction analysis in anion-exchange purification. A separate in-depth characterization method facilitates purity analysis and detailed impurity profiling of oligonucleotide samples. Both methods incorporate (semi-)automated data processing and reporting tools to deliver fast and high-quality results, supporting the early development of oligonucleotide therapeutics.
{"title":"Rapid and flexible analytical characterization of oligonucleotides in early drug discovery","authors":"Kathrin Stavenhagen , Manasses Jora , Carina Leandersson , Rebecca Rae , Julien Bourquin , Vahid Golghalyani , Werngard Czechtizky , Tomas Leek","doi":"10.1016/j.chroma.2025.466517","DOIUrl":"10.1016/j.chroma.2025.466517","url":null,"abstract":"<div><div>Antisense oligonucleotides and small interfering RNA are effective RNA-targeting technologies to modulate gene expression. Early drug discovery programs involve extensive oligonucleotide synthesis and testing in <em>in vitro</em> and <em>in vivo</em> assays, demanding fast and flexible analytical characterization for assessing the oligonucleotide purity prior to testing. Rapid methods and data analysis represent a bottleneck in delivering high-throughput analytical solutions. Here, we present two liquid chromatography-mass spectrometry methods for oligonucleotide molecular weight confirmation, purity assessment and impurity profiling applicable across early drug discovery. A novel high-throughput screening method enables rapid purity checks of hundreds of oligonucleotide samples a day and can be utilized for fraction analysis in anion-exchange purification. A separate in-depth characterization method facilitates purity analysis and detailed impurity profiling of oligonucleotide samples. Both methods incorporate (semi-)automated data processing and reporting tools to deliver fast and high-quality results, supporting the early development of oligonucleotide therapeutics.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1765 ","pages":"Article 466517"},"PeriodicalIF":4.0,"publicationDate":"2025-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145511399","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}
Pub Date : 2025-11-04DOI: 10.1016/j.chroma.2025.466518
Huiyang Wang , Guanxi Ye , Qihui Zhang , Zongrui Li , Hui Yang , Ying Zhou , Senhao Xu , Jingxin Wang , Yunjiang Yu
Human hair serves as a validated non-invasive biomonitoring tool for evaluating human exposure to specific emerging pollutants. In the present study, an effective and reliable method for the simultaneous determination of 24 phthalate diesters (PAEs) and monoesters (mPAEs) as well as 36 per- and polyfluoroalkyl substances (PFASs) has been developed and validated in human hair based on ultra-performance liquid chromatography coupled with electrospray tandem mass spectrometry (UPLC-ESI-MS/MS). Sample preparation protocols include washing and grinding of hair, ultrasonic solvent extraction, and dispersive solid-phase extraction (d-SPE) cleanup. The proposed method was verified through blank and matrix spiking analysis. The recoveries of the analytes ranged from77 % to 117 % for PAEs and mPAEs with relative standard deviations (RSDs) below 20 %, and from 83 % to 117 % for PFASs with RSDs below 20 %. All the internal standards were recovered between 82±2 % to 105±5 %. The limits of detection (LODs) of PAEs and mPAEs were from 0.042 to 24 ng/g and from 0.002 to 1.25 ng/g for PFASs. The developed method was applied to measure target analytes in actual hair samples which exhibited that DEHP and MEHP emerged as the predominant PAEs and mPAEs compounds while PFOA and PFOS were the most abundant PFASs. The developed method indicated robust performance in hair analysis of PFASs and PAEs for biomonitoring.
