Stationary Phase For High-performance Liquid Chromatography Research Articles

Glutathione-functionalized covalent organic frameworks@silica as a hydrophilic-hydrophobic balanced mixed-mode stationary phase for highly efficient separation of compounds with a wide range of polarity

BackgroundCovalent organic frameworks (COFs) are a highly promising stationary phase for high-performance liquid chromatography (HPLC), but the separation of polar compounds is limited by their low hydrophilicity. Therefore, it is crucial to develop novel COFs-based stationary phases with balanced hydrophilicity-hydrophobicity for the efficient separation of different polar compounds. ResultsIn this paper, glutathione (GSH)-functionalized COFs@silica microspheres (GSH-COFs@SiO2) were synthesized via a two-step, post-synthesis modification strategy. The COFs particles was constructed onto silica surface by the covalent conjugation of 1,3,5-tris(4-aminophenyl)benzene and 2,5-divinylterephthalaldehyde. GSH containing abundant –NH2 and –COOH groups was bonded onto the surface of COFs@SiO2 to further enhance hydrophilicity. The resulting GSH-COFs@SiO2 exhibited balanced hydrophilicity-hydrophobicity and can be used in hydrophilic/reversed-phase liquid chromatography modes through multiple retention mechanisms. Consequently, a variety of compounds with different polarity, including nucleosides/bases, benzoic acids, anilines, phenols, alkylbenzenes and polycyclic aromatic hydrocarbons, were well separated with ideal resolution, satisfactory column efficiency and good peak shapes. Furthermore, this novel column exhibited remarkable column stability, as evidenced by intra-day relative standard deviations of 0.08 %–0.18 % for retention time and 0.45 %–1.47 % for peak area. Significance and noveltyThis work demonstrates the superior hydrophilic-hydrophobic selectivity of GSH-COFs@SiO2 stationary phases towards compounds with a wide range of polarity and provides a very facile and easily popularized post-synthetic modification route for hydrophilic-hydrophobic balanced COFs-based HPLC stationary phases.

A Validated Chiral Chromatography Method for Enantiomeric Separation of Pomalidomide in Human Plasma.

In the present work, new chiral stationary phase high-performance liquid chromatography (CSP-HPLC) method was established and validated for the quantification of pomalidomide (PMD) enantiomers in human plasma. The chromatographic enantiomeric separation was achieved on a Daicel-CSP, Chiralpack IA 4.6 × 250mm, 5μm; because of its advantages of high degree of retention, high resolution capacity, better reproducibility, ability to produce lower back pressure and low degree of tailing. The mobile phase was maintained as methanol: glacial acetic acid (499.50ml:50μL). Ultraviolet wavelength for detection was 220nm. PMD enantiomer-I and enantiomer-II were separated at 8.83 and 15.34min, respectively. Limit of detection and limit of quantification for each enantiomer and the calibration curve of standard PMD was linear in range between 10-5,000ngmL-1. The method was validated according to The International Council for Harmonisation of Technical Requirements of Pharmaceuticals for Human Use (ICH(Q2R1)) specific guidelines. We found no interference peak with PMD chromatogram obtained. This is a simple, reliable and specific method for detection and quantification of enantiomer of PMD in human plasma sample.

Metal/covalent organic framework@silica double-layer core–shell composite: A high-performance liquid chromatographic stationary phase

Nowadays, metal/covalent organic frameworks (MOFs/COFs) have shown bright application prospects in the field of chromatographic separation due to their unique porous characteristics. However, chromatographic stationary phases prepared with only a single MOF or COF material have a few inherent defects, which keep them from satisfying the actual application fully. In contrast, MOF/COF composites have more superior properties that are more favorable for separation. Therefore, this study innovatively proposed a new strategy to apply MOF/COF modified silica composites for high performance liquid chromatography (HPLC). To be specific, using ethylenediamine as a linker, Zn-based MOF and imine-based COF were modified on the aminated silica gel layer by layer, and a MOF-5/COF@silica bilayer core–shell composite was successfully prepared. The composite of MOF and COF can complement each other in realizing the advantages of the two materials. The research results show that the MOF-5/COF composite can provide a large number of active sites with a variety of interaction forces for the analytes. The MOF-5/COF@silica stationary phase can separate a wide range of hydrophilic and hydrophobic substances, which can also be used for the effective detection of environmental hormones in lake water samples. This study verifies the feasibility of preparing MOF and COF co-modified silica gel composite to improve the separation performance of stationary phase, and demonstrates the superiority of the double-layer porous core–shell structure for efficient HPLC analysis. This provides a new idea for the further development and application of porous materials in the field of HPLC.

