Open-tubular Capillary Electrochromatography Column Research Articles

Chiral covalent organic framework-based open tubular capillary electrochromatography column for enantioseparation of selected amino acids and pesticides.

A novel chiral covalent organic framework (CCOF) was synthesized with an imine covalent organic framework TpBD (synthesized via Schiff-base reaction between phloroglucinol (Tp) and benzidine (BD)) modified using (1S)-(+)-10-camphorsulfonyl chloride as chiral ligand by chemical bonding method for the first time, and was characterized by X-ray diffraction, Fourier-transform infrared spectra, X-ray photoelectron spectroscopy, nitrogen adsorption/desorption, thermogravimetry analysis, and zeta-potential. The results revealed that the CCOF had good crystallinity, high specific surface area and good thermal stability. Then, the CCOF was employed as stationary phase in open-tubular capillary electrochromatography (OT-CEC) column (the CCOF-bonded OT-CEC column) for enantioseparation of 21 single chiral compounds (12 natural amino acids including acidic, neutral and basic, 9 pesticides including herbicides, insecticides and fungicides) and simultaneous enantioseparation of mixture amino acids and pesticides with similar structures or properties. Under the optimized CEC conditions, all the analytes reached the baseline separation with high resolutions of 1.67-25.93 and selectivity factors of 1.06-3.49 in 8min. Finally, the reproducibility and stability of the CCOF-bonded OT-CEC column were measured. Relative standard deviations (RSDs) of retention time and separation efficiency were 0.58-4.57% and 1.85-4.98%, and not obviously changed after 150 runs. These results demonstrate that COFs-modified OT-CEC explore a promising method to separate chiral compounds.

Preparation of β-cyclodextrin covalent organic framework-immobilized poly(glycidyl methacrylate) nanoparticle-coated open tubular capillary electrochromatography column for chiral separation.

A new enantioselective open-tubular capillary electrochromatography was developed employing poly(glycidyl methacrylate) nanoparticles/β-cyclodextrin covalent organic frameworks chemically immobilized on the inner wall of the capillary as a stationary phase. A pretreated silica-fused capillary reacted with 3-aminopropyl-trimethoxysilane followed by poly(glycidyl methacrylate) nanoparticles and β-cyclodextrin covalent organic frameworks through a ring-opening reaction. The resulting coating layer on the capillary was characterized by scanning electron microscopy and Fourier transform infrared spectroscopy. The electroosmotic flow was studied to evaluate the variation of the immobilized columns. The chiral separation performance of the fabricated capillary columns was validated by the analysis of the four racemic proton pump inhibitors including lansoprazole, pantoprazole, tenatoprazole, and omeprazole. The influences of bonding concentration, bonding time, bonding temperature, buffer type and concentration, buffer pH, and applied voltage on the enantioseparation of four proton pump inhibitors were investigated. Good enantioseparation efficiencies were achieved for all enantiomers. In the optimum conditions, the enantiomers of four proton pump inhibitors were fully resolved within 10min with high resolutions of 9.5-13.9. The column-to-column and inter- to intra-day repeatability of the fabricated capillary columns through relative standard deviation were found better than 9.54%, exhibiting satisfactory stability and repeatability of the fabricated capillary columns.

Preparation of open tubular capillary column covalently coated with polystyrene sulfonate with 4,4'-azobis (4-cyanopentanoyl chloride) as polymerization initiator for electrochromatographic separation of alkaloids, sulfonamides and peptides.

An open tubular capillary electrochromatography column covalently bonded with polystyrene sulphonate was prepared via in situ polymerization using functionalized azo-initiator 4,4'-Azobis(4-cyanopentanoyl chloride). Scanning electron, fluorescence and atomic force microscopy techniques showed the formation of a relatively rough layer of polymer. In addition, -CN and C = O stretching vibrations from infrared spectroscopy proved the successful immobilization of the azo-initiator through covalent bonding and X-rayPhotoelectronSpectroscopy confirmed the elemental composition of the formed polymer layer. The prepared column was found to be appropriate for small and medium-sized molecules separation. Compared to bare fused silica capillary column higher selectivity and resolution were obtained for the separation of alkaloids, sulfonamides and peptides as a result of the electrostatic and pi-pi stacking interactions between the small organic molecules and the coated column without compromising the electroosmotic flow mobility. Separation efficiency was also increased compared to the bare capillary for the separation of alkaloids (about 1.5 times). Moreover, intraday, inter day, intra batch and inter batch relative standard deviation values of retention time and peak area of peptides were within 2% and 10%, respectively, indicating a good repeatability of the column preparation procedure. The developed method for the covalent bonding of polymers through a functionalized azo-initiator could represent a promising stable method for the preparation of open tubular column. This article is protected by copyright. All rights reserved.