{"title":"Development of an effective method for biomonitoring of per- and polyfluoroalkyl substances and phthalate diesters and monoesters in human hair using liquid chromatography coupled with tandem mass spectrometry","authors":"Huiyang Wang , Guanxi Ye , Qihui Zhang , Zongrui Li , Hui Yang , Ying Zhou , Senhao Xu , Jingxin Wang , Yunjiang Yu","doi":"10.1016/j.chroma.2025.466518","DOIUrl":"10.1016/j.chroma.2025.466518","url":null,"abstract":"<div><div>Human hair serves as a validated non-invasive biomonitoring tool for evaluating human exposure to specific emerging pollutants. In the present study, an effective and reliable method for the simultaneous determination of 24 phthalate diesters (PAEs) and monoesters (mPAEs) as well as 36 per- and polyfluoroalkyl substances (PFASs) has been developed and validated in human hair based on ultra-performance liquid chromatography coupled with electrospray tandem mass spectrometry (UPLC-ESI-MS/MS). Sample preparation protocols include washing and grinding of hair, ultrasonic solvent extraction, and dispersive solid-phase extraction (d-SPE) cleanup. The proposed method was verified through blank and matrix spiking analysis. The recoveries of the analytes ranged from77 % to 117 % for PAEs and mPAEs with relative standard deviations (RSDs) below 20 %, and from 83 % to 117 % for PFASs with RSDs below 20 %. All the internal standards were recovered between 82±2 % to 105±5 %. The limits of detection (LODs) of PAEs and mPAEs were from 0.042 to 24 ng/g and from 0.002 to 1.25 ng/g for PFASs. The developed method was applied to measure target analytes in actual hair samples which exhibited that DEHP and MEHP emerged as the predominant PAEs and mPAEs compounds while PFOA and PFOS were the most abundant PFASs. The developed method indicated robust performance in hair analysis of PFASs and PAEs for biomonitoring.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1765 ","pages":"Article 466518"},"PeriodicalIF":4.0,"publicationDate":"2025-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145511416","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}
Pub Date : 2025-11-03DOI: 10.1016/j.chroma.2025.466511
Fan Wang , Rongkang Mu , Xin Chen , Xiaoying Yin , Fangyu Zhou , Yiren Song , Kexin Guo , Xuan Mao , Haochen Zhai , Tonglin Fan , Zhangyi Chen , Bingxing Zhang , Bingqi Zhu , Peilin Yu , Zhihao Xu , Jian Wang , Liya Hong
The presence of polymerized impurities in carbapenem antibiotics poses a public health risk due to their potential to induce passive cutaneous allergic reactions. Previous literature has shown that isomeric impurities can exhibit different biological properties and toxicological profiles. Therefore, studying the polymerized and isomeric impurities of carbapenem antibiotics is highly significant. However, these impurities have received relatively little attention in previous studies. Conversely, current studies on the structural elucidation of pharmaceutical impurities often neglect to investigate formation mechanisms. They also neglect the in-depth analysis of MS/MS fragment ion disparities, which can result in incorrect structural attribution. In order to characterize impurities and elucidate polymerization mechanisms, this study has developed an integrated analytical strategy incorporating multidimensional chromatographic separation, high-resolution mass spectrometry, theoretical calculations, and an automated analytical tool assisted in the auxiliary identification. HPSEC and RP-HPLC methods with novel separation principles and two-dimensional chromatographic methods (UHPLC-Q-TOF MS and 2D HPSEC × LC-Q-Exactive MS) were established to resolve polymerized species in biapenem and meropenem. This enabled the systematic separation and identification of 15 impurities (including 11 polymerized variants) through multistage MSⁿ fragment analysis in dual ionization modes. On this basis, the possible dimerization mechanism of the dimeric isomers was elucidated via theoretical calculations. This allowed both the dominant polymerization sites and the dimerization reaction to be recognized. Additionally, our self-developed automated analytical tool assisted in the auxiliary identification of dimeric isomer configurations in actual samples. The dimer configurations identified from the LC-MS/MS results were consistent with the dominant dimer configurations revealed by theoretical calculations. This work establishes a self-validating methodology that integrates “multidimensional separation–high-resolution MSn characterization –mechanistic validation- automated analysis tool auxiliary identification analysis,” for the precise characterization of polymerized impurities.
{"title":"From reaction energetics to automated analysis tool auxiliary identification: A self-validating multidimensional liquid chromatography- mass spectrometry framework for characterizing polymerized impurities in carbapenem antibiotics","authors":"Fan Wang , Rongkang Mu , Xin Chen , Xiaoying Yin , Fangyu Zhou , Yiren Song , Kexin Guo , Xuan Mao , Haochen Zhai , Tonglin Fan , Zhangyi Chen , Bingxing Zhang , Bingqi Zhu , Peilin Yu , Zhihao Xu , Jian Wang , Liya Hong","doi":"10.1016/j.chroma.2025.466511","DOIUrl":"10.1016/j.chroma.2025.466511","url":null,"abstract":"<div><div>The presence of polymerized impurities in carbapenem antibiotics poses a public health risk due to their potential to induce passive cutaneous allergic reactions. Previous literature has shown that isomeric impurities can exhibit different biological properties and toxicological profiles. Therefore, studying the polymerized and isomeric impurities of carbapenem antibiotics is highly significant. However, these impurities have received relatively little attention in previous studies. Conversely, current studies on the structural elucidation of pharmaceutical impurities often neglect to investigate formation mechanisms. They also neglect the in-depth analysis of MS/MS fragment ion disparities, which can result in incorrect structural attribution. In order to characterize impurities and elucidate polymerization mechanisms, this study has developed an integrated analytical strategy incorporating multidimensional chromatographic separation, high-resolution mass spectrometry, theoretical calculations, and an automated analytical tool assisted in the auxiliary identification. HPSEC and RP-HPLC methods with novel separation principles and two-dimensional chromatographic methods (UHPLC-Q-TOF MS and 2D HPSEC × LC-Q-Exactive MS) were established to resolve polymerized species in biapenem and meropenem. This enabled the systematic separation and identification of 15 impurities (including 11 polymerized variants) through multistage MSⁿ fragment analysis in dual ionization modes. On this basis, the possible dimerization mechanism of the dimeric isomers was elucidated via theoretical calculations. This allowed both the dominant polymerization sites and the dimerization reaction to be recognized. Additionally, our self-developed automated analytical tool assisted in the auxiliary identification of dimeric isomer configurations in actual samples. The dimer configurations identified from the LC-MS/MS results were consistent with the dominant dimer configurations revealed by theoretical calculations. This work establishes a self-validating methodology that integrates “multidimensional separation–high-resolution MS<sup>n</sup> characterization –mechanistic validation- automated analysis tool auxiliary identification analysis,” for the precise characterization of polymerized impurities.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1765 ","pages":"Article 466511"},"PeriodicalIF":4.0,"publicationDate":"2025-11-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145475389","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}
Pub Date : 2025-11-03DOI: 10.1016/j.chroma.2025.466514
Diana R. Cunha , Shu-An Hsieh , M. Beatriz Quinaz , Marcela A. Segundo , Jared L. Anderson
The increasing global demand for biopharmaceuticals calls for scalable and cost-efficient purification strategies. Downstream processing remains a major challenge, accounting for up to 80 % of total production costs and high solvent-related energy consumption. In this context, the present work investigates aqueous biphasic systems (ABS) based on deep eutectic solvents (DES), to reduce both purification costs and the environmental impact of purification methodologies. A series of DESs composed of tetraalkylammonium-based salts and choline chloride were prepared, characterized, and evaluated for their efficiency in extracting γ-globulins, employed here as monoclonal antibody (mAb) surrogates.
The system was validated for rituximab (RTX) extraction in PBS (pH 7.4) with extraction efficiencies > 87.0 % and relative standard deviations < 6.4 %, demonstrating the accuracy and repeatability of the method. The [N4444+][Cl−] : 3[Gly]-based ABS also enabled efficient RTX extraction directly from cell culture media, highlighting the DES-based ABS potential as a sustainable alternative for downstream biopharmaceutical purification.
{"title":"Aqueous biphasic systems based on deep eutectic solvents for extraction of biopharmaceuticals in downstream processing","authors":"Diana R. Cunha , Shu-An Hsieh , M. Beatriz Quinaz , Marcela A. Segundo , Jared L. Anderson","doi":"10.1016/j.chroma.2025.466514","DOIUrl":"10.1016/j.chroma.2025.466514","url":null,"abstract":"<div><div>The increasing global demand for biopharmaceuticals calls for scalable and cost-efficient purification strategies. Downstream processing remains a major challenge, accounting for up to 80 % of total production costs and high solvent-related energy consumption. In this context, the present work investigates aqueous biphasic systems (ABS) based on deep eutectic solvents (DES), to reduce both purification costs and the environmental impact of purification methodologies. A series of DESs composed of tetraalkylammonium-based salts and choline chloride were prepared, characterized, and evaluated for their efficiency in extracting γ-globulins, employed here as monoclonal antibody (mAb) surrogates.</div><div>The system was validated for rituximab (RTX) extraction in PBS (pH 7.4) with extraction efficiencies > 87.0 % and relative standard deviations < 6.4 %, demonstrating the accuracy and repeatability of the method. The [N<sub>4444</sub><sup>+</sup>][Cl<sup>−</sup>] : 3[Gly]-based ABS also enabled efficient RTX extraction directly from cell culture media, highlighting the DES-based ABS potential as a sustainable alternative for downstream biopharmaceutical purification.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1765 ","pages":"Article 466514"},"PeriodicalIF":4.0,"publicationDate":"2025-11-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145494192","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}
Pub Date : 2025-11-02DOI: 10.1016/j.chroma.2025.466512
Jian Zhang , Joseph Chamieh , Peter Hesemann , Hervé Cottet
Coatings with zwitterionic compounds have been developed as antifouling surface to reduce or eliminate nonspecific adsorption to the solid/liquid interface. In this work, zwitterionic silica (SBSi) coating was modified on fused silica capillary for the first time by silanization (covalent bonding). The self-assembled SBSi coating was characterized using X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM) and contact angle. A comparative study of electroosmotic flow (EOF) with bare fused silica capillary showed that the SBSi coating could provide effective suppression of EOF at pH 5.5. CE separations of three intact proteins (lysozyme, ribonuclease A, and myoglobin) were performed at pH 3.5 using different voltages ranging between 10 kV and 30 kV. Accordingly, the coating performances were assessed by plotting the plate height H versus the analyte velocity u, leading to similar performances as compared with polyelectrolyte multilayers (SMIL) coatings in similar conditions.
{"title":"Evaluation of zwitterionic silica monolayer as new coating for the separation of intact proteins by capillary electrophoresis","authors":"Jian Zhang , Joseph Chamieh , Peter Hesemann , Hervé Cottet","doi":"10.1016/j.chroma.2025.466512","DOIUrl":"10.1016/j.chroma.2025.466512","url":null,"abstract":"<div><div>Coatings with zwitterionic compounds have been developed as antifouling surface to reduce or eliminate nonspecific adsorption to the solid/liquid interface. In this work, zwitterionic silica (SBSi) coating was modified on fused silica capillary for the first time by silanization (covalent bonding). The self-assembled SBSi coating was characterized using X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM) and contact angle. A comparative study of electroosmotic flow (EOF) with bare fused silica capillary showed that the SBSi coating could provide effective suppression of EOF at pH 5.5. CE separations of three intact proteins (lysozyme, ribonuclease A, and myoglobin) were performed at pH 3.5 using different voltages ranging between 10 kV and 30 kV. Accordingly, the coating performances were assessed by plotting the plate height <em>H</em> versus the analyte velocity <em>u</em>, leading to similar performances as compared with polyelectrolyte multilayers (SMIL) coatings in similar conditions.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1765 ","pages":"Article 466512"},"PeriodicalIF":4.0,"publicationDate":"2025-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145511362","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}
Pub Date : 2025-11-02DOI: 10.1016/j.chroma.2025.466513
Mathew Sara, Jürgen Gailer
Toxic metal(loid)s including arsenic, cadmium, mercury, and nickel currently contaminate ∼15 % of agricultural soils globally, which compromises food safety and chronically exposes millions of people to these inorganic pollutants. While these and other persistent inorganic pollutants are known to be absorbed into the human bloodstream, including that of babies, children and pregnant women to some degree, the resulting health ramifications remain poorly defined. To this end, a better understanding of the bioinorganic chemistry of toxic metal species in the bloodstream is critical, but hampered by a lack of appropriate tools. To gain insight, LC-methods offer the capability to observe the on-column complex formation when metal species are injected with ligands being dissolved in the mobile phase and to determine the stability of medicinal drugs to establish their shelf-life. Since these observations can also be made by employing an appropriate pH 7.4 mobile phase buffer (i.e. at near physiological conditions), LC-methods can be transformed into a powerful bioinorganic research tool. Considering that toxic metal species must be absorbed into the bloodstream before they can cause harm, one of the biggest problems in the postgenomic world is to better understand their exposure-response relationship. To this end, we will highlight how LC-methods can be tailored to address pertinent bioinorganic chemistry questions at the blood-organ nexus. By providing recent examples we will demonstrate that the application of LC-methods can provide fundamentally new insight into the exposure-response relationship of toxic metal species in humans.
{"title":"Tailoring LC-based metallomics methods to disentangle the bioinorganic chemistry of toxic mercury and cadmium species at the blood-organ nexus","authors":"Mathew Sara, Jürgen Gailer","doi":"10.1016/j.chroma.2025.466513","DOIUrl":"10.1016/j.chroma.2025.466513","url":null,"abstract":"<div><div>Toxic metal(loid)s including arsenic, cadmium, mercury, and nickel currently contaminate ∼15 % of agricultural soils globally, which compromises food safety and chronically exposes millions of people to these inorganic pollutants. While these and other persistent inorganic pollutants are known to be absorbed into the human bloodstream, including that of babies, children and pregnant women to some degree, the resulting health ramifications remain poorly defined. To this end, a better understanding of the bioinorganic chemistry of toxic metal species in the bloodstream is critical, but hampered by a lack of appropriate tools. To gain insight, LC-methods offer the capability to observe the on-column complex formation when metal species are injected with ligands being dissolved in the mobile phase and to determine the stability of medicinal drugs to establish their shelf-life. Since these observations can also be made by employing an appropriate pH 7.4 mobile phase buffer (i.e. at near physiological conditions), LC-methods can be transformed into a powerful bioinorganic research tool. Considering that toxic metal species must be absorbed into the bloodstream before they can cause harm, one of the biggest problems in the postgenomic world is to better understand their exposure-response relationship. To this end, we will highlight how LC-methods can be tailored to address pertinent bioinorganic chemistry questions at the blood-organ nexus. By providing recent examples we will demonstrate that the application of LC-methods can provide fundamentally new insight into the exposure-response relationship of toxic metal species in humans.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1765 ","pages":"Article 466513"},"PeriodicalIF":4.0,"publicationDate":"2025-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145493818","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}
Pub Date : 2025-11-01DOI: 10.1016/j.chroma.2025.466510
Xuan Li , Tao Jiang , Honghui Cheng , Xu Wang , Lili Lan , Guoxiang Sun
Objective
To overcome the limitations of single-component quality control in traditional Chinese medicine (TCM) formulations, this study developed an integrated chemical-antioxidant activity evaluation strategy for assessing the quality consistency of Kangshou Pills (KSPs).
Methods
Thirty production batches of KSPs were analyzed using six-wavelength fusion fingerprinting (SWFFP). Chemical consistency was evaluated using the systematic quantitative fingerprint method (SQFM). Additionally, the dissolution profiles of overall compounds were determined by ultraviolet full-wavelength dissolution fingerprinting, and subsequently analyzed using the f2 factor method and dissolution-systematically quantified fingerprint method (DSQFM). The dissolution amount of 2,3,5,4′-tetrahydroxystyrene-2-O-β-d-glucoside (THSG) was measured by HPLC to validate the UV-based results. Furthermore, antioxidant activity was determined using the 1,1-diphenyl-2-picryl-hydrazyl radical (DPPH) radical scavenging assay. Partial least squares (PLS) regression was employed to establish spectrum-effect relationships between chromatographic data and antioxidant efficacy.
Results
SWFFP identified 26 common peaks with high batch-to-batch consistency (Sm > 0.97; 88.2 < Pm< 121.6 %), as confirmed by SQFM parameters and multivariate analysis. Notably, twenty-four batches showed good dissolution consistency (f2 > 50; 90 % < Pdm < 110 %). THSG dissolution profiles showed excellent correlation between predicted and measured values (r > 0.98). PLS and additional analyses demonstrated that most compounds in KSPs possess antioxidant activity, including two quantitative markers: THSG and emodin.
Conclusion
The established multi-dimensional approach successfully integrates chemical fingerprinting with antioxidant activity evaluation, thereby providing a scientifically robust framework for quality assessment of KSPs. This strategy enables comprehensive "quality-efficacy" evaluation and offers a reproducible model for quality standardization of TCM formulations.
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Pub Date : 2025-11-01DOI: 10.1016/j.chroma.2025.466508
Pingping Zhu, Amy C. Harms, Pascal Maas, Manisha Bakas, Julia Josette Whien, Anne-Charlotte Dubbelman, Thomas Hankemeier
Matrix effect is a well-known issue affecting accuracy and repeatability in metabolomics studies using liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS). Post-column infusion of standards (PCIS) is a promising strategy to monitor and correct matrix effect but has been rarely reported in untargeted metabolomics. The major challenges lie in selecting appropriate PCISs and identifying the most suitable PCIS to correct the matrix effect experienced by each feature. In this study, we aim to present a method for selecting suitable PCISs for matrix effect compensation based on the artificial matrix effect (MEart) created by post-column infusion of compounds that disrupt the ESI process. Our hypothesis is that the suitable PCIS for a given analyte can be identified by comparing the PCISs’ ability in MEart compensation. We evaluated this approach using 19 stable-isotopically labeled (SIL) standards spiked in plasma, urine, and feces. PCISs selected based on MEart were compared to those selected by biological matrix effect (MEbio), with 17 out of 19 SIL standards (89 %) showing consistent PCIS selection, demonstrating the effectiveness of MEart in identifying suitable PCISs. Applying MEart-selected PCISs to correct for the MEbio resulted in improved MEbio for most of the SILs affected by matrix effect and maintained MEbio for those experiencing no matrix effect. We demonstrated the efficacy of MEart in selecting suitable PCISs for MEbio correction within an LC-PCIS-MS method. Importantly, since MEart can be assessed for any detected feature, its application holds great potential for identifying suitable PCISs for matrix effect correction in untargeted metabolomics.
{"title":"Matrix effects in untargeted LC-MS metabolomics: From creation to compensation with post-column infusion of standards","authors":"Pingping Zhu, Amy C. Harms, Pascal Maas, Manisha Bakas, Julia Josette Whien, Anne-Charlotte Dubbelman, Thomas Hankemeier","doi":"10.1016/j.chroma.2025.466508","DOIUrl":"10.1016/j.chroma.2025.466508","url":null,"abstract":"<div><div>Matrix effect is a well-known issue affecting accuracy and repeatability in metabolomics studies using liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS). Post-column infusion of standards (PCIS) is a promising strategy to monitor and correct matrix effect but has been rarely reported in untargeted metabolomics. The major challenges lie in selecting appropriate PCISs and identifying the most suitable PCIS to correct the matrix effect experienced by each feature. In this study, we aim to present a method for selecting suitable PCISs for matrix effect compensation based on the artificial matrix effect (ME<sub>art</sub>) created by post-column infusion of compounds that disrupt the ESI process. Our hypothesis is that the suitable PCIS for a given analyte can be identified by comparing the PCISs’ ability in ME<sub>art</sub> compensation. We evaluated this approach using 19 stable-isotopically labeled (SIL) standards spiked in plasma, urine, and feces. PCISs selected based on ME<sub>art</sub> were compared to those selected by biological matrix effect (ME<sub>bio</sub>), with 17 out of 19 SIL standards (89 %) showing consistent PCIS selection, demonstrating the effectiveness of ME<sub>art</sub> in identifying suitable PCISs. Applying ME<sub>art</sub>-selected PCISs to correct for the ME<sub>bio</sub> resulted in improved ME<sub>bio</sub> for most of the SILs affected by matrix effect and maintained ME<sub>bio</sub> for those experiencing no matrix effect. We demonstrated the efficacy of ME<sub>art</sub> in selecting suitable PCISs for ME<sub>bio</sub> correction within an LC-PCIS-MS method. Importantly, since ME<sub>art</sub> can be assessed for any detected feature, its application holds great potential for identifying suitable PCISs for matrix effect correction in untargeted metabolomics.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1765 ","pages":"Article 466508"},"PeriodicalIF":4.0,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145475391","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}
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