Pub Date : 2026-01-26DOI: 10.1016/j.chroma.2026.466749
Yaming Sun , Shiqi Chai , Junnan Li , Chenchen Song , Hailiang Zhao , Lijun He , Dongmei Liu
Phthalate acid esters (PAEs) are ubiquitous pollutants in food, necessitating accurate detection to ensure food quality and safety. Herein, a novel MOF-199/graphene aerogel composite-derived porous carbon material (MGC) was successfully fabricated and applied as a solid-phase microextraction (SPME) coating for the efficient extraction and enrichment of trace PAEs from condiment. The MGC coating exhibits high enrichment capacity for PAEs, attributed to its hydrophobic carbon skeleton and abundant mesoporous structure. A sensitive analytical method based on headspace SPME coupled with gas chromatography-flame ionization detection was established using the MGC coated fiber. The method achieved a good linear response, low limits of detection (0.005–0.1 μg L−1), satisfactory recoveries (71.9 %-122.6 %). The intra-fiber relative standard deviations are between 0.1 % and 7.9 %, and those for inter-fiber are from 4.8 % to 11.2 %. The successful determination of PAEs in soy sauce and monosodium glutamate confirms the feasibility and practicality of the method. This work offers a novel analytical method with exceptional sensitivity and reliability, while also expanding the application of porous carbon in SPME.
{"title":"MOF-199/graphene aerogel-derived porous carbon as solid-phase microextraction coating for the highly sensitive determination of phthalate acid esters in condiments","authors":"Yaming Sun , Shiqi Chai , Junnan Li , Chenchen Song , Hailiang Zhao , Lijun He , Dongmei Liu","doi":"10.1016/j.chroma.2026.466749","DOIUrl":"10.1016/j.chroma.2026.466749","url":null,"abstract":"<div><div>Phthalate acid esters (PAEs) are ubiquitous pollutants in food, necessitating accurate detection to ensure food quality and safety. Herein, a novel MOF-199/graphene aerogel composite-derived porous carbon material (MGC) was successfully fabricated and applied as a solid-phase microextraction (SPME) coating for the efficient extraction and enrichment of trace PAEs from condiment. The MGC coating exhibits high enrichment capacity for PAEs, attributed to its hydrophobic carbon skeleton and abundant mesoporous structure. A sensitive analytical method based on headspace SPME coupled with gas chromatography-flame ionization detection was established using the MGC coated fiber. The method achieved a good linear response, low limits of detection (0.005–0.1 μg L<sup>−1</sup>), satisfactory recoveries (71.9 %-122.6 %). The intra-fiber relative standard deviations are between 0.1 % and 7.9 %, and those for inter-fiber are from 4.8 % to 11.2 %. The successful determination of PAEs in soy sauce and monosodium glutamate confirms the feasibility and practicality of the method. This work offers a novel analytical method with exceptional sensitivity and reliability, while also expanding the application of porous carbon in SPME.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1770 ","pages":"Article 466749"},"PeriodicalIF":4.0,"publicationDate":"2026-01-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146090512","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 : 2026-01-24DOI: 10.1016/j.chroma.2026.466745
Cameron DiSpirito, Siddharth Parasnavis, Matthew Aspelund, Steven M Cramer
During synthesis of IgG-like bispecific antibodies (bsAbs), product-related impurity formation via the mispairing of heavy chains and light chains (LCs) is common despite protein engineering efforts to mitigate these species. These mispaired impurities have similar biophysical characteristics to the target bsAb and are often difficult to remove with conventional platform processes. In this paper we demonstrate the utility of Capto MMC ImpRes for removing ½ IgG, homodimer and LC-mispair product-related impurities from an asymmetric IgG-like bsAb. Screening experiments from pH 5.0 to 9.5 showed that the ½ IgG and homodimer impurities eluted at pH 9.5, while the LC-mispair could be removed from the bsAb at pH 5.5 using a linear salt gradient. These results were then employed as a guide to propose a single column, multi-pH step gradient process for the removal of all product-related impurities. The final process included loading at pH 7.5 followed by elution of the homodimer and ½ IgG impurities from the column with a pH 9.5 wash. The column pH was then reduced to 5.5 and the LC-mispair impurity was subsequently removed during a 1 M NaCl step. Finally, a pH 8.0 with 1 M NaCl eluent was employed to recover the bound bsAb. This one step MMCEX polishing process resulted in an increased purity of the bsAb from 67.3% to 94.5% with a 72% overall recovery of the bsAb. These results demonstrate the utility of single column multimodal chromatographic processes when operated at multiple pHs for the removal of challenging product-related impurities from bsAbs.
