J Sep Sci [JOURNAL]
Sun
200 papers
Microwave-Assisted Meta-Chloroperoxybenzoic Acid Epoxidation for Qualitative and Quantitative Analysis of Unsaturated Lipid Double-Bond Positional Isomers by Liquid Chromatography-Tandem Mass Spectrometry.
Liu D, Chen H, Huang Y … +1 more , Zhou T
J Sep Sci · 2026 Jun · PMID 42240584 · Publisher ↗
Unsaturated lipid double-bond (C═C) positional isomers are closely related to lipid structure and biological function, but their reliable qualitative and quantitative analysis remains challenging by conventional liquid c...
Unsaturated lipid double-bond (C═C) positional isomers are closely related to lipid structure and biological function, but their reliable qualitative and quantitative analysis remains challenging by conventional liquid chromatography-mass spectrometry (LC-MS) methods. In this study, we developed a microwave-assisted meta-chloroperoxybenzoic acid (m-CPBA) epoxidation method coupled with LC-tandem MS for the identification and quantification of unsaturated lipid C═C positional isomers. The reaction conditions were optimized to facilitate the formation of fully epoxidized products, simplify the chromatographic peak profiles, and enhance their suitability for targeted quantitative analysis. The developed method showed good analytical performance, including satisfactory linearity (r > 0.99), precision (relative standard deviation <15%), recovery (75.5%-106.5%), and acceptable matrix effects (86.0%-117.3%). The method was successfully applied to rat brain tissue and plasma samples from a rat depression model, enabling the quantification of multiple unsaturated lipids and the characterization of their C═C positional isomers in complex biological matrices. These results demonstrate that microwave-assisted m-CPBA epoxidation provides a rapid, efficient, and practical strategy for LC-MS-based analysis of unsaturated lipid isomers.
Hydrogen-bonded Organic Frameworks Decorated Polymeric High Internal Phase Emulsions as Adsorbents for Solid-Phase Extraction of Estrogens.
Wei J, Yu S, Ruan Y … +4 more , Wang H, Xie M, Ruan G, Huang Y
J Sep Sci · 2026 Jun · PMID 42237672 · Publisher ↗
Due to lipid-rich and severe matrix interference, the accurate determination of trace estrogens in cosmetics is highly challenging. Although solid-phase extraction (SPE) is widely used to mitigate the matrix interference...
Due to lipid-rich and severe matrix interference, the accurate determination of trace estrogens in cosmetics is highly challenging. Although solid-phase extraction (SPE) is widely used to mitigate the matrix interference, conventional adsorbents are often limited to cosmetic matrices. To this end, polymeric high internal phase emulsions modified with a hydrogen-bonded organic framework (polyHIPEs@HOF) were used as an adsorbent for the SPE of five estrogenic compounds. Benefiting from the hypercrosslinked architecture with interconnected macroporosity of polyHIPEs and hydrophilicity of HOF, the synthesized composites were suitable for SPE of estrogen in a cosmetic sample. Under optimized conditions, polyHIPEs@HOF enabled the rapid extraction of five estrogens from complex cosmetic samples, achieving adsorption equilibrium within 4 min, and the extraction recoveries exceeded 78.28%. The adsorption capacities for estrogens ranged from 8.44 to 15.39 mg g, and the adsorption involved multiple interactions (hydrogen bonding, π-π stacking, and hydrophobic effects). Furthermore, the polyHIPEs@HOF composite was able to eliminate the interference of the complex cosmetic matrix, and the extraction efficiency remained stable over 10 consecutive reuse cycles. These results confirm that the polyHIPEs@HOF composite is a highly efficient and reusable adsorbent for the extraction of estrogens in complex sample matrices.
Capillary Electrophoresis Immunoassay for Detection of Anti-insulin Antibodies.
James VD, Cunningham SK, Roper MG
J Sep Sci · 2026 Jun · PMID 42220215 · Full text
Type 1 diabetes mellitus is an autoimmune disorder primarily affecting children and adolescents. Diagnostic techniques for autoimmune diseases typically examine the amounts of autoantibodies (AAbs) and are performed usin...
