Reversed-phase Capillary Electrochromatography Research Articles

Poly(carboxyethyl acrylate-co-ethylene glycol dimethacrylate) precursor monolith with bonded anthracenyl ligands for use in reversed-phase capillary electrochromatography based on hydrophobic and π-π interactions

In this research report, the post polymerization functionalization (PPF) of a carboxyethyl acrylate (CEA)-co-ethylene glycol dimethacrylate (EDMA) [poly-CEA-co-EDMA)] precursor monolith with 2-aminoanthracene was carried out in the presence of an organic solvent soluble carbodiimide, namely N,N´-dicyclohexylcarbodiimide (DCC), yielding the so-called anthracenyl-poly-CEA-co-EDMA monolith. This novel monolith proved to be an excellent monolithic stationary for reversed-phase capillary electrochromatography (RP-CEC) with hydrophobic and π-π interactions of a wide range of nonpolar solutes including those bearing aryl functional groups in their structures such as polycyclic aromatic hydrocarbons (PAHs), toluene derivatives and aniline derivatives as well as solutes carrying in their structures electron withdrawing substituents such as dinitrophenyl-amino acids (DNP-AAs) and di-DNP-AAs. The retention behaviors of the just mentioned solutes obtained on the anthracenyl-poly-CEA-co-EDMA monolithic column were compared to those obtained on octadecyl-poly-CEA-co-EDMA monolithic column prepared from the same carboxy-precursor monolith. The results demonstrated the superiority of anthracenyl column over the octadecyl column for the separation and enhanced selectivity for aromatic solutes since it provides not only hydrophobic interactions but also π-π interactions with aromatic nonpolar solutes.

Reversed-phase capillary electrochromatography of pre-column derivatized mono- and oligosaccharides with three different ultraviolet absorbing tags

In this research report, an in house developed octadecyl monolithic (ODM) column has been exploited in the reversed-phase capillary electrochromatography (RP-CEC) of precolumn derivatized mono- and oligosaccharides with three different tagging agents, namely 1-naphthylamine (1-NA), 2-aminoanthracene (2-AA) and 3-amino-2,7-naphthalenedisulfonic acid (ANDSA). These three derivatizing agents, which differed in their charges, nonpolar characters and optical absorption properties, led to different RP-CEC elution patterns and UV detection signals. In fact, the limit of detection of the derivatized sugars were 50 µM for the ANDSA- and 1-NA-sugar derivatives and 35 µM for the 2-AA-sugar derivatives due to the presence of three fused aromatic rings in 2-AA versus 2 fused rings in the 1-NA and ANDSA tags. Furthermore, while the longer ANDSA-oligosaccharides eluted later than the shorter ones and the ANDSA-monosaccharides, 1-NA- and 2-AA-sugar derivatives necessitated the presence of borate ions at alkaline pH in the mobile phase to form in situ charged derivatives to facilitate their separation by RP-CEC, and the elution order was the reversal of that observed with the ANDSA-sugar derivatives; that is the mono- eluted later than the larger size oligosaccharides. In addition, plots of log tR vs. number of glucose residues (nGlc) for derivatized glucose and maltooligosaccharides yielded straight lines with slopes representing log η where η is the retention time modulus (i.e., ratio of retention time of two neighboring derivatives differing in one glucosyl residue). In the case of 1-NA and 2-AA derivatives, η was smaller than unity while it was greater than unity in the case of ANDSA-sugar derivatives because the elution occurred in the order of decreasing size of the homologous sugar derivatives in the former than in the later derivatives. The prepared ODM column was stable for more than a month of continuous use, a fact that allowed a good repeatability for intraday and interday analyzes.