{"title":"A single column multimodal cation exchange process for removal of half antibody, homodimer and light chain mispaired product-related impurities from a bispecific antibody.","authors":"Cameron DiSpirito, Siddharth Parasnavis, Matthew Aspelund, Steven M Cramer","doi":"10.1016/j.chroma.2026.466745","DOIUrl":"https://doi.org/10.1016/j.chroma.2026.466745","url":null,"abstract":"<p><p>During synthesis of IgG-like bispecific antibodies (bsAbs), product-related impurity formation via the mispairing of heavy chains and light chains (LCs) is common despite protein engineering efforts to mitigate these species. These mispaired impurities have similar biophysical characteristics to the target bsAb and are often difficult to remove with conventional platform processes. In this paper we demonstrate the utility of Capto MMC ImpRes for removing ½ IgG, homodimer and LC-mispair product-related impurities from an asymmetric IgG-like bsAb. Screening experiments from pH 5.0 to 9.5 showed that the ½ IgG and homodimer impurities eluted at pH 9.5, while the LC-mispair could be removed from the bsAb at pH 5.5 using a linear salt gradient. These results were then employed as a guide to propose a single column, multi-pH step gradient process for the removal of all product-related impurities. The final process included loading at pH 7.5 followed by elution of the homodimer and ½ IgG impurities from the column with a pH 9.5 wash. The column pH was then reduced to 5.5 and the LC-mispair impurity was subsequently removed during a 1 M NaCl step. Finally, a pH 8.0 with 1 M NaCl eluent was employed to recover the bound bsAb. This one step MMCEX polishing process resulted in an increased purity of the bsAb from 67.3% to 94.5% with a 72% overall recovery of the bsAb. These results demonstrate the utility of single column multimodal chromatographic processes when operated at multiple pHs for the removal of challenging product-related impurities from bsAbs.</p>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1771 ","pages":"466745"},"PeriodicalIF":4.0,"publicationDate":"2026-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146099749","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 : 2026-01-23DOI: 10.1016/j.chroma.2026.466736
Jia Chen , Run-qin Wang , Bei-bei Guo , Li Yang , Shuang Hu
A magnetic mesoporous silica (SBA-15)-based dispersive solid phase microextraction was first proposed and introduced for simultaneous analysis of four cardiac glycosides (including deslanoside, lanatoside C, digoxin, and digitoxin) in human plasma coupled with HPLC-UV. In this study, magnetic C18-modified SBA-15 (SBA-15/C18/Fe3O4) was used as the extractant, which was prepared and characterized using a suite of techniques including Fourier transform infrared spectroscopy, transmission electron microscopy, N2 adsorption-desorption, vibrating sample magnetometer, scanning electron microscopy, energy-dispersive X-ray spectroscopy and X-ray photoelectron spectroscopy. The influencing parameters of the extraction were investigated and optimized. The adsorption process of SBA-15/C18/Fe3O4 followed the Langmuir isotherm and pseudo-second-order kinetic models. The adsorbent also exhibited excellent thermal and chemical stability, alongside good reusability. Under the optimum experimental conditions, the proposed method demonstrated satisfactory performance with good linearity (8-3200 ng/mL for deslanoside and 1.6-640 ng/mL for lanatoside C, digoxin and digitoxin with the correlation coefficients no less than 0.9987), high precision (0.8%<RSD<9.7%) and accuracy (-10.3%<RE<8.4%), and acceptable extraction recoveries (56.5%-71.0%). In addition, an absence of matrix effect was observed for the proposed method when applied to human plasma, while satisfactory lower limits of quantification (8 ng/mL for deslanoside and 1.6 ng/mL for lanatoside C, digoxin and digitoxin) and stability were obtained. Overall, owing to its simplicity, efficiency, quickness and eco-friendliness, the developed method serves as a promising alternative for determining cardiac glycosides in complex biological matrices.