Homochiral iron-based γ-cyclodextrin metal-organic framework for stereoisomer separation in the open tubular capillary electrochromatography

Metal organic frameworks (MOFs), as a novel separation mediums, have in recent years attracted wide consideration in capillary electrochromatography (CEC). However, the instability of frameworks limited the application of the MOF-based chiral separation capillary. Herein, Iron-based γ-cyclodextrin metal-organic framework (Fe-CD-MOF) with mesoporous was developed as the chiral stationary phase in open tubular capillary electrochromatography column (OT-CEC) for the stereoisomer separation of fourteen chiral drugs and one chiral alcohol. The capillary column with MOF coating exhibited the best performance (resolution value: 17.07) for anisodamine in any MOF-based capillary column that has never been reported. Moreover, the effect of pH, buffer concentration, and methanol content has been investigated. The novel coated capillary was also applied for the quantitative analysis of a real sample. The Fe-CD-MOF coated capillary showed outstanding repeatability and stability, with the satisfactory relative standard deviations (RSDs) for intra-day, inter-day, and column-to-column. Finally, the enantioseparation mechanism of the Fe-CD-MOF coated capillary was evaluated by adsorption kinetic experiments. In summary, this work indicated the novel and renewable Fe-CD-MOF was a potential chiral stationary phase in CEC chiral separation.

A covalent organic framework for chiral capillary electrochromatography using a cyclodextrin mobile phase additive.

Covalent organic frameworks (COFs) have been recognized as promising solid phases in capillary electrochromatography (CEC). Imine-based COF-coated open-tubular CEC column (COF TpBD-coated OT column) was prepared and characterized by X-ray diffraction (XRD), Fourier-transform infrared (FT-IR) spectra, thermogravimetric analysis (TGA), nitrogen adsorption/desorption (Brunauer-Emmett-Teller [BET]), and scanning electron microscopy (SEM). The results showed that the column efficiency was up to 26,776 plate/m, and the thickness of stationary phase was about 6.00 μm for the column prepared under the optimal conditions. Enantioseparation of 15 kinds of the single chiral compounds (histidine, arginine, lysine, leucine, threonine, methionine, valine, aspartic acid and glutamic acid, fipronil, diclofop, imazamox, quizalofop-p, imazethapyr, and acephate) and 3 kinds of mixed amino acids racemaces (valine, methionine, and glutamic acid) were performed with three methods: capillary electrochromatography with COF TpBD-coated OT column (Method 1), CEC with COF TpBD-coated OT column as the separation channels, and capillary electrophoresis (CE) with HP-β-CD as the chiral mobile phase additive (Method 2) and CE with HP-β-CD as the chiral mobile phase additive (Method 3). Separation efficiency and chiral selectivity of Method 2 was best among the three methods. Under the optimal separation conditions of Method 2, all the enantiomers reached the baseline separation regardless of the single chiral compounds or the mixed amino acids. Relative standard deviation (RSDs) of the mean column efficiency for reproducibility and stability was in the range of 0.46-1.49%. This combination of CEC and CE has great potential for use in chiral separation.

A Novel Chiral Covalent Organic Framework in Open Tubular Capillary Electrochromatography Column for Rapid and Simultaneous Enantioseparation of Amino Acids and Pesticides

A Novel Chiral Covalent Organic Framework in Open Tubular Capillary Electrochromatography Column for Rapid and Simultaneous Enantioseparation of Amino Acids and Pesticides

Preparation of a polydopamine/β-cyclodextrin coated open tubular capillary electrochromatography column and application for enantioseparation of five proton pump inhibitors.