Poly(carboxyethyl acrylate‐co‐ethylene glycol dimethacrylate) precursor monolith with bonded octadecyl ligands for use in reversed‐phase capillary electrochromatography

A carboxy precursor monolithic column, namely poly(carboxy ethyl acrylate-co-ethylene glycol dimethacrylate) was first produced in a 100μm i.d. fused-silica capillary and subsequently surface bonded with n-octadecyl (C18 ) ligands by a post-polymerization functionalization process with octadecylamine in the presence of N,N´-dicyclohexylcarbodiimide. The bonding of octadecyl ligands was achieved via an amide linkage between the carboxy functions of the precursor monolith and the amino group of the octadecylamine compound. The resulting C18 monolith exhibited a very low electroosmotic flow (EOF), a fact that required the incorporation of small amounts of 2-acrylamido-2-methylpropane sulfonic acid (AMPS) in the polymerization solution to produce a precursor monolith with fixed negative charges of sulfonate groups. This may indicate that the conjugation of the carboxy functions with octadecylamine occurred to a large extent so that the amount of residual carboxy functions was sparsely dispersed and not enough to produce a desirable EOF. The EOF velocity of the C18 column having fixed negative charges provided by the incorporated AMPS increased with increasing ACN content of the mobile phase signaling an increased binding of mobile phase ions to the polar amide linkages near the monolithic surface, and a decreased viscosity of the mobile phase, both of which would result in increased EOF velocity. The C18 monolithic column constituted a novel nonpolar sorbent for reversed-phase capillary electrochromatography for nonpolar solutes, e.g., alkylbenzenes, alkylphenyl ketones, and polyaromatic hydrocarbons, and slightly polar compounds including phenol and chlorophenols. The C18 monolithic column exhibited relatively high selectivity toward chlorophenols differing by one chloro substituent.

Preparation of Benzyl Quinine-modified Monolithic Column for Reversed-phase Capillary Electrochromatography.

N-Benzylquininium chloride is a versatile functional monomer with quinoline and benzyl groups, which is beneficial for reversed-phase chromatography. In this study, a novel monolithic column with reversed-phase mode was synthesized using N-benzylquininium chloride as the monomer and 3-(acryloyloxy)-2-hydroxypropyl methacrylate as the cross-linker in a binary porogenic solvent consisting of PEG 400 and a 0.05 M sodium hydroxide aqueous solution. The alkaline solution were found to be useful for the improvement of the mechanical stability of the porous monoliths. The monolithic column showed excellent reversed-phase selectivity and various compounds, such as alkylbenzenes, phenols and polycyclic aromatic hydrocarbons, were separated successfully. The highest column efficiency was 1.75 × 105 N m-1. The relative standard deviations of the migration time for thiourea and four alkylbenzenes were all less than 5.0%, which indicates the monolithic column has good stability. The application of the monolithic column for the analysis of polycyclic aromatic hydrocarbons in spiked lake water samples illustrated its great potential for practical application.

Ionic liquid-copolymerized monolith incorporated with zeolitic imidazolate framework-8 as stationary phases for enhancing reversed phase selectivity in capillary electrochromatography

A novel ionic liquid (1-allyl-methylimidazolium chloride, AlMeIm+Cl−) polymer monolith poly(ionic liquid-co-ethylene dimethacrylate) incorporated with zeolitic imidazolate framework-8 (ZIF-8-poly(IL-co-EDMA)) was firstly synthesized as stationary phases of monolithic column for capillary electrochromatography by one-step copolymerization. Incorporation of ZIF-8 into ionic liquid polymer monolith evidently enhanced the separation selectivity for four alkylbenzenes in reversed phase capillary electrochromatography (CEC), due to the synergistic effect derived from the same imidazole ring structure of ionic liquid and organic ligands of ZIF-8. Meanwhile, electroosmotic flow (EOF) was generated by ionic liquid in a wide range of pH values from 2.0 to 12.0. The resultant monolithic columns were characterized by scanning electron microscopy (SEM) and Fourier transform infrared (FT-IR). The results indicated that the prepared monolithic columns had good permeability and mechanism stability. The resultant monolithic columns were applied for the separation of neutral compounds, anilines and phenols. The highest column efficiency was 2.07 × 105 plates m−1 (theoretical plates, N) for toluene. Under optimal conditions, reproducibility was obtained with relative standard deviations (RSDs) of the retention time for run-to-run, day-to-day, column-to-column and batch-to-batch were in the range of 1.58 − 3.19%, 1.92 − 3.87%, 3.84 − 4.96% and 2.63 − 4.33%, respectively. Incorporation ZIF-8 into ionic liquid polymer monolith was a promising way for the application of new materials in the fabrication of novel monolithic columns.

Monolithic capillary columns consisting of poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) and their diol derivatives with incorporated hydroxyl functionalized multiwalled carbon nanotubes for reversed-phase capillary electrochromatography.

Two types of monolithic stationary phases with incorporated hydroxyl functionalized multiwalled carbon nanotubes (OH-MWCNTs) were introduced and evaluated, namely, the poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) monolith, denoted as poly(GMA-co-EDMA), and a diol derivative of the poly(GMA-co-EDMA) monolith. The diol derivative monolith was obtained by subjecting the poly(GMA-co-EDMA) monolith with physically incorporated OH-MWCNTs to an acid treatment with 0.1 M sulfuric acid at a moderate temperature of 50 °C for a total of 7.5 h. Also, the poly(GMA-co-EDMA) monolith with both physically and covalently incorporated OH-MWCNTs was prepared by subjecting the physically incorporated monolithic column to a Lewis acid catalyzed reaction in the presence of BF3 in order to react some of the OH-MWCNTs with the epoxy rings of the poly(GMA-co-EDMA) monolith. In all cases, the OH-MWCNTs were subjected to high power sonication at an output power of 10 W for 15 min with the aim of better dispersing the incorporated nanotubes into the monoliths under investigation. In fact, high power sonication yielded columns with a relatively higher plate count (∼2 fold increase) when compared to low power sonication. While the incorporation of OH-MWCNTs into the poly(GMA-co-EDMA) monolith acted as an amendment boosting the nonpolar character of the monolith and providing additional π-π interactions, the diol derivative monolith with its polar backbone character acted nearly as a support for the OH-MWCNT stationary phase giving rise to a carbon nanotube sorbent providing hydrophobic and π-π interactions via the incorporated OH-MWCNTs. These two kinds of columns were evaluated using alkylbenzenes, toluene derivatives, aniline compounds, phenols and polyaromatic hydrocarbons.

Novel Monolithic Stationary Phase with Surface-Grafted Triphenyl Selector for Reversed-Phase Capillary Electrochromatography

A triphenylmethylamine-functionalized monolithic capillary column was newly designed for reversed-phase capillary electrochromatographic applications. Incorporation of the three phenyl rings-containing selector (also referred to as trityl selector) was achieved through post-polymerization functionalization of a generic monolithic matrix bearing nucleophilic-sensitive hydroxysuccinimide moieties. Such a 3D polymer matrix was obtained through UV-induced in situ free radical copolymerization of N-acryloxysuccinimide and ethylene dimethacrylate. The separation properties of the trityl monolithic capillary column were initially evaluated vis-a-vis polycyclic aromatic hydrocarbons, as model hydrophobic compounds, and compared to the separation ability of a benzylamine-functionalized monolithic capillary column prepared using the same generic monolithic matrix. Electrochromatographic separation of phenols and anilines was also considered, and our preliminary results suggest the occurrence of hydrophobic interactions due to the aromatic and non-polar nature of the surface-grafted trityl selector. The triphenyl monolithic capillary column exhibited relative standard deviation values (% RSD) below 4.1 % for the here-studied chromatographic parameters, namely, retention factor, selectivity, resolution, and efficiency.

Retention-time prediction for polycyclic aromatic compounds in reversed-phase capillary electro-chromatography.

Log Po/w based models are often used for the retention time prediction of reversed phase liquid chromatography. Here, we present the investigation of the applicability of log Po/w based retention time predictions for the separation in capillary electro-chromatography (CEC). A test set of five polycyclic aromatic hydrocarbons was separated using two different stationary phases with three different mobile phases each. The resulting retention times were correlated with the experimental log Po/w values as well as with calculated log Po/w values. The used methods include quantitative structure property relationship (QSPR) models as well as molecular dynamic methods such as the linear interaction energy (LIE) or the Bennett acceptance ratio (BAR). The results indicate that rigorous simulation models are capable of accurately reproducing experimental results and that the electrophoretic mobility of analytes in CEC separations leads to significant deviations in the retention time prediction.

Preparation and evaluation of o-phenanthroline immobilized on a hybrid silica monolith modified with ionic liquids for reversed-phase pressurized capillary electrochromatography.

A novel o-phenanthroline-immobilized ionic-liquid-modified hybrid monolith for capillary electrochromatography was synthesized based on chloropropyl-silica, which was prepared by the in situ polymerization of tetramethoxysilane and 3-chloropropyltrimethoxysilane via a sol-gel process. The morphology of the hybrid monolith was characterized by scanning electron microscopy, and relatively stable anodic electroosmotic flow was observed under a broad pH ranged from pH 3.0 to 9.0. The separation mechanism was investigated by separating four neutral molecules (toluene, dimethylformamide, formamide, and thiourea). The obtained hybrid monolith possessed an obviously reversed-phase retention mechanism, but when the acetonitrile content in the mobile phase was >90% v/v, a weak hydrophilic mechanism was observed on the resultant o-phenanthroline-modified chloropropyl-silica hybrid monolith. The reproducibility of the column was also investigated by measuring relative standard deviations of the migration time for four neutral molecules. Relative standard deviations of run to run (n = 3), day to day (n = 3), and column to column (n = 3) were in the range of 0.4-0.7, 0.9-2.1, and 1.4-3.3%, respectively. Basic separations of various polar analytes including phenols and aromatic amines were successfully achieved.

A polymer monolith for hydrophilic and dynamically surfactant-modified reversed-phase capillary electrochromatography

A stable poly(META-co-PETA) monolith was fabricated and employed for hydrophilic interaction (HI) and dynamically-modified reversed-phase (RP) electrochromatography. Detailed effects on the column characteristics including the permeability, swelling propensity factor, retention factor, EOF and column efficiency were investigated. Studies on the column stability and heat dissipation were also implemented. The interrupt problem caused by joule heating was eliminated and the satisfactory characteristics in terms of the uniform morphology, good reproducibility and multiple retention modes were gained. Based on the obtained monolith, versatile separation interactions including HI, RP, HI-strong anion exchange (HI-SAX) and RP-SAX were evaluated and utilized for capillary electrochromatography (CEC). Efficient separations of phenols and nucleic acid bases and nucleosides were achieved with HI-CEC. With an easy dynamical modification of sodium dodecylsulfate (SDS), efficient separations of alkyl benzenes, polycyclic aromatic hydrocarbons, estrogens and basic anilines were also achieved with RP or RP-SAX. It presented a facile polymer monolith for hydrophilic and dynamically SDS-modified RP modes in CEC.

Preparation and evaluation of ionic liquid-gold nanoparticles functionalized silica monolithic column for capillary electrochromatography

This paper describes the development of silica monolithic column modified with ionic liquids-gold nanoparticles (ILs-GNPs) for capillary electrochromatography (CEC). The novel ILs (1-methyl-2-mercapto-3-butylimidazolium bromide) were synthesized and used to modify GNPs functionalized silica monolithic column via the formation of a Au-S bond. The morphology of the GNPs and ILs-GNPs functionalized silica (ILs-GNPs-silica) monolithic column were characterized by transmission electron microscopy and scanning electron microscope, respectively. A cathodic electroosmotic flow was observed at pH above 6.4 on ILs-GNPs-silica monolithic column, which was reversed at acidic pH. The electrochromatographic performance of ILs-GNPs-silica monolithic column was evaluated by separation of different kinds of analytes such as hydrophobic, polar and basic compounds. The ILs-GNPs-silica monolithic column displayed enhanced hydrophobic retention characteristics in the separation of five hydrophobic n-alkylbenzenes when compared to the ILs bonded silica monolithic column. The column efficiencies for the n-alkylbenzenes were from 62,000 to 110,000 N m(-1). The ILs-GNPs-silica monolithic column exhibited reversed-phase electrochromatographic behavior toward neutral solutes. Separation of polar compounds was demonstrated on ILs-GNPs-silica monolithic column in reversed-phase CEC mode using high aqueous mobile phases. The relatively good peak shape and high separation efficiency on ILs-GNPs-silica monolithic column was obtained for basic solutes when compared to silica monolithic column modified GNPs.

Controlling retention, selectivity and magnitude of EOF by segmented monolithic columns consisting of octadecyl and naphthyl monolithic segments – applications to RP‐CEC of both neutral and charged solutes

Monolithic capillaries made of two adjoining segments each filled with a different monolith were introduced for the control and manipulation of the electroosmotic flow (EOF), retention and selectivity in reversed phase-capillary electrochromatography (RP-CEC). These columns were called segmented monolithic columns (SMCs) where one segment was filled with a naphthyl methacrylate monolith (NMM) to provide hydrophobic and π-interactions, while the other segment was filled with an octadecyl acrylate monolith (ODM) to provide solely hydrophobic interaction. The ODM segment not only provided hydrophobic interactions but also functioned as the EOF accelerator segment. The average EOF of the SMC increased linearly with increasing the fractional length of the ODM segment. The neutral SMC provided a convenient way for tuning EOF, selectivity and retention in the absence of annoying electrostatic interactions and irreversible solute adsorption. The SMCs allowed the separation of a wide range of neutral solutes including polycyclic aromatic hydrocarbons (PAHs) that are difficult to separate using conventional alkyl-bonded stationary phases. In all cases, the k' of a given solute was a linear function of the fractional length of the ODM or NMM segment in the SMCs, thus facilitating the tailoring of a given SMC to solve a given separation problem. At some ODM fractional length, the fabricated SMC allowed the separation of charged solutes such as peptides and proteins that could not otherwise be achieved on a monolithic column made from NMM as an isotropic stationary phase due to the lower EOF exhibited by this monolith.

Two-dimensional strong cation-exchange liquid chromatography/reversed-phase pressurized capillary electrochromatography for separation of complex samples

A 2-D separation platform was constructed using micro strong cation-exchange liquid chromatography (μ-SCXLC) and reversed-phase pressurized capillary electrochromatography (RP-pCEC) for the analysis of complex samples. Samples were fractionated by the first-dimension μ-SCXLC with a linear solvent gradient and then injected into the second-dimension RP-pCEC for further separation. The μ-SCXLC/RP-pCEC 2-D system with three separation mechanisms, namely strong cation-exchange, reversed-phase chromatography and electrophoresis, provided high selectivity, high resolution and high peak capacity compared to one-dimensional chromatographic approaches. Separation effectiveness of this 2-D system was demonstrated by the analysis of different kinds of complex samples, such as traditional Chinese medicine Cortex Phellodendri, bovine serum albumin (BSA) tryptic digest and real serum tryptic digest. A theoretical peak capacity of approximately 1200 was achieved, which proves its promising potential for the separation and analysis of complex samples.

Preparation and Characterization of Gold Nanoparticle-Modified Silica Monolith for Capillary Electrochromatography

Abstract A novel silica monolithic stationary phase functionalized with octadecanethiol gold nanoparticles (GNPs) was prepared for capillary electrochromatography (CEC). The bare silica monolith was prepared by a sol-gel process and then modified chemically with 3-mercaptopropyltrimethoxysilane, followed by immobilization of GNPs and modification by octadecanethiol. The relationship between plot of plate height and the linear velocity on octadecanethiol GNPs-modified silica monolith (Silica-GNPs-C18) was investigated, and a minimum plate height of about 7.8 μm for toluene was obtained. The electroosmotic flow characteristics of the Silica-GNPs-C18 and bare silica monolith (Silica) were studied by varying the percentage of organic modifier in buffer and buffer pH. To evaluate the column performance, hydrophobic alkylbenzenes as model compounds were tested on the prepared Silica, thiol-modified silica monolith (Silica-SH), and Silica-GNPs-C18. The electrochromatographic retention behavior of alkylbenzenes on Silica-GNPs-C18 column was also investigated. The experiment results showed that Silica-GNPs-C18 column exhibits typical reversed-phase electrochromatographic behavior toward neutral solutes. Reversed-phase CEC separations of four selected polar phenols were also demonstrated on Silica-GNPs-C18 column using mobile phase containing 15% methanol.

Naphthyl methacrylate‐phenylene diacrylate‐based monolithic column for reversed‐phase capillary electrochromatography via hydrophobic and π interactions

A neutral naphthyl methacrylate-phenylene diacrylate-based monolith (NPM) was introduced for RP-CEC of various neutral and charged solute probes via hydrophobic and π interactions. The NPM column was prepared by the in situ polymerization of naphthyl methacrylate as the functional monomer and 1,4-phenylene diacrylate (PDA) as the crosslinker in a ternary porogenic solvent containing cyclohexanol, dodecanol and water. The NPM column exhibited cathodal EOF despite the fact that it was devoid of any fixed charges. NPM exhibited stronger EOF than its counterpart naphthyl methacrylate monolith (NMM) made from the in situ polymerization of naphthyl methacrylate and trimethylolpropane trimethacrylate (TRIM). As for NMM, it is believed that the EOF arises from the adsorption of mobile phase ions onto the monolith surface. The higher EOF exhibited by NPM may be attributed to the acrylate nature of PDA as compared to the methacrylate nature of TRIM, and therefore PDA has a higher binding capacity for mobile phase ions due to its higher polarity than TRIM. The adsorption of mobile phase ions together with the additional π interactions offered by the aromatic rings of the NPM matrix modulated solute retention and separation selectivity. The applications of NPM were demonstrated by the separation of a wide range of small and large solutes including peptides, tryptic peptide maps and proteins.

A novel, neutral hydroxylated octadecyl acrylate monolith with fast electroosmotic flow velocity and its application to the separation of various solutes including peptides and proteins in the absence of electrostatic interactions

A neutral hydroxylated octadecyl monolith (ODM-OH) for reversed-phase capillary electrochromatography has been developed. The ODM-OH was prepared by the in situ polymerization of octadecyl acrylate and pentaerythritol triacrylate (PETA) in a ternary porogenic solvent. Pentaerythritol triacrylate possesses a hydroxyl functional group, which imparts the monolith with a hydrophilic group, thus the acronym ODM-OH. The ODM-OH column exhibited cathodal EOF over a wide range of pH and ACN concentration in the mobile phase despite the fact that it was devoid of any fixed charges. This ODM-OH monolith exhibited stronger EOF than its counterpart the ODM made from the in situ polymerization of octadecyl acrylate and trimethylolpropane trimethacrylate. Similar to ODM, it is believed that the EOF was due to the adsorption of ions from the mobile phase onto the surface of the monolith thus imparting the neutral monolithic column the zeta potential necessary to support the EOF. The higher EOF exhibited by ODM-OH was due to the presence of polar OH groups on its surface, which would favor stronger adsorption of ions from the mobile phase. The wide applications of the neutral ODM-OH column were demonstrated in the separation of a wide range of small and large solutes. As a typical result, the ODM-OH was able to separate proteins quite rapidly yielding 200,000 plates/m.

Enantioseparation on cellulose dimethylphenylcarbamate-modified zirconia monolithic columns by reversed-phase capillary electrochromatography

This work reports the preparation of monolithic zirconia chiral columns for separation of enantiomeric compounds by capillary electrochromatography (CEC). Using sol–gel technology, a porous monolith having interconnected globular-like structure with through-pores is synthesized in the capillary column as a first step in the synthesis of monolithic zirconia chiral capillary columns. In the second step, the surface of the monolith is modified by coating with cellulose tris(3,5-dimethylphenylcarbamate) (CDMPC) as the chiral stationary phase to obtain a chiral column (CDMPCZM). The process of the preparation of the zirconia monolithic capillary column was investigated by varying the concentrations of the components of the sol solution including polyethylene glycol, water and acetic acid. CDMPCZM is mechanically stable and no bubble formation was detected with the applied current of up to 30 μA. The enantioseparation behavior of the CDMPCZM columns was investigated by separating a set of 10 representative chiral compounds by varying the applied voltage and pH and organic composition of the aqueous organic mobile phases.