Type 1 diabetes mellitus is an autoimmune disorder primarily affecting children and adolescents. Diagnostic techniques for autoimmune diseases typically examine the amounts of autoantibodies (AAbs) and are performed using enzyme-linked immunosorbent assays and radioimmunoassays. Insulin autoantibody (IAA) is one of the earliest biomarkers in juveniles prone to type 1 diabetes; therefore, the development of highly sensitive assays could enhance early diagnostic capabilities. To this end, we developed a capillary electrophoresis laser-induced fluorescence noncompetitive immunoassay for the detection of monoclonal anti-insulin Abs as a first step toward the development of an IAA assay. In this study, a fluorescently labeled insulin was used as an affinity probe to target anti-insulin Abs. In the presence of an Ab, the separated mixture showed two distinct peaks correlating to bound and free labeled insulin. Sample incubation and separation parameters were optimized, and produced a detection limit (LOD) of 1.5 ng/mL (∼10 pM) of Ab. To further demonstrate the utility of the assay, calibration in a serum matrix was also performed and showed a linear response. The assay was also tested using a polyclonal Ab, which resulted in a dose-dependent decrease in the labeled insulin peak, producing an LOD of 170 ng/mL (∼1 nM). This robust system provides a foundation for further development of an IAA assay.
Systematic Chemical Profiling of Allii macrostemonis Bulbus Based on Ultra-High-Performance Liquid Chromatography-Quadrupole-Exactive Orbitrap Mass Spectrometry.
Yan SL, Li Q, Tan JJ … +3 more , Wen W, Zeng ZH, Wang LP
J Sep Sci · 2026 Jun · PMID 42212502 · Publisher ↗
This study aimed to systematically characterize the chemical composition and develop a reliable strategy for identifying compounds in Allii macrostemonis Bulbus (AMB). This study employed an efficient, highly sensitive,...
This study aimed to systematically characterize the chemical composition and develop a reliable strategy for identifying compounds in Allii macrostemonis Bulbus (AMB). This study employed an efficient, highly sensitive, and high-resolution analytical approach utilizing ultra-high-performance liquid chromatography-Quadrupole-Exactive Orbitrap mass spectrometry with parallel reaction monitoring scanning. A total of 130 compounds were preliminarily discovered in the AMB. The 130 compounds included 77 steroidal saponins, 23 nitrogen-containing compounds, 14 organic acids, 14 amino acids, and two other compounds, by comparing with reference substances, secondary characteristic ion fragments, public databases, and literature. Among these, A total of 77 compounds were reported for the first time. This result significantly expands the knowledge of the chemical constituents of AMB and contributes to the understanding of its active constituents for quality control.
Magnetic Boric Acid-activated Biochar Adsorbents for Efficient Extraction of Chloramphenicol in Honey Samples.
Tang A, Cao Y, Xiao X … +2 more , Guo M, Lin B
J Sep Sci · 2026 Jun · PMID 42212490 · Publisher ↗
Single biochar materials have limitations, including limited adsorption capacity, poor selectivity, and poor recyclability. Some activation methods are needed to modify them to improve their performance. Boric acid modif...
Single biochar materials have limitations, including limited adsorption capacity, poor selectivity, and poor recyclability. Some activation methods are needed to modify them to improve their performance. Boric acid modification offers unique advantages, including increased specific surface area, optimized pore structure, introduction of oxygen-containing functional groups, and improved yield. Herein, a magnetic boric acid-activated biochar was developed as an adsorbent for magnetic solid-phase extraction in combination with liquid chromatography for the efficient extraction and sensitive determination of trace amounts of chloramphenicol in honey samples. The developed method showed good linearity in the range of 0.1-500 µg·L with R of 0.9999, limit of detection (signal-to-noise ratio [S/N] = 3) of 0.01 µg·L, and limit of quantification (S/N = 10) of 0.03 µg·L. The spiked recovery rate ranged from 82.0% to 107.8%, and the relative standard deviation ranged from 0.5% to 10.0%.
An Effective and Systematic Strategy for Metabolic Profiling of Golvatinib In Vitro by Combining UHPLC-MS/MS and UHPLC-Q-Orbitrap-HRMS.
Cao L, Zhang T, Zhu Y … +2 more , Wang X, Chang J
J Sep Sci · 2026 Jun · PMID 42210592 · Publisher ↗
Golvatinib is a tyrosine kinase inhibitor in development for treating solid tumors such as advanced hepatocellular carcinoma. The present study aimed to clarify the metabolic fates of golvatinib using human and rat liver...