Thermodynamic Characteristics and Selectivity of the Liquid-Phase Adsorption of Aromatic Compounds on Hypercrosslinked Polystyrene Networks with Ultimate-High Crosslinking Densities by Data of Liquid Chromatography.

This study delves into the thermodynamics of liquid-phase adsorption on hypercrosslinked polystyrene networks (HPSNs), widely recognized for their distinct structure and properties. Despite the considerable progress in HPSN synthesis and characterization, gaps persist regarding the chromatographic retention mechanism, thermodynamics of adsorption, and their impact on the adsorption selectivity, especially in the case of networks with ultra-high crosslinking densities (up to 500%). Utilizing high-performance liquid chromatography (HPLC), we have explored, for the first time, the thermodynamic intricacies of liquid-phase adsorption onto HPSNs crosslinked in the entire range of the crosslinking degree from 100 to 500%. Our findings reveal the dependences of thermodynamic characteristics and selectivity of adsorption on the crosslinking degree, textural features, and liquid-phase composition in the region of extremely low adsorbent surface coverages (Henry's range). We have detected that, in the case of HPSNs, the dependence of the thermodynamic characteristics of adsorption on the liquid-phase composition is different than for classical HPLC stationary phases. Moreover, we scrutinize the impact of the molecular structure of the studied aromatic compounds on the thermodynamic characteristics and selectivity of the liquid-phase adsorption on HPSNs. Investigating liquid-phase adsorption selectivity, we demonstrate the pivotal role of π-π interactions in separating aromatic compounds on HPSNs. Eventually, we unveil that the thermodynamic characteristics of adsorption peculiarly depend on the crosslinking degree due to the profound impact of the crosslinking on the electronic density in benzene rings in HPSNs, whereas the separation throughput peaks for the polymer with a 500% crosslinking degree, attributed to its exceptionally rigid network structure, moderate swelling and micropore volume, and minimum specific surface area.

Comparison of Chromatographic Stationary Phases Using a Bayesian-Based Multilevel Model.

Herein, we used a Bayesian multilevel model of chromatographic retention to compare five reversed-phase high-performance liquid chromatography stationary phases: XBridge Shield RP18, XTerra MS C18, XBridge Phenyl, XBridge C8, and Xterra MS C8. For this, we used a large data set of retention times collected using chromatographic techniques coupled with mass spectrometry. The experiments were conducted in gradient mode for an initial mixture of 300 small analytes for a wide range of pH values in methanol and acetonitrile at two temperatures and for three gradient durations. Our analysis was based on a mechanistic model derived from the principles and fundamentals of liquid chromatography and utilized previously reported chromatographic parameters. The data and model were used to characterize the between-column differences in the chromatographic parameters of the neutral, acidic, and basic analytes. The analysis provides an interpretable summary of stationary-phase properties that can be used in decision-making, i.e., finding the best chromatographic conditions using limited experimental data. The proposed approach is an interesting alternative to the existing approaches used to compare chromatographic stationary phases.

Immobilization of two dendritic organic phases onto silica and their molecular shape recognition for polycyclic aromatic hydrocarbons, tocopherols and carotenoid isomers

BackgroundMolecular shape selectivity, based on the size and shape parameters of the molecule, such as length and planarity, is a separation process that can be used for compounds with restricted shapes, such as isomers. The separation of geometric isomers is challenging because these compounds have similar physicochemical properties but differ slightly in molecular shape. The ability to separate and quantify these isomers is important in high performance liquid chromatography (HPLC), which is one of the most widely used techniques in separation science today, because the shape of the molecule has a strong influence on biological processes. ResultsWe prepared symmetrical discoidal dendrimeric organomolecule gelators (GSDM) and o-phenylenediamine-derived low-molecular-weight dendrimeric organomolecule gelators (G1) and bonded them to silica surfaces. The dendritic organic compound-grafted silica (SiO2@GSDM and SiO2@G1) was used as HPLC stationary phases for the separation of shape-restricted isomers of polycyclic aromatic hydrocarbons (PAHs), carotenoids and tocopherols. The two phases exhibit a very high molecular shape selectivity compared to the commercially available alkyl phases. There are differences in molecular shape selectivity between the two stationary phases. Changes in the chemical structure of dendritic organic compounds can alter the orientation of the molecules, as well as changes in the molecular recognition ability. It was found that SiO2@GSDM has high molecular linear selectivity for PAHs at different temperatures, even at 50 °C. The planar selectivity of SiO2@GSDM was better for triphenylene and o-terphenyl benzenes compared to SiO2@G1. SignificanceThis separation behavior may be attributed to the combined effect of weak interaction centers, which allowed the effective separation of bioactive and shape-restricted isomers through multiple interactions. Furthermore, SiO2@GSDM showed better separation of tocopherols and carotenoids, suggesting that the backbone and ordered structure of organic molecular gelators is an effective way to improve the shape selectivity of the molecules, whereas the molecular orientation of the functional groups influences the separation mechanism of the shape-restricted isomers.

Construction of MOFs@COFs composite material as stationary phase for efficient separation of diverse organic compounds

BackgroundThe development of efficent chromatographic stationary phases (SP) with mixed-mode or multiple interactions in high-performance liquid chromatography (HPLC) for the separation of complex samples is a challenging task. Metal organic frameworks (MOFs)-based SP can provide desired multiple interactions and enable the separation of a diverse range of solutes, but have limitations of low column efficiency and poor stability. ResultsHerein, the hybrid MOFs@Covalent organic frameworks (COFs) materials were used as SP in HPLC due to their synergistic structural features. The SiO2@NH2-UiO-66@CTF SP was synthesized by integration of NH2-UiO-66 and covalent triazine framework (CTF) onto silica surface. Due to the unique structure of SiO2@NH2-UiO-66@CTF with hierarchical-pores, this column showed higher column efficiency (up to 49,369 plates m−1 for alkylbenzenes) than the reported columns packed with MOFs-based SP. Owing to the Zr4+-N coordination bonding between CTF and NH2-UiO-66, the structural stability of SiO2@NH2-UiO-66@CTF can be improved. Furthermore, this new column exhibited remarkable column stability with relative standard deviation of retention time of <0.40% after 400 injections. With the combined advantages of multifunctional properties, high column efficiency, and good stability, SiO2@NH2-UiO-66@CTF SP showed excellent selectivity for the separation of a variety of hydrophobic, aromatic, heteroatomic, and hydrophilic analytes. Significance and noveltyThis work not only offers a promising SP with multiple retention mechanisms for HPLC, but also provides an efficient strategy for development of high column efficiency MOFs-based SP with good stability. Moreover, the MOFs@COFs hybrid materials were expanded in application area through this study, and the research results can also afford the foundation for further explore its structural characteristics.

Development progress of stationary phase for supercritical fluid chromatography and related application in natural products

Supercritical fluid chromatography (SFC) is an environment-friendly and efficient column chromatography technology that was developed to expand the application range of high performance liquid chromatography (HPLC) using a supercritical fluid as the mobile phase. A supercritical fluid has a temperature and pressure that are above the critical values as well as relatively dynamic characteristics that are between those of a gas and liquid. Supercritical fluids combine the advantages of high solubility and diffusion, as their diffusion and viscosity coefficients are equivalent to those of a gas, while maintaining a density that is comparable with that of a liquid. Owing to the remarkable compressibility of supercritical fluids, analyte retention in SFC is significantly influenced by the density of the mobile phase. Thus, the column temperature and back pressure are crucial variables that regulate analyte retention in SFC. Increasing the back pressure can increase the density and solubility of the mobile phase, leading to reductions in retention time. The column temperature can affect selectivity and retention, and the degree to which different analytes are affected by this property varies. On the one hand, increasing the temperature reduces the density of the mobile phase, thereby extending the retention time of the analytes; on the other hand, it can also increase the energy of molecules, leading to a shorter retention time of the analyte on the stationary phase. CO2, the most widely employed supercritical fluid to date, presents moderate critical conditions and, more importantly, is miscible with a variety of polar organic solvents, including small quantities of water. In comparison with the mobile phases used in normal-phase liquid chromatography (NPLC) and reversed-phase liquid chromatography (RPLC), the mobile phase for SFC has a polarity that can be extended over a wide range on account of its extensive miscibility. The compatibility of the mobile phase determines the diversity of the stationary phase. Nearly all stationary phases for HPLC, including the nonpolar stationary phases commonly used for RPLC and the polar stationary phases commonly used for NPLC, can be applied to SFC. Because all stationary phases can use the same mobile-phase composition, chromatographic columns with completely different polarities can be employed in SFC. The selectivity of SFC has been effectively expanded, and the technique can be used for the separation of diverse analytes ranging from lipid compounds to polar compounds such as flavonoids, saponins, and peptides. The choice of stationary phase has a great impact on the separation effect of analytes in SFC. As new stationary phases for HPLC are constantly investigated, specialized stationary phases for SFC have also been continuously developed. Researchers have discovered that polar stationary phases containing nitrogen heterocycles such as 2-EP and PIC are highly suitable for SFC because they can effectively manage the peak shape of alkaline compounds and provide good selectivity in separating acidic and neutral compounds.The development of various stationary phases has promoted the applications of SFC in numerous fields such as pharmaceuticals, food production, environmental protection, and natural products. In particular, natural products have specific active skeletons, multiple active groups, and excellent biological activity; hence, these materials can provide many new opportunities for the discovery of novel drugs. According to reports, compounds related to natural products account for 80% of all commercial drugs. However, natural products are among the most challenging compounds to separate because of their complex composition and low concentration of active ingredients. Thus, superior chromatographic methods are required to enable the qualitative and quantitative analysis of natural products. Thanks to technological improvements and a good theoretical framework, the benefits of SFC are gradually becoming more apparent, and its use in separating natural products is expanding. Indeed, in the past 50 years, SFC has developed into a widely used and efficient separation technology. This article provides a brief overview of the characteristics, advantages, and development process of SFC; reviews the available SFC stationary phases and their applications in natural products over the last decade; and discusses prospects on the future development of SFC.

Research progress on the construction and applications of metal-organic frameworks in chromatographic stationary phases

Metal-organic frameworks (MOFs) are a class of porous crystalline materials composed of metal centers or clusters assembled with organic ligands. These materials possess excellent properties, such as large surface areas, high porosities, uniform pore sizes, and diverse structures. Thus, MOFs have been widely applied in various fields, including catalysis, adsorption, sensing, sample pretreatment, and chromatographic separation. The applications of MOFs as stationary phases for chromatographic separation and analysis have attracted considerable attention from the research community in recent years. Compared with traditional chromatographic stationary phases, such as mesoporous silica, nanoparticles, and porous layers, MOFs possess flexible and tunable pore sizes and structures, thereby enabling precise control over their intermolecular interactions. Furthermore, the wide range of functional ligands and topologies of MOFs could potentially facilitate the separation and analysis of complex samples. These unique advantages render MOFs highly suitable for constructing novel chromatographic stationary phases.This article focuses primarily on the construction methods of MOFs as chromatographic stationary phases, and provides an overview of the latest research advancements in their applications in several chromatographic separation techniques such as high performance liquid chromatography (HPLC), gas chromatography (GC), and capillary electrochromatography (CEC). The existing methods for the preparation and construction of MOFs-based chromatographic stationary phases are classified and evaluated. The construction methods for MOFs as stationary phases for HPLC mainly include filling, precursor-doped polymerization, and post-modification. The construction methods for MOFs as stationary phases for GC predominantly include in situ growth, static coating, and dynamic coating. The stationary phases for CEC can be categorized into packed columns, monolithic columns, and open-tubular columns. Compared with monolithic and packed columns, open-tubular CEC (OT-CEC) offers numerous advantages, including a more flexible and convenient preparation method, enhanced compatibility with various separation media, and higher separation efficiency. Consequently, OT-CEC has emerged as an important method for investigating the preparation of stationary phases for CEC. Several methods such as physical adsorption, covalent attachment, and electrostatic interactions have been developed for the preparation and modification of MOFs-based CEC stationary phases, and extensive studies have been conducted to optimize the performance and applications of MOFs in OT-CEC. However, the existing methods for constructing MOFs-based chromatographic stationary phases present certain limitations. Therefore, the selection of the appropriate MOFs, optimization of their preparation methods, and examination of their performance in different separation modes have become the focus of intensive research.This review also summarizes the different analytical targets (e. g., chiral small molecules, biomacromolecules, and nonchiral molecules) and corresponding separation effects achieved using various MOFs-based chromatographic stationary phases. Finally, future studies focusing on the development of MOFs as chromatographic separation media are discussed. Overall, this review provides a valuable reference for the rational construction and practical applications of advanced MOFs-based chromatographic stationary phases.

Synthesis of hierarchically porous zirconium-based metal-organic framework@silica core-shell stationary phase through etching strategy for liquid chromatography

Metal organic frameworks (MOFs) show promise to be employed as stationary phase for high performance liquid chromatography (HPLC), however, the microporous structures of MOFs seriously restrict the diffusion and mass transfer of solute molecules, leading to a low column efficiency. In this paper, the fabrication of hierarchically porous UiO-66@SiO2 (HP- UiO-66@SiO2) core-shell microspheres via H2O2 etching has been proposed as a viable approach to enhance the separation performance of MOFs-based columns for HPLC. Through the direct treatment of the preliminary prepared UiO-66@SiO2 microspheres with H2O2 etching, HP-UiO-66@SiO2 core-shell microspheres were successfully synthesized with an enlarged pore size of up to 9 nm, facilitating efficient mass transfer in chromatographic separation. The prepared HP-UiO-66@SiO2 core-shell microspheres were then explored as stationary phase in HPLC to separate the nonpolar alkyl benzene homologues, the polar aromatic alcohol homologues and the xylene isomers. The results indicated that the baseline separations of these solutes were achieved successfully with narrow peak width and higher resolution than the UiO-66@SiO2 column. The HP-UiO-66@SiO2 column exhibited superior separation performance, reaching a maximum plate number of 134,459/m for fluorene, and showing good reproducibility. As a result, this template-free approach suggests that the fabrication of hierarchically porous MOFs@silica core-shell microspheres is a successful approach to enhance the column efficiency of MOFs-based columns in HPLC.

Homochiral organic molecular cage RCC3-R-modified silica as a new multimodal and multifunctional stationary phase for high-performance liquid chromatography.

In this work, homochiral reduced imine cage was covalently bonded to the surface of the silica to prepare a novel high-performance liquid chromatography stationary phase, which was applied for the multiple separation modes such as normal phase, reversed-phase, ion exchange, and hydrophilic interaction chromatography. The successful preparation of the homochiral reduced imine cage bonded silica stationary phase was confirmed by performing a series of methods including X-ray photoelectron spectroscopy, thermogravimetric analysis, and infrared spectroscopy. From the extracted results of the chiral resolution in normal phase and reversed-phase modes, it was demonstrated that seven chiral compounds were successfully separated, among which the resolution of 1-phenylethanol reached the value of 3.97. Moreover, the multifunctional chromatographic performance of the new molecular cage stationary phase was systematically investigated in the modes of reversed-phase, ion exchange, and hydrophilic interaction chromatography for the separation and analysis of a total of 59 compounds in eight classes. This work demonstrated that the homochiral reduced imine cage not only achieved multiseparation modes and multiseparation functions performance with high stability, but also expanded the application of the organic molecular cage in the field of liquid chromatography.

Naphthalimide-Annulated [n]Helicenes: Red Circularly Polarized Light Emitters

Two [n]heliceno-bis(naphthalimides) 1 and 2 (n = 5 and 6, respectively) where two electron-accepting naphthalimide moieties are attached at both ends of helicene core were synthesized by effective two-step strategy, and their enantiomers could be resolved by chiral stationary-phase high-performance liquid chromatography (HPLC). The single-crystal X-ray diffraction analysis of enantiopure fractions of 1 and 2 confirmed their helical structure, and together with experimental and calculated circular dichroism (CD) spectra, the absolute configuration was unambiguously assigned. Both 1 and 2 exhibit high molar extinction coefficients for the S0-S1 transition and high fluorescence quantum yields (73% for 1 and 69% for 2), both being outstanding for helicene derivatives. The red circularly polarized luminescence (CPL) emission up to 615 nm for 2 with CPL brightness (BCPL) up to 66.5 M-1 cm-1 demonstrates its potential for applications in chiral optoelectronics. Time-dependent density functional theory (TD-DFT) calculations unambiguously showed that the large transition magnetic dipole moment |m| of 2 is responsible for its high absorbance dissymmetry (gabs) and luminescence dissymmetry (glum) factor.

Silica microparticles modified with ionic liquid bonded chitosan as hydrophilic moieties for preparation of high-performance liquid chromatographic stationary phases.

Two novel stationary phases, 1-(4-bromobutyl)-3-methylimidazolium bromide bonded chitosan modified silica and 1-(4-bromobutyl)-3-methylimidazolium bromide bonded chitosan derivatized calix[4]arene modified silica stationary phase, were synthesized using 1-(4-bromobutyl)-3-methylimidazolium bromide bonding chitosan as a polarity regulator solvingthe limitation of the strong hydrophobicity of calixarene in the application of hydrophilic field. The resulting materials were characterized by solid-state nuclear magnetic resonance, Fourier-transform infrared spectra, scanning electron microscopy, elemental analysis, and thermogravimetric analysis. Based on the hydrophilicity endowed by 1-(4-bromobutyl)-3-methylimidazolium bromide bonded chitosan, the retention mode of ILC-Sil and ILCC4-Sil could be effectively switched from the hydrophilic mode to a hydrophilic/hydrophobic mixed mode and could simultaneously provide various interactions with solutes, including hydrophilic, π-π, ion-exchange, inclusion, hydrophobic, and electrostatic interactions. On the basis of these interactions, successful separation and higher shape selectivity were achieved among compounds that vary in polarity under both reverse-phase and hydrophilic interactive liquid chromatography conditions. Moreover, the ILCC4-Sil was successfully applied tothe determination of morphine in actual samples using solid-phase extraction and mass spectrometry. The LOD and LOQ were 15 pg/mL and 54 pg/mL, respectively. This work presents an exceptionally flexible adjustment strategy for the retention and selectivity of a silica stationary phase by tuning the modification group.

Synthesis and Stereodynamic and Emission Properties of Dissymmetric Bis-Aryl Carbazole Boranes and Identification of a CPL-Active B–C Atropisomeric Compound

We synthesized bis-aryl carbazole borane derivatives having emissive properties and axial chirality. The resolution of a thermally stable atropisomeric pair (compound 1b), due to a B-C chiral axis, was achieved by chiral stationary-phase high-performance liquid chromatography (CSP-HPLC). Complete photophysical properties of all compounds were measured and simulated by time-dependent density functional theory (TD-DFT) calculations of the complete fluorescence cycle, and circularly polarized luminescence spectra were obtained for the atropisomers of compound 1b, whose absolute configuration was derived using a TD-DFT simulation of the electronic circular dichroism (ECD) spectra.

Fluoro-Functionalized Spherical Covalent Organic Frameworks as a Liquid Chromatographic Stationary Phase for the High-Resolution Separation of Organic Halides.

The development of novel stationary phases with specific functionality is of great importance in chromatographic separation. Herein, we fabricated fluoro-functionalized spherical covalent organic frameworks (SF-COFs) via a bottom-up strategy as stationary phases for high-performance liquid chromatography (HPLC). Benefiting from the significant monodispersity, narrow size distribution, and high fluorine content, the SF-COFs packed column showed high column efficiency and excellent resolution for the separation of the organic fluorides involving polyfluorobenzenes, polychlorobenzenes, polybromobenzenes, perfluoroalkyl methacrylates, and halogenated trifluorotoluenes, which cannot be separated on the fluorine-free spherical covalent organic frameworks packed column. Especially, the column efficiency of 20 100-38 500 plates/m was obtained for polyfluorobenzenes, and the relative standard deviations of the retention time for continuous 10 separations of polychlorobenzenes and polybromobenzenes were less than 0.98%. Furthermore, the prepared SF-COFs packed column showed overwhelming superiority in the separation of organic halides compared with commercial C18 and pentafluorophenyl (PFP) packed columns. In addition, the compounds with different hydrophobicity or aromatic ring structure were also successfully separated on the SF-COFs packed column. This work extended the application of spherical COFs and provided a new way to introduce specific functional groups into the COF-based stationary phase for HPLC.

Direct electrochemical monitoring of in vitro glucuronidation metabolism by the hydrophobic selectivity of lipophilic micelles

Glucuronidation metabolism plays an important role in the detoxification and clearance of lipophilic drugs. High-performance liquid chromatography (HPLC) is the most common technology for the separation and analysis of drug metabolism. Inspired by the hydrophobic selectivity of the alkyl bonded silica stationary phase in HPLC, we prepared the functional membrane consisting of lipophilic micelles and mesoporous silica (designated as LM@MSM) modified on indium tin oxide (ITO) electrode, and performed a detailed investigation of its permselectivity towards different anthraquinones (AQs) and their glucuronide metabolites. We demonstrated that the diffusion flux of these AQ analytes was relevant to their chemical structure and partition coefficient, thus leading to the hydrophobic selectivity of the LM@MSM. This selectivity was further supported by the retention behavior of these AQ analytes in HPLC based on the hydrophobic-subtraction model. Consequently, the developed LM@MSM sensor was successfully applied to the direct and rapid electrochemical monitoring of in vitro glucuronidation metabolism of lipophilic AQs with high selectivity and sensitivity.

Synthesis and application of chitosan thiourea derivatives as chiral stationary phases in HPLC

Chitosan 2-thiourea derivatives with various substituents, including 3-(methylthio)propyl, phenyl, octyl and ethoxycarbonyl, at the 2-position of the glucosamine skeleton were prepared via isothiocyanates with the above substituents. The obtained chitosan 2-thiourea derivatives without ethoxycarbonyl were then esterified to develop a new series of chitosan 2-thiourea-3,6-diphenylcarbamate derivatives. The enantioseparation properties of the obtained chitosan derivatives were examined by high-performance liquid chromatography (HPLC). These results demonstrated that these chitosan 2-[3-(methylthio)propylthiourea]-3,6-diphenylcarbamate derivatives showed attractive chiral recognition abilities, especially for dihydropyridine calcium antagonist racemates. This result was probably attributed to the fact that the 2-thiourea substituents of this series of chitosan derivatives, as well as the 3,6-phenylcarbamate substituents, provided more favorable sites, which evidently enhanced the interactions between the enantiomers and the chitosan derivatives. The mechanism involved in the enantioseparation of the chitosan 2-[3-(methylthio)propylthiourea]-3,6-diphenylcarbamate derivatives was further discussed by molecular docking simulation.

Separation of anilines by a covalent triazine-triphenyl polymer as a stationary phase for their normal-phase and reverse-phase determination by high-performance liquid chromatography (HPLC)

A covalent triazine-triphenyl polymer (CTPCC-TP) was examined as a novel column packing material for high-performance liquid chromatography (HPLC). The separation performance of CTPCC-TP was characterized using anilines as the analytes in reverse-phase (RP) and normal-phase (NP) modes. Various retention mechanisms, such as hydrogen-bonding, π-π, and hydrophobic-hydrophobic interactions, endowed CTPCC-TP with a hydrophilic-hydrophobic balance performance, demonstrating promise for applications as a stationary phase for HPLC in normal-phase and reversed-phase. The CTPCC-TP column showed good reproducibility with relative standard deviations from 0.28 to 0.93%, 1.73 to 3.12%, 1.57 to 3.69%, and 1.36 to 2.54% for retention time, peak height, peak area, and peak width at half maximum, respectively. In the normal and reverse phase modes, a thermodynamically spontaneous transfer process is demonstrated through negative ΔG values controlled by negative ΔH and ΔS values during the chromatographic separation. Excellent chemical and solvent stabilities, and the hydrophilic-hydrophobic balance performance make CTPCC-TP a good candidate for use in HPLC.

Understanding the Working Mechanism of the Novel HKUST-1@BPS Composite Materials as Stationary Phases for Liquid Chromatography.

Composite materials have been used based on coordination polymers or microporous metal-organic frameworks (MOFs) combined with mesoporous matrices for adsorption-related techniques, which enable outflanking some adverse phenomena manifested during pristine components operation and enhance the performance and selectivity of the resulting materials. In this work, for the first time, the novel HKUST-1@BPS composites synthesized by the microwave-assisted (MW) technique starting from microporous HKUST-1 (Cu3(btc)2) MOF and biporous silica matrix (BPS) with bimodal mesopore size distribution were comparatively studied as materials for liquid-phase adsorption techniques utilizing the high-performance liquid chromatography (HPLC) method and benzene as a model adsorbate. It was established that the studied HKUST-1@BPS composites can function as stationary phases for HPLC, unlike the pristine HKUST-1 and bare BPS materials, due to the synergetic effect of both components based on the preliminary enhanced adsorbate mass transfer throughout the silica mesopores and, subsequently, its penetrating into HKUST-1 micropores. The suggested mechanism involves the initial deactivation of open metal Cu2+ sites in the HKUST-1 framework structure by isopropanol molecules upon adding this polar component into the mobile phase in the region of the isopropanol concentration of 0.0 to 0.2 vol.%. Thereafter, at the medium range of varying the isopropanol concentration in the eluent of 0.2 to 0.3 vol.%, there is an expansion of the previously inaccessible adsorption centers in the HKUST-1@BPS composites. Subsequently, while further increasing the isopropanol volume fraction in the eluent in the region of 0.3 to 5.0 vol.%, the observed behavior of the studied chromatographic systems is similar to the quasi-normal-phase HPLC pattern. According to the obtained thermodynamic data, benzene adsorption into HKUST-1 micropores from solutions with a vol.% of isopropanol in the range of 0.4 to 5.0 follows the unique entropy-driven mechanism previously described for the MIL-53(Al) framework. It was found that HKUST-1 loading in the composites and their preparation conditions have pronounced effects on their physicochemical properties and adsorption performance, including the adsorption mechanism.

Separating and purifying of Panax notoginseng saponins using a rosin-based polymer-bonded with silica as a high-performance liquid chromatography stationary phase

In this study, methyl methacrylate and fumaropimaric acid tris (ethylene glycol) ester were used as functional monomer and cross-linker, respectively, to prepare a rosin-based polymer-bonded with silica as a high-performance liquid chromatography stationary phase via a coating–bonding method. The produced stationary phase was characterized by thermogravimetric analysis, Fourier transform infrared spectroscopy, and microporous physical adsorption, and the specific surface area and particle size distribution were calculated. The results showed that the stationary phase particles have good sphericity, having a particle size of approximately 4.81 µm, specific surface area of 280 m2·g−1, and average pore diameter of 6.76 nm, and were nontoxic. Further, column evaluation revealed that the produced stationary phase exhibited typical reversed-phase chromatographic behavior, hydrophobicity, good reproducibility and strong steric selectivity. Thus, the resin was used for separating and purifying Panax notoginseng saponins, and the effects of the chromatographic conditions on separating Panax notoginseng saponins were investigated; moreover, the thermodynamic parameters of column separation were studied. Finally, Panax notoginseng saponins purification using the new stationary phase resulted an increase in purity from 45.27% to 85.08%.