{"title":"Magnetic C18-modified mesoporous silica as an efficient adsorbent for dispersive solid phase microextraction of cardiac glycosides in human plasma","authors":"Jia Chen , Run-qin Wang , Bei-bei Guo , Li Yang , Shuang Hu","doi":"10.1016/j.chroma.2026.466736","DOIUrl":"10.1016/j.chroma.2026.466736","url":null,"abstract":"<div><div>A magnetic mesoporous silica (SBA-15)-based dispersive solid phase microextraction was first proposed and introduced for simultaneous analysis of four cardiac glycosides (including deslanoside, lanatoside C, digoxin, and digitoxin) in human plasma coupled with HPLC-UV. In this study, magnetic C<sub>18</sub>-modified SBA-15 (SBA-15/C<sub>18</sub>/Fe<sub>3</sub>O<sub>4</sub>) was used as the extractant, which was prepared and characterized using a suite of techniques including Fourier transform infrared spectroscopy, transmission electron microscopy, N<sub>2</sub> adsorption-desorption, vibrating sample magnetometer, scanning electron microscopy, energy-dispersive X-ray spectroscopy and X-ray photoelectron spectroscopy. The influencing parameters of the extraction were investigated and optimized. The adsorption process of SBA-15/C<sub>18</sub>/Fe<sub>3</sub>O<sub>4</sub> followed the Langmuir isotherm and pseudo-second-order kinetic models. The adsorbent also exhibited excellent thermal and chemical stability, alongside good reusability. Under the optimum experimental conditions, the proposed method demonstrated satisfactory performance with good linearity (8-3200 ng/mL for deslanoside and 1.6-640 ng/mL for lanatoside C, digoxin and digitoxin with the correlation coefficients no less than 0.9987), high precision (0.8%<RSD<9.7%) and accuracy (-10.3%<RE<8.4%), and acceptable extraction recoveries (56.5%-71.0%). In addition, an absence of matrix effect was observed for the proposed method when applied to human plasma, while satisfactory lower limits of quantification (8 ng/mL for deslanoside and 1.6 ng/mL for lanatoside C, digoxin and digitoxin) and stability were obtained. Overall, owing to its simplicity, efficiency, quickness and eco-friendliness, the developed method serves as a promising alternative for determining cardiac glycosides in complex biological matrices.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1771 ","pages":"Article 466736"},"PeriodicalIF":4.0,"publicationDate":"2026-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146071099","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 : 2026-01-23DOI: 10.1016/j.chroma.2026.466737
Weiman Zhao, Longchan Liu, Wenxiang Fan, Ziwei Li, Haizhen Zhang, Linnan Li, Zhengtao Wang, Li Yang
Ginsenosides are a class of natural glycosidic compounds characterized by structural diversity and significant bioactivities, mainly derived from Panax genus plants. However, there are still challenges in achieving efficient and green separation of ginsenosides from complex plant matrices. Traditional methods often suffer from high consumption of organic solvents, poor selectivity, and complex purification processes. In this study, a boronic acid-functionalized dendritic mesoporous silica nanomaterial was constructed based on the reversible covalent interaction between boronic acid ligands and the cis-diol motifs in ginsenosides. It was used as the solid phase extraction adsorbent for the separation and purification of ginsenosides. Taking the decoction of Panax notoginseng stems and leaves as an example, the adsorption performance and impurity removal efficiency of this strategy were systematically evaluated. The key extraction parameters were optimized by single-factor experiments and Box-Behnken response surface methodology. Quantitative analysis of nine major ginsenosides using HPLC-CAD demonstrated that the developed strategy outperformed traditional macroporous resin, achieving enrichment fold of 3.4-4.0, which were nearly 2.0 times higher than those of the resin, along with adsorption recoveries of 93.5-98.7% and desorption recoveries of 88.9–97.9%. The extracted sample was further analyzed by UPLC-Q-TOF-MS. 127 ginsenosides were successfully identified, and the interference signals of non-ginsenosides were markedly reduced. The greenness evaluation by Analytical GREEnness metric and Green Analytical Procedure Index indicated that the boron affinity strategy had significant advantages in terms of environmental friendliness and process sustainability. The solid phase extraction technique based on boron-affinity materials provides a promising approach for the efficient, large-scale and green preparation of ginsenosides and other cis-diol natural products.
{"title":"A green and efficient boronate affinity solid-phase extraction strategy for ginsenosides enrichment based on dendritic mesoporous silica","authors":"Weiman Zhao, Longchan Liu, Wenxiang Fan, Ziwei Li, Haizhen Zhang, Linnan Li, Zhengtao Wang, Li Yang","doi":"10.1016/j.chroma.2026.466737","DOIUrl":"10.1016/j.chroma.2026.466737","url":null,"abstract":"<div><div>Ginsenosides are a class of natural glycosidic compounds characterized by structural diversity and significant bioactivities, mainly derived from <em>Panax</em> genus plants. However, there are still challenges in achieving efficient and green separation of ginsenosides from complex plant matrices. Traditional methods often suffer from high consumption of organic solvents, poor selectivity, and complex purification processes. In this study, a boronic acid-functionalized dendritic mesoporous silica nanomaterial was constructed based on the reversible covalent interaction between boronic acid ligands and the cis-diol motifs in ginsenosides. It was used as the solid phase extraction adsorbent for the separation and purification of ginsenosides. Taking the decoction of <em>Panax notoginseng</em> stems and leaves as an example, the adsorption performance and impurity removal efficiency of this strategy were systematically evaluated. The key extraction parameters were optimized by single-factor experiments and Box-Behnken response surface methodology. Quantitative analysis of nine major ginsenosides using HPLC-CAD demonstrated that the developed strategy outperformed traditional macroporous resin, achieving enrichment fold of 3.4-4.0, which were nearly 2.0 times higher than those of the resin, along with adsorption recoveries of 93.5-98.7% and desorption recoveries of 88.9–97.9%. The extracted sample was further analyzed by UPLC-Q-TOF-MS. 127 ginsenosides were successfully identified, and the interference signals of non-ginsenosides were markedly reduced. The greenness evaluation by Analytical GREEnness metric and Green Analytical Procedure Index indicated that the boron affinity strategy had significant advantages in terms of environmental friendliness and process sustainability. The solid phase extraction technique based on boron-affinity materials provides a promising approach for the efficient, large-scale and green preparation of ginsenosides and other cis-diol natural products.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1770 ","pages":"Article 466737"},"PeriodicalIF":4.0,"publicationDate":"2026-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146083672","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 : 2026-01-23DOI: 10.1016/j.chroma.2026.466735
Andrea Castellaneta, Ilario Losito, Claudio Crisau, Arnold Zöldhegyi, Zuzana Hrušovská, Hans-Jürgen Rieger, Imre Molnár, Tommaso R I Cataldi
Liquid chromatography-mass spectrometry (LC-MS) is an attractive alternative to GC-MS for oxysterol (OS) analysis, as it eliminates the need for chemical derivatization. However, in-source fragmentation during electrospray ionization (ESI) or atmospheric pressure chemical ionization (APCI), usually adopted for LC-MS analysis of OS, may lead to the generation of isomeric ions from non-isomeric precursors. This prevents a complete unambiguous identification along the MS dimension and makes chromatographic separation a fundamental step. In this work, analytical design space modelling empowered by a software-based modelling tool (DryLab®) was employed to optimize the reversed-phase LC separations of ten clinically relevant OS, including isomeric ring-oxidized and side chain-oxidized sterols. Three stationary phases, octadecyl (C18), cyanopropyl (ES-CN), and pentafluorophenyl (F5), were compared in terms of separation efficacy, using HPLC columns of identical geometry (150 × 2.1 mm, 2.7 μm) packed with core-shell particles. Gradient steepness, column temperature, and composition of the organic modifier (mixture of methanol and acetonitrile) in the mobile phase were selected as critical method parameters for successive construction of the design spaces on the selected stationary phases. All the resulting optimized methods resolved positional isomers and diastereomers, excepting 25(R/S)-26-hydroxycholesterol epimers. The C18 stationary phase (at 52 °C) offered high selectivity but demanded the longest run times (ca. 80 min). ES-CN (at 50 °C) and F5 (at 25 °C) stationary phases enabled faster effective runs (< 45 min), with ES-CN also reducing retention/chemical artifacts that are presumably triggered by π-π binding interactions with the stationary phase.
液相色谱-质谱法（LC-MS）是一种有吸引力的替代气相色谱-质谱法分析氧甾醇（OS），因为它消除了化学衍生化的需要。然而，通常用于OS LC-MS分析的电喷雾电离（ESI）或大气压化学电离（APCI）过程中的源内破碎可能导致非异构前体生成异构离子。这阻止了沿着质谱维度进行完全明确的鉴定，并使色谱分离成为一个基本步骤。在这项工作中，利用基于软件的建模工具（DryLab®）进行分析设计空间建模，以优化10种临床相关OS的反相LC分离，包括异构体环氧化和侧链氧化甾醇。采用相同几何形状（150 × 2.1 mm, 2.7 μm）填充核壳颗粒的高效液相色谱柱，比较了十八烷基（C18）、氰丙基（ES-CN）和五氟苯基（F5）三种固定相的分离效果。选择梯度陡度、柱温和流动相中有机改性剂（甲醇和乙腈混合物）的组成作为在选定的固定相上连续构建设计空间的关键方法参数。除25(R/S)-26-羟基胆固醇外显体外显体外，所有优化方法均可分离位置异构体和非对映体。C18固定相（52°C）具有高选择性，但需要最长的运行时间（约80分钟）。ES-CN（50°C）和F5（25°C）固定相使有效运行速度更快（< 45分钟），ES-CN还减少了可能由π-π结合与固定相相互作用引发的保留/化学伪影。
{"title":"Overcoming challenges in LC separation of oxysterols through guided design space modelling: A comparison of three stationary phases.","authors":"Andrea Castellaneta, Ilario Losito, Claudio Crisau, Arnold Zöldhegyi, Zuzana Hrušovská, Hans-Jürgen Rieger, Imre Molnár, Tommaso R I Cataldi","doi":"10.1016/j.chroma.2026.466735","DOIUrl":"https://doi.org/10.1016/j.chroma.2026.466735","url":null,"abstract":"<p><p>Liquid chromatography-mass spectrometry (LC-MS) is an attractive alternative to GC-MS for oxysterol (OS) analysis, as it eliminates the need for chemical derivatization. However, in-source fragmentation during electrospray ionization (ESI) or atmospheric pressure chemical ionization (APCI), usually adopted for LC-MS analysis of OS, may lead to the generation of isomeric ions from non-isomeric precursors. This prevents a complete unambiguous identification along the MS dimension and makes chromatographic separation a fundamental step. In this work, analytical design space modelling empowered by a software-based modelling tool (DryLab®) was employed to optimize the reversed-phase LC separations of ten clinically relevant OS, including isomeric ring-oxidized and side chain-oxidized sterols. Three stationary phases, octadecyl (C18), cyanopropyl (ES-CN), and pentafluorophenyl (F5), were compared in terms of separation efficacy, using HPLC columns of identical geometry (150 × 2.1 mm, 2.7 μm) packed with core-shell particles. Gradient steepness, column temperature, and composition of the organic modifier (mixture of methanol and acetonitrile) in the mobile phase were selected as critical method parameters for successive construction of the design spaces on the selected stationary phases. All the resulting optimized methods resolved positional isomers and diastereomers, excepting 25(R/S)-26-hydroxycholesterol epimers. The C18 stationary phase (at 52 °C) offered high selectivity but demanded the longest run times (ca. 80 min). ES-CN (at 50 °C) and F5 (at 25 °C) stationary phases enabled faster effective runs (< 45 min), with ES-CN also reducing retention/chemical artifacts that are presumably triggered by π-π binding interactions with the stationary phase.</p>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1770 ","pages":"466735"},"PeriodicalIF":4.0,"publicationDate":"2026-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146111677","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 : 2026-01-23DOI: 10.1016/j.chroma.2026.466746
Daniel Frerichs , Alexandra Höltzel , Andreas Steinhoff , Fabrice Gritti , Kevin D. Wyndham , Thomas H. Walter , Ulrich Tallarek
As a first step towards predicting retention for mixed-mode liquid chromatography (MMLC) separations of small, hydrophilic, ionizable analytes, we created a slit-pore model of a silica-based, endcapped, reversed-phase/anion-exchange (RP/AEX) stationary phase and performed molecular dynamics simulations of its solvation by a water‒acetonitrile (W‒ACN) mobile phase under conditions where the AEX function is neutral or charged. In the latter case, sodium and chloride ions were added as co-ions and counterions, respectively, to the 60/40 (v/v) W/ACN mobile phase. The silica surface functionalization, which modeled existing column technology, yielded a unique surface topology, where bonded-phase islands formed by octadecylsilyl chains around a central tertiary alkylamine group are interspersed with hydroxylated silica patches that are exposed to the mobile phase. The two MMLC systems were analyzed regarding the solvent density, structure, and orientation with respect to the density distribution of hydrophobic and hydrophilic bonded-phase elements (alkyl groups and nitrogen atoms, respectively) at different distances from the silica surface. Effects originating from the surface functionalization were captured by comparison with a conventional RPLC stationary phase, whereas effects originating from the presence of charge were quantified by comparing the two simulated MMLC systems (neutral vs. charged surface). The analysis of ion density distributions and ion contact profiles, in particular, suggested that the surface topology of the MMLC stationary phase shields the AEX function from contact with hydrophilic counterions and favors the accumulation of analyte compounds with both hydrophobic and hydrophilic structural elements.
