Steroid hormones are important trace bioactive compounds found in food, environmental, biological, and regulatory samples. However, their accurate analysis remains challenging due to their low concentration, strong hydrophobicity, highly similar structures, and susceptibility to interference from complex matrices. Molecularly imprinted polymers contain binding sites designed to complement target molecules in size, shape, and functional-group arrangement and have been applied to the selective enrichment and detection of steroid hormones. This review examines steroid hormone molecularly imprinted polymers with emphasis on hormone structure, template and monomer selection, cross-linkers, porogens, and surface or magnetic imprinting strategies. Furthermore, it summarizes the integrated applications of molecularly imprinted polymers with electrochemistry, optics, quartz crystal microbalances, surface plasmon resonance, photonic crystals, and wearable sensing platforms, and reviews their current applications in environmental monitoring, food safety analysis, clinical testing, and biomonitoring. Key priorities are rational imprint design, matrix-compatible interfaces, and standardized performance evaluation to support the use of steroid hormone molecularly imprinted polymers in complex samples.
Deciphering the pharmacodynamic material basis of classical traditional Chinese medicine (TCM) formulas remains a critical challenge, owing to their multicomponent nature, multitarget interactions, and intricate in vivo metabolic fates. To address this issue, a metabolism-oriented strategy was established and applied to Wuao Sanzi Decoction (WASZD), a classic TCM formula for respiratory diseases such as asthma. Ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry was used for systematic analysis of the chemicalome, in vivo prototypes, metabolites, and tissue distribution of WASZD. A total of 200 chemical components were identified in vitro, including 102 flavonoids, 29 triterpenoids, 10 steroids, 14 organic acids, and 45 other compounds. Further, 91 prototypes and 212 metabolites in vivo were characterized in biological samples of rats. Key metabolic pathways involved phase I reactions (reduction, oxidation, and demethylation), phase II reactions (glucuronidation, methylation, and sulfation), and gut microbiota-mediated ring cleavage of flavonoids, which generated bioactive small-molecule phenolic acids. Tissue distribution analysis revealed that prototypes and metabolites were predominantly enriched in the ileum, colon, liver, kidney, and lung, with flavonoids and their metabolites identified as the most abundant components, suggesting they are the core bioactive candidates of WASZD. This study not only putatively clarifies the pharmacodynamic material basis of WASZD but also establishes a reproducible technical framework for decoding the complex in vivo behavior of multi-component TCM formulas, facilitating their modernization and rational clinical application.
"Wen Tong Plaster" is renowned for its therapeutic effects of warming meridians, dispelling cold, and promoting blood circulation. However, its chemical composition and active pharmacological substances have yet to be fully elucidated. In this study, the chemical components of "Wen Tong Plaster" cataplasm and the transdermal components in vivo and in vitro were comprehensively analyzed by mass spectrometry targeted quantification method, and the release kinetics of the main transdermal components in vitro were evaluated, and the concentration-time variation of key active transdermal components in vivo was studied, finally molecular docking technology was used to predict the mechanism of action of key bioactive transdermal components. Using UHPLC-Q-Exactive Orbitrap MS, a total of 129 plant chemical components were found in the "Wen Tong Plaster" cataplasm extract, with 41 components detected as in vitro transdermal compounds. In subcutaneous tissue and uterine tissue, 53 and 36 in vivo transdermal components were characterized, respectively. The release kinetics of "Wen Tong Plaster" cataplasm were evaluated by tetrahydropalmatine, corydaline, and dihydrotanshinone I, three marker components, and fitted to a first-order kinetic model based on integrated release kinetics. After "Wen Tong Plaster" cataplasm administration, tetrahydropalmatine, corydaline, dihydrotanshinone I, and α-cyperone were accurately quantified in the subcutaneous tissue, plasma, and uterus tissue by UHPLC-QqQ-MS/MS, while the concentration-time changes of them in the subcutaneous tissue, plasma, and uterus tissue were investigated. The molecular docking results indicated that all four components exhibited favorable binding affinity with the targets, particularly demonstrating strong binding capacity to the cytochrome P450 1A1 target, suggesting it as a potential therapeutic target for "Wen Tong Plaster" cataplasm. In summary, the study revealed the pharmacodynamic material basis and in vitro release kinetics of "Wen Tong Plaster" cataplasm, providing a theoretical foundation and data support for further research on the mechanism of action and clinical application.