An open tubular capillary electrochromatography column was prepared by immobilizing β-cyclodextrin on the inner wall of pretreated capillary via noncovalent adsorption of polydopamine. The resulting coating layer on the capillary was characterized by scanning electron microscopy and Fourier transform infrared spectroscopy. Electroosmotic flow was studied to evaluate the variation of the immobilized columns. The prepared columns showed good chiral separation performance toward five proton pump inhibitors including lansoprazole, pantoprazole, tenatoprazole, rabeprazole, and omeprazole. The influences of β-cyclodextrin concentration, coating time, buffer pH, buffer concentration, and applied voltage on separation were investigated. In the optimum conditions, the enantiomers of five analytes were fully resolved within 15min with high resolutions of 4.57 to 8.13. The method was extensively validated in terms of accuracy, precision, and linearity and proved to be robust. The relative standard deviation values for migration times and peak areas of the analytes representing intraday and interday were less than 1.9 and 3.6%, respectively. Further, the polydopamine/β-cyclodextrin coated capillary column could be successively used over 100 runs without showing significant decrease in the separation efficiency.

Solvothermal-assisted in situ rapid growth of octadecylamine functionalized polydopamine-based permanent coating as stationary phase for open-tubular capillary electrochromatography.

In recent years, mussel-inspired polydopamine (PDA) based materials have been widely used as stationary phases for open-tubular capillary electrochromatography (OT-CEC) because of their various excellent properties. Nevertheless, the traditional synthesis routes of functionalized PDA-based capillary columns usually are time-consuming and limited in aqueous solutions. Herein, we report a facile and rapid route to prepare octadecylamine (ODA) functionalized PDA coated OT-CEC columns in organic solvents via a novel one-step in situ solvothermal-assisted coating strategy. Through this developed solvothermal-assisted approach, the growth rate of ODA/PDA coating was significantly speeded up and their hybrid coating process on the capillary inner surface could be rapidly completed in 60min. The successful preparation of the solvothermal-assisted ODA/PDA hybrid coating were systematically characterized and confirmed by several methods. The influence of the preparation parameters on the formation of hybrid coating and the separation ability of the ODA/PDA modified columns were systematically explored. Consequently, the high-efficiency baseline separation of four kinds of neutral, acidic and basic analytes were achieved based on the ODA/PDA modified columns. The repeatability of the solvothermal-assisted ODA/PDA coated column was also studied, and the relative standard deviations for intra-day, inter-day and column-to-column were all less than 5%. Additionally, the solvothermal-assisted ODA/PDA modified column exhibited good stability and long lifetime.

Preparation of a thiols β-cyclodextrin/gold nanoparticles-coated open tubular column for capillary electrochromatography enantioseparations.

Inspired by the distinct chemical and physical properties of nanoparticles, here a novel open-tubular capillary electrochromatography column was prepared by electrostatic assembly of poly(diallydimethylammonium chloride) onto the inner surface of a fused-silica capillary, followed by self-adsorption of negatively charged SH-β-cyclodextrin/gold nanoparticles. The formation of the SH-β-cyclodextrin/gold nanoparticles coated capillary was confirmed and characterized by scanning electron microscopy and energy dispersive spectrometry. The results of scanning electron microscopy and energy dispersive spectrometry studies indicated that SH-β-cyclodextrin/gold nanoparticles were successfully coated on the inner wall of the capillary column. The performance of the SH-β-cyclodextrin/gold nanoparticles coated capillary was validated by the analysis of six pairs of chiral drugs, namely zopiclone, carvedilol, salbutamol, terbutaline sulfate, phenoxybenzamine hydrochloride, and ibuprofen. Satisfactory enantioseparation results were achieved, confirming the use of gold nanoparticles as the support could enhance the phase ratio of the open-tubular capillary column. Additionally, the stability and reproducibility of the SH-β-cyclodextrin/gold nanoparticles coated capillary column were also investigated. Then, this proposed method was well validated with good linearity (≥0.999), recovery (90.0-93.5%) and repeatability, and was successfully used for enantioseparation of ibuprofen in spiked plasma samples, which indicated the new column's potential usage in biological analysis.