{"title":"Molecular dynamics simulations of mixed-mode chromatography: Generation and solvation of a silica-based, reversed-phase/anion-exchange stationary phase","authors":"Daniel Frerichs , Alexandra Höltzel , Andreas Steinhoff , Fabrice Gritti , Kevin D. Wyndham , Thomas H. Walter , Ulrich Tallarek","doi":"10.1016/j.chroma.2026.466746","DOIUrl":"10.1016/j.chroma.2026.466746","url":null,"abstract":"<div><div>As a first step towards predicting retention for mixed-mode liquid chromatography (MMLC) separations of small, hydrophilic, ionizable analytes, we created a slit-pore model of a silica-based, endcapped, reversed-phase/anion-exchange (RP/AEX) stationary phase and performed molecular dynamics simulations of its solvation by a water‒acetonitrile (W‒ACN) mobile phase under conditions where the AEX function is neutral or charged. In the latter case, sodium and chloride ions were added as co-ions and counterions, respectively, to the 60/40 (v/v) W/ACN mobile phase. The silica surface functionalization, which modeled existing column technology, yielded a unique surface topology, where bonded-phase islands formed by octadecylsilyl chains around a central tertiary alkylamine group are interspersed with hydroxylated silica patches that are exposed to the mobile phase. The two MMLC systems were analyzed regarding the solvent density, structure, and orientation with respect to the density distribution of hydrophobic and hydrophilic bonded-phase elements (alkyl groups and nitrogen atoms, respectively) at different distances from the silica surface. Effects originating from the surface functionalization were captured by comparison with a conventional RPLC stationary phase, whereas effects originating from the presence of charge were quantified by comparing the two simulated MMLC systems (neutral vs. charged surface). The analysis of ion density distributions and ion contact profiles, in particular, suggested that the surface topology of the MMLC stationary phase shields the AEX function from contact with hydrophilic counterions and favors the accumulation of analyte compounds with both hydrophobic and hydrophilic structural elements.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1770 ","pages":"Article 466746"},"PeriodicalIF":4.0,"publicationDate":"2026-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146090509","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}
A novel high-performance liquid chromatography (HPLC) approach, termed the phase-separation mode, was developed by employing a phase-separation multiphase flow (PSMF) as the mobile phase. Unlike conventional modes, this system exploits two coexisting liquid phases that generate distinct liquid–liquid interfaces in packed columns. In this study, we established a theoretical framework for predicting analyte elution times in the phase-separation mode and validated them experimentally. A theoretical equation was derived on the basis of the proposed separation mechanism, incorporating capacity factors between the stationary and mobile phases as well as between the two mobile phases, in addition to their respective hold-up times. The model was applied using an octadecylsilyl (ODS)-modified silica column and biphasic water/acetonitrile/ethyl acetate mixtures as eluents, with naphthalene derivatives serving as analytes. The calculated elution times showed good agreement with experimental values in most solvent systems. Some deviations were observed when the minor phase might behave as a non-continuous liquid domain near the particle surface. These deviations were not simple errors but provided supplementary mechanistic insights we discussed. The present work establishes a mechanistic and theoretical basis for the design and evaluation of chromatographic separations in the phase-separation mode using PSMF.
{"title":"Phase-separation mode in HPLC: Theoretical prediction and experimental evaluation of elution times","authors":"Yoshiki Hiramatsu , Takeshi Iharada , Kazuhiko Tsukagoshi","doi":"10.1016/j.chroma.2026.466734","DOIUrl":"10.1016/j.chroma.2026.466734","url":null,"abstract":"<div><div>A novel high-performance liquid chromatography (HPLC) approach, termed the phase-separation mode, was developed by employing a phase-separation multiphase flow (PSMF) as the mobile phase. Unlike conventional modes, this system exploits two coexisting liquid phases that generate distinct liquid–liquid interfaces in packed columns. In this study, we established a theoretical framework for predicting analyte elution times in the phase-separation mode and validated them experimentally. A theoretical equation was derived on the basis of the proposed separation mechanism, incorporating capacity factors between the stationary and mobile phases as well as between the two mobile phases, in addition to their respective hold-up times. The model was applied using an octadecylsilyl (ODS)-modified silica column and biphasic water/acetonitrile/ethyl acetate mixtures as eluents, with naphthalene derivatives serving as analytes. The calculated elution times showed good agreement with experimental values in most solvent systems. Some deviations were observed when the minor phase might behave as a non-continuous liquid domain near the particle surface. These deviations were not simple errors but provided supplementary mechanistic insights we discussed. The present work establishes a mechanistic and theoretical basis for the design and evaluation of chromatographic separations in the phase-separation mode using PSMF.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1770 ","pages":"Article 466734"},"PeriodicalIF":4.0,"publicationDate":"2026-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146083741","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}
Supercritical fluid chromatography coupled to high-resolution mass spectrometry (SFCHRMS) is gaining attention for non-target screening (NTS), yet systematic evaluations of data-processing workflows remain scarce. Here, we present the first comprehensive comparison between a component-profile (CP) approach based on Region of Interest analysis combined with Component Detection Algorithm and Multivariate Curve Resolution (ROI-CODA-MCR) and a feature-profile (FP) workflow, MS-DIAL. Using extracted ion chromatograms (EICs) as ground truth, both workflows were optimized and benchmarked for standard detection, peak intensity reliability, blank filtering performance, and suspect screening for wastewater samples. Both detected a comparable number of standards (33–35 out of 37) but ROI-derived intensities closely matched EICs (average ratio ≈1, RSD = 5.8%), while MS-DIAL intensities deviated substantially (up to two orders of magnitude, RSD >200%). In non-target analysis, MS-DIAL produced ≈1000 features with many false positives, while ROI-CODA-MCR yielded ≈200 components with lower blank contributions and superior resolution of in-source fragments and adducts. MS-DIAL detected more suspect compounds, highlighting a higher risk of false negatives in the CP approach, particularly for low-intensity or sparse signals. Our results demonstrate that SFCHRMS datasets, often including intense background signals, challenge key assumptions underlying both FP and CP workflows and necessitate careful parameter optimization.
{"title":"Comparative assessment of feature and component profiling workflow for supercritical fluid chromatography – high resolution mass spectrometry analysis","authors":"Samuele Pellacani , Giorgio Tomasi , Gerrit Renner , Selina Tisler","doi":"10.1016/j.chroma.2026.466730","DOIUrl":"10.1016/j.chroma.2026.466730","url":null,"abstract":"<div><div>Supercritical fluid chromatography coupled to high-resolution mass spectrometry (SFC<img>HRMS) is gaining attention for non-target screening (NTS), yet systematic evaluations of data-processing workflows remain scarce. Here, we present the first comprehensive comparison between a component-profile (CP) approach based on Region of Interest analysis combined with Component Detection Algorithm and Multivariate Curve Resolution (ROI-CODA-MCR) and a feature-profile (FP) workflow, MS-DIAL. Using extracted ion chromatograms (EICs) as ground truth, both workflows were optimized and benchmarked for standard detection, peak intensity reliability, blank filtering performance, and suspect screening for wastewater samples. Both detected a comparable number of standards (33–35 out of 37) but ROI-derived intensities closely matched EICs (average ratio ≈1, RSD = 5.8%), while MS-DIAL intensities deviated substantially (up to two orders of magnitude, RSD >200%). In non-target analysis, MS-DIAL produced ≈1000 features with many false positives, while ROI-CODA-MCR yielded ≈200 components with lower blank contributions and superior resolution of in-source fragments and adducts. MS-DIAL detected more suspect compounds, highlighting a higher risk of false negatives in the CP approach, particularly for low-intensity or sparse signals. Our results demonstrate that SFC<img>HRMS datasets, often including intense background signals, challenge key assumptions underlying both FP and CP workflows and necessitate careful parameter optimization.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1769 ","pages":"Article 466730"},"PeriodicalIF":4.0,"publicationDate":"2026-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146045905","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 : 2026-01-22DOI: 10.1016/j.chroma.2026.466728
Yilin Liu , Juan Wen , Xu Zhang , Xiao Lin , Zhuangzhuang Feng , Dejun Bao , Xiaojian Hu , Ying Zhu
Endocrine disrupting chemicals (EDCs) are a class of emerging contaminants that have raised significant public health concerns. Human co-exposure to mixtures of specific EDCs, such as phthalate and bisphenols, and the associated health risks remain poorly understood. To support biomonitoring and epidemiological studies on these compounds, there is a critical need for sensitive and reliable methods for their quantification in human specimens. Herein, we developed and validated an analytical method based on ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) for the simultaneous determination of 15 phthalate metabolites and 9 bisphenols in human serum. A systematic comparison between liquid-liquid extraction (LLE) and high-efficiency matrix removal (HMR) clean-up approach revealed that HMR offered advantages in terms of extraction efficiency and matrix effect reduction, and was ultimately selected as the final clean-up method for this study. The developed method demonstrated satisfactory performance, with recoveries ranging from 81 % to 115 %, with the relative standard deviations (RSD) below 18 %. The calibration curves established using isotope-labeled internal standards exhibited good linearity, with correlation coefficients (R2) consistently above 0.997. The limits of detection (LODs) and limits of quantification (LOQs) were determined to be in the range of 0.01 ng/mL to 0.2 ng/mL and 0.04 ng/mL to 0.7 ng/mL, respectively. Both intra-day and inter-day precision were satisfactory, with RSDs values ranging from 4.2 % to 13 % and 6.9 % to 14 %, respectively. This sensitive analytical method was successfully applied to the determination of target analytes in human serum samples from 15 pregnant women, demonstrating its applicability for monitoring of these emerging contaminants in human serum.
{"title":"Determination of 15 phthalates metabolites and 9 bisphenols in human serum by using high-performance liquid chromatography coupled with tandem mass spectrometry","authors":"Yilin Liu , Juan Wen , Xu Zhang , Xiao Lin , Zhuangzhuang Feng , Dejun Bao , Xiaojian Hu , Ying Zhu","doi":"10.1016/j.chroma.2026.466728","DOIUrl":"10.1016/j.chroma.2026.466728","url":null,"abstract":"<div><div>Endocrine disrupting chemicals (EDCs) are a class of emerging contaminants that have raised significant public health concerns. Human co-exposure to mixtures of specific EDCs, such as phthalate and bisphenols, and the associated health risks remain poorly understood. To support biomonitoring and epidemiological studies on these compounds, there is a critical need for sensitive and reliable methods for their quantification in human specimens. Herein, we developed and validated an analytical method based on ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) for the simultaneous determination of 15 phthalate metabolites and 9 bisphenols in human serum. A systematic comparison between liquid-liquid extraction (LLE) and high-efficiency matrix removal (HMR) clean-up approach revealed that HMR offered advantages in terms of extraction efficiency and matrix effect reduction, and was ultimately selected as the final clean-up method for this study. The developed method demonstrated satisfactory performance, with recoveries ranging from 81 % to 115 %, with the relative standard deviations (RSD) below 18 %. The calibration curves established using isotope-labeled internal standards exhibited good linearity, with correlation coefficients (<em>R<sup>2</sup></em>) consistently above 0.997. The limits of detection (LODs) and limits of quantification (LOQs) were determined to be in the range of 0.01 ng/mL to 0.2 ng/mL and 0.04 ng/mL to 0.7 ng/mL, respectively. Both intra-day and inter-day precision were satisfactory, with RSDs values ranging from 4.2 % to 13 % and 6.9 % to 14 %, respectively. This sensitive analytical method was successfully applied to the determination of target analytes in human serum samples from 15 pregnant women, demonstrating its applicability for monitoring of these emerging contaminants in human serum.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1769 ","pages":"Article 466728"},"PeriodicalIF":4.0,"publicationDate":"2026-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146023325","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 : 2026-01-22DOI: 10.1016/j.chroma.2026.466726
Yue Deng , Shuang Feng , Xiaoyan Zhang , Jiarui Zhang , Jiale Liu , Xinyue Zhou , Shimeng Fu , Yong Zhang , Lei Yin , Meiyun Shi
This study presents an innovative analytical approach utilizing ultra-performance convergence chromatography tandem mass spectrometry (UPCC-MS/MS) for the systematic separation and quantification of 20 oligomers (n = 9–28) of polyethylene glycol 1000 (PEG1K) in rat urine. A key innovation involves the use of ammonium adducts ( [M+NH₄]⁺ and [M + 2NH₄]²⁺) as precursor ions, significantly enhancing sensitivity and selectivity, particularly for higher oligomers (n > 21), where doubly charged species predominated. Coupled with a post-column compensation strategy, ionization efficiency was improved by 1–10-fold. Chromatographic separation was achieved under isocratic conditions using a green mobile phase based on supercritical CO₂ with a methanol–formic acid co-solvent, enabling efficient baseline resolution within a short runtime while minimizing organic solvent consumption. The method was validated per FDA guidelines, demonstrating excellent linearity, precision, and accuracy. Its applicability was confirmed through excretion kinetics studies in rats tail vein injection of PEG1K, revealing rapid renal clearance with 23% of the dose excreted within 4 h and cumulative excretion reaching 64% at 48 h. Green analytical chemistry metric assessments (BAGI and GEMAM) highlighted the method's superior environmental profile. This work establishes a sensitive, efficient, and sustainable platform for polymer pharmacokinetic research.
{"title":"High-throughput and green UPCC-MS/MS assay for simultaneous quantification of 20 oligomers of PEG1K coupled with ammonium adduct and post-column infusion strategies to enhance sensitivity","authors":"Yue Deng , Shuang Feng , Xiaoyan Zhang , Jiarui Zhang , Jiale Liu , Xinyue Zhou , Shimeng Fu , Yong Zhang , Lei Yin , Meiyun Shi","doi":"10.1016/j.chroma.2026.466726","DOIUrl":"10.1016/j.chroma.2026.466726","url":null,"abstract":"<div><div>This study presents an innovative analytical approach utilizing ultra-performance convergence chromatography tandem mass spectrometry (UPCC-MS/MS) for the systematic separation and quantification of 20 oligomers (<em>n</em> = 9–28) of polyethylene glycol 1000 (PEG1K) in rat urine. A key innovation involves the use of ammonium adducts ( [M+NH₄]⁺ and [M + 2NH₄]²⁺) as precursor ions, significantly enhancing sensitivity and selectivity, particularly for higher oligomers (<em>n</em> > 21), where doubly charged species predominated. Coupled with a post-column compensation strategy, ionization efficiency was improved by 1–10-fold. Chromatographic separation was achieved under isocratic conditions using a green mobile phase based on supercritical CO₂ with a methanol–formic acid co-solvent, enabling efficient baseline resolution within a short runtime while minimizing organic solvent consumption. The method was validated per FDA guidelines, demonstrating excellent linearity, precision, and accuracy. Its applicability was confirmed through excretion kinetics studies in rats tail vein injection of PEG1K, revealing rapid renal clearance with 23% of the dose excreted within 4 h and cumulative excretion reaching 64% at 48 h. Green analytical chemistry metric assessments (BAGI and GEMAM) highlighted the method's superior environmental profile. This work establishes a sensitive, efficient, and sustainable platform for polymer pharmacokinetic research.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1770 ","pages":"Article 466726"},"PeriodicalIF":4.0,"publicationDate":"2026-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146090510","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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