Pub Date : 2026-01-14DOI: 10.1016/j.chroma.2026.466707
Vidya Annavarapu, Michael G. Bartlett
Oligonucleotide therapeutics are an emerging class of drugs, but their development is challenged by difficulties in efficient extraction and quantification, primarily due to their polarity and susceptibility to degradation by nucleases. Solid phase extraction (SPE) is one of the methods used for oligonucleotide extraction, offering sensitivity along with selectivity. Surfactant-based buffer has traditionally been used in SPE extraction of oligonucleotides, but there is now a growing focus on exploring alternatives to this method. In this study, we investigated the use of a non-surfactant buffer pretreatment module for SPE extraction of oligonucleotides using both WAX-based and mixed-mode microplates. The methods for both mixed-mode and WAX-based SPE microplates were validated and applied in an in-vitro stability study. Our study showed good recovery and easy sample preparation when using the non-surfactant buffer pretreatment module, across both microplate types. The surfactant-based buffer (SB) used with proteinase K also gave a better recovery when applied to the WAX-based SPE microplate.
{"title":"Comparative evaluation of protein digestion and solid-phase extraction strategies for oligonucleotide quantification in plasma using LC-MS/MS","authors":"Vidya Annavarapu, Michael G. Bartlett","doi":"10.1016/j.chroma.2026.466707","DOIUrl":"10.1016/j.chroma.2026.466707","url":null,"abstract":"<div><div>Oligonucleotide therapeutics are an emerging class of drugs, but their development is challenged by difficulties in efficient extraction and quantification, primarily due to their polarity and susceptibility to degradation by nucleases. Solid phase extraction (SPE) is one of the methods used for oligonucleotide extraction, offering sensitivity along with selectivity. Surfactant-based buffer has traditionally been used in SPE extraction of oligonucleotides, but there is now a growing focus on exploring alternatives to this method. In this study, we investigated the use of a non-surfactant buffer pretreatment module for SPE extraction of oligonucleotides using both WAX-based and mixed-mode microplates. The methods for both mixed-mode and WAX-based SPE microplates were validated and applied in an in-vitro stability study. Our study showed good recovery and easy sample preparation when using the non-surfactant buffer pretreatment module, across both microplate types. The surfactant-based buffer (SB) used with proteinase K also gave a better recovery when applied to the WAX-based SPE microplate.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1768 ","pages":"Article 466707"},"PeriodicalIF":4.0,"publicationDate":"2026-01-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146008387","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 : 2026-01-14DOI: 10.1016/j.chroma.2026.466711
Devendra Kumar , Neerja Trivedi
Antibody-drug conjugates (ADCs) are complex biotherapeutics that combine the selectivity of monoclonal antibodies with the potency of cytotoxic payloads. Their structural heterogeneity, arising from variable conjugation sites, drug-to-antibody ratios (DAR), linker stability, and post-translational modifications, presents significant challenges for characterization and quality control. Multi-Attribute Methods (MAM), based on high-resolution mass spectrometry, have emerged as powerful tools to simultaneously identify and quantify critical quality attributes at multiple levels of analysis, including intact protein, subunits, and peptides. Compared with traditional assays such as capillary electrophoresis-sodium dodecyl sulfate, hydrophobic interaction chromatography, and Ion exchange chromatography, MAM offers higher specificity, sensitivity, and the ability to detect unexpected modifications in a single workflow. This review summarizes recent advances in the application of MAM to ADC development, highlighting strategies for monitoring DAR distribution, conjugation site occupancy, and stability of linkers and payloads. We also discuss challenges in method qualification, data handling, and regulatory acceptance, along with the potential of automation and machine learning to support large-scale implementation. Finally, we explore the future outlook of MAM as an integrated platform method for ADCs characterization and quality control, with emphasis on its role in regulatory submissions and quality control release testing.
