J Chromatogr A [JOURNAL]
Sun
200 papers
Chromatographic resolution and reliable quantification of eleven endogenous and microbiota-derived tryptophan metabolites in human plasma by HPLC-MS/MS.
Tkachenko K, Laroni A, Ponassi M … +3 more , Maioli D, Rosano C, Profumo A
J Chromatogr A · 2026 Jul · PMID 42061175 · Publisher ↗
Tryptophan metabolism plays a central role in host-microbiota interactions and immune regulation, yet the simultaneous quantification of metabolites from the kynurenine, serotonin, and indole pathways remains analyticall...
Tryptophan metabolism plays a central role in host-microbiota interactions and immune regulation, yet the simultaneous quantification of metabolites from the kynurenine, serotonin, and indole pathways remains analytically challenging due to their diverse physicochemical properties and wide dynamic concentration ranges. Here, we report an efficient, derivatization-free HPLC-MS/MS workflow for the targeted quantification of eleven key tryptophan-derived metabolites in human plasma. The method employs a biphenyl stationary phase to achieve robust chromatographic resolution of structurally similar analytes, including compounds known to co-elute in C18 stationary phase, improving chromatographic selectivity and minimizing co-elution of structurally related analytes. Sample preparation requires only 50 µL of plasma and relies on a simple protein-precipitation step, allowing high throughput. The method demonstrated strong performance in terms of sensitivity, accuracy, and reproducibility, supported by extensive internal standardization. The combination of multi-pathway coverage, analytical cycle, and suitability for human plasma positions this workflow as a valuable tool for monitoring gut-related metabolic signatures in both research studies and potential clinical applications.
Magnetic ionic liquid-functionalized graphene for DNA extraction and authentication of beef products.
Tang X, Chen Y, Ding X
J Chromatogr A · 2026 Jul · PMID 42061174 · Publisher ↗
Food authentication, particularly the identification of meat species, relies on the efficient extraction of high-quality DNA from complex matrices. Conventional magnetic solid-phase extraction materials typically require...
Food authentication, particularly the identification of meat species, relies on the efficient extraction of high-quality DNA from complex matrices. Conventional magnetic solid-phase extraction materials typically require multistep synthesis to coat functional layers onto an iron oxide core, which complicates their preparation and limits practical application. To address this challenge, we developed a magnetic ionic liquid-functionalized graphene adsorbent (G-MIL) that integrates the magnetic component and the functional modifier into a single material, thereby eliminating the need for a separate magnetic core and simplifying the synthesis process. This G-MIL material enabled the development of a rapid, one-step DNA extraction method that combines vortex-assisted dispersion and magnetic separation, effectively integrating isolation, enrichment, and purification without tedious centrifugation steps. The key extraction parameters were systematically optimized. The method demonstrated excellent repeatability (RSD = 2.15%, n = 3) and high selectivity for DNA in the presence of interfering proteins and amino acids. Most importantly, this approach was successfully applied to the authentication of beef products, reliably distinguishing pure beef from adulterated counterparts. This work not only presents a novel and efficient adsorbent for DNA extraction but also provides a robust and straightforward platform for enhancing traceability and ensuring authenticity in the food supply chain.
Mechanistic insights into the separation of xylose and arabinose on calcium-form ion-exchange resin.
Kou J, Lin K, Tong B … +7 more , Xiang H, Wang H, Zhuang W, Yang P, Zhu C, Ying H, Wu J
J Chromatogr A · 2026 Jul · PMID 42054786 · Publisher ↗
The separation of structurally similar monosaccharides remains a key challenge in biomass processing and carbohydrate purification. In this study, the adsorption and chromatographic separation of xylose and l-arabinose o...
The separation of structurally similar monosaccharides remains a key challenge in biomass processing and carbohydrate purification. In this study, the adsorption and chromatographic separation of xylose and l-arabinose on a calcium-form cation-exchange resin (UBK555) were systematically investigated through equilibrium experiments, fixed-bed chromatography, and multiscale theoretical analysis. Adsorption isotherms obtained at 25 °C showed excellent linearity (R² > 0.99), indicating Henry-type adsorption behavior within the studied concentration range. Under single-component conditions, arabinose exhibited a higher Henry constant than xylose, suggesting a stronger intrinsic affinity toward Ca²⁺ sites. In binary systems, however, an asymmetric cooperative effect was observed: increasing xylose concentration significantly enhanced arabinose adsorption, whereas arabinose exerted negligible influence on xylose adsorption. To quantitatively describe this phenomenon, a cooperative transport-dispersive model (CTDM) incorporating a xylose-dependent adsorption parameter was developed. The model accurately reproduced breakthrough profiles under various feed compositions, linking cooperative adsorption equilibrium with dynamic chromatographic behavior. Density functional theory (DFT), Crystal Orbital Hamilton Population (COHP), and X-ray photoelectron spectroscopy (XPS) revealed the molecular origin of this behavior. Arabinose formed stronger bidentate coordination with Ca²⁺ sites, while xylose interacted through weaker monodentate coordination. Co-adsorbed xylose stabilized arabinose adsorption via enhanced hydrogen bonding and cooperative electronic interactions rather than competitive displacement. These results revealed a non-competitive cooperative adsorption mechanism between structurally similar pentoses and provide a theoretical basis for optimizing calcium-based chromatographic separations.
Opposing effects of arginine on Fc- and VH3-mediated antibody-Protein A interactions.
Wan T, Wang H, Chen Y … +3 more , Chen J, Fei L, Li Y
J Chromatogr A · 2026 Jul · PMID 42054785 · Publisher ↗
MabCaptureC is a new Protein A resin recently launched by Thermo Fisher. In addition to Fc-binding, MabCaptureC also possesses VH3-binding capability. In consistency with previous reports, we found that elution of a VH3-...
MabCaptureC is a new Protein A resin recently launched by Thermo Fisher. In addition to Fc-binding, MabCaptureC also possesses VH3-binding capability. In consistency with previous reports, we found that elution of a VH3-containing antibody from MabCaptureC requires conditions harsher than those normally used due to the existence of VH3-mediated binding in addition to the regular Fc-mediated binding. In addition, even though arginine is widely known for having a promoting effect on antibody elution from Protein A columns, at 100 mM it unexpectedly suppressed elution of the VH3-containing antibody from this resin, reducing the yield from 79.3% to 4.6%. A comprehensive study using multiple commercial Protein A resins with distinct binding specificities (i.e., Fc-binding only, Fc- and VH3-binding, and VH3-binding only) revealed that the suppressive effect of arginine on elution is specific to VH3-containing antibody from Protein A resins with VH3-binding capability. Results from the current study strongly suggest that arginine exhibits an opposing effect on Fc- and VH3-mediated binding: it weakens the former while strengthens the latter. Leveraging the opposing effect of arginine on Fc-binding and VH3-binding, we successfully separate a VH3 mAb from a non-VH3 mAb using MabCaptureC with arginine-containing elution buffer. Furthermore, by applying a similar approach, we effectively removed the homodimer byproducts associated with the production of an asymmetric bispecific antibody (bsAb) whose two parental mAbs' VH regions have different origins (i.e., VH1 and VH3), which demonstrates the practical value of the current finding.
A validated chiral LC-MS/MS method for enantioselective quantification of aromatic lactic acids in human faeces.
Gaafar AM, Stolberg-Mathieu G, Roager HM … +1 more , La Barbera G
J Chromatogr A · 2026 Jul · PMID 42048934 · Publisher ↗
Bifidobacterium, a key genus of the infant gut microbiome, produces d- and l- enantiomers of aromatic lactic acids that may influence early-life immune development through stereochemistry-dependent biological activity. N...
Bifidobacterium, a key genus of the infant gut microbiome, produces d- and l- enantiomers of aromatic lactic acids that may influence early-life immune development through stereochemistry-dependent biological activity. No validated analytical methods currently enable their accurate enantio‑separation and quantification in human biological samples. We report the first validated, targeted, liquid chromatography-mass spectrometry method using a chiral column for the enantioselective separation and quantification of d- and l- forms of phenyllactic acid (PLA), 4-hydroxyphenyllactic acid (4OH-PLA), and indolelactic acid (ILA) in faecal samples. The method achieves baseline separation of all enantiomers within 10 min, with resolution values of 2.66 (PLA), 1.77 (4OH-PLA), and 2.42 (ILA). Solid-phase extraction reduces matrix effects (>80 %) and improves analyte recovery (>80 %). Limits of quantification range from 2.9 to 6.7 ng mL, and calibration curves show excellent linearity (R² > 0.99). Inter- and intra-day precision expressed as %RSD are < 15 % for most analytes. The method was successfully applied to infant faecal samples, enabling sensitive and stereospecific quantification of aromatic lactic acids, thus establishing a foundation for exploring the biological relevance of these enantiomers in infant health.
Design and evaluation of a dicationic ionic liquid-embedded C18 stationary phase for mixed-mode chromatography.
Hu Y, Duan X, Li Q … +4 more , Wang L, Song Y, He L, Zhao W
J Chromatogr A · 2026 Jul · PMID 42048933 · Publisher ↗
A novel mixed-mode stationary phase, Sil-DIL-C18, was successfully developed by covalently embedding a dicationic dipyridinium ionic liquid within the C18 chains grafted to the silica. The chromatographic retention was s...
A novel mixed-mode stationary phase, Sil-DIL-C18, was successfully developed by covalently embedding a dicationic dipyridinium ionic liquid within the C18 chains grafted to the silica. The chromatographic retention was systematically evaluated under varying mobile-phase composition, salt concentration, and pH. Compared with conventional single-mode Extend-C18 and HILIC Plus columns, Sil-DIL-C18 exhibited markedly superior resolution of both hydrophobic solutes (alkylbenzenes and PAHs) and polar/ionizable compounds (nucleosides, sulfonamides, alkaloids, organic acids, and antipyretic analgesics). This superior versatility was achieved by simply tuning the mobile-phase composition to activate reversed-phase, hydrophilic, or ion-exchange mechanisms on the same column. The embedded dipyridinium dicationic moieties not only conferred good compatibility with highly aqueous mobile phases but also improved peak symmetry for basic analytes, thereby overcoming major limitations of traditional alkyl-bonded phases. Retention mechanisms were quantitatively elucidated using Abraham's solvation parameter model and Tanaka tests, confirming the synergistic contribution of multi-modal interactions. The new column also demonstrated excellent retention repeatability. Owing to these advantages, Sil-DIL-C18 was successfully applied to the determination of acetaminophen in commercial Tylenol tablets.
Impact of sample preparation strategies on GC-MS- and UHPLC-QTOF-MS-based metabolomic profiling of Eryngium biebersteinianum Nevski.
Alimzhanova M, Meirbekov N, Syrgabek Y … +3 more , Tugelbayeva L, Batyrbekova S, López-Serna R
J Chromatogr A · 2026 Jul · PMID 42044604 · Publisher ↗
Sample preparation plays an important role in plant metabolomics, as the extraction strategy determines metabolite coverage and analytical sustainability. This study analyses the influence of green and conventional extra...
Sample preparation plays an important role in plant metabolomics, as the extraction strategy determines metabolite coverage and analytical sustainability. This study analyses the influence of green and conventional extraction techniques on the metabolomic fingerprint of Eryngium biebersteinianum Nevski. To verify the validity of the method, four extraction methods such as subcritical CO₂ extraction, ultrasound-assisted extraction, heat reflux extraction, and Soxhlet extraction were compared in terms of metabolite recovery and chemical selectivity. The analysis was performed to achieve comprehensive profiling by combining GC-MS and UHPLC-QTOF-MS. The results show that multivariate chemometric analysis revealed a pronounced extraction-dependent separation of samples. The data obtained indicate that subcritical CO₂ extraction and ultrasound-assisted extraction favored the recovery of lipophilic compounds, whereas heat reflux extraction and Soxhlet extraction preferentially extracted more polar metabolites. This study shows that the complementary nature of analytical platforms enabled an expanded metabolomic coverage. Furthermore, greenness of sample preparation was evaluated using the AGREEprep metric. It has been found that ultrasound-assisted extraction and subcritical CO₂ extraction offered improved sustainability compared to conventional extraction methods. From the research that has been carried out, it is possible to conclude that extraction strategy plays an important role in shaping metabolomic fingerprints. Consequently, the proposed method can be readily used in practice for extraction-oriented metabolomics in sustainable analytical chemistry.
Super critical method transfer: Inter-manufacturer qualification and comparison of three SFC instruments.
Legelli M, Wirtz M, Lamotte S
J Chromatogr A · 2026 Jul · PMID 42044603 · Publisher ↗
In contrast to liquid chromatography (LC), in supercritical fluid chromatography (SFC) small changes in instrument setup and configuration can affect solvent density, which causes selectivity changes. Therefore, method t...
In contrast to liquid chromatography (LC), in supercritical fluid chromatography (SFC) small changes in instrument setup and configuration can affect solvent density, which causes selectivity changes. Therefore, method transfer is less trivial than in LC. Moreover, depending on the manufacturer, different modes of operation will affect the results. For this research, methods were developed on a Waters UPC² instrument and replicated on an SFC system of Shimadzu, namely Nexera UC, and Agilent's 1260 Infinity II SFC, respectively. The comparison of the three instruments based on a systematic qualification was the basis for subsequent method transfer. Gradient tests to characterise gradient accuracy and precision, as well as to determine the dwell volume and pump performance of each system, were carried out. Further, the injection reproducibility was evaluated, which lead to insights into injection modes of each system and the effect of different manufacturers. As the Agilent system employs feed injection of samples, and the Shimadzu instrument is capable of co-injection approaches, investigations into minimising solvent strength effects were of particular interest. Following these experiments, a method for the separation of configurational isomers of vitamin A acetate was transferred and evaluated on all three instruments. Despite the inherently lower robustness of SFC compared to LC, the results presented here demonstrate the feasibility of method transfer while also showcasing characteristics of the SFC technique. Moreover, this research shall give an overview of the instrument options and how the differences in setup affect the separation.