In situ one-pot synthesis of polydopamine/octadecylamine co-deposited coating in capillary for open-tubular capillary electrochromatography

Mussel-inspired polydopamine (PDA) based materials are attractive as stationary phase for open-tubular capillary electrochromatography (OT-CEC) due to their many fascinating properties. However, all of the existing strategies for fabricating PDA based OT-CEC columns are limited in aqueous solutions. Consequently, it is a challenge work to directly immobilize the hydrophobic functional materials onto the inner wall of PDA modified capillary. Herein, by using the organic amine-inducing co-deposition strategy, a novel preparative method was developed for in situ one-pot synthesis of PDA/octadecylamine (ODA) co-deposited coating inside capillary as OT-CEC stationary phase. The formation and morphology of the PDA/ODA co-deposited coating were characterized by field emission scanning electron microscopy, atomic force microscope, attenuated total reflectance Fourier transform infrared spectroscopy and contact angle measurements. The separation performance of the fabricated PDA/ODA modified columns was validated by the separation of alkylbenzenes and steroids, which could achieve baseline separation with high separation efficiency. Their separation was found to follow the reversed phase chromatographic retention mechanism. The co-deposited column showed good stability and long lifetime. The repeatability of the PDA/ODA co-deposited column was also evaluated, with the relative standard deviations for intra-day and inter-day runs less than 5% and column-to-column runs less than 6%.

Sol-gel technique for the preparation of β-cyclodextrin gold nanoparticles as chiral stationary phase in open-tubular capillary electrochromatography.

A novel open-tubular capillary electrochromatography column coated with β-cyclodextrin was prepared using the sol-gel technique. In the sol-gel approach, owing to the three-dimensional network of sol-gel and the strong chemical bond between the stationary phase and the surface of capillary columns, good chromatographic characteristics and unique selectivity in separating enantiomers were shown. The influences of capillary inner diameter, coating time, organic modifier, buffer pH, and buffer concentration on separation were investigated. The sol-gel-coated β-cyclodextrin column has shown improved enantioseparation efficiency of chlorphenamine, brompheniramine, pheniramine, zopiclone in comparison with the sol-gel matrix capillary column. The migration time relative standard deviation of the separation of the enantiomers was less than 0.89% over five runs and 2.9% from column to column. This work confirmed that gold nanoparticles are promising electrochromatographic support to enhance the phase ratio of open-tubular capillary electrochromatography column in capillary electrochromatography.

Covalent bonding of Schiff base network-1 as a stationary phase for capillary electrochromatography

Covalent organic frameworks (COFs), featuring low densities, high surface areas, and good thermal and chemical stabilities, are gradually attracting interest in the field of analytical chemistry. A type of microporous polymer network material named Schiff base network-1 (SNW-1) was introduced into a capillary column through covalent bonding. The obtained SNW-1-coated capillary column was characterized by thermogravimetric analysis, scanning electron microscopy, Fourier transform infrared spectroscopy, and X-ray diffraction. Then, the SNW-1-coated capillary column was successfully utilized for the open-tubular capillary electrochromatography (OT-CEC) separation of sulfonamides, cephalosporins, amino acids and parabens. The fabricated capillary column showed good separation efficiency (Rs > 1.4), stability and reproducibility (relative standard deviation (RSD) < 5.88%). To the best of our knowledge, this is the first report of a covalent bonding strategy to bond an SNW material to a capillary column for OT-CEC.

Capillary electrochromatography using knitted aromatic polymer as the stationary phase for the separation of small biomolecules and drugs

Hypercrosslinked polymers (HCPs) are currently receiving great attention due to their unique characteristics and potential uses in diverse areas. However, the field of HCPs for open-tubular capillary electrochromatography (OT-CEC) separations has not been explored. Here, a knitted aromatic polymer (KAP) was in-situ grown on the inner wall of the capillary column for OT-CEC for the first time. The silylating reagent containing phenyl was served as monomer, immobilized on the inner wall of the capillary column, and then KAPs-modified capillary column was prepared through in-situ hypercrosslinking reaction. The surface structure and morphology of KAPs-modified capillary column was characterized by Fourier transform infrared spectra (FT-IR) and scanning electron microscopy (SEM). The prepared capillary columns showed good separation performance for neutral compounds, small biomolecules, such as nucleosides, amino acids, small peptides, and non-steroidal anti-inflammatory drugs, sulfa drugs. In addition, the KAPs-modified capillaries showed good reproducibility, with relative standard deviations for intra-day, inter-day and column-to-column runs less than 1.59%, 2.55%, and 5.19% respectively. The strategy of in-situ immobilization of KAPs provides a new approach for the application of the material in the analytical fields.