{"title":"Multi-attribute methods in antibody-drug conjugate characterization and quality control: Opportunities, challenges, and regulatory outlook","authors":"Devendra Kumar , Neerja Trivedi","doi":"10.1016/j.chroma.2026.466711","DOIUrl":"10.1016/j.chroma.2026.466711","url":null,"abstract":"<div><div>Antibody-drug conjugates (ADCs) are complex biotherapeutics that combine the selectivity of monoclonal antibodies with the potency of cytotoxic payloads. Their structural heterogeneity, arising from variable conjugation sites, drug-to-antibody ratios (DAR), linker stability, and post-translational modifications, presents significant challenges for characterization and quality control. Multi-Attribute Methods (MAM), based on high-resolution mass spectrometry, have emerged as powerful tools to simultaneously identify and quantify critical quality attributes at multiple levels of analysis, including intact protein, subunits, and peptides. Compared with traditional assays such as capillary electrophoresis-sodium dodecyl sulfate, hydrophobic interaction chromatography, and Ion exchange chromatography, MAM offers higher specificity, sensitivity, and the ability to detect unexpected modifications in a single workflow. This review summarizes recent advances in the application of MAM to ADC development, highlighting strategies for monitoring DAR distribution, conjugation site occupancy, and stability of linkers and payloads. We also discuss challenges in method qualification, data handling, and regulatory acceptance, along with the potential of automation and machine learning to support large-scale implementation. Finally, we explore the future outlook of MAM as an integrated platform method for ADCs characterization and quality control, with emphasis on its role in regulatory submissions and quality control release testing.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1768 ","pages":"Article 466711"},"PeriodicalIF":4.0,"publicationDate":"2026-01-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146028098","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 : 2026-01-14DOI: 10.1016/j.chroma.2026.466708
Tomás M. Mac Loughlin , Marcos Navarro , Marianela Ramos , Daiana D. Dell' Arciprete , C. Felipe Herrera Niño , F. Manuel Flores , Ma. Leticia Peluso
Glyphosate (GLY) and its main degradation product aminomethylphosphonic acid (AMPA), together with the chemically related phosphinic acid herbicide glufosinate (GLU), are widely used agrochemicals. Their reliable quantification in groundwater remains analytically challenging due to complexation with divalent cations and matrix-dependent ionization. This work developed and validated a groundwater-adapted LC-MS/MS method using FMOC derivatization and isotopically labeled glyphosate (GLY*) as internal standard. The new protocol (SUBTE, from SUBTERRANEAN) introduces an acidification in the presence of excess EDTA to disrupt metal complexes prior to derivatization and was benchmarked against a conventional borate-buffer workflow (BORAX). Normalized calibration (analyte/GLY*) was linear from 0.5 to 20 µg·L−1 (R2 ≥ 0.99) and origin-compatible. Operational LOD/LOQ in matrix were 0.003/0.01 µg·L−1 for GLY, 0.01/0.04 for GLU, and 0.02/0.06 for AMPA, meeting precision criteria (≤20 % RSD at the lowest level). Across real groundwater samples spanning wide conductivity and Ca+2 + Mg+2 ranges (n = 50), SUBTE achieved mean recoveries of ∼96 % compared with ∼70 % for BORAX, demonstrating effective mitigation of divalent-cation artefacts and enhanced robustness in hard waters. No analyte exceeded the detection limit in any well at the time of sampling, consistent with the low-mobility, Ca-rich geo-pedologic setting of the Pampean plain. The SUBTE workflow thus provides a validated, matrix-resilient approach for routine monitoring of GLY, AMPA, and GLU in groundwater, ensuring reliable quantification under variable hydrogeochemical conditions.
{"title":"Groundwater-adapted FMOC-LC-MS/MS for glyphosate, glufosinate and AMPA: EDTA-assisted preconditioning and application in Buenos Aires wells","authors":"Tomás M. Mac Loughlin , Marcos Navarro , Marianela Ramos , Daiana D. Dell' Arciprete , C. Felipe Herrera Niño , F. Manuel Flores , Ma. Leticia Peluso","doi":"10.1016/j.chroma.2026.466708","DOIUrl":"10.1016/j.chroma.2026.466708","url":null,"abstract":"<div><div>Glyphosate (GLY) and its main degradation product aminomethylphosphonic acid (AMPA), together with the chemically related phosphinic acid herbicide glufosinate (GLU), are widely used agrochemicals. Their reliable quantification in groundwater remains analytically challenging due to complexation with divalent cations and matrix-dependent ionization. This work developed and validated a groundwater-adapted LC-MS/MS method using FMOC derivatization and isotopically labeled glyphosate (GLY*) as internal standard. The new protocol (SUBTE, from SUBTERRANEAN) introduces an acidification in the presence of excess EDTA to disrupt metal complexes prior to derivatization and was benchmarked against a conventional borate-buffer workflow (BORAX). Normalized calibration (analyte/GLY*) was linear from 0.5 to 20 µg·L<sup>−1</sup> (R<sup>2</sup> ≥ 0.99) and origin-compatible. Operational LOD/LOQ in matrix were 0.003/0.01 µg·L<sup>−1</sup> for GLY, 0.01/0.04 for GLU, and 0.02/0.06 for AMPA, meeting precision criteria (≤20 % RSD at the lowest level). Across real groundwater samples spanning wide conductivity and Ca<sup>+2</sup> + Mg<sup>+2</sup> ranges (<em>n</em> = 50), SUBTE achieved mean recoveries of ∼96 % compared with ∼70 % for BORAX, demonstrating effective mitigation of divalent-cation artefacts and enhanced robustness in hard waters. No analyte exceeded the detection limit in any well at the time of sampling, consistent with the low-mobility, Ca-rich geo-pedologic setting of the Pampean plain. The SUBTE workflow thus provides a validated, matrix-resilient approach for routine monitoring of GLY, AMPA, and GLU in groundwater, ensuring reliable quantification under variable hydrogeochemical conditions.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1768 ","pages":"Article 466708"},"PeriodicalIF":4.0,"publicationDate":"2026-01-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146028176","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 : 2026-01-12DOI: 10.1016/j.chroma.2026.466706
Min Zhu , Long-yu Zhu , Lu-yan An , Ju Wang , Jun-qin Qiao , Han-yue Yang , Wei-juan Zheng , Hong-zhen Lian
Research on G-quadruplexes (G4s)-RNA helicase associated with AU-rich element (RHAU) interaction has facilitated G4s-targeted therapeutic development. Despite this progress, the interaction between dimeric G-quadruplexes (d-G4s) and RHAU remain less explored compared to monomeric structures. Developing convenient and visual methods to elucidate the interaction mechanisms between different d-G4s structures and RHAU can provide new insights into binding modes, thus aiding in the design of molecular tools targeting d-G4s. In this study, we combined molecular dynamics (MD) simulations with size-exclusion chromatography (SEC) and isothermal titration calorimetry (ITC) to investigate structural dependencies in d-G4s-RHAU interactions for the first time. Diverse d-G4s included hybrid non-parallel (d-24TTG), intramolecular tandem parallel (dAGRO100, GGA8), interlocking parallel (93del), and intermolecular stacked parallel (T30695, T30177) structures were analyzed. MD simulations revealed that the distinct SEC retention behaviors in d-G4s-RHAU interactions were mainly governed by binding site accessibility, inter-site steric hindrance, and binding free energy gradients, trends supported by ITC measured affinities. Furthermore, energy decomposition analysis identified Glu26 in RHAU as a critical residue contributing to electrostatic interactions. Mutating to Arg26 substantially decreased the binding free energy (-94.05 ± 0.41 kJ/mol), emphasizing its functional importance. Thus, this work demonstrates that MD simulations are indispensable for revealing the causes of experimental phenomena and understanding the mechanisms underlying chromatographic behavior. This combined strategy not only discerns interaction patterns stemming from structural diversity but facilitates the rapid screening of G4s-targeting molecules.
g -四重复合物(G4s)-RNA解旋酶与富au元素（RHAU）相互作用的研究促进了G4s靶向治疗的开发。尽管取得了这些进展，但与单体结构相比，二聚体g -四聚物（d-G4s）和RHAU之间的相互作用仍然较少被探索。开发方便直观的方法来阐明不同d-G4s结构与RHAU之间的相互作用机制，可以为研究结合模式提供新的见解，从而有助于设计针对d-G4s的分子工具。在这项研究中，我们首次将分子动力学（MD）模拟与尺寸排除色谱（SEC）和等温滴定量热法（ITC）相结合，研究了d-G4s-RHAU相互作用的结构依赖性。分析了不同结构的d-G4s，包括杂化非平行（d-24TTG）、分子内串联平行（dAGRO100、GGA8）、互锁平行（93del）和分子间堆叠平行（T30695、T30177）。MD模拟表明，d-G4s-RHAU相互作用中不同的SEC保留行为主要受结合位点可达性、位点间空间位阻和结合自由能梯度的控制，这一趋势得到了ITC测量亲和性的支持。此外，能量分解分析确定了RHAU中的Glu26是参与静电相互作用的关键残基。突变Arg26显著降低了结合自由能（-94.05±0.41 kJ/mol），强调了其功能的重要性。因此，这项工作证明了MD模拟对于揭示实验现象的原因和理解色谱行为的机制是必不可少的。这种组合策略不仅可以识别源于结构多样性的相互作用模式，还可以促进g4s靶向分子的快速筛选。
{"title":"Molecular dynamics simulations elucidate the structural determinants of size-exclusion chromatography behavior in dimeric G-quadruplex-RHAU","authors":"Min Zhu , Long-yu Zhu , Lu-yan An , Ju Wang , Jun-qin Qiao , Han-yue Yang , Wei-juan Zheng , Hong-zhen Lian","doi":"10.1016/j.chroma.2026.466706","DOIUrl":"10.1016/j.chroma.2026.466706","url":null,"abstract":"<div><div>Research on G-quadruplexes (G4s)-RNA helicase associated with AU-rich element (RHAU) interaction has facilitated G4s-targeted therapeutic development. Despite this progress, the interaction between dimeric G-quadruplexes (<span>d</span>-G4s) and RHAU remain less explored compared to monomeric structures. Developing convenient and visual methods to elucidate the interaction mechanisms between different <span>d</span>-G4s structures and RHAU can provide new insights into binding modes, thus aiding in the design of molecular tools targeting <span>d</span>-G4s. In this study, we combined molecular dynamics (MD) simulations with size-exclusion chromatography (SEC) and isothermal titration calorimetry (ITC) to investigate structural dependencies in <span>d</span>-G4s-RHAU interactions for the first time. Diverse <span>d</span>-G4s included hybrid non-parallel (<span>d</span>-24TTG), intramolecular tandem parallel (dAGRO100, GGA8), interlocking parallel (93del), and intermolecular stacked parallel (T30695, T30177) structures were analyzed. MD simulations revealed that the distinct SEC retention behaviors in <span>d</span>-G4s-RHAU interactions were mainly governed by binding site accessibility, inter-site steric hindrance, and binding free energy gradients, trends supported by ITC measured affinities. Furthermore, energy decomposition analysis identified Glu26 in RHAU as a critical residue contributing to electrostatic interactions. Mutating to Arg26 substantially decreased the binding free energy (-94.05 ± 0.41 kJ/mol), emphasizing its functional importance. Thus, this work demonstrates that MD simulations are indispensable for revealing the causes of experimental phenomena and understanding the mechanisms underlying chromatographic behavior. This combined strategy not only discerns interaction patterns stemming from structural diversity but facilitates the rapid screening of G4s-targeting molecules.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1769 ","pages":"Article 466706"},"PeriodicalIF":4.0,"publicationDate":"2026-01-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145973921","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}
Analyzing intact proteins by capillary electrophoresis (CE) is challenging but highly valuable, as it holds potential for future clinical applications via the separation and quantification of protein biomarkers. To achieve this, the development of CE methods requires improvements in separation efficiency, reproducibility, and reduction of sample adsorption on the capillary wall. Double-chained surfactant coatings were described as potential candidate leading to high separation efficiency. In this work, a capillary coating based on dioctadecyldimethylammonium bromide (DODAB), a double-chained cationic surfactant, was investigated by assessing the critical parameters affecting DODAB-vesicle preparation and coating performances. These parameters include temperature & sonication of the coating solution, nature of the coating buffer, and capillary inner diameter. The plate height (H) of proteins as a function of their linear migration velocity (u) was systematically plotted to evaluate the separation performances under different kinetic conditions by applying different separation voltages. DODAB coating showed high surface homogeneity leading to low slope (p) of the H vs u curve (2.23 ± 0.99 ms in average on 3 different capillaries) on 5 model basic proteins (lysozyme, carbonic anhydrase, ribonuclease A, β-lactoglobulin A and myoglobin) using 2 M acetic acid, pH 2.2, as background electrolyte in a 40 cm total length and 25 μm inner diameter capillary. Excellent separation performances were achieved reaching up to 606,000 ± 27,000 plates/m at −12 kV for lysozyme, with excellent repeatability (RSD on migration times = 0.21 %).