Porous organic polymers driven by bis-urea anion receptor sites for adsorption of perfluoroalkyl carboxylic acids.
Lin Q, Sun W, Shen JW … +2 more , Wei Y, Wang C
J Chromatogr A · 2026 Jul · PMID 42035509 · Publisher ↗
Perfluoroalkyl carboxylic acids (PFCAs), a representative class of per- and polyfluoroalkyl substances (PFASs), have drawn increasing attention due to their extreme persistence and potential health risks. Conventional ad...
Perfluoroalkyl carboxylic acids (PFCAs), a representative class of per- and polyfluoroalkyl substances (PFASs), have drawn increasing attention due to their extreme persistence and potential health risks. Conventional adsorbents for adsorption of PFCAs mainly rely on hydrophobic and electrostatic interactions. Herein, we developed an "anion-receptor-site engineering" strategy by polycondensing methylidynetri-p-phenylene triisocyanate with 3,3'-diaminobenzidine to fabricate a porous organic polymer BD(NH)-POP with ortho-phenylene bis-urea linkages. The adsorbent was systematically evaluated for adsorption of C4-C10 PFCAs, covering removal efficiency, interference response, regeneration, and trace-level enrichment. BD(NH)-POP features a mesoporous structure (∼13.47 nm), hydrophobic interface (contact angle ∼126.5°), and tunable surface charge (pH≈4.42), affording rapid kinetics and high adsorption capacity. Competitive experiments revealed that oxygen-containing anions (e.g., carboxylates) strongly inhibit PFCA uptake, indicating that directional, multipoint hydrogen bonding between the bis-urea N-H motifs and carboxylate headgroups plays a key role, while hydrophobic and electrostatic interactions provide synergistic enhancement. The adsorbent can be regenerated through an alkaline organic solvent plus high-concentration urea system and exhibits robust enrichment capability at trace levels (0.1-20 ng/mL, R = 0.963-0.999). This work proposes and validates a "headgroup anchoring-hydrophobic stabilization" adsorption framework, offering guidance for molecular recognition-based POP design for PFASs remediation.
Evaluation of retention and selectivity for positional isomers in hydrophilic interaction liquid chromatography using a quantitative retention assessment approach.
Muscatiello D, Kleiner D, Guo Y
J Chromatogr A · 2026 Jul · PMID 42035508 · Publisher ↗
Hydrophilic interaction chromatography has been shown to be able to separate positional isomers on polar stationary phases. However, detailed mechanisms for positional isomer separation are not clearly understood due to...
Hydrophilic interaction chromatography has been shown to be able to separate positional isomers on polar stationary phases. However, detailed mechanisms for positional isomer separation are not clearly understood due to the lack of methodologies that can differentiate different mechanisms (i.e., partitioning, adsorption, electrostatic interactions) involved in HILIC. In addition, the driving force for selectivity has not been investigated due to unclear retention mechanisms. In this study, we have applied the quantitative retention assessment methodology to evaluate the retention mechanisms and separation of various positional isomers of hydroxybenzenes and dihydroxybenzoic acids. The study results indicate that adsorption is the main retention mechanism for hydroxybenzene isomers, but partitioning becomes more significant at high ammonium acetate concentrations. For dihydroxybenzoic acid isomers, partitioning is the main retention mechanism in the entire range of ammonium acetate concentration used in this study and adsorption is a minor mechanism. On the positively charged stationary phase, electrostatic attraction makes a moderate contribution to the observed retention at low salt concentration. On the negatively charged stationary phase, electrostatic repulsion significantly reduces retention below 10 mM ammonium acetate. Quantitative assessment of retention mechanisms makes it feasible to identify the driving force for selectivity for the positional isomers. Mechanistic-based evaluation indicates that adsorption underscores the selectivity for resorcinol/catechol and phloroglucinol/1,2,4-trihydroxybenzene isomer pairs, but partitioning drives the selectivity for 1,2,4-trihydroxybenzene and pyrogallol. Selectivity for dihydroxybenzoic acid isomers is driving primarily by adsorption on the negatively charged zwitterionic phase; however, electrostatic repulsion plays a significant role at low ammonium acetate concentrations. The driving force for selectivity on the positively charged mixed-mode phase is not clear, but adsorption is likely to play an important role. In addition, the log P data for these isomers found in public databases is often based on calculation and it is difficult to use to evaluate the relative polarity of these isomers. The partitioning coefficients measured in this study can potentially be used for this purpose.
A novel purification strategy based on functionalized melamine sponge for determining multiple pesticides in complex-matrix American Ginseng.
Huang H, Zheng X, Li L … +3 more , Yang D, Liu S, Wang Y
J Chromatogr A · 2026 Jul · PMID 42035507 · Publisher ↗
This study addresses the complicated pretreatment steps in pesticide residue detection of American Ginseng by introducing a simplified QuEChERS purification method. The core of this method is a novel composite material (...
This study addresses the complicated pretreatment steps in pesticide residue detection of American Ginseng by introducing a simplified QuEChERS purification method. The core of this method is a novel composite material (PSA/WT@MeS) made by loading PSA and multi-walled carbon nanotubes onto a three-dimensional melamine sponge. This material effectively removes multiple interferents from complex matrices. When used in the QuEChERS workflow, it allows purification and separation of American Ginseng samples in a single step, greatly streamlining the traditional process. Combined with gas chromatography-mass spectrometry (GC-MS), 12 common pesticides in American Ginseng were measured. The method showed good linearity (R² > 0.99) for all 12 pesticides across the 1-20 µg/kg range. Detection limits ranged from 0.01 to 0.12 µg/kg, and quantification limits from 0.03 to 0.40 µg/kg. Matrix effects were kept within ±20%. At three spiked levels, average recoveries ranged from 87.8% to 109.8%, with relative standard deviations below 9.9%. This approach offers faster pretreatment and better purification. It provides a practical option for detecting pesticide residues in American Ginseng and other complex matrices.
Analytical challenges and occurrence of antibiotics in biosolids from municipal sewage treatment plants in North Spain.
Barros A, Castro G, Ramil M … +1 more , Rodríguez I
J Chromatogr A · 2026 Jul · PMID 42033873 · Publisher ↗
Antibiotics are among the most concerning pharmaceutical contaminants released from municipal sewage treatment plants (STPs), occurring both in treated effluents and in dewatered biosolids. This study examines key analyt...
Antibiotics are among the most concerning pharmaceutical contaminants released from municipal sewage treatment plants (STPs), occurring both in treated effluents and in dewatered biosolids. This study examines key analytical challenges during the determination of ten antibiotics in sewage wastes using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Furthermore, results of their distribution in dewatered biosolids from different STPs are presented. Fluoroquinolones were identified as the most problematic compounds due to sorption on glassware, strong interaction with sample matrix, and signal suppression effects during LC-MS/MS analysis. Optimized extraction, based on sonication of freeze-dried samples with a buffered acetonitrile-water solution (pH 4.4, 1:1), yielded average recoveries between 74% and 108%, with moderate variability across sludge types. During simultaneous quantification of multiclass antibiotics, fluoroquinolones exhibited moderate to high signal attenuation depending on the matrix. For this group of compounds, signal suppression could be mitigated by fractionating extracts using mixed-mode (reversed phase and cation exchange) sorbents. Combined with isotopically labelled surrogate standards, solvent based calibration enabled accurate quantification of all targeted compounds, achieving limits of quantification below 5 ng g⁻¹. Azithromycin, clarithromycin, norfloxacin, ciprofloxacin and ofloxacin were ubiquitous in dewatered biosolids, with median concentrations ranging from 7 ng g (clarithromycin) to 1761 ng g (ofloxacin). A mass balance assessment of emissions through treated wastewater and biosolids highlighted azithromycin, ciprofloxacin, norfloxacin, and ofloxacin as the priority antibiotics for monitoring in final dewatered solid waste streams (biosolids) from STPs.
Affinity membranes in downstream bioprocessing: From chemical design rules to ligand engineering.
Gao P, Smet M, Dehaen W … +2 more , Zydney AL, Yang X
J Chromatogr A · 2026 Jul · PMID 42033872 · Publisher ↗
Affinity membrane chromatography (MC) offers a compelling alternative to resin-based separations by coupling convective transport with molecularly selective ligand-protein recognition. Despite decades of development, aff...
Affinity membrane chromatography (MC) offers a compelling alternative to resin-based separations by coupling convective transport with molecularly selective ligand-protein recognition. Despite decades of development, affinity MC has achieved only limited performance gains, largely because ligand immobilization and functional-layer construction remain chemically underdefined. Binding capacity and selectivity are still governed by empirically chosen reaction pathways rather than predictive design principles. This review reframes affinity MC through the lens of chemical design rules, elucidating how interfacial reaction mechanisms, functional-layer architecture, and ligand molecular structure collectively control affinity performance. Guided by the aim to overcome the selectivity-permeability trade-off, we analyze how immobilization chemistries dictate ligand orientation, accessibility, and alkaline stability, how spatial organization within functional layers regulates effective binding, and how rational ligand engineering can overcome intrinsic limitations. Importantly, we highlight how controlling reaction orthogonality and interfacial kinetics enables the construction of chemically defined affinity layers that are compatible with high-surface-area membrane architectures. By integrating advances in surface chemistry, polymer reaction engineering, and biomolecular recognition, we establish a set of chemical design rules that shift affinity MC from empirical optimization toward rational materials design. This framework provides a foundation for developing next-generation affinity membranes with enhanced capacity, durability, and translational relevance in downstream bioprocessing.
Mechanism of peak fronting in ion exclusion chromatography.
Novič M, Haddad PR
J Chromatogr A · 2026 Jul · PMID 42033871 · Publisher ↗
Ion exclusion chromatography (IEC) is a distinctive separation technique for weakly acidic analytes that can use pure water as the eluent. Under these conditions, analytes are eluted with a fronted peak shape. A thorough...
Ion exclusion chromatography (IEC) is a distinctive separation technique for weakly acidic analytes that can use pure water as the eluent. Under these conditions, analytes are eluted with a fronted peak shape. A thorough investigation of the conversion of IEC columns (fully functionalised 8% crosslinked styrene-divinylbenzene copolymer substrate) from the aqueous phase to the appropriate analyte form and back to the water form revealed that the IEC column capacity is dynamic. Specifically, a certain quantity of analyte is retained by the stationary phase only when a specific amount of the same analyte is present in the mobile phase. This characteristic is referred to as the dynamic column capacity. Moreover, dynamic column capacity is influenced by both the type and concentration of the analyte. The studied stationary phase shows linear concentration-dependent dynamic capacity, which was found to be significantly greater for aromatic and heteroaromatic organic acids in comparison to those of aliphatic organic acids. Through rigorous experimentation, it has been determined that the dynamic column capacity is the primary factor governing the amount of dynamically retained fraction of the analyte by the stationary phase as the injected sample progresses along the column. Consequently, characteristic peak fronting is generated, with the peak initiating at the column void volume. The resultant peak fronting curves (for differing concentrations of the selected analyte) follow the same fronting curve up to the inflection point where the individual concentration reach steady-state level. As the IEC column exhibits a dynamic column capacity specific to each analyte, the profile of the analyte in the eluent at the column exit is determined by the characteristics of each active component, as exemplified here by a single organic acid. Additionally, it was observed that owing to this phenomenon, an identical mass of injected analyte (generated by using increased volumes with decreased concentration of the analyte) results in identical chromatographic peaks.
Retention modeling of oligonucleotides on an amide-based HILIC column: A descriptor-driven approach.
Maurer J, Addepalli B, Gritti F … +3 more , Lauber MA, Guillarme D, Fekete S
J Chromatogr A · 2026 Jul · PMID 42030656 · Publisher ↗
Hydrophilic interaction chromatography (HILIC) has recently gained attention as a powerful tool for the analysis of nucleic acid-based therapeutics, offering high resolving power and excellent compatibility with mass spe...
Hydrophilic interaction chromatography (HILIC) has recently gained attention as a powerful tool for the analysis of nucleic acid-based therapeutics, offering high resolving power and excellent compatibility with mass spectrometry. However, the successful development of HILIC methods requires a clear understanding of the underlying retention mechanisms. Despite numerous studies, the respective contributions of hydrophilic partitioning, polar interactions, and ionic interactions to oligonucleotide retention remain poorly understood. In this work, the role of hydrogen bonding, dipole-dipole interactions, solvophobic effects, and ionic interactions in governing oligonucleotide retention have been systematically investigated. A series of targeted design-of-experiments studies, performed on a representative panel of samples to investigate the impact of oligonucleotide structure on retention, provided a refined framework for interpreting HILIC separations of oligonucleotides. All experiments were performed on an amide-bonded stationary phase, and the proposed descriptor-based retention model applies specifically to this surface chemistry. Our results demonstrate that solvophobic effects and hydrogen bonding are the dominant drivers of retention, whereas hydrophilic partitioning contributes negligibly under practical conditions. This mechanistic insight has important consequences for method development: introducing protic solvents into the mobile phase, conditions that would disrupt the water-rich layer, substantially increases selectivity and enables the resolution of closely related oligonucleotide species. Altogether, these findings shift the conceptual basis of HILIC for nucleic acids from a hydrophilic partitioning/interaction model toward a retention mechanism best described as a solvophobic-interaction‑dominated HILIC regime, in which protic solvents assist selectivity through H‑bond competition. This new perspective provides a more accurate foundation for designing, optimizing, and interpreting HILIC methods for oligonucleotide analysis.
A compact visible-light spectrometric detector for capillary electrophoresis.
Rütter V, Kaljurand M, Gorbatsova J … +3 more , Pardy T, Scheler O, Mazina-Šinkar J
J Chromatogr A · 2026 Jul · PMID 42030655 · Publisher ↗
This article presents a compact and portable capillary electrophoresis (CE) detector that uses a light-emitting diode (LED) for excitation and a Hamamatsu C12880MA mini spectrometer for detecting substances that absorb v...
This article presents a compact and portable capillary electrophoresis (CE) detector that uses a light-emitting diode (LED) for excitation and a Hamamatsu C12880MA mini spectrometer for detecting substances that absorb visible light. The detector features a robust design, including an adjustable slit that allows for precise optical alignment with the capillary detection window, thereby reducing interference from scattered LED light. All parts are housed within a custom 3D-printed enclosure. Unlike standard portable CE detectors, this device captures full absorbance spectra for better compound identification. The custom instrument separated food colorants with dye detection limits near 40 μg/mL. Results were linear from 20-1000 μg/mL, with correlation coefficients over 0.999 and peak height RSDs of 5-8%.
Evaluation of chromatographic resins for high efficacy separation of radium from chemical analogues.
Coupannec M, Kmak KN, Despotopulos JD … +1 more , Sudowe R
J Chromatogr A · 2026 Jul · PMID 42030654 · Publisher ↗
The separation of Radium (Ra) from interfering chemical analogues is of major importance for both the accurate measurement of Ra-226 in environmental samples and the preparation of radium targets destined for medical iso...
The separation of Radium (Ra) from interfering chemical analogues is of major importance for both the accurate measurement of Ra-226 in environmental samples and the preparation of radium targets destined for medical isotope production. The adsorption behavior of Ra, Barium (Ba), and Strontium (Sr) was investigated on multiple crown-ether-based proprietary extraction chromatographic resins developed by TrisKem Int. Among these, the TK101 resin, characterized by a novel room-temperature ionic liquid (RTIL) dual extractant-solvent system, demonstrated enhanced affinity and selectivity, particularly at low acid concentrations, achieving a D of 3477 ± 73 for Ra in 0.04 M HNO₃, whereas the maximum Ra D obtained with the traditional Sr resin in 8 M HNO was only 12.33 ± 0.4. In 0.04 M HNO, a Ra distribution ratio (D) of 776 was achieved within 1 min, indicating rapid uptake kinetics. The adsorption capacities were estimated at 19.6 ± 0.1 mg/g for Ba and 12.0 ± 0.2 mg/g for Sr. However, competing alkali metals significantly reduced alkaline earth metal uptake, indicating that the TK101 resin is best suited for radium purification following a preconcentration step to minimize matrix effects in high salinity samples. Dynamic column studies confirmed the strong chromatographic performance of the TK101 resin, with no significant Ba or Sr breakthrough in the Ra fraction and a Ra recovery of 87 ± 2.8%. These results demonstrate the potential of the TK101 resin for selective radium separation in analytical and radiochemical applications.
Analysis of phenotypic quality differences between green and purple stems of Paris. polyphylla var. yunnanensis based on ATR-FTIR, LC-MS/MS and HPLC.
Ma H, Zhu X, Wang Y
J Chromatogr A · 2026 Jul · PMID 42030653 · Publisher ↗
Paris polyphylla var. yunnanensis is a perennial herb with rhizome as medicinal part. However, the existing research on its above-ground parts is still insufficient, especially the biological phenotypic evaluation and qu...
Paris polyphylla var. yunnanensis is a perennial herb with rhizome as medicinal part. However, the existing research on its above-ground parts is still insufficient, especially the biological phenotypic evaluation and quality evaluation that can directly reflect the pharmacodynamic effect is still unclear, which limits the in-depth exploration of its medicinal value. To this end, this study characterizes the spectral differences between distinct parts of two phenotypes of P. polyphylla var. yunnanensis using attenuated total reflection fourier transform infrared (ATR-FTIR) spectra, obtaining functional group information related to the chemical constituents of P. polyphylla var. yunnanensis. Comprehensive study on the overall chemical characteristics of different parts of P. polyphylla var. yunnanensis. Subsequently, liquid chromatography-tandem mass spectrometry (LC-MS/MS) was then used to target differences in flavonoid metabolites. Through distinctive metabolite analysis, We found that Isorhamnetin-3-O-neohesperidoside was significantly upregulated in the PS (Purple stem of P. polyphylla var. yunnanensis) group compared to the GS (Green stem of P. polyphylla var. yunnanensis) group, and it was the only unique differential metabolite present exclusively in both the PS and GS groups when compared to other groups. In the KEGG analysis, the AMPK signaling pathway was uniquely enriched in both the PS and GS groups. To understand the quality differences between the two phenotypes, we determined the content of nine saponins in different parts of P. polyphylla var. yunnanensis using high-performance liquid chromatography (HPLC). The results showed that the groups with higher total saponin content among all groups were GL (Green leaf of P. polyphylla var. yunnanensis) and PL (Purple leaf of P. polyphylla var. yunnanensis), both of which were derived from the above ground parts. Meanwhile, Polyphyllin I exhibited higher levels in the PR (Purple rhizome of P. polyphylla var. yunnanensis) and GR (Green rhizome of P. polyphylla var. yunnanensis) groups. Finally, we combined ATR-FTIR and HPLC to construct a PLSR quantitative prediction model, achieving rapid evaluation of three quality control indicators.
Development of a poly(2,6-diaminopyridine)-coated magnetic porous carbon nanosorbent for simultaneous dispersive micro-solid phase extraction of buprenorphine and naloxone from biological fluids.
Mohammadi F, Dadfarnia S, Haji Shabani AM … +1 more , Asgharinezhad AA
J Chromatogr A · 2026 Jul · PMID 42030652 · Publisher ↗
The design of efficient, environmentally friendly, and stable nanosorbents capable of simultaneous extraction of analytes with diverse chemical properties remains a significant challenge in analytical chemistry. This stu...
The design of efficient, environmentally friendly, and stable nanosorbents capable of simultaneous extraction of analytes with diverse chemical properties remains a significant challenge in analytical chemistry. This study reports the synthesis and application of a novel poly(2,6-diaminopyridine)-coated magnetic porous carbon (MPC@P2,6-DAP) material, derived from ZIF-67 via high-temperature pyrolysis, as an effective nanosorbent for dispersive micro-solid phase extraction (D-µSPE) of buprenorphine and naloxone from biological matrices. The physicochemical characteristics of the synthesized sorbent were comprehensively evaluated using scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), and vibrating sample magnetometry (VSM). Optimal extraction conditions were established at pH 8.7, a sorption time of 8 min, 30 mg sorbent dosage, 5% (w/v) NaCl concentration, and elution with 122 µL of ACN-MeOH (1:1 v/v). Under these conditions, detection limits of 0.10 and 0.15 µg L⁻¹, with linear dynamic ranges of 0.4 -750 and 0.5-750 µg L⁻¹ for buprenorphine and naloxone were obtained, respectively. The method exhibited excellent intra-day and inter-day precisions, with relative standard deviations of ≤ 6.0% and ≤ 10.2%, respectively. Application to biological samples demonstrated satisfactory recoveries between 95.0 and 101.8%, confirming the suitability of the sorbent for rapid, sensitive, and green extraction of target opioids.
Evaluation and application of a chlorobenzimidazole bonded stationary phase for polyphenols separation in supercritical fluid chromatography.
Song C, Feng J, Xia D … +6 more , Jin G, Yu D, Liu Y, Liu Y, Guo Z, Liang X
J Chromatogr A · 2026 Jul · PMID 42025607 · Publisher ↗
Polyphenols are widely present in natural products and exhibit significant biological activities. However, their structural similarity and diversity pose challenges to efficient separation. Supercritical fluid chromatogr...
Polyphenols are widely present in natural products and exhibit significant biological activities. However, their structural similarity and diversity pose challenges to efficient separation. Supercritical fluid chromatography (SFC) is a promising separation technique, yet commercial stationary phases lack sufficient selectivity due to limited functional groups. Therefore, there is an urgent need to develop multifunctional stationary phases to meet the separation requirements of complex polyphenol samples. This study included the design and synthesis of a new multifunctional bonded silica stationary phase (CMBZ), which incorporates hydroxyl, thioether, benzene ring, imidazole, and chlorine groups. Evaluation was performed using fifty-three structurally diverse polyphenolic standards, and the results demonstrated that CMBZ exhibited a more versatile retention mechanism and superior selectivity compared to commercial SFC columns. This research presents a promising approach for the effective separation of polyphenols. More significantly, it presents innovative pathways for research on novel SFC bonded stationary phases.
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