In situ synthesis of the imine-based covalent organic framework LZU1 on the inner walls of capillaries for electrochromatographic separation of nonsteroidal drugs and amino acids

The covalent organic framework LZU1 (Lan Zhou University-1) is one of the imine-based covalent organic frameworks (COFs) that possesses attractive properties such as structural regularity, stability and good porosity. This work reports on the in-situ synthesis of COF-LZU1 on the inner walls of capillary column for open-tubular capillary electrochromatography (OT-CEC). The fused-silica capillary was first modified with 3-aminopropyltriethoxysilane and glutaraldehyde. Epitaxial growth of COF-LZU1 on the inner walls was accomplished by Schiff base reaction. The formation of COF-LZU1 was confirmed by SEM and FT-IR. The COF-LZU1 coating increases the interactions between analytes and coating, which remarkably improves the CEC separation selectivity of neutral analytes, amino acids and nonsteroidal anti-inflammatory drugs. The COF-LZU1-modified column exhibits good stability and repeatability. The precisions (relative standard deviations) for intra-day, inter-day and column-to column are <1.6%, 5.6% and 6.8%, respectively. Hence, the imine-based COFs represent attractive stationary phase for application in CEC separations.

High Efficiency Robust Open Tubular Capillary Electrochromatography Column for the Separation of Peptides

High Efficiency Robust Open Tubular Capillary Electrochromatography Column for the Separation of Peptides

Preparation of a β-Cyclodextrin-Based Open-Tubular Capillary Electrochromatography Column and Application for Enantioseparations of Ten Basic Drugs.

An open-tubular capillary electrochromatography column was prepared by chemically immobilized β-cyclodextrin modified gold nanoparticles onto new surface with the prederivatization of (3-mercaptopropyl)-trimethoxysilane. The synthesized nanoparticles and the prepared column were characterized by transmission electron microscopy, scanning electron microscopy, infrared spectroscopy and ultraviolet visible spectroscopy. When the column was employed as the chiral stationary phase, no enantioselectivity was observed for ten model basic drugs. So β-cyclodextrin was added to the background electrolyte as chiral additive to expect a possible synergistic effect occurring and resulting in a better separation. Fortunately, significant improvement in enantioselectivity was obtained for ten pairs of drug enantiomers. Then, the effects of β-cyclodextrin concentration and background electrolyte pH on the chiral separation were investigated. With the developed separation mode, all the enantiomers (except for venlafaxine) were baseline separated in resolutions of 4.49, 1.68, 1.88, 1.57, 2.52, 2.33, 3.24, 1.63 and 3.90 for zopiclone, chlorphenamine maleate, brompheniramine maleate, dioxopromethazine hydrochloride, carvedilol, homatropine hydrobromide, homatropine methylbromide, venlafaxine, sibutramine hydrochloride and terbutaline sulfate, respectively. Further, the possible separation mechanism involved was discussed.

Open tubular capillary column for the separation of cytochrome C tryptic digest in capillary electrochromatography.

A silica capillary of 50 μm internal diameter and 500 mm length (416 mm effective length) was chemically modified with 4-(trifluoromethoxy) phenyl isocyanate in the presence of dibutyl tin dichloride as catalyst. Sodium diethyl dithiocarbamate was reacted with the terminal halogen of the bound ligand to incorporate the initiator moiety, and in situ polymerization was performed using a monomer mixture of styrene, N-phenylacrylamide, and methacrylic acid. The resultant open tubular capillary column immobilized with the copolymer layer was used for the separation of tryptic digest of cytochrome C in capillary electrochromatography. The sample was well eluted and separated into many components. The elution patterns of tryptic digest of cytochrome C were studied with respect to pH and water content in the mobile phase. This preliminary study demonstrates that open tubular capillary electrochromatography columns with a modified copolymer layer composed of proper nonpolar and polar units fabricated by reversible addition-fragmentation transfer polymerization can be useful as separation media for proteomic analysis.

Enhancement of capillary electrochromatographic separation performance by conductive polymer in a layer-by-layer fabricated graphene stationary phase

In this work, we fabricated a novel graphene-based capillary column for open-tubular capillary electrochromatography (OT-CEC) by a layer-by-layer strategy. To immobilize graphene onto the inner surface of silica capillary, a bio-inspired method was first used to functionalize the capillary surface with a layer of polydopamine (PDA). Graphene oxide (GO) