Sol-gel stationary phases for capillary electrochromatography

A review is presented on the current state of the art and future trends in the development of sol-gel stationary phases for capillary electrochromatography (CEC). The design and synthesis of stationary phases with prescribed chromatographic and surface charge properties represent challenging tasks in contemporary CEC research. Further developments in CEC as a high-efficiency liquid-phase separation technique will greatly depend on new breakthroughs in the area of stationary phase development. The requirements imposed on CEC stationary phase performance are significantly more demanding compared with those for HPLC. The design of CEC stationary phase must take into consideration the structural characteristics that will provide not only the selective solute/stationary phase interactions leading to chromatographic separations but also the surface charge properties that determine the magnitude and direction of the electroosmotic flow responsible for the mobile phase movement through the CEC column. Therefore, the stationary phase technology in CEC presents a more complex problem than in conventional chromatographic techniques. Different approaches to stationary phase development have been reported in contemporary CEC literature. The sol-gel approach represents a promising direction in this important research. It is applicable to the preparation of CEC stationary phases in different formats: surface coatings, micro/submicro particles, and monolithic beds. Besides, in the sol-gel approach, appropriate sol-gel precursors and other building blocks can be selected to create a stationary phase with desired structural and surface properties. One remarkable advantage of the sol-gel approach is the mild thermal conditions under which the stationary phase synthesis can be carried out (typically at room temperature). It also provides an effective pathway to integrating the advantageous properties of organic and inorganic material systems, and thereby enhancing and fine-tuning chromatographic selectivity of the created hybrid organic-inorganic stationary phases. This review focuses on recent developments in the design, synthesis, characterization, properties, and applications of sol-gel stationary phases in CEC.     

Evaluation of an amide‐based stationary phase for supercritical fluid chromatography

A relatively new stationary phase containing a polar group embedded in a hydrophobic backbone (i.e., ACE ® C18‐amide) was evaluated for use in supercritical fluid chromatography. The amide‐based column was compared with columns packed with bare silica, C₁₈ silica, and a terminal‐amide silica phase. The system was held at supercritical pressure and temperature with a mobile phase composition of CO₂ and methanol as cosolvent. The linear solvation energy relationship model was used to evaluate the behavior of these stationary phases, relating the retention factor of selected probes to specific chromatographic interactions. A five‐component test mixture, consisting of a group of drug‐like molecules was separated isocratically. The results show that the C₁₈‐amide stationary phase provided a combination of interactions contributing to the retention of the probe compounds. The hydrophobic interactions are favorable; however, the electron donating ability of the embedded amide group shows a large positive interaction. Under the chromatographic conditions used, the C₁₈‐amide column was able to provide baseline resolution of all the drug‐like probe compounds in a text mixture, while the other columns tested did not.     

Classification of special octadecyl-bonded phases by the carotenoid test

Amongst the numerous base-deactivated ODS phases obtained by increasing the bonding density or/and by efficient endcapping treatments, some particular stationary phases have been developed, to limit the additional interactions of basic compounds with residual silanols, to work at extreme pH or with rich water mobile phases. Horizontal polymeric phases, sterically protected ones, hybrid silicas, propylene bridge, are particularly used for this purpose. Octadecyl chains with embedded polar groups and hydrophilic endcapping are also used in this goal. The properties of these phases were studied with a simple test consisting in the injection of carotenoid pigments in Subcritical Fluid Chromatography. The molecules used and the nature of the mobile phase allow the determination of hydrophobicity, polar site accessibility and type or/and bonding density of the stationary phases. Whatever the type of the phases, the particular stationary phases do not show any remarkable property, in comparison to other base-deactivated C18-bonded phases. On the other hand, embedded and polar-endcapped phases display a specific behaviour in regard of hydrophilic interactions, which are highlighted by the absence of water in the subcritical fluid. Additional properties of these phases are described, such as steric recognition and retention performances. As expected, polar-embedded phases are less retentive than classical ODS ones, but are sometimes able to provide greater steric recognition. On the other hand, the polar-endcapped phases display greater hydrophobicity than polar-embedded ones. From a simple classification diagram based on chromatographic properties, differences can be noticed between the polar-embedded groups (amide, carbamate, ether, sulfonamide) and between embedded and endcapped phases. Surprising behaviours are also noticed for some on the tested phases.     

