Selectivity Of Stationary Phases Research Articles

Novel PEI/Zein core–shell composite as mixed-mode stationary phase for high performance liquid chromatography

Based on the adhesion of polyethyleneimine (PEI), a novel PEI/zein co-modified core-shell stationary phase (PEI/Zein@SiO2) was prepared by doping zein to form a composite modification layer. The stationary phase achieved effective separation of nucleosides, bases and antibiotics in hydrophilic interaction mode on account of the hydrophilic groups of composite coating. With the hydrophobicity of zein, the flavones could be separated in reversed-phase mode. In short, the separation and analysis of hydrophilic/hydrophobic compounds were accomplished excellently by the PEI/Zein@SiO2 column with mixed double mode. The prepared chromatographic stationary phase not only avoided the dissolution of zein, but also covered the strong adsorption of some analytes caused by silica hydroxyl groups on the surface of silica spheres. The morphological structure and specific surface area of the material were reflected by various characterization techniques. Hydrophilic/hydrophobic compounds were used as tested analytes to research separation performance and retention mechanisms of PEI/Zein@SiO2 column. The stability and reproducibility of the PEI/Zein@SiO2 stationary phase were satisfied. Therefore, the modification of zein could improve the separation selectivity of stationary phase effectively for complex samples, which had the potential to be one of the significant potential application materials in stationary phase packing.

Insights into the selectivity of polar stationary phases based on quantitative retention mechanism assessment in hydrophilic interaction chromatography

Hydrophilic interaction chromatography (HILIC) offers different selectivity than reversed-phase liquid chromatography (RPLC). However, our knowledge of the driving force for selectivity is limited and there is a need for a better understanding of the selectivity in HILIC. Quantitative assessment of retention mechanisms makes it possible to investigate selectivity based on understanding the underlying retention mechanisms. In this study, selected model compounds from the Ikegami selectivity tests were evaluated on different polar stationary phases. The study results revealed significant insights into the selectivity in HILIC. First, hydroxy and methylene selectivity is driven by hydrophilic partitioning; but surface adsorption for 2-deoxyuridine or 5-methyluridine reduces the selectivity factor. Furthermore, the retention of 2-deoxyuridine or 5-methyluridine by surface adsorption in combination with the phase ratio explain the difference in hydroxy or methylene selectivity observed among different stationary phases. Investigations on xanthine positional isomers (1-methylxanthine/3-methylxanthine, theophylline/theobromine) indicate that isomeric selectivity is controlled by surface adsorption; however, hydrophilic partitioning may contribute to resolution by enhancing overall retention. In addition, two pairs of nucleoside isomers (adenosine/vidarabine, 2′-deoxy and 3′-deoxyguanosine) provide an example that isomeric selectivity can also be controlled by hydrophilic partitioning if their partitioning coefficients are significantly different in HILIC. Although more data is needed, the current study provides a mechanistic based understanding of the selectivity in HILIC and potentially a new way to design selectivity tests.

The Effect of the Stationary Phase on Resolution in the HPLC-Based Separation of Racemic Mixtures Using Vancomycin as a Chiral Selector: A Case Study with Profen Nonsteroidal Anti-Inflammatory Drugs

Chiral resolution is a technique of choice, making it possible to obtain asymmetric and enantiomerically pure compounds from a racemic mixture. This study investigated the behavior of vancomycin when used as a chiral additive in high-performance liquid chromatography (HPLC) to separate enantiomers of nonsteroidal anti-inflammatory drugs (NSAIDs), including ketoprofen, ibuprofen, flurbiprofen, and naproxen enantiomeric impurities. We compared two achiral stationary phases (C18 and NH2) to assess the impact of mobile phase composition and stationary phase on the vancomycin retention time in the racemic resolution of drug enantiomers. Our results demonstrated the successful enantioseparation of all drugs using vancomycin in the mobile phase (phosphate buffer 0.05 M/2-propanol, 50/50) with an NH2 column. This enhanced separation on the NH2 column resulted from the chromatography system’s efficiency and vancomycin dimers’ stereoselective interaction on the NH2 surface. This study underscores the importance of stationary phase selection in the chiral resolution of NSAIDs with vancomycin as a chiral additive. It offers valuable insights for future research and development of NSAID chiral separation methods, highlighting potential vancomycin applications in this context.

Changes in the cis-trans isomer selectivity of a reversed-phase liquid chromatography column during use with acidic mobile phase conditions

Reversed-phase liquid chromatography (RPLC) is the analytical tool of choice for monitoring process-related organic impurities and degradants in pharmaceutical materials. Its popularity is due to its general ease-of-use, high performance, and reproducibility in most cases, all of which have improved as the technique has matured over the past few decades. Nevertheless, in our work we still occasionally observe situations where RPLC methods are not as robust as we would like them to be in practice due to variations in stationary phase chemistry between manufactured batches (i.e., lot-to-lot variability), and changes in stationary phase chemistry over time. Over the last three decades several models of RPLC selectivity have been developed and used to quantify and rationalize the effects of numerous parameters (e.g., effect of bonded phase density) on separation selectivity. The Hydrophobic Subtraction Model (HSM) of RPLC selectivity has been used extensively for these purposes; currently the publicly available database of column parameters contains data for 750 columns. In this work we explored the possibility that the HSM could be used to better understand the chemical basis of observed differences in stationary phase selectivity when they occur – for example, lot-to-lot variations or changes in selectivity during column use. We focused our attention on differences and changes in the observed selectivity for a pair of cis-trans isomers of a pharmaceutical intermediate. Although this is admittedly a challenging case, we find that the observed changes in selectivity are not strongly correlated with HSM column parameters, suggesting that there is a gap in the information provided by the HSM with respect to cis-trans isomer selectivity specifically. Further work with additional probe molecules showed that larger changes in cis-trans isomer selectivity were observed for pairs of molecules with greater molecular complexity, compared to the selectivity changes observed for simpler molecules. These results do not provide definitive answers to questions about the chemical basis of changes in stationary phase chemistry that lead to observed differences in cis-trans isomer selectivity. However, the results do provide important insights about the critical importance of molecular complexity when choosing probe compounds and indicate opportunities to develop improved selectivity models with increased sensitivity for cis-trans isomer selectivity.

Selectivity optimization in liquid chromatography via stationary phase tuning.

In striving for the best possible separation, the selectivity of stationary phases as an optimization parameter is often underestimated although there are many ways to influence this powerful tool. This review serves to provide an insight into the various ways of adapting the selectivity of a separation in liquid chromatography. Approaches via temperature and flow rate tuning are discussed as a basis followed by focusing on the stationary phase as the superior optimization parameter. Highly selective stationary phases hereby provide an advantage for groups of similar analytes. For more complex mixtures, separations can be improved using mixed-mode technologies where different retention mechanisms are combined. Serial coupling, mixed-bed columns, and stationary phase optimized selectivity liquid chromatography provide solutions to various degrees. Finally, the advantages of stationary phase tuning over adaption of mobile phase and/or temperature are presented in terms of optimum application range.

Aroma Compounds in Essential Oils: Analyzing Chemical Composition Using Two-Dimensional Gas Chromatography-High Resolution Time-of-Flight Mass Spectrometry Combined with Chemometrics.

Analyzing essential oils is a challenging task for chemists because their composition can vary depending on various factors. The separation potential of volatile compounds using enantioselective two-dimensional gas chromatography coupled with high-resolution time-of-flight mass spectrometry (GC×GC-HRTOF-MS) with three different stationary phases in the first dimension was evaluated to classify different types of rose essential oils. The results showed that selecting only ten specific compounds was enough for efficient sample classification instead of the initial 100 compounds. The study also investigated the separation efficiencies of three stationary phases in the first dimension: Chirasil-Dex, MEGA-DEX DET-β, and Rt-βDEXsp. Chirasil-Dex had the largest separation factor and separation space, ranging from 47.35% to 56.38%, while Rt-βDEXsp had the smallest, ranging from 23.36% to 26.21%. MEGA-DEX DET-β and Chirasil-Dex allowed group-type separation based on factors such as polarity, H-bonding ability, and polarizability, whereas group-type separation with Rt-βDEXsp was almost imperceptible. The modulation period was 6 s with Chirasil-Dex and 8 s with the other two set-ups. Overall, the study showed that analyzing essential oils using GC×GC-HRTOF-MS with a specific selection of compounds and stationary phase can be effective in classifying different oil types.

Effect of stereoconfiguration of aromatic ligands on retention and selectivity of terphenyl isomer-bonded stationary phases

The structure of ligands has a significant influence on the separation properties of alkyl and aromatic phases in reversed-phase liquid chromatography. Compared with alkyl phases, the effect of stereoconfiguration of aromatic ligands on the retention and selectivity of stationary phases has rarely been addressed. To illustrate the issue, three terphenyl isomer-bonded stationary phases were prepared via the coupling chemistry of isocyanate with terphenyl amine isomers, 3,4-diphenylaniline, 2,4-diphenylaniline and 4-amino-p-terphenyl, respectively. The retention behaviors of stationary phases were assessed in terms of retention strength, selectivity, hydrophobic and π-π interaction by five kinds of solutes. It is found that the selectivity towards the solutes is slightly larger on the branched m-terphenyl-bonded phase (m-π3) than o-terphenyl-bonded phase (o-π3) but is significantly improved on the chain p-terphenyl-bonded phase (p-π3). The results can be interpreted by the ease self-adjustment of the conformation of the chain p-terphenyl ligand and the smaller steric effect of p-π3 towards the insertion of solutes into the ligand brushes. In addition, the p-π3 yields excellent selective separation towards aromatic solutes. These findings are of significance in the design of aromatic stationary phases.

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.

End-modification of poly(ether-carbonate) copolymer by adamantane cages: An effective approach for improving the selectivity of gas chromatographic stationary phases

This work reports the investigation of a new poly(propylene-carbonate) copolymer terminated by the adamantane cages (APPC) as the stationary phase for gas chromatographic (GC) analyses. In GC, the selectivity of a stationary phase is the key factor that governs the column separation performance for analytes, particularly those of high similarity in structures and properties. As such, we employed more than a dozen of isomer mixtures of separation difficulty for investigating the separation performance of the APPC column, involving the isomers of alkanes, alkylbenzenes, halobenzenes phenols and anilines. Meanwhile, the column coated with poly(propylene carbonate) diol (PPCD) differing from APPC only in their terminal groups and two commercial columns coated with polyethylene glycol (PEG) and polysiloxane, respectively, were used as the reference columns. The separation results evidenced the distinctly advantageous performance of the APPC column over the reference columns. Moreover, the APPC column showed excellent repeatability and reproducibility with the relative standard deviation (RSD) values over the range of 0.01%–0.04% for run-to-run, 0.15%–0.28% for day-to-day and 3.4%–3.9% for column-to-column (n = 4). Its application to GC–MS analysis of the verbena essential oil demonstrated its separation advantages for a wide range of components in practical samples. Up to date, the adamantyl-terminated poly(ether-carbonate) copolymers have not been reported in any fields. Its high-resolution performance demonstrates the feasibility of adamantyl-terminated block copolymers as highly selective stationary phases for GC analyses, which offers a vast room for fundamental researches and applications.

Impact of mobile phase composition on reverse-phase separation of polar basic compounds

Method development for determination of polar basic compounds on ion-suppression reverse-phase liquid chromatography, requires appropriate selection of stationary phase and composition and pH value of the mobile phase. The addition of the organic solvent in aqueous buffers with defined pH value, results in shift of the pH value of the mobile phase, resulting in significant change in chromatographic retention and selectivity. The aim of this study was to evaluate the impact of the variation of pH value of the mobile phase and percentage of the organic solvent, on the chromatographic separation of basic compound (octenidine dihydrochloride). The design of experiment was used for the optimization of the chromatographic conditions and selection of the most suitable mobile phase composition. The optimal composition of the mobile phase, providing satisfactory chromatographic performance based on ion-suppression reverse-phase chromatography, included 0.1M citric acid/0.2M sodium dihydrogen phosphate, pH =2.5, combined with 70% v/v organic solvent-acetonitrile. The suitability of the method was confirmed by satisfactory results for the system suitability parameters, specificity, linearity and precision. Key Words: polar basic compounds, ion-suppression, pH value, composition of mobile phase

Does High Polarity Mean High Retention on Stationary Phases in Gas Chromatography?

Stationary phase chemistry, polarity, and selectivity have been of ongoing interest since the inception of gas chromatography (GC) in the 1950s. In the early days when most analyses were performed on packed columns, there were hundreds of stationary phase materials available. Today, with modern capillary columns, most GC analyses are performed with a few stationary phases, with a wide array of choices for specialty applications. Stationary phases are often classified using the broad term polarity, with polar stationary phases recommended for separating polar analytes and nonpolar stationary phases recommended for nonpolar analytes. In this installment, we examine the idea of stationary phase polarity. We examine the assumptions inherent in the most popular stationary phase polarity evaluating systems—McReynolds constants and the polarity scale. We see that high polarity does not always mean greater retention or higher selectivity.

Selection of Stationary Phase and Mobile Phase in High Performance Liquid Chromatography

Selection of ideal mobile phase and stationary phase is very important to get accurate separation of mixtures or impurities as a whole. There indeed are parameters which are explained in this article, important to be kept in mind while a method development of HPLC method. But as such there is no list of such parameters and their accurate limits can be applied while method development. In this article, there have been mention of certain parameters which are generally looked for, and not exact, but optimal application of such has been discussed. When we talk about stationary phase, parameters which we tend to optimize very often include mostly commonly the pH of the analyte as well the mobile phase used and stability of the column packing material over a range of temperature. Choosing of stationary phase greatly depends on the nature of analyte. Stationary phase will depend and vary simultaneously if analyte is lipophilic or an ionic compound. In order to increase separation and increase mass transfer there have been new addition to column packing material apart from the widely used silica. New advances result into greater column loading, better flow, reduced plate height and reduced back pressure. Leaving alone stationary phase, mobile phase selection is also a task of its own. Very commonly used mobile phase solvents include acetonitrile, methanol, THF, water and buffer of salts like acetate, phosphate, etc. Apart from these traditional choices, there have been use of pressurized hot water and ionic liquids as mobile phases which are termed as “Environmental-friendly” because these solvents are devoid of any organic counterpart or organic modifier in them, which produces harmful fumes on getting heated. Apart from the environmentally friendly mobile phase options, we have some specific chemicals to be used when chemical property of the analyte is specific.

Ingenious introduction of aminopropylimidazole to tune the hydrophobic selectivity of dodecyl-bonded stationary phase for environmental organic pollutants

Nowadays, the efficient detection of environmental organic pollutants has been a research focus in the field of chromatographic analysis. In this work, aminopropylimidazole was designedly introduced to adjust the hydrophobicity of the long-chain alkyl group, and a novel chromatographic stationary phase for the separation and analysis of environmental organic pollutants was specially prepared. Specifically, the waste aminopropyl silica gel (5 μm) was first recycled as the raw material, which was calcined at high temperature and then acidified with hydrochloric acid solution to obtain the activated silica gel. Afterwards, a new type of aminopropylimidazole/dodecyl dual-functionalized silica stationary phase (Sil-API-DD) was synthesized via one-pot reaction of the activated silica gel with 3-chloropropyltrimethoxysilane, dodecyltrimethoxysilane and 1-(3-aminopropyl)imidazole. The separation performance of Sil-API-DD was evaluated using environmental organic pollutants as the model analytes, and the results showed that an effective separation of seven alkylphenols, five bisphenols, six phthalates, seven steroid hormones and six benzophenones in reversed-phase chromatography was achieved. Moreover, the developed Sil-API-DD was found to exhibit more flexible selectivity and superior separation efficiency compared with the commercial C18 column under the tested conditions, suggesting a good application prospect.

Advanced Development of Supercritical Fluid Chromatography in Herbal Medicine Analysis.

The greatest challenge in the analysis of herbal components lies in their variety and complexity. Therefore, efficient analytical tools for the separation and qualitative and quantitative analysis of multi-components are essential. In recent years, various emerging analytical techniques have offered significant support for complicated component analysis, with breakthroughs in selectivity, sensitivity, and rapid analysis. Among these techniques, supercritical fluid chromatography (SFC) has attracted much attention because of its high column efficiency and environmental protection. SFC can be used to analyze a wide range of compounds, including non-polar and polar compounds, making it a prominent analytical platform. The applicability of SFC for the separation and determination of natural products in herbal medicines is overviewed in this article. The range of applications was expanded through the selection and optimization of stationary phases and mobile phases. We also focus on the two-dimensional SFC analysis. This paper provides new insight into SFC method development for herbal medicine analysis.

A natural deep eutectic solvent - protonated L-proline-xylitol - based stationary phase for gas chromatography

The paper presents a new kind of stationary phase for gas chromatography based on deep eutectic solvents (DES) in the form of a mixture of L-proline (protonated with hydrochloric acid) as a hydrogen bond acceptor (HBA) and xylitol as a hydrogen bond donor (HBD) in a molar ratio of HBA:HBD 5:1. DES immobilized on a silanized chromatographic support was tested by gas chromatography (GC) in order to determine its resolving power for volatile organic compounds. Studies have demonstrated the suitability of this type of DES as a stationary phase for GC. The Rohrschneider-McReynolds constants were determined for the synthesized DES, revealing that it is a polar stationary phase (Σ(ΔI) = 1717). The selectivity of DES is influenced by the presence of hydroxyl groups in the xylitol structure capable of forming hydrogen bonds of a donor nature and the proton acceptor properties of the protonated L-proline structure. The presence of additional interactions is brought about by the presence of the carboxyl group. As a result, the retention of alcohols is several times higher (200-670%) than the expected value based on their boiling points. A significant increase in retention (400-970%) was also found for pyridine derivatives. The developed DES stationary phase is characterized by very good repeatability of retention and stability (up to 140°C). The efficiency of the prepared columns (6300-11300 theoretical plates/m) and the selectivity of the DES stationary phase are competitive with the commercial products.

Synthesis of C8F13-SiO2 stationary phase for chromatographic separation of highly polar compounds

Up to now, the chromatographic separation and analysis of highly polar compounds are still challenging because of low retention ability and selectivity of conventional stationary phases (SPs). Developing new SPs to achieve high-resolution and high-efficiency separation of polar compounds is of great importance for health and medicine, food safety, environmental protection, etc.. Herein, an influential multi-step modification strategy (modification with 1H,1H,2H,2H-perfluorooctyltriethoxysilane on the silica twice and end-capped with trimethylchlorosilane once) is introduced to prepare C8F13-SiO2 SP. The physical properties of C8F13-SiO2 SP, including morphologies, pore structures and carbon content were fully characterized by various instrumental techniques. The packed HPLC column with C8F13-SiO2 SP exhibited high column efficiency, good peak shape (tailing factors for most analytes are between 0.9 and 1.3), and resolution for simultaneous separation of ginsenosides, sulfonamides, nucleosides, and benzoylurea insecticides (BUs); meanwhile, the retention mechanism of typical reversed-phase liquid chromatography (RPLC) mode was observed. A column efficiency of 30,220 plates/m for separation of ginsenoside Re was obtained. Furthermore, the resulting columns benefit from the advantages of supreme reproducibility, repeatability and stability. Importantly, the chromatographic properties (e.g. column efficiencies, chromatographic peak shapes, retention ability and resolution) of the C8F13-SiO2 SP for the polar analytes were superior to those of the commercial SPs (including commercial amino-modified silica SP, diol-modified silica SP and commercial PFP-SiO2 SP). Thus, the C8F13-SiO2 SP is able to satisfy the strict requirements of highly polar compounds separation.

An Assessment of Stationary Phase Selectivity in SFC

Supercritical fluid chromatography (SFC) has seen a recent resurgence in interest following investment in the development of instrument technology by numerous instrument manufacturers. Increased focus on sustainability in chromatographic science, coupled with the orthogonality to reversed phase HPLC, is likely to further drive the uptake of SFC in many sectors. As with any form of chromatography, optimizing separation selectivity is a key variable in providing adequate resolution and accurate identification and quantification of target analytes. Stationary phase chemistry can be readily exploited to substantially alter the separation selectivity obtained. This article examines and characterizes the selectivity differences offered by three prototype SFC phases.

Effect of phenyl numbers in polyphenyl ligand on retention properties of aromatic stationary phases

Aromatic phase, as one type of reversed-phase stationary phases, shows complementary selectivity to the n-alkyl counterparts especially for certain challenging separation tasks. However, effect of phenyl numbers in aromatic ligands on retention behaviors has rarely been addressed compared with the alkyl stationary phases. To illustrate the issue, a series of polyphenyl stationary phases were facially prepared via the coupling chemistry of isocyanate with amine, including aniline (π1), 4-aminobiphenyl (π2), 4-amino-p-terphenyl (π3) and [1,1′:4′,1′':4′',1′''-quaterphenyl]-4-amine (π4), respectively. The chromatographic behaviors of the new stationary phases as well as the traditional C18 were systematically compared in terms of retention mode, hydrophobic and aromatic selectivity, shape selectivity and π-π interaction by various analytes, including alkylbenzenes, polycyclic aromatic hydrocarbons congeners and substituted benzenes with electron-withdrawing groups. Due to the homologous structure of four polyphenyl ligands, the hydrophobic selectivity, aromatic selectivity and shape selectivity of stationary phases increase with phenyl numbers in the bonded polyphenyl ligands, whereas the increment becomes insignificant from U-π3 to U-π4. This phenomenon is explained by the insertion degree of analytes in the polyphenyl ligand brushes. Compared with the homemade C18, the polyphenyl phases indicate insignificant changes of shape selectivity with temperature. Notably, the new polyphenyl phases demonstrate the great selective separation towards the electron-deficient compounds through the π-π interaction. These findings make up for the understanding of the retention behavior of aromatic stationary phases.

Contribution of ionic interactions to stationary phase selectivity in hydrophilic interaction chromatography.

We compared the separation selectivities of 19 different hydrophilic interaction chromatography columns. The stationary phases included underivatized silica and hybrid particles, cyano‐bonded silica, materials with neutral ligands such as amide, diol, pentahydroxy, and urea, zwitterionic sorbents, and mixed‐mode materials with amine functionalities. A set of 77 small molecules was used to evaluate the columns. We visualized the retention behavior of the different columns using retention time correlation plots. The analytes were classified as cations, anions, or neutral based on their estimated charge under the separation conditions. This involved adjusting the dissociation constants of the analytes for the acetonitrile content of the mobile phase and experimentally determining the pH of the mobile phase containing 70% acetonitrile. The retention correlation plots show that the selectivity differences strongly depended on ionic interactions. Comparisons of the neutral stationary phases (e.g., diol vs. amide) showed more similar selectivity than did comparisons of neutral columns versus columns with cation or anion exchange activity (bare silica or amine columns, respectively). The zwitterionic columns did not behave as perfectly neutral. The correlation plots indicated that they exhibited either cation or anion exchange activity, although to a lesser degree than the silica and amine‐containing stationary phases.

Characterization and applications of a trioctyl(3/4-vinylbenzyl)phosphonium stationary phase for use in capillary liquid chromatography

The morphology, composition, and selectivity of a silica-based monolithic stationary phase, grafted by a layer of trioctyl(3/4-vinylbenzyl)phosphonium chloride ([P888VBn]Cl), is presented. The results of elemental analysis confirmed that the prepared stationary phase contains 38.8 at.% of silicon, 60.2 at.% of carbon, and 1.0 at.% of phosphorus. Capillary columns (150 × 0.1 mm) for liquid chromatography were evaluated using alkylbenzenes, monosubstituted benzenes, polyaromatic compounds, substituted benzene regioisomers, and aromatic carboxylic and amino acids. The prepared ionic liquid (IL)-based stationary phase exhibits hydrophobic, hydrophilic, and electrostatic interactions, as confirmed by experiments on the evaluation of the effect of the mobile phase composition (content of acetonitrile and ammonium formate) on the isocratic chromatographic separation. Thus, the IL-based capillary column demonstrates a unique separation selectivity compared to Phenyl-, C8-, and C18-stationary phases, and high efficiency for an expanding number of structurally diverse compounds.