Chronic fatigue syndrome (CFS) is a complex condition that requires effective intervention and treatment. Buqi-Zhitong-Decoction (BQZTD), an empirical formula, has been used clinically to treat CFS and has shown good clinical results. However, the underlying bioactive components and mechanism of action for its treatment of CFS remain unclear. This study is based on the serum pharmacochemistry method, utilizing LC-MS/MS technology for detection and analysis of BQZTD water extracts and their components entering the bloodstream. A total of 98 compounds were preliminarily identified from the BQZTD. Furthermore, 44 ingredients were detected in rat serum, including 19 prototype components and 25 metabolite components. The results of network pharmacology analysis indicated that the BQZTD might improve CFS by regulating immune, inflammatory, neuroendocrine system, and energy metabolism-related pathways. In addition, Prostaglandin-Endoperoxide Synthase 2 (PTGS2) plays a central role in the regulatory network. Molecular docking simulations with six potential active components in serum revealed that all of them exhibit high affinity for PTGS2, with compound Isoformononetin showing the strongest binding affinity. This study represents the first comprehensive identification of the chemical components, targets, and signaling pathways of BQZTD, clarifying its potential mechanism of action in treating CFS. The research findings provide a reference basis for pharmacodynamic studies and quality control.
Infliximab (IFX) is a cornerstone biologic for pediatric inflammatory bowel disease, where therapeutic drug monitoring (TDM) is essential but complicated by inter-platform variability. Systematic comparison of automated immunoassays with reference methods is therefore warranted. This study aimed to validate an in-house chemiluminescence immunoassay (CLIA) for IFX quantification and evaluate its agreement with liquid chromatography-tandem mass spectrometry (LC-MS/MS). A CLIA kit was validated by assessing the limit of blank, specificity, linearity, accuracy, precision, matrix equivalence, and stability. IFX concentrations in 52 pediatric plasma samples were measured by CLIA and a reference LC-MS/MS method. Comparability was evaluated using linear regression, Passing-Bablok regression, and Bland-Altman analysis. CLIA showed excellent performance (lower limit of quantification: 0.640 µg/mL; specificity <10.0% interference; reportable range: 0.640-462 µg/mL). LC-MS/MS yielded higher concentrations than the CLIA (median: 6.25 vs. 2.87 µg/mL). Strong correlation was observed (r = 0.9101) with no significant deviation from linearity (Passing-Bablok, p > 0.05). However, Bland-Altman revealed substantial mean relative bias (67.9%) and poor categorical agreement (52% concordance) within the therapeutic range (3-7 µg/mL), indicating non-interchangeability. We validated a rapid, automated CLIA suitable for high-throughput TDM. Despite strong correlation, significant quantitative differences preclude direct result substitution, underscoring the need for method-specific therapeutic thresholds.
Bamboo juice is a traditional Chinese herbal medicine and drink, and bamboo juice oral liquid (BJOL) is widely marketed in China for clearing away heat, resolving phlegm, and relieving cough. However, the chemical composition has not been fully characterized. This study employed UHPLC-Q-Exactive Orbitrap MS combined with molecular networking to comprehensively characterize the chemical constituents of BJOL. Mass spectrometric data were acquired in both positive and negative ion modes, and the raw data were uploaded to Global Natural Products Social Molecular Networking to construct molecular networking based on the similarity of MS2 fragmentation patterns. Compound identification was further achieved by comparison with reference standards, database searching, literature verification, and analysis of fragmentation pathways. A total of 121 compounds were identified, including 48 phenylpropanoids, 12 flavonoids, 28 organic acids, 9 amino acids and nucleosides, and 24 other constituents. Among them, 44 compounds were reported in BJOL for the first time. Thus, the UHPLC-Q-Exactive Orbitrap MS strategy established in this study can be applied to the chemical characterization of BJOL, and also provides a reference for investigating its pharmacodynamic components and analyzing constituents of other traditional Chinese medicine preparations.
Mangiferin (MGF), a bioactive C-glucosylxanthone predominantly found in Mangifera indica L., has attracted increasing interest due to its recognized antioxidant and pharmacological properties. In this study, an aqueous ethanolic maceration process was systematically optimized for MGF recovery from mango plant biomass using a Taguchi experimental design followed by Box-Behnken response surface methodology. The optimized conditions of 39.9°C, 8 h, and 80% v/v ethanol yielded 41.60 mg/g dry weight of MGF, confirming the robustness of the predictive model. Biochemical profiling of the optimized extract revealed 4.36 mg gallic acid equivalents/g dry weight of total phenolics, 73.12 mg/g of reducing sugars, and low protein content, indicating a phytochemical matrix enriched in glycosylated phenolics. Antioxidant evaluation by the 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay demonstrated 87.74% radical inhibition at 400 µg/mL and a 50% inhibitory concentration of 225 µg/mL, reflecting strong radical scavenging capacity. Greenness assessment using the Analytical GREEnness metric approach software yielded a score of 0.72, validating the environmental compatibility of the proposed method. The results demonstrate that statistically guided aqueous ethanolic maceration represents an efficient, scalable, and sustainable platform for the recovery of MGF-rich extracts with potential pharmaceutical, nutraceutical, and cosmetic applications.
Huangqi Shengmai Yin (HQSMY) displays notable cardioprotective properties. Nevertheless, the principal pharmacodynamic constituents underpinning this efficacy have not yet been fully clarified. To tackle this issue, an integrative workflow coupling UHPLC‑Q‑TOF‑MS/MS fingerprinting, chemometrics, and spectrum‑effect correlations was constructed for the systematic discovery of cardioprotective markers. Fifty-one components from 10 batches of HQSMY samples were characterized by UHPLC‑Q‑TOF‑MS/MS in conjunction with the GNPS molecular networking platform. Subsequently, chromatographic fingerprints were established, with 22 and 29 characteristic common peaks assigned in negative and positive ion modes, respectively. Interbatch consistency was assessed via chemometric methods. Spectrum‑effect relationship analyses were conducted through Pearson correlation, gray relational analysis, and OPLS regression, revealing that formononetin, astrapterocarpan, astragaloside II, schisandrin, and biochanin A exhibited highly significant positive correlations with H9c2 cardiomyocyte protective activity. All these compounds attained gray relational scores exceeding 0.7, implying strong associations with cardioprotective action. These five components were then chosen from the candidate markers for subsequent functional validation using the CCK-8 assay, which yielded EC50 values of 18.60 ± 3.15 µM (biochanin A), 26.42 ± 4.78 µM (formononetin), 92.93 ± 19.14 µM (astragaloside II), 70.85 ± 13.26 µM (schisandrin), and 72.73 ± 14.05 µM (astrapterocarpan). The cardioprotective effect of astragaloside II, schisandrin, and astrapterocarpan in H2O2-induced H9c2 injury model were reported for the first time. This integrated analytical strategy not only pinpoints the primary cardioprotective components of HQSMY but also furnishes a dependable methodological framework for quality evaluation and pharmacodynamic material research.
Baicalin (BA), a key flavonoid isolated from Scutellariae Radix (SR), possesses diverse pharmacological activities encompassing anticancer, anti-inflammatory, and hepatoprotective effects. In order to achieve rapid separation and enrichment of BA from complex matrix samples, core-shell magnetic molecularly imprinted polymers (MMIPs) were fabricated using BA as the template molecule and Fe3O4 nanoparticles as the magnetic core. Subsequently, key preparation parameters, including the ratio of functional monomers to cross-linking agents, imprinting time, and the dosage of BA were systematically optimized. Under the optimized conditions, the prepared MMIPs exhibited rapid binding kinetics and a remarkable maximum adsorption capacity of 18.21 mg/g, accompanied by an imprinting factor (IF) of 3.919. Furthermore, the developed MMIPs were successfully employed for the selective extraction and determination of BA from SR. Specifically, the average recoveries of BA at three spiked concentrations (5.00, 11.20, and 20.00 µg/mL) were 107.0%, 92.7%, and 105.2%, respectively, with corresponding relative standard deviations (RSDs) of 5.31%, 1.65%, and 3.28% (n = 3). Overall, this work provides a novel and efficient adsorbent for trace flavonoids enrichment from complex herbal samples, offering a promising pretreatment strategy for active ingredient analysis in traditional Chinese medicine research.
Direct oral anticoagulants (DOACs), including apixaban, dabigatran, edoxaban, and rivaroxaban, may require quantitative measurement in selected clinical situations despite fixed-dose administration. This study developed and validated a quick, easy, cheap, effective, rugged, and safe (QuEChERS)-assisted liquid chromatography-tandem mass spectrometry method for their simultaneous determination in citrate plasma and quantitative dried plasma spot (qDPS) samples. Chromatographic separation was achieved on a Kinetex PS C18 column using a 4.0 min gradient, followed by positive electrospray ionization and scheduled multiple reaction monitoring. Sample preparation was optimized by comparing acetonitrile (ACN) protein precipitation, unbuffered and citrate-buffered QuEChERS extraction salts, and dispersive solid-phase extraction clean-up using primary-secondary amine, C18, EMR Lipid, Z-Sep+, or their combinations. The final procedure comprised ACN extraction with MgSO4, NaCl, and Bond Elut EMR Lipid, followed by clean-up with MgSO4, primary-secondary amine, and Z-Sep+. ACN provided the most consistent recovery across analytes, concentration levels, and matrices. The method was validated in accordance with International Council for Harmonization M10 using commercial plasma calibrators and independently prepared quality-control samples. Accuracy and precision met the predefined acceptance criteria in both matrices. Mean recovery ranged from 84.2% to 91.2% in citrate plasma and from 79.6% to 87.8% in dried plasma samples. Internal standard-normalized matrix factors ranged from 0.87 to 0.98, indicating limited ion suppression after the optimized clean-up procedure. External verification using materials from two independent proficiency-testing providers showed satisfactory agreement with assigned target values, with all results falling within the providers' acceptance ranges. The method provides a robust QuEChERS-assisted workflow for the quantitative determination of DOACs in conventional citrate plasma and qDPS samples. Its clinical interchangeability between matrices requires further evaluation using paired patient samples.
Swertia purpurascens is an important ethno-medicinal herb used for treating fever and colds. Despite its significance, the phytochemical profile of this species remains unexplored. This study aims to conduct a phytochemical investigation, develop a green and reliable analytical method assessing 11 specialized metabolites of S. purpurascens, and identify their suitable biological targets via network pharmacology. Among isolated metabolites, 3-hydroxybenzyl-β-d-glucopyranoside (1) has been reported for the first time from the Swertia genus, whereas compounds gentiopicroside (2), 1-hydroxy-3,5-dimethoxyxanthone (5), and methylswertianin (6) have been isolated for the first time from S. purpurascens. The developed method showed good linearity, sensitivity, precision, accuracy, and a positive green analytical profile. Results revealed that the stem part was the major source of metabolites (Yield: 125.70 ± 1.79 mg/g), compared with the other analyzed parts (flowers, leaves, and roots). Compounds 2 (0.72%-6.94%), 9 (0.08%-5.61%), and 1 (0.07%-2.71%) were quantified as the major compounds among different plant parts. Furthermore, network pharmacology analysis predicted the potential therapeutic relevance of these compounds with suggested pathways related to diabetes and cancer diseases. This study represents the first attempt to simultaneously quantify 11 specialized metabolites of S. purpurascens and to perform a network pharmacology-based analysis, thereby enriching its phytochemical understanding.
Fluorinated liquid crystal monomers (FLCMs) are widely used in various electronic materials owing to their outstanding optoelectronic properties. However, their persistence, bioaccumulation, and toxicity, such as endocrine disruption, in the environment have rendered them emerging contaminants of significant concern. Switchable natural deep eutectic solvents (SNADESs) are prepared from natural compounds and their metabolites. These solvents possess the characteristics of green solvents while overcoming many limitations of conventional organic solvents, and have thus been widely adopted in the analysis and detection of environmental pollutants. In this study, a pH-SNADES was utilized as the extraction medium. Combined with ultrasound-assisted homogeneous liquid-liquid microextraction, a method for the extraction of FLCMs from environmental water samples was established, and the target analytes were qualitatively and quantitatively determined by gas chromatography-mass spectrometry. The results demonstrated that five of these compounds exhibited excellent linearity in the range of 0.5-500 ng mL-1, while the other two (4-(3,4-difluorophenyl)-4'-pentyl-1,1'-bi(cyclohexyl) and 3,4,5-trifluoro-4'-(4-pentylcyclohexyl)biphenyl) showed linearity in the range of 1-500 ng mL-1, with limits of detection ranging from 0.001 to 0.167 ng mL-1 and limits of quantification ranging from 0.003 to 0.556 ng mL-1. Applied to real water samples, the method yielded spiked recoveries of 80.79%-125.42%, relative standard deviations of 0.09%-5.14%, and enrichment factors of 8-26, with its greenness and applicability further validated by three greenness assessment tools. The proposed method is efficient, environmentally friendly, and features mild reaction conditions and simple operation, providing technical support for the detection of FLCMs in environmental water samples.
Crataegi Folium (CF), a traditional Chinese medicine widely used for promoting blood circulation and regulating lipid metabolism, contains diverse bioactive constituents with potential health-promoting effects. However, the rapid localization and identification of its functional compounds remain challenging due to the complexity of its chemical constituents. This study aimed to establish an integrated approach using high-performance thin-layer chromatography (HPTLC) coupled with bioactivity assay and mass spectrometry (MS) for the rapid screening and identification of pancreatic lipase inhibitory and antibacterial constituents in CF. Two HPTLC developing systems with different polarity ranges were optimized to comprehensively separate the chemical constituents, enabling effect-directed analysis. The results revealed that the pancreatic lipase inhibitory activity originated primarily from low-polarity triterpenoids. One active zone, consisting of co-migrating isomers, was successfully separated by in-situ iodine derivatization and characterized as maslinic acid and corosolic acid through comparison with reference standards. Another active zone was identified as arjunolic acid by using the exclusion method. Based on the Vibrio fischeri bioassay, six observed V. fischeri bioluminescence-inhibitory zones were mainly associated with high-polarity compounds, among which five were identified as isoquercitrin, hyperoside, chlorogenic acid, vitexin-2''-O-rhamnoside, and vitexin-4''-O-glucoside. Our findings provide new insights into bioactive constituents of CF, and demonstrate that HPTLC-bioautography-MS is an efficient and integrated platform for the rapid discovery and identification of markers for quality assessment of bioactive compounds in complex botanical matrices.
Accurate estimation of water solubility is crucial for chemical screening, yet traditional predictive tools such as the General Solubility Equation (GSE) require melting point and octanol-water partition coefficient data and often struggle with weak electrolytes. This study presents two highly efficient reversed-phase liquid chromatography (RPLC) correlation strategies evaluated, comprising 49 binary solvent systems on three stationary phases. First, simple, melting point, octanol-water partition coefficient, and descriptor-free, isocratic models were explored to estimate the water solubility of neutral organic compounds using RPLC retention as the sole input. This approach demonstrated superior predictive accuracy of 0.307 log units and a more uniform error distribution across both non-electrolytes and weak electrolytes than the more conventional GSE. Second, to improve the predictive accuracy of estimating water solubility, RPLC retention and melting point data were explored. Predictive accuracy improved notably at higher organic volume fractions (50%-70% v/v) across all evaluated stationary phases. The correlation model comprising 70% (v/v) acetonitrile on an XTerra MS C18 stationary phase yielded a coefficient of determination of 0.962, a standard deviation of the model fit of 0.276, and an average absolute error value of 0.222. Together, these complementary chromatographic modelling strategies offer powerful, high-throughput alternatives for rapid solubility screening: providing either a direct, descriptor-free approach for compounds lacking structural information or a highly precise RPLC model approach combining retention and melting point data that minimizes prediction errors below 0.3 log units.
The recycling of waste lead-acid batteries and the sensitive detection of trace p-phenylenediamine (PPD) antioxidants represent two pressing challenges in resource recovery and environmental safety. Herein, an integrated solution was proposed by synthesizing a novel lead-based metal-organic framework (WLP-Pb-MOF) from waste lead paste (WLP) as a sustainable metal source. The material served as a coating for a solid-phase microextraction (SPME) fiber. When coupled with gas chromatography-mass spectrometry, it enabled a highly sensitive method for determining three PPDs in water samples. The unique slit-shaped pore architecture and hydrophobic surface of WLP-Pb-MOF facilitated efficient enrichment through synergistic interactions, including size-sieving, hydrophobic partitioning, π-π stacking, and hydrogen bonding. This synergy resulted in extraction efficiencies significantly outperforming commercial SPME fibers. The method demonstrated good linearity from 5 to 2000 ng L-1, with detection limits of 0.29-1.60 ng L-1. In real water samples, the method achieved spiked recoveries of 91.3%-108.5%, with relative standard deviations below 8.78%. These results confirmed its accuracy and practical reliability. This work not only presents a viable strategy for the value-added recycling of waste lead but also provides a robust analytical platform for monitoring trace organic pollutants, effectively bridging sustainable resource recovery with advanced environmental monitoring.
Nardostachys jatamansi is an endangered aromatic plant that produces a chemically complex and diverse array of secondary metabolites. In this study, we aimed to comprehensively characterize the chemical diversity of N. jatamansi by liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based metabolite annotation. A metabolite annotation workflow combining feature detection, library matching, MetFrag-based in silico fragmentation, SIRIUS fragmentation tree analysis, manual MS/MS interpretation, molecular networking, chemical classification, and structural validation of selected compounds was applied to methanolic extracts of N. jatamansi. Using this integrated strategy, 111 peaks were structurally annotated, and an additional 13 peaks were assigned to tentative structural level annotations based on MS/MS fragment similarity in a molecular network, giving a total of 124 peaks classified at the chemical structural level. The annotated metabolites mainly comprised iridoids, lignans, sesquiterpenoids, sesquiterpenoid hybrids, monoterpenoids, flavonoids, and other constituents such as fatty acids and phenylpropanoids, revealing the remarkable chemical diversity of N. jatamansi. The annotation performance of each metabolite annotation platform was systematically compared in terms of coverage, efficiency, and chemical class level specificity, revealing that these platforms provided complementary annotations whose combined use with molecular networking greatly enhanced metabolite coverage and structural interpretation. This integrated LC-MS/MS and multi-platform workflow extends current knowledge of the chemical diversity of N. jatamansi.
Niaoduqing Granules (NDQG) is a Chinese patent medicine preparation composed of more than 10 herbal ingredients and is widely used in the clinical treatment of chronic kidney disease. Nevertheless, comprehensive and systematic investigations into the chemical constituents and pharmacodynamic material basis of NDQG remain insufficient. Herein, a binary chromatographic fingerprinting strategy was developed for the systematic characterization of the chemical constituents in NDQG using both acidic and alkaline mobile phase systems. Independent collection and combined analysis of acidic/alkaline mobile phases enabled the effective separation of compounds with significant polarity differences and varying chromatographic properties in NDQG, resulting in good peak shapes. Using ultra-high-performance liquid chromatography-Q-Exactive Orbitrap mass spectrometry, this study identified 187 chemical components from the NDQG preparation (58 compounds were qualitatively confirmed by reference standards). The prototype components distributed to plasma and kidney tissues in C57BL/6 mice following oral administration of NDQG were further investigated. 40 components were detected in the blood circulation, and 36 were identified in kidney tissues. Using a cobalt chloride-induced Human Kidney-2 (HK-2) cell injury model, further activity screening of 28 candidate compounds revealed that nine compounds (formononetin [FMN], ononin [ONO], liquiritin, ferulic acid, quercetin, rosmarinic acid [ROA], paeoniflorin, calycosin [CS], and salvianolic acid B [SAB]) effectively alleviated HK-2 cell injury. Among them, FMN, ONO, ROA, CS, and SAB significantly inhibited the mRNA expression of both tumor necrosis factor-α and interleukin-18, thereby exerting favorable anti-inflammatory effects. These components may act as important lead compounds, providing valuable implications for drug discovery and development. In summary, combining component characterization and activity screening, this study not only provides key data for the quality control of NDQG but also furnishes a scientific basis and novel insights for clarifying its pharmacodynamic material basis and supporting the subsequent development of innovative drugs.
Polymerizable zwitterionic liquids (ZILs) were investigated for the development of thermally stable wall-coated open-tubular (WCOT) stationary phases for gas chromatography (GC) in the separation of volatile carboxylic acids (VCAs). Three imidazolium-based ZILs containing alkenyl and acrylate functional groups were synthesized and evaluated based on their polymerization and used as separation media. Among the evaluated chemical structures, only the acrylate ZIL demonstrated the ability to undergo free radical polymerization and was subsequently employed in the preparation of chromatographic columns. WCOT columns were prepared by the static coating method followed by in-situ polymerization inside the GC capillary columns, resulting in chromatographic efficiencies up to 3300 plates m-1. The resulting stationary phase enabled separation of VCAs under isothermal conditions at 100°C and exhibited reduced bleed (from 6.92-7.80 pA at 40°C to 154.70-157.57 pA at 250°C) compared to a corresponding non-polymerizable ZIL (from 7.58 pA at 40°C to 396.38 pA at 250°C), indicating improved thermal stability of the stationary phase upon polymerization. However, thermal treatment up to 250°C resulted in no chromatographic retention of benzyl alcohol, suggesting that the stationary phase undergoes an increase in rigidity under these conditions. To address this limitation, poly(ethylene glycol) methacrylate (PEGMA) was incorporated as a co-monomer at 10%, 20%, 25%, and 40% weight fractions to introduce flexibility within the polymeric network. With increasing PEGMA content, progressively narrower peaks of benzyl alcohol were observed after thermal treatment (3.3, 1.10, 0.81, and 0.194 min for 10%, 20%, 25%, and 40% weight fractions, respectively), indicating improved mass transfer within the stationary phase. These results demonstrate the feasibility of polymerizable ZIL stationary phases for GC and provide insight into the role of polymer composition in controlling chromatographic performance after thermal treatment.
This review examines recent advances in the use of natural deep eutectic solvents (NaDES) for natural product sample preparation, with emphasis on extraction, purification, analytical compatibility, and sustainability. NaDES emerge as versatile, tunable, and biocompatible media that enhance the recovery and stability of diverse phytochemicals through extensive hydrogen‑bond networks and customizable polarity. The review summarizes current knowledge on NaDES formation principles, key physicochemical properties, and classification and evaluates their performance across conventional and advanced extraction techniques, including ultrasound, microwave, and pressure‑assisted processes. Evidence shows that NaDES frequently outperform traditional organic solvents in selectivity, extraction efficiency, and compound preservation. Their integration into purification workflows, chromatographic and spectroscopic analysis, and sensor platforms highlights growing analytical relevance. Sector‑specific applications in food, cosmetics, and pharmaceuticals illustrate the potential for direct formulation, improved bioactivity, and enhanced functional properties. Sustainability assessments using green metrics demonstrate that NaDES‑based methods reduce solvent consumption, waste generation, and environmental impact. Remaining challenges include viscosity management, removal strategies, and regulatory acceptance. Overall, NaDES represent a transformative class of green solvents that advances natural product research by combining efficiency, analytical compatibility, and strong alignment with green chemistry principles.
This work presents a fast and straightforward chromatographic method with fluorescence detection to quantify glufosinate in soil solutions. The method is based on the reaction between the primary amine group of glufosinate and o-phthaldialdehyde with 2-mercaptoethanol, producing a stable fluorescent derivative in under 2 min, which remains stable for more than 70 min (relative standard deviation [RSD] = 3.6%). The method does not require automated timing systems. Glyphosate, which lacks a primary amine, does not interfere, and potential interferences from aminomethyl phosphonic acid and ammonium are effectively circumvented by chromatographic separation, as their indole derivatives are well resolved from the glufosinate derivative. An isocratic chromatographic run using 20:80 methanol:piperazine-N,N'-bis(2-ethanesulfonic acid) buffer at 1.0 mL/min lasts 5 min. The method showed linearity over the concentration range of 0.10 and 20 µmol/L, with detection and quantification limits of 0.015 and 0.050 µmol/L, respectively. Calibration curve reproducibility is high (slope RSD < 2.1%, n = 5). The method was effectively applied to study glufosinate adsorption in an Oxisol, revealing a maximum adsorption capacity of 2.5 ± 0.2 µmol/g, significantly lower than that reported for glyphosate.