The synthesis of chloropropyl-functionalized silica hybrid monolithic column with modification of N,N-dimethyl-N-dodecylamine for capillary electrochromatography separation

A chloropropyl-functionalized silica (CP-silica) hybrid monolithic column was synthesized within the confines of a capillary via the sol–gel process using tetramethoxysilane (TMOS) and (3-chloropropyl)-trimethoxysilane (CPTMS) as the precursors. The resulting CP-silica hybrid monolith inside the capillary showed homogeneous macroporous morphology and was well attached to the inner wall of the capillary. The obtained CP-silica hybrid monolithic capillary column demonstrated the inherent hydrophobic property and could be applied as a reversed-phase stationary phase in CEC directly. Due to the great chemical reactivity of the incorporated chloro groups on the hybrid silica monolithic matrix, the chloropropyl moieties on the surface of the hybrid silica monolith matrix could be conveniently further modified by a tertiary amine of N,N-dimethyl-N-dodecylamine (DMDA) via the nucleophilic substitution reaction at 70 °C to introduce a dodecyl groups (C12) onto the CP-silica hybrid monolithic matrix. The resulting C12-silica hybrid monolithic column not only demonstrated the significantly enhanced hydrophobic property in the separation of alkylbenzenes in reversed-phase capillary electrochromatography (RP-CEC), but also the strong electroosmotic flow (EOF) in a wide pH range. Five alkylbenzenes could be baseline separated in 3 min with column efficiency ranging from 189 700 to 221 000 N/m with a 70% ACN running buffer in CEC.

Enantioseparation of Neutral Compounds on a Quinine Carbamate-Immobilized Zirconia in Reversed-Phase Capillary Electrochromatography

chiral analytes in HPLC and recently in CEC. In this work we report enantio-separation of non-acidic chiral analytes on a quinine carbamate-immobilized zirconia (QNZ) in reversed-phase (RP) CEC. Influences of pH, composition of the buffer, acetonitrile content and the applied voltage on enantioseparation were examined. Enantiomers of the analytes investigated are well separated in acetonitrile/phosphate buffer mobile phases. Separation data on QNZ were compared to those on QN-bonded silica (QNS). Retention was longer but better enantioselectivity and resolution were obtained on QNZ than QNS.

Neutral octadecyl monolith for reversed phase capillary electrochromatography of a wide range of solutes

A neutral octadecyl monolithic (ODM) column for RP capillary electrochromatography (RP-CEC) has been developed. The ODM column was prepared by the in situ polymerization of octadecyl acrylate (ODA) as the monomer and trimethylolpropanetrimethacrylate (TRIM) as the crosslinker, in a ternary porogenic solvent containing cyclohexanol, ethylene glycol, and water. The ODM column exhibited cathodal EOF over a wide range of pH and ACN concentration in the mobile phase despite the fact that it was devoid of any fixed charges. It is believed that the EOF is due to the adsorption of ions from the mobile phase onto the surface of the monolith thus imparting to the neutral ODM column the zeta potential necessary to support the EOF required for mass transport across the monolithic column. Furthermore, the adsorption of mobile phase ions to the neutral monolith modulated solute retention and affected the separation selectivity. The wide applications of the neutral ODM column were demonstrated by its ability to separate a wide range of small and large solutes, both neutral and charged. While the separation of the neutral solutes was based on RP retention mechanism, the charged solutes were separated on the basis of their electrophoretic mobility and hydrophobic interaction with the C18 ligands of the stationary phase. As a typical result, the neutral monolithic column was able to separate peptides quite rapidly with a separation efficiency of nearly 200,000 plates/m, and this efficiency was exploited in tryptic peptide mapping of standard proteins, e. g., lysozyme and cytochrome C, by isocratic elution.