{"title":"Critical parameters for achieving high efficiency and reproducible double-chained cationic surfactant coatings for protein separation by capillary electrophoresis","authors":"Chirapha Prakobdi , Laurent Leclercq , Phoonthawee Saetear , Hervé Cottet","doi":"10.1016/j.chroma.2026.466705","DOIUrl":"10.1016/j.chroma.2026.466705","url":null,"abstract":"<div><div>Analyzing intact proteins by capillary electrophoresis (CE) is challenging but highly valuable, as it holds potential for future clinical applications via the separation and quantification of protein biomarkers. To achieve this, the development of CE methods requires improvements in separation efficiency, reproducibility, and reduction of sample adsorption on the capillary wall. Double-chained surfactant coatings were described as potential candidate leading to high separation efficiency. In this work, a capillary coating based on dioctadecyldimethylammonium bromide (DODAB), a double-chained cationic surfactant, was investigated by assessing the critical parameters affecting DODAB-vesicle preparation and coating performances. These parameters include temperature & sonication of the coating solution, nature of the coating buffer, and capillary inner diameter. The plate height (<em>H</em>) of proteins as a function of their linear migration velocity (<em>u</em>) was systematically plotted to evaluate the separation performances under different kinetic conditions by applying different separation voltages. DODAB coating showed high surface homogeneity leading to low slope (<em>p</em>) of the <em>H</em> vs <em>u</em> curve (2.23 ± 0.99 ms in average on 3 different capillaries) on 5 model basic proteins (lysozyme, carbonic anhydrase, ribonuclease A, β-lactoglobulin A and myoglobin) using 2 M acetic acid, pH 2.2, as background electrolyte in a 40 cm total length and 25 μm inner diameter capillary. Excellent separation performances were achieved reaching up to 606,000 ± 27,000 plates/m at −12 kV for lysozyme, with excellent repeatability (RSD on migration times = 0.21 %).</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1768 ","pages":"Article 466705"},"PeriodicalIF":4.0,"publicationDate":"2026-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146034307","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 : 2026-01-10DOI: 10.1016/j.chroma.2026.466703
FeiFei Dai , Ruoyu Zhang , Dandan Nie , Shuqing Ma , Yurong Qin
A quantitative method for determining 69 odorants in water was developed using arrow-type headspace solid-phase microextraction (SPME) coupled with gas chromatography-tandem mass spectrometry (GC–MS/MS). Key extraction parameters, including extraction and desorption temperatures and times, NaCl addition, fiber type, and rotation speed, were systematically optimized. Under the optimized conditions, target compounds were efficiently enriched, separated on a polar chromatographic column, detected in selected reaction monitoring (SRM) mode, and quantified using external calibration. The method demonstrated high sensitivity, with limits of detection (LODs) ranging from 0.015 to 75 ng/L, and 55 compounds exhibiting LODs below 10 ng/L. All analytes showed excellent linearity (R2 > 0.995) within their respective ranges. Precision was satisfactory, with relative standard deviations (RSDs) between 1.10 % and 17.85 %, including 59 compounds with RSDs <10 %. Average recoveries at three spiking levels ranged from 80 % to 120 %. This automated and environmentally friendly method offers high sensitivity and accuracy, meeting the analytical requirements for complex odorants in water.
{"title":"Development of an arrow solid-phase microextraction GC–MS/MS method for the determination of 69 odorants in water","authors":"FeiFei Dai , Ruoyu Zhang , Dandan Nie , Shuqing Ma , Yurong Qin","doi":"10.1016/j.chroma.2026.466703","DOIUrl":"10.1016/j.chroma.2026.466703","url":null,"abstract":"<div><div>A quantitative method for determining 69 odorants in water was developed using arrow-type headspace solid-phase microextraction (SPME) coupled with gas chromatography-tandem mass spectrometry (GC–MS/MS). Key extraction parameters, including extraction and desorption temperatures and times, NaCl addition, fiber type, and rotation speed, were systematically optimized. Under the optimized conditions, target compounds were efficiently enriched, separated on a polar chromatographic column, detected in selected reaction monitoring (SRM) mode, and quantified using external calibration. The method demonstrated high sensitivity, with limits of detection (LODs) ranging from 0.015 to 75 ng/L, and 55 compounds exhibiting LODs below 10 ng/L. All analytes showed excellent linearity (R<sup>2</sup> > 0.995) within their respective ranges. Precision was satisfactory, with relative standard deviations (RSDs) between 1.10 % and 17.85 %, including 59 compounds with RSDs <10 %. Average recoveries at three spiking levels ranged from 80 % to 120 %. This automated and environmentally friendly method offers high sensitivity and accuracy, meeting the analytical requirements for complex odorants in water.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1769 ","pages":"Article 466703"},"PeriodicalIF":4.0,"publicationDate":"2026-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145964651","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}
In this work, an environmentally method was proposed for the trace determination of benzoylureas by high performance liquid chromatography system after micro-solid phase extraction method. Sugarcane bagasse-based biochar alginate beads as were employed as sorbent material. The synthesized material was characterized. The univariate optimization approach was performed to determine optimum conditions for both preparation of sugarcane bead adsorbents and micro-solid phase extraction procedure. Under optimal conditions, the limit of detection was found to be in the range of 1.0 – 2.0 μg L–1 and the linearity was obtained between 3.0 and 400 μg L–1 with coefficient of determination greater than 0.99. The applicability of the investigated method was evaluated with recovery studies by spiking experiments in tea samples. The acceptable recovery results between 72.1 and 121.1 % by applying matrix matching calibration showed that the proposed analytical approach is feasible for the determination of benzoylurea insecticides in tea matrices.
{"title":"Eco-friendly sugarcane bagasse-based biochar alginate beads as green sorbents for rapid and highly efficient benzoylurea insecticides extraction in tea samples","authors":"Kittiya Sakhaem , Sujittra Aintharaprasert , Dararat Saenkam , Phatchara Rattanaphonsaen , Wannipha Khiaophong , Yanawath Santaladchaiyakit , Jitlada Vichapong","doi":"10.1016/j.chroma.2026.466701","DOIUrl":"10.1016/j.chroma.2026.466701","url":null,"abstract":"<div><div>In this work, an environmentally method was proposed for the trace determination of benzoylureas by high performance liquid chromatography system after micro-solid phase extraction method. Sugarcane bagasse-based biochar alginate beads as were employed as sorbent material. The synthesized material was characterized. The univariate optimization approach was performed to determine optimum conditions for both preparation of sugarcane bead adsorbents and micro-solid phase extraction procedure. Under optimal conditions, the limit of detection was found to be in the range of 1.0 – 2.0 μg L<sup>–1</sup> and the linearity was obtained between 3.0 and 400 μg L<sup>–1</sup> with coefficient of determination greater than 0.99. The applicability of the investigated method was evaluated with recovery studies by spiking experiments in tea samples. The acceptable recovery results between 72.1 and 121.1 % by applying matrix matching calibration showed that the proposed analytical approach is feasible for the determination of benzoylurea insecticides in tea matrices.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1769 ","pages":"Article 466701"},"PeriodicalIF":4.0,"publicationDate":"2026-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145964714","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 : 2026-01-10DOI: 10.1016/j.chroma.2026.466702
Chiara De Luca , Chiara Nosengo , Amirmohammad Faraji Shovey , Antonio Ricci , Martina Catani , Alberto Cavazzini , Fabrice Gritti , Simona Felletti
This research examined the kinetic behavior of two chiral probes (FMOC-Alanine and Haloxyfop) using a zwitterionic teicoplanin-based column and an aqueous-methanol mobile phase. More specifically, experimental results, supported by literature data measured using the same elution mode, indicate that the presence of a water layer adsorbed on the particle surface hinders analyte surface diffusion leading to localized adsorption. This shows cascading effects on longitudinal diffusion, solid–liquid mass transfer resistance and eddy dispersion, which become analyte-independent. The only difference in the measured plate heights is therefore given only by slow adsorption–desorption kinetics, confirming an important correlation between analyte structure and the extent of band broadening in chiral chromatography.
The findings reported in this study can be expanded to other molecules, suggesting that similar kinetic behavior could be expected if the same experimental conditions are maintained (e.g., same column manufacturer, silica particles, and elution mode). This would help accelerating the method development and the understanding of both the chiral recognition mechanism and the sources of band broadening occurring in chiral chromatography.
{"title":"Exploring adsorption–desorption kinetics and eddy dispersion in enantioselective hydrophilic interaction chromatography using teicoplanin-based chiral stationary phase","authors":"Chiara De Luca , Chiara Nosengo , Amirmohammad Faraji Shovey , Antonio Ricci , Martina Catani , Alberto Cavazzini , Fabrice Gritti , Simona Felletti","doi":"10.1016/j.chroma.2026.466702","DOIUrl":"10.1016/j.chroma.2026.466702","url":null,"abstract":"<div><div>This research examined the kinetic behavior of two chiral probes (FMOC-Alanine and Haloxyfop) using a zwitterionic teicoplanin-based column and an aqueous-methanol mobile phase. More specifically, experimental results, supported by literature data measured using the same elution mode, indicate that the presence of a water layer adsorbed on the particle surface hinders analyte surface diffusion leading to localized adsorption. This shows cascading effects on longitudinal diffusion, solid–liquid mass transfer resistance and eddy dispersion, which become analyte-independent. The only difference in the measured plate heights is therefore given only by slow adsorption–desorption kinetics, confirming an important correlation between analyte structure and the extent of band broadening in chiral chromatography.</div><div>The findings reported in this study can be expanded to other molecules, suggesting that similar kinetic behavior could be expected if the same experimental conditions are maintained (e.g., same column manufacturer, silica particles, and elution mode). This would help accelerating the method development and the understanding of both the chiral recognition mechanism and the sources of band broadening occurring in chiral chromatography.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1769 ","pages":"Article 466702"},"PeriodicalIF":4.0,"publicationDate":"2026-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145973877","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 : 2026-01-10DOI: 10.1016/j.chroma.2026.466704
Alexandre Prefontaine , Justin Liba , Vic Spicer , Suvrat Chowdhary , Christin Treiber-Kleinke , Vladimir Kubyshkin , Beate Koksch , Ned Budisa , Oleg V. Krokhin
We investigated the reversed-phase chromatographic behaviour of peptides carrying nine non-canonical proline analogues using LC-MS/MS bottom-up proteomics analysis of Escherichia coli (E.coli) following co-translational incorporation. Of particular interest were the epimers 4R- and 4S-fluoroproline. On average, peptide interactions with the hydrophobic C18 phase increased in the order: 4R-Flp < Pro < 4S-Flp. The mean retention shifts were found to be -0.11 and +0.06% acetonitrile for 4R- and 4S-Flp, respectively (0.1% formic acid eluent additive). This diminutive average difference in hydrophobic properties belies the unpredictable chromatographic behaviour of modified peptides. The incorporation of either fluorinated epimer, each relatively amphipathic, can make certain peptides more hydrophobic, while for others, the peptide becomes more hydrophilic. Incorporation of a single 4R-fluoroproline produces a shift ranging from (2.18 to +2.55) %ACN, while 4S-fluoroproline produces shifts ranging from (-4.82 to +4.67) %ACN on the RP HPLC elution scale. In addition to common composition-dependent factors affecting peptide retention, such as peptide length, we identified several sequence-specific features that explain deviations from the average retention characteristics. These include nearest neighbour effects and amphipathic helicity. Other analogues investigated encompassed diverse chemical features, including hydroxylation (4R-hydroxyproline), dehydrogenation (3,4-dehydroproline), ring expansion (pipercolic acid), heteroatom inclusion (3-thia-proline & 4-thia-proline) and bridge ring structures (4,5-trans-methanoproline & 4,5-cis-methanoproline).
{"title":"Reversed-phase chromatography of peptides carrying non-canonical proline analogues, with special focus on 4R/4S-fluoroproline","authors":"Alexandre Prefontaine , Justin Liba , Vic Spicer , Suvrat Chowdhary , Christin Treiber-Kleinke , Vladimir Kubyshkin , Beate Koksch , Ned Budisa , Oleg V. Krokhin","doi":"10.1016/j.chroma.2026.466704","DOIUrl":"10.1016/j.chroma.2026.466704","url":null,"abstract":"<div><div>We investigated the reversed-phase chromatographic behaviour of peptides carrying nine non-canonical proline analogues using LC-MS/MS bottom-up proteomics analysis of <em>Escherichia coli</em> (<em>E.coli)</em> following co-translational incorporation. Of particular interest were the epimers 4<em>R</em>- and 4<em>S</em>-fluoroproline. On average, peptide interactions with the hydrophobic C18 phase increased in the order: 4<em>R</em>-Flp < Pro < 4<em>S</em>-Flp. The mean retention shifts were found to be -0.11 and +0.06% acetonitrile for 4<em>R</em>- and 4<em>S</em>-Flp, respectively (0.1% formic acid eluent additive). This diminutive average difference in hydrophobic properties belies the unpredictable chromatographic behaviour of modified peptides. The incorporation of either fluorinated epimer, each relatively amphipathic, can make certain peptides more hydrophobic, while for others, the peptide becomes more hydrophilic. Incorporation of a single 4<em>R</em>-fluoroproline produces a shift ranging from (2.18 to +2.55) %ACN, while <em>4S-</em>fluoroproline produces shifts ranging from (-4.82 to +4.67) %ACN on the RP HPLC elution scale. In addition to common composition-dependent factors affecting peptide retention, such as peptide length, we identified several sequence-specific features that explain deviations from the average retention characteristics. These include nearest neighbour effects and amphipathic helicity. Other analogues investigated encompassed diverse chemical features, including hydroxylation (4<em>R</em>-hydroxyproline), dehydrogenation (3,4-dehydroproline), ring expansion (pipercolic acid), heteroatom inclusion (3-thia-proline & 4-thia-proline) and bridge ring structures (4,5-<em>trans</em>-methanoproline & 4,5-<em>cis</em>-methanoproline).</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1769 ","pages":"Article 466704"},"PeriodicalIF":4.0,"publicationDate":"2026-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145973922","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 : 2026-01-08DOI: 10.1016/j.chroma.2026.466683
Ashraf Ali , Ya Song , Yongxing Hu , Sarah Alharthi , Eman Y. Santali , Wenjie Zhao
Mixed-mode stationary phases enhance separation flexibility for complex samples by integrating multiple retention mechanisms in high performance liquid chromatography (HPLC). In this study, a novel mixed-mode stationary phase was prepared by functionalizing sub-1 µm porous silica monolith particles (pore size ∼ 35 nm) with 4-vinyl-1,3-sulfopropyl pyridinium betaine (VSPB) and octadecyl silane (C18). The stationary phase (SMP-C18-SB) was characterized by X-ray photoelectron spectroscopy (XPS), scanning electron microscopy, transmission electron microscopy, solid-state 13C NMR spectroscopy, Brunauer-Emmett-Teller (BET) and Barrett-Joyner-Halenda (BJH) analysis. The resultant stationary phase was packed into stainless-steel column (150×4.6 mm) by slurry packing method with sequential pressure of 80-100 MPA. The SMP-C18-SB column demonstrated excellent separation performance for alkylbenzenes, nucleosides, zwitterionic compounds, peptides, and proteins, facilitated by hydrophobic, hydrogen bonding, and hydrophilic interactions from C18 and SB groups. SMP-C18-SB column exhibited superior separation performance in both reversed-phase and hydrophilic interaction modes. Moreover, the column back pressure was quite low owing to the irregular shape of SMPs. The SMP-C18-SB column also exhibited excellent repeatability, with intra-day relative standard deviations (RSDs) of <0.11% for retention time and <1.02% for peak area (n = 10 each) after nearly 1000 consecutive injection. Owing to its versatile mixed-mode capability, the SMP-C18-SB column could be used for the separation of complex mixtures containing polar, non-polar and zwitteionic compounds.
{"title":"Preparation of a novel C18-sulfobetaine stationary phase based on silica monolith particles for multimodal high performance liquid chromatography","authors":"Ashraf Ali , Ya Song , Yongxing Hu , Sarah Alharthi , Eman Y. Santali , Wenjie Zhao","doi":"10.1016/j.chroma.2026.466683","DOIUrl":"10.1016/j.chroma.2026.466683","url":null,"abstract":"<div><div>Mixed-mode stationary phases enhance separation flexibility for complex samples by integrating multiple retention mechanisms in high performance liquid chromatography (HPLC). In this study, a novel mixed-mode stationary phase was prepared by functionalizing sub-1 µm porous silica monolith particles (pore size ∼ 35 nm) with 4-vinyl-1,3-sulfopropyl pyridinium betaine (VSPB) and octadecyl silane (C18). The stationary phase (SMP-C18-SB) was characterized by X-ray photoelectron spectroscopy (XPS), scanning electron microscopy, transmission electron microscopy, solid-state 13C NMR spectroscopy, Brunauer-Emmett-Teller (BET) and Barrett-Joyner-Halenda (BJH) analysis. The resultant stationary phase was packed into stainless-steel column (150×4.6 mm) by slurry packing method with sequential pressure of 80-100 MPA. The SMP-C18-SB column demonstrated excellent separation performance for alkylbenzenes, nucleosides, zwitterionic compounds, peptides, and proteins, facilitated by hydrophobic, hydrogen bonding, and hydrophilic interactions from C18 and SB groups. SMP-C18-SB column exhibited superior separation performance in both reversed-phase and hydrophilic interaction modes. Moreover, the column back pressure was quite low owing to the irregular shape of SMPs. The SMP-C18-SB column also exhibited excellent repeatability, with intra-day relative standard deviations (RSDs) of <0.11% for retention time and <1.02% for peak area (n = 10 each) after nearly 1000 consecutive injection. Owing to its versatile mixed-mode capability, the SMP-C18-SB column could be used for the separation of complex mixtures containing polar, non-polar and zwitteionic compounds.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1769 ","pages":"Article 466683"},"PeriodicalIF":4.0,"publicationDate":"2026-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145973924","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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