金属有机框架色谱固定相的研究进展

金属有机框架(MOFs)是由金属离子或金属簇与有机配体通过配位作用自组装形成的一类新型多孔材料. MOFs具有独特的拓扑结构、丰富的孔隙结构、可调的孔道尺寸、巨大的比表面积以及灵活的表面修饰等特征,是色谱分离领域颇受关注的一类新型固定相. 综述了近几年MOFs材料作为固定相在气相色谱、液相色谱及手性拆分等领域应用的研究进展,展现MOFs材料在色谱分离领域的优异性能和应用潜力,并对MOFs材料在色谱固定相领域今后的发展进行了展望.     

金属有机骨架及其复合材料在分离领域中的应用进展

金属有机骨架（MOFs）是一类由无机金属离子与有机配体自组装形成的新型有机-无机杂化多孔材料,因具有比表面积超高、结构多样、热稳定性良好、孔道尺寸和性质可调等优势,在分离领域表现出重要的应用价值。然而,采用传统方法制备的MOFs多为粒径在微米或亚微米尺度的晶体,且颗粒形貌不规则,因此限制了MOFs在样品前处理和色谱固定相等领域的应用和发展。构建基于MOFs的复合材料是弥补MOFs应用缺陷的一项有效措施,有望在保留MOFs优越的分离特性的同时,引入基体材料的特定性能。该文简要综述了近年来MOFs及其复合材料在吸附、样品前处理和色谱固定相等分离领域中的应用进展,并对MOFs在分离科学中的应用前景做出展望。     

Hydride-Based Separation Materials for High Performance Liquid Chromatography and Open Tubular Capillary Electrochromatography

 Silica hydride is a recent development in chromatographic support materials for high performance liquid chromatography (HPLC) where hydride groups replace 95% of the silanols on the surface. This conversion changes many of the fundamental properties of the material as well as the bonded stationary phases that are the result of further chemical modification of the hydride surface. Some unique chromatographic properties of hydride-based phases are described as well as some general application areas where these bonded materials may be used in preference to or have advantages not available from typical stationary phases. The fabrication, properties and applications of etched chemically modified capillaries for electrophoretic analysis are also reviewed. It is shown that the etching process creates a surface that is fundamentally different than a bare fused silica capillary. The new surface matrix produces unique electroosmotic flow properties and is more compatible with basic and biological compounds. After chemical modification of the surface, the bonded organic moiety (stationary phase) contributes to the control of migration of solutes in the capillary. Both electrophoretic and chromatographic processes take place in the etched chemically modified capillaries leading to a variety of experimental variables that can be used to optimize separations. A number of examples of separations on these capillaries are described.     

Thermally responsive chromatographic materials using functional polymers

Functional polymers that respond to small changes in environmental stimuli with large changes in their structure and properties are often called "intelligent" polymers. We have modified material surfaces with such polymers and used them in separation systems. Silica beads were modified with the temperature-responsive polymer poly(N-isopropylacrylamide) (PNIPAAm). PNIPAAm-grafted surfaces exhibited temperature-driven alterations of hydrophilic-hydrophobic surface-properties. Using this feature, PNIPAAm and related temperature-responsive polymers have been used to generate temperature-sensitive stationary phases for chromatographic separations. We attached several different functional polymers, including temperature- and pH-responsive polymers, to silica beads. These temperature-responsive stationary phases are useful in development of separation methods since adjusting the temperature represents an extra tool for optimizing the selectivity. Applications of thermally responsive columns for separations in the HPLC mode are demonstrated.     

Chromatographic characterization of hydrophilic interaction liquid chromatography stationary phases: hydrophilicity, charge effects, structural selectivity, and separation efficiency

Fourteen commercially available particle-packed columns and a monolithic column for hydrophilic interaction liquid chromatography (HILIC) were characterized in terms of the degree of hydrophilicity, the selectivity for hydrophilic-hydrophobic substituents, the selectivity for the regio and configurational differences in hydrophilic substituents, the selectivity for molecular shapes, the evaluation of electrostatic interactions, and the evaluation of the acidic-basic nature of the stationary phases using nucleoside derivatives, phenyl glucoside derivatives, xanthine derivatives, sodium p-toluenesulfonate, and trimethylphenylammonium chloride as a set of samples. Principal component analysis based on the data of retention factors could separate three clusters of the HILIC phases. The column efficiency and the peak asymmetry factors were also discussed. These data on the selectivity for partial structural differences were summarized as radar-shaped diagrams. This method of column characterization is helpful to classify HILIC stationary phases on the basis of their chromatographic properties, and to choose better columns for targets to be separated. Judging from the retention factor for uridine, these HILIC columns could be separated into two groups: strongly retentive and weakly retentive stationary phases. Among the strongly retentive stationary phases, zwitterionic and amide functionalities were found to be the most selective on the basis of partial structural differences. The hydroxyethyl-type stationary phase showed the highest retention factor, but with low separation efficiency. Weakly retentive stationary phases generally showed lower selectivity for partial structural differences.     

Chromatographic classification and comparison of commercially available reversed-phase liquid chromatographic columns containing polar embedded groups/amino endcappings using principal component analysis

Polar embedded phases have become increasingly popular in liquid chromatography (LC) analysis. These phases can produce diverse chromatographic selectivities as a result of their differing base silica, the type of polar embedded group (i.e. amide, urea, carbamate, ether or sulphonamide moieties) and the length of the alkyl ligand. Four column characterization protocols, using differing test probes, have been used to characterize 18 of these phases together with 17 alkyl phases (some of which contained novel polar endcapping, i.e. amino), which have been evaluated using principal component analysis (PCA). PCA provided graphical comparisons of the differences/similarities between these phases and between their corresponding C-alkyl, amino endcapped and enhanced polar selectivity phases.     

Comparison of C18 silica bonded phases selectivity in micellar liquid chromatography

The paper describes a new test designed in micellar LC (MLC) to compare the commercial C18 stationary phase properties. This test provides the total hydrophobicity, hydrophilicity, steric selectivity, hydrogen bonding, and ion‐exchange capacity properties calculation of the ODS stationary phases. Both the test compounds and chromatographic separation conditions choice for column characterization in MLC are detailed. The chromatographic performance of several stationary phases that are used in MLC was evaluated with specific chromatographic test comprising nine test compounds, possessing different physico‐chemical properties, which were injected on different supports with two micellar mobile phases: one at pH 7.0 (0.075 mol/L SDS and 1.5% v/v 1‐pentanol), and other at pH 2.7 (0.075 mol/L SDS and 1.5% v/v 1‐pentanol adjusted to pH by TFA). Fundamental column chromatographic properties were obtained under these conditions and were treated by hierarchical cluster analysis. From the results of cluster analysis, two closely related groups of columns are distinguished, and it was shown that the chosen column characteristic parameters allow characterizing both sorbent and micellar chromatographic system properties. Eleven columns were analyzed by this test, which allows a comparison of columns with the aim of the selection of suitable and analogous column for the analysis with MLC.     

Stationary phases for thin-layer chromatography

This paper presents a review of the literature concerning development of the stationary phases for thin-layer chromatography (TLC) in the last ten years. The silica gel remains the most important adsorbent for TLC separation. The kinetic properties of the silica-gel thin layer and the new TLC plates have been presented. Other materials used as stationary phase were alumina, zirconium oxide, Florisil, and ion-exchanger. Chemically new bonded stationary phase development is also discussed. The improvement of the separations of some organic mixtures by impregnation of silica gel, cellulose, or polyamide plates (with transition metal ions and silver salts) and their applications is presented. The impregnation of the thin layer with organic stationary phase and inclusion complexes is another method used for the enhancement of the separation efficiences. Another modality to improve the selectivity in TLC using ion-pairing as reagent of impregnation is described as well. The actual state of chiral separation by TLC is discussed with concrete references to recent advances in chiral stationary phases. The use of nonpolar chemically bonded stationary phases impregnated with transitional metal ions is presented as chiral stationary phases. The cellulose, modified cellulose, chitin, chitosan, and their derivatives are presented and their potential for the analysis of the racemates is discussed. The cyclodextrines and macrocyclic antibiotics were used with very good results for enantiomeric separation by TLC. A new separation approach with molecular imprinting polymers was reported as a chiral stationary phase in TLC. The examples provide a wide range of structural types that can be readily resolved enantiomerically by TLC.     

Phenyl‐bonded stationary phases—The influence of polar functional groups on retention and selectivity in reversed‐phase liquid chromatography

The chromatographic properties of four phenyl‐bonded phases with different structures were studied. The columns used were packed with a stationary phase containing a phenyl ring attached to the silica surface using different types of linkage molecules. As a basic characteristic of the bonded phases, the hydrophobicity and silanol activity (polarity) were investigated. The presence of the polar amino and amide groups in the structure of the bonded ligand strongly influences the polarity of the bonded phase. Columns were compared according to methylene selectivity using a series of benzene homologues and according to their shape and size selectivity using polycyclic aromatic hydrocarbons. The measurements were done using methanol/water and acetonitrile/water mobile phases. The presented results show that the presence of polar functional groups in the ligand structure strongly influences the chromatographic properties of the bonded phase.     

Characterization of monolithic columns for HPLC

Monolithic stationary phases and columns have rapidly become highly popular separation media for liquid chromatography, in spite of their recent discovery. However, their most important features have not yet been completely clarified. A complete understanding of their performance and of their intrinsic characteristics will require the systematic acquisition of many series of reliable experimental data and their consistent analysis from different points of view. Progress in their design and production requires now that the chromatographic behavior of monolithic columns be studied in close connection with their physico-chemical and structural properties. The main goal of this review is to summarize fundamental information on some physico-chemical and chromatographic characteristics of monolithic stationary phases and columns for RPLC. The material reviewed deals only with silica-based monolithic columns. First, structural information on the porosities and the size of the pores in monolithic columns is reported. Second, results of chromatographic studies that deal with the characterization of monolithic columns are summarized. Third, results of detailed studies made on the adsorption equilibrium and the surface heterogeneity of monolithic stationary phases are presented. Finally, results on the mass transfer kinetics in monolithic columns derived from the applications of the classical random-walk model and of the moment theory to a new model of the monolith are discussed.     

Unveiling the Role of Hydrogen Bonds in Luminescent N-Annulated Perylene Liquid Crystals

We report the liquid-crystalline (LC) and luminescent properties of a series of N-annulated perylenes ( 1 – 4 ) in whose molecular structures amide and ester groups alternate. We found that the LC properties of these compounds not only depend on the number of hydrogen-bonding units, but also on the relative position of the amide linkers in the molecule. The absence of amide groups in compound 1 leads to no LC properties, whereas four amide groups induce the formation of a wide temperature range columnar hexagonal phase in compound 4 . Remarkably, compound 3 , with two amide groups in the inner part of the structure, stabilizes the columnar LC phases better than its structural isomer 2 , with the amide groups in the outer part of the molecule. Similarly, we found that only compounds 1 and 2 , which have no hydrogen bonding units in the inner part of the molecule, exhibit luminescence vapochromism upon exposure to organic solvent vapors.     

新型碳纳米管色谱固定相制备的研究进展

碳纳米管（CNTs）作为一种新型的功能材料,具有优异的物理、化学和机械性能,已经在分析化学领域得到了广泛的关注和应用。通过填充法或原位化学气相沉积法,可制备CNTs气相色谱固定相;将CNTs沉积在硅胶微球或有机聚合物基质微球表面,可制备填充式CNTs液相色谱固定相;通过包埋共聚法将CNTs嵌入聚合物整体柱内,可制备毛细管CNTs液相色谱整体柱。本文主要综述了近年来CNTs（单壁碳纳米管和多壁碳纳米管）用于色谱固定相制备的研究现状,包括气相色谱及液相色谱,并对该领域今后的发展进行展望。     

Retention pattern profiling of fungal metabolites on mixed-mode reversed-phase/weak anion exchange stationary phases in comparison to reversed-phase and weak anion exchange separation materials by liquid chromatography-electrospray ionisation-tandem mass spectrometry

Retention properties of 79 fungal metabolites (including neutral, acidic, basic, and amphoteric compounds) were evaluated on distinct mixed-mode reversed-phase/weak anion exchange (RP/WAX)-type stationary phases by liquid chromatography-electrospray ionisation-tandem mass spectrometry (LC-ESI-MS/MS) in gradient as well as in isocratic elution mode. The RP/WAX separation materials were prepared by functionalising thiol-modified silica with N-(10-undecenoyl)-3-aminoquinuclidine and N-(10-undecenoyl)-3-alpha-aminotropane, respectively. To evaluate complementarity in chromatographic selectivity the physico-chemically heterogeneous solute set was analysed also on a RP phase (C(18)), an amino-type WAX phase, and a commercially available RP/WAX-like mixed-mode phase. Analytes may interact with the RP/WAX ligands via (attractive/repulsive) ionic, RP-like hydrophobic, as well as hydrophilic (HILIC) retention mechanisms. Individual interactive increments were found to be basically controlled by the nature and amount of organic modifier, pH value of eluent, and ionic strength of buffer additives. It could be demonstrated that RP/WAX columns offer the potential to separate compounds by exploiting a combination of various chromatographic interaction modes, which is not accessible with conventional RP and WAX columns. Such multi-modal properties increase both versatility and degrees of freedom for adjustment of chromatographic selectivity. For example, highly polar mycotoxins such as moniliformin were well retained on RP/WAX-type phases without compromising RP-selectivity for neutral (e.g. aflatoxins) and most basic solutes (e.g. epimer separation of ergot alkaloids) under fully MS-compatible conditions like a hydro-organic eluent with acetonitrile as organic modifier and an acetic acid/ammonium acetate buffer. Flexibility of the employed mixed-mode separation materials may be of value particularly for LC-ESI-MS/MS-based bioanalytics involving analytes with widely varying physico-chemical properties or applications prone to matrix effects.     

Hydride-based silica stationary phases for HPLC: fundamental properties and applications

Silica hydride is a recent development in chromatographic support materials for HPLC where hydride groups replace 95% of the silanols on the surface. This conversion changes many of the fundamental properties of the material as well as the bonded stationary phases that are the result of further chemical modification of the hydride surface. The general approach for fabricating the silica hydride and subsequent bonded phases is reviewed. Properties of the silica hydride surface are compared to those of the standard material obtained in the preparation of most commercial HPLC stationary phases. Some unique chromatographic properties of hydride-based phases are described as well as some general application areas where these bonded materials may be used in preference to or have advantages not available from typical stationary phases.     

Propylamido-Modified C18 Reversed-Phase for High-Performance Liquid Chromatography

The synthesis and chromatographic properties of a propylamido-modified C18 stationary phase are described. The propylamido-modified C18 phase was prepared by bonding stearic acid to 3-aminopropyl-silica gel. The synthesis was simple and reproducible through amide formation by using N-hydroxysuccinimide and dicyclohexylcarbodiimide. This prepared stationary phase exhibited excellent abilities to separate polycyclic aromatic hydrocarbons (PAHs), polysiloxanes and phenols by HPLC. The chromatographic results show that both the C3 moiety and the amido-group in this phase contribute significantly to the HPLC separation process. The chromatographic behavior of the prepared phase and commercial C18 phases was also compared in analyses of PAHs and phenols. Although the commercial C18 phases have been extensively used, both the unique selectivity and the highly reproducible synthesis of our prepared phase make it an excellent complement to ordinary C18 phases.     

Synthesis and characterization of new organometallic complexes bonded stationary phases for high-performance liquid chromatography

Summary The preparation, characterization and potential liquid chromatographic applications of various organometallic iron complexes silica stationary phases are presented. These new supports are synthesized by covalently linking ferrocene, as well as some of its cationic derivatives, to appropriately derivatized silica support matrices. These columns exhibit moderate to high selectivity towards the separation of polycyclic aromatic hydrocarbons (PAHs). A charge transfer retention mechanism has been proposed. A comparison with a reference stationary phase, 3,5-dinitrobenzamide (DNB), to quantify the acceptor power of the new stationary bonded phases, is also reported. Finally, the effect of varying the derivatives of the bonded metallocene on PAHs retention is discussed.