Golvatinib is a tyrosine kinase inhibitor in development for treating solid tumors such as advanced hepatocellular carcinoma. The present study aimed to clarify the metabolic fates of golvatinib using human and rat liver microsomes. For this purpose, a comprehensive and systematic workflow was established for investigating the in vitro metabolic fates. Separation was performed on a Waters ACQUITY BEH C column with a mobile phase consisting of 0.1% formic acid in water and acetonitrile. Detection was carried out by multiple reaction monitoring mode using the transitions m/z 634.3→184.2 for golvatinib and m/z 650.3→200.2 for golvatinib N-oxide. This method exhibited excellent linearity across a concentration range of 1.0-2000 nM (r > 0.995). The validated assay was subsequently applied to evaluate the metabolic stability of golvatinib. The results revealed that golvatinib underwent rapid metabolism in rat liver microsomes (t = 13.76 ± 0.79 min), while its turnover was considerably slower in human liver microsomes (t = 56.93 ± 3.17 min). Furthermore, twelve metabolites were identified using Orbitrap high-resolution mass spectrometry (HRMS) via accurate mass determination and fragmentation analysis. Major metabolic pathways of golvatinib encompass oxygenation, N-demethylation, N-oxidation, N-hydroxylation, dealkylation, oxidative deamination, and oxidative defluorination. Further study demonstrated that CYP3A4 was the principal enzyme involved in metabolizing golvatinib. To the best of our knowledge, this is the first report combining ultra-high-performance liquid chromatography-tandem MS (UHPLC-MS/MS) with UHPLC-Quadrupole-Orbitrap-HRMS for profiling golvatinib metabolism in vitro, thereby laying a foundation for subsequent pharmacokinetic study.
Analysis of Chemical Components in Tangerine Peel Flower Tea by Ultra-Performance Liquid Chromatography-Q-Exactive Orbitrap Tandem Mass Spectrometry and High-Performance Liquid Chromatography-Photodiode Array.
Deng X, Zeng Y, Deng J … +8 more , Peng M, Chen B, Xu J, Wang Y, Wei M, Cai Y, Li L, Zheng G
J Sep Sci · 2026 May · PMID 42204832 · Publisher ↗
Tea has long been consumed in China as a significant part of wellness practices. Tangerine Peel Flower Tea, meticulously developed as a beverage with throat-moistening and liver-protecting functions, is formulated with t...
Tea has long been consumed in China as a significant part of wellness practices. Tangerine Peel Flower Tea, meticulously developed as a beverage with throat-moistening and liver-protecting functions, is formulated with tangerine peel and licorice as its core ingredients, supplemented with other medicinal and edible herbs such as chrysanthemum and jujube. However, a comprehensive understanding of its chemical composition remains to be elucidated. In this study, the chemical component of Tangerine Peel Flower Tea was comprehensively analyzed using ultra-performance liquid chromatography-Q-Exactive Orbitrap tandem mass spectrometry. A total of 134 components were preliminarily identified through the integration of chromatographic retention time, m/z values, and secondary fragments. These encompassed 80 flavonoids and their glycosides, 16 terpenoids, eight phenylpropanoids, five alkaloids, and 25 other components. Additionally, a high-performance liquid chromatography-photodiode array (HPLC-PDA) was established to enable the concurrent quantification of six main constituents in 20 batches of Tangerine Peel Flower Tea. The results showed that the contents of the six components ranked from high to low as follows: hesperidin (4.164-10.822 mg·g), glycyrrhizic acid (1.463-5.953 mg·g), nobiletin (0.831-1.637 mg·g), tangeretin (0.558-1.160 mg·g), chlorogenic acid (0.456-1.037 mg·g), and ferulic acid (0.084-0.208 mg·g). The established HPLC method demonstrated acceptable feasibility and reliability through methodological validation. This study promotes the quality standardization of Tangerine Peel Flower Tea and supports the further development and utilization of tangerine peel.
Amino Acid Oligomer Brushes as Chiral Stationary Phases: Effects of Side-Chain Architecture and Mobile-Phase Modifiers on Enantioseparation Behavior.
Cai C, Cao M, Zhang B … +1 more , Ke Y
J Sep Sci · 2026 May · PMID 42178869 · Publisher ↗
In this study, a series of oligomer-brush chiral stationary phases was developed to systematically investigate the influence of amino acid side chains on enantioseparation. Through a structure-preserving "grafting-to" st...
In this study, a series of oligomer-brush chiral stationary phases was developed to systematically investigate the influence of amino acid side chains on enantioseparation. Through a structure-preserving "grafting-to" strategy, three well-defined chiral oligomers derived from l-proline, l-phenylalanine, and l-leucine were covalently immobilized onto silica microspheres. The enantioseparation capabilities of these chiral stationary phases were comprehensively evaluated using 27 representative racemates under normal-phase conditions. The results revealed that the side-chain architecture plays a decisive role in the chiral recognition microenvironment. The stationary phase with a rigid pyrrolidine ring (CSP-P(Pro-An)) demonstrated the broadest applicability for bulky, conformationally restricted analytes and was the only stationary phase capable of resolving the tested axially chiral metal complexes. The stationary phase with aromatic side chains (CSP-P(Phe-An)) showed enhanced selectivity for specific π-rich analytes, while the stationary phase with a flexible isobutyl side chain (CSP-P(Leu-An)) provided the highest separation factors for several 3,5-dinitrobenzoyl-derivatized amino acids. Furthermore, an investigation into the influence of mobile-phase modifiers found that, compared to other aprotic modifiers such as ethyl acetate, tetrahydrofuran, and dichloromethane, methyl tert-butyl ether provided a more optimal balance between retention and enantioselectivity. This is attributed to the steric shielding effect produced by the tert-butyl group of methyl tert-butyl ether, which minimizes its competitive interactions for essential hydrogen-bonding sites. These results indicate that both the side-chain structure and the choice of mobile-phase modifier are key parameters for tailoring the performance of amino acid-based oligomer-brush chiral stationary phases.
Development of a Sustainable and Practical Natural Deep Eutectic Solvent-based Ultrasound-assisted Microextraction Method for Determining Fipronil and Its Degradation Products in Brewed Coffee.
Bierhals NS, Castro ES, Monteiro SA … +2 more , Lando VR, Merib J
J Sep Sci · 2026 May · PMID 42175847 · Publisher ↗
Fipronil and its degradation products, such as fipronil sulfide and fipronil desulfinyl, exhibit toxicological concerns and require reliable analytical methods for monitoring in complex matrices such as brewed coffee. Th...
Fipronil and its degradation products, such as fipronil sulfide and fipronil desulfinyl, exhibit toxicological concerns and require reliable analytical methods for monitoring in complex matrices such as brewed coffee. This study aimed to develop a novel ultrasound-assisted liquid-liquid microextraction approach using a natural deep eutectic solvent (NADES), followed by high-performance liquid chromatography-diode array detection, for the determination of these compounds in brewed coffee. The optimized extraction parameters obtained through a central composite design consisted of a sample volume of 4.2 mL, 130 µL of NADES as extraction solvent, and an extraction time of 9.2 min. The method was validated, achieving coefficients of determination (R) higher than 0.9973 and limits of detection and quantification of 4.5 and 15.0 µg L, respectively, for all analytes. Intraday precision ranged from 6.1% to 13.7%, interday precision from 12.1% to 17.6%, and accuracy (relative recoveries) from 95.8% to 109.3%. The method featured low solvent consumption, reduced waste generation, and high analytical performance, offering a sustainable alternative for monitoring these compounds. High sustainability and practicality were demonstrated by scores of 8.11 and 67.5, obtained using the Sample Preparation Metric of Sustainability and the Blue Applicability Grade Index, respectively. Finally, the method was successfully applied to seven brewed coffee samples purchased in Porto Alegre, Brazil, confirming its applicability as a green workflow in food quality monitoring.
Synergistic Mixed-Micelle Extraction Coupled With Cloud Point Enrichment for Simultaneous Recovery of Camptothecin Alkaloids and Hyperoside From Camptotheca acuminata Seeds.
Zhou J, Qu B, Zhang Z … +7 more , Gao X, Li X, Liang J, Fan X, Yuan X, Fu Y, Gu C
J Sep Sci · 2026 May · PMID 42175839 · Publisher ↗
An integrated and eco-friendly method for the simultaneous extraction and enrichment of camptothecin (CPT) alkaloids and hyperoside (HYP), including CPT, 10-hydroxycamptothecin (HCPT), 10-methoxycamptothecin (MCPT), and...
An integrated and eco-friendly method for the simultaneous extraction and enrichment of camptothecin (CPT) alkaloids and hyperoside (HYP), including CPT, 10-hydroxycamptothecin (HCPT), 10-methoxycamptothecin (MCPT), and HYP from Camptotheca acuminata seeds, was developed using mixed-micelle extraction and enrichment by cloud point property (MME-ECP). A mixed-surfactant system, optimally formulated with 4% Triton X-114 and 4% Genapol X-080, demonstrated superior extraction efficiency for the four target compounds when synergistically combined with ultrasonic assistance. The cloud point behavior of the surfactants facilitated the enrichment of the four compounds. Various experimental conditions were investigated to evaluate and optimize the extraction and preconcentration process. Under optimal conditions, the extraction yields of CPT, HCPT, MCPT, and HYP reached 1007.62, 1408.61, 268.45, and 157.67 µg/g, respectively. Compared to traditional methanol/ethanol extraction, the mixed-micelle method demonstrates superior performance, especially over ethanol extraction, with an extraction yield enhancement of 40.20%-168.81%. Furthermore, the surfactants were successfully recycled from the extraction medium using AB-8 macroporous adsorption resin, achieving high recovery rates of 83.32% for T-114 and 87.36% for GX-080. Overall, this study highlights the integrated MME-ECP method as an efficient, economical, and eco-friendly approach for extracting and enriching the key active compounds from C. acuminata seeds. This method holds great promise as an innovative extraction technique, with considerable potential for large-scale industrial use.
A Modified QuEChERS Combined With HPLC-MS/MS for the Determination of Emamectin Benzoate, Spinetoram, and Their Major Metabolites in Eggplant Field Samples.
Pei T, Wang X, Hu J
J Sep Sci · 2026 May · PMID 42153591 · Publisher ↗
To control thrips in eggplant cultivation, a new pesticide formulation (9% emamectin benzoate·spinetoram microemulsion) is under registration in China. However, the residues of emamectin benzoate (EMB), spinetoram, and p...
To control thrips in eggplant cultivation, a new pesticide formulation (9% emamectin benzoate·spinetoram microemulsion) is under registration in China. However, the residues of emamectin benzoate (EMB), spinetoram, and prime metabolites of spinetoram in eggplant remained unclear. In this study, field trials were performed in accordance with good agricultural practice across eight provinces of China. The pesticide formulation was sprayed once at 13.5 g a.i./ha, and the recommended pre-harvest interval was 7 days. For the simultaneous determination of EMB, spinetoram, and its metabolites in eggplant, a modified quick, easy, cheap, effective, rugged, and safe method combined with high-performance liquid chromatography tandem mass spectrometry was developed and validated. The combination of 25 mg of primary-secondary amine, 25 mg of Nano-ZrO, and 5 mg of multi-walled carbon nanotubes was applied to purify the eggplant extracts. Method validation demonstrated excellent linearity (r ≥ 0.9991), acceptable accuracy with recoveries of 82.7%-102.6%, precision (relative standard deviations ≤7.7%), sensitivity (limits of detection, 6.59 × 10-3.06 × 10 ng; limits of quantification, 0.0025-0.01 mg/kg), and matrix effects (≤25.2%). When harvested after the recommended 7-day interval, the residues of EMB (<0.01 mg/kg) and spinetoram (≤0.025 mg/kg) in eggplant were lower than the maximum residue limits in China. The residues of metabolites N-formyl-XDE-175-J and N-demethyl-XDE-175-J in eggplant were lower than the limits of quantification (0.01 mg/kg). This study offers recommendations for the effective use of this pesticide formulation on eggplant.
Elucidating the Material Basis and Mechanism of Shaoyao Gancao Decoction Against Alcoholic Liver Disease via Ultra-High-Performance Liquid Chromatography-Mass Spectrometry and Network Pharmacology.
Li H, Pi M, Hu J … +4 more , Liang Y, Wu Q, Guo Y, Li B
J Sep Sci · 2026 May · PMID 42138455 · Publisher ↗
Alcoholic liver disease (ALD) is a common hepatic disorder. Shaoyao Gancao Decoction (SGD) can be used to treat ALD, but its material basis and mechanism of action are not yet fully understood. In this study, ultra-high-...
Alcoholic liver disease (ALD) is a common hepatic disorder. Shaoyao Gancao Decoction (SGD) can be used to treat ALD, but its material basis and mechanism of action are not yet fully understood. In this study, ultra-high-performance liquid chromatography-mass spectrometry was employed to identify 49 components in Paeoniae Radix Alba extract and 101 components in Glycyrrhizae Radix et Rhizoma, respectively. Furthermore, 147 components were identified in the SGD extract, primarily including flavonoids, terpenoids, phenolics, coumarins, and organic acids. Further analysis detected 14 blood-absorbed components in the plasma of administered rats. Network pharmacology results suggest that SGD may exert its therapeutic effects against ALD by regulating pathways related to inflammation, metabolism, and cell survival, thereby influencing biological processes such as cell proliferation, inflammatory responses, and substance metabolism. Additionally, by integrating six network analysis algorithms, isoliquiritigenin was identified as a core component, and prostaglandin-endoperoxide synthase 2 (PTGS2) as a core target. Molecular dynamics simulations verified that isoliquiritigenin and the PTGS2 protein can bind stably, providing a solid structural basis for its potential mechanism of action. This study comprehensively elucidates the material basis of SGD and preliminarily explores its potential mechanisms, providing a foundation and reference for further research on this formula.
Fabrication of Novel Magnetic Crown Ether-based Microporous Organic Network for Efficient Magnetic Solid-Phase Extraction of Polar Chlorophenols From Environmental Water Samples.
Xu CY, Meng XY, Cui YY … +1 more , Yang CX
J Sep Sci · 2026 May · PMID 42117751 · Publisher ↗
Chlorophenols, polar priority pollutants with carcinogenic, teratogenic, and mutagenic effects, are ubiquitous in environmental waters and pose risks to aquatic organisms and human health. However, their presence at trac...
Chlorophenols, polar priority pollutants with carcinogenic, teratogenic, and mutagenic effects, are ubiquitous in environmental waters and pose risks to aquatic organisms and human health. However, their presence at trace levels and high polarity hinders their selective enrichment and accurate quantification. To address this, we ingeniously devised and constructed a novel crown ether functionalized magnetic microporous organic network (FeO@CE-MON) as an efficient adsorbent for magnetic solid-phase extraction (MSPE) of trace chlorophenols from complex aqueous matrices. Synthesized by coupling brominated 18-crown-6 ether with aromatic alkynyl groups, FeO@CE-MON integrates the conjugated MON framework, molecular recognition capability of crown ether cavities, and magnetic responsiveness of FeO nanoparticles, achieving selective chlorophenols enrichment via hydrophobic, π-π stacking, and hydrogen bonding interactions. After systematically optimizing key MSPE parameters, a sensitive MSPE-high-performance liquid chromatography-ultraviolet method was established, featuring low detection limits (0.3-0.5 µg L), a wide linear range (1.0-500 µg L), excellent reproducibility, and superior enrichment efficiency. This is the first example of fabricating crown ether-based MMON for MSPE, providing a rapid and sensitive method for monitoring trace chlorophenols in complex environmental waters. It also expands MON monomer diversity and highlights the potential of combining supramolecule crown ether with porous MONs in sample pretreatment.
Systematic Analysis of Chemical Constituents in Shenzhuo Decoction Based on Iterative Ultra-High Performance Liquid Chromatography Coupled With Quadrupole Time-of-Flight Tandem Mass Spectrometry and Multidimensional Data Annotation.
Yang Z, Zou D, Zhu J … +7 more , Huang Q, Liu S, Wang X, He X, Wang X, Huang Y, He F
J Sep Sci · 2026 May · PMID 42116565 · Publisher ↗
Shenzhuo Decoction (SZT), a classical traditional Chinese medicine (TCM) formula from Synopsis of the Golden Chamber (Jinkui Yaolue), demonstrates clinical efficacy in treating ankylosing spondylitis (AS); however, its s...
Shenzhuo Decoction (SZT), a classical traditional Chinese medicine (TCM) formula from Synopsis of the Golden Chamber (Jinkui Yaolue), demonstrates clinical efficacy in treating ankylosing spondylitis (AS); however, its systematic chemical profiling remains insufficient. This study established an integrated strategy for chemical constituent analysis of SZT, combining iterative ultra-high performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UHPLC-Q-TOF MS/MS) with multidimensional data annotation. The iterative UHPLC-Q-TOF MS/MS technology employed 20 eV/40 eV dual collision energies, along with three iterative scans and a dynamic exclusion method. The multidimensional data annotation for SZT analysis was derived from an internal database containing 997 constituents, combined with Progenesis QI and SIRIUS. A total of 130 constituents were identified from SZT using the above methods. Compared with a conventional LC-MS identification method (single 20 eV collision energy, no iterative scanning), the iterative strategy identified an additional 33 constituents from SZT, indicating a 34% improvement in detection sensitivity. More importantly, network pharmacology validation revealed that three of the additionally identified constituents (medicarpin, licochalcone A, and acetoxy-methylisoflavone) exhibited high anti-AS pharmacological activity (ranking among the top ten in the 130-component scoring values of SZT), further demonstrating that constituents identified via the iterative strategy more comprehensively reflect the material basis of SZT's pharmacological effects. This strategy establishes a foundation for research into the chemical constituents of SZT and provides a reliable paradigm for complex TCM profiling.
Design, Fabrication, and Application of a Chiral Cyclodextrin Stationary Phase for Enantiomeric Separation and Analysis.
Lv L, Qi C, Yang J … +5 more , Yin Y, Wang Y, Fang Z, Li M, Li Q
J Sep Sci · 2026 May · PMID 42116564 · Publisher ↗
Cyclodextrin-based chiral stationary phases (CSPs) have attracted considerable interest in the field of enantiomeric separation due to their chiral recognition properties. However, the design and preparation of cyclodext...
Cyclodextrin-based chiral stationary phases (CSPs) have attracted considerable interest in the field of enantiomeric separation due to their chiral recognition properties. However, the design and preparation of cyclodextrin-based CSPs with satisfactory application prospects are necessary for further exploration. Herein, we report an effective method for fabricating β-cyclodextrin-derived CSP by chemical bonding of 3,5-dimethylphenylcarbamate-mono-6-ethylenediamine-β-cyclodextrin (i.e., DMPCCB) onto silica microspheres via the diamine linker. After structure characterization, the performances in enantiomeric separation were assessed using chiral drugs, including proton pump inhibitors, flavanones, and dihydropyridine calcium channel blockers, under the reversed-phase mode through high-performance liquid chromatography (HPLC). As a result, DMPCCB chirally recognized the tested analytes, where 11 out of 15 analytes were baseline enantioseparated. The promising enantioselectivity was attributed to the introduction of methyl groups on DMPCCB, resulting in flexible host-guest interactions that enhanced chiral recognition for the tested analytes. Under this background, a novel enantioselective HPLC-tandem mass spectrometry method based on DMPCCB chiral column was developed and validated for the determination of naringenin enantiomers in rat plasma, and successfully applied to a pharmacokinetics study. Findings showed a significant enantioselective absorption and elimination for naringenin enantiomers in both normal and acute alcoholic liver injury rats. This work provided a new idea for the preparation of cyclodextrin-based CSPs, making it a promising application in the analysis of chiral drugs.
Correction to "Simultaneous Determination of 10 Fat-Soluble Vitamins by Ultra-Performance Liquid Chromatography in Multivitamins With Minerals Capsules".
J Sep Sci · 2026 May · PMID 42112747 · Publisher ↗
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Volatilomic Differentiation of Protected-Origin Italian Balsamic Vinegars by HS-SPME-GC×GC-TOFMS.
Malcangi S, Polidoro A, Romagnoli M … +2 more , Cavazzini A, Franchina FA
J Sep Sci · 2026 May · PMID 42112742 · Full text
Balsamic vinegars produced under Protected Designation of Origin (PDO) and Protected Geographical Indication (PGI) schemes differ substantially in raw materials, processing intensity, and aging regimes. Although these di...
Balsamic vinegars produced under Protected Designation of Origin (PDO) and Protected Geographical Indication (PGI) schemes differ substantially in raw materials, processing intensity, and aging regimes. Although these differences are expected to shape their volatile composition, a comprehensive chemical characterization capable of reliably discriminating the two denominations remains analytically challenging. In this study, headspace solid-phase microextraction combined with comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (HS-SPME-GC×GC-TOFMS) was used to characterize PDO and PGI balsamic vinegars and to evaluate the feasibility of volatilomic differentiation between the investigated products. Extraction conditions were comparatively assessed using a pooled quality-control sample by evaluating two SPME fiber coatings (PDMS/DVB and DVB/CAR/PDMS) and a high ionic-strength condition (40% w/v NaCl); the DVB/CAR/PDMS fiber combined with salting-out provided the highest analytical response and was selected for subsequent analyses. After peak detection and alignment, 1397 features were initially detected and reduced to 227 after blank and artifact filtering. Mann-Whitney U testing with false discovery rate correction identified 100 statistically significant features, of which 67 were putatively identified. Multivariate analysis revealed a clear separation between PDO and PGI samples: analyzed PDO vinegars were enriched in furanoids, lactones, esters, aldehydes, acids, phenols, ketones, and alcohols, whereas PGI samples showed comparatively higher levels of terpenoids. These chemical trends align with the technological differences between the two production systems. Mapping volatile features to literature-reported odor descriptors suggested that the higher abundance of furanoids, lactones, and related compounds in the PDO samples may contribute to sweeter, caramel-like, creamy, balsamic, woody, and floral nuances. Overall, the integration of HS-SPME-GC×GC-TOFMS with statistical filtering and multivariate analysis demonstrates the feasibility of a volatilomic analytical workflow capable of differentiating selected PDO and PGI products, and establishes a methodological framework for future large-scale studies on authenticity and quality control.
Establishment of Non-Target Screening and Quantitative Analysis Strategy Based on UHPLC Coupled With Orbitrap Exploris 240 and QqQ Mass Spectrometry to Improve the Quality Control of Ziyu Ointment.
Liao M, Hu J, Xi Z … +4 more , Guan S, Gao J, Cheng J, He T
J Sep Sci · 2026 May · PMID 42112725 · Publisher ↗
Ziyu ointment (ZYO) is a commonly used, approved ingredient in traditional Chinese medicine (TCM) medication for treating superficial swelling and ulcers. However, due to limited methodologies, more accurate and selectiv...
Ziyu ointment (ZYO) is a commonly used, approved ingredient in traditional Chinese medicine (TCM) medication for treating superficial swelling and ulcers. However, due to limited methodologies, more accurate and selective qualitative and quantitative analysis methods for its active ingredients have not yet been established. This impedes both the full clinical development and application of ZYO as well as the clarification of its pharmacological mechanism. In this research, a rapid and sensitive UHPLC-Orbitrap Exploris MS method with higher-energy collisional dissociation (HCD) was established for identifying ZYO's chemical components. A total of 167 chemical components were found, including 34 carboxylic acids and esters, 32 phenylpropanoids, 27 terpenoids, 18 flavonoids, 12 naphthoquinones, 11 phthalides, 8 phenolic acids, 5 phenols, 3 steroids, 2 quinones, 2 alkaloids, and 13 miscellaneous compounds. Among these, UHPLC-QqQ-MS/MS was used to simultaneously determine 12 analytes (either with high contents or significant bioactivities), namely, (1) loganin, (2) sweroside, (3) ferulic acid, (4) senkyunolide I, (5) bergapten, (6) ziyuglycoside II, (7) imperatorin, (8) shikonin, (9) ligustilide, (10) isoimperatorin, (11) β-hydroxyisovalerylshikonin, and (12) levistilide A. The foundation for future use and quality assurance of the active ingredients in ZYO was established by this study.
Surface-Grafted Polyamide Stationary Phase on poly(styrene-divinylbenzene) Microspheres for Hydrophilic Interaction Chromatography.
Yang M, Yu Z, Zhang F … +2 more , Jin R, Yang B
J Sep Sci · 2026 May · PMID 42112721 · Publisher ↗
An amide-functionalized stationary phase for hydrophilic interaction chromatography was developed by surface-confined grafting of N-vinylformamide onto poly(styrene-divinylbenzene) microspheres. The grafting reaction was...
An amide-functionalized stationary phase for hydrophilic interaction chromatography was developed by surface-confined grafting of N-vinylformamide onto poly(styrene-divinylbenzene) microspheres. The grafting reaction was restricted to the microparticle surface by selecting a polar monomer (N-vinylformamide) and a non‑swelling reaction solvent (ethanol), preventing monomer penetration into the polymer matrix. The resulting stationary phase exhibited typical HILIC retention behavior and provided effective separation of polar model analytes, including nucleosides, amino acids, and vitamins, with a plate count of up to 41 980 plates per meter for cytosine. The phase demonstrated excellent hydrolytic stability across a wide pH range (2-10) and extremely low column bleed. Under gradient elution with up to 50% aqueous content, baseline drift was negligible-comparable to that of a blank, and significantly lower than that of commercial silica‑based amide columns.
A Protocol for Characterizing Comprehensive Two-Dimensional Liquid Chromatography Systems.
Aebischer MK, Pardon M, Gadot C … +10 more , Arena K, Carstensen N, Laemmerhofer M, Dugo P, Cacciola F, Mondello L, Sarrut M, Cabooter D, Heinisch S, Guillarme D
J Sep Sci · 2026 May · PMID 42112711 · Full text
Comprehensive two-dimensional liquid chromatography (LC × LC) is an effective strategy to increase peak capacity and improve separation performance for complex samples, offering a valuable alternative to conventional one...
Comprehensive two-dimensional liquid chromatography (LC × LC) is an effective strategy to increase peak capacity and improve separation performance for complex samples, offering a valuable alternative to conventional one-dimensional liquid chromatography (1D-LC). However, LC × LC method development and optimization remain challenging due to the interconnection of numerous parameters within and between dimensions. Both thermodynamic factors (e.g., stationary phase, mobile phase, temperature) and kinetic factors (e.g., gradient profile, sampling rate, column geometry, flow rates) strongly influence the analytical performance. To achieve effective method optimization, two-dimensional liquid chromatography (2D-LC) instrumentation-related parameters must first be characterized, as they directly influence both method development and overall performance. In this work, we propose a practical guide (protocol) for comprehensive 2D-LC system characterization. For clarity and ease of implementation, all calculations associated with this protocol can be performed using the Characterization module of the 2D-LC Smart Calculator software, freely accessible online. The protocol usability was validated through inter-laboratory testing. The impact of instrumental features on LC × LC performance was then assessed by in silico evaluation of the theoretical LC × LC performance of the characterized systems. The results clearly demonstrate the benefits of the proposed protocol, emphasizing the importance of system-specific characterization for reliable LC × LC implementation. In addition, practical recommendations are provided to help users optimize their instruments when certain parameters are not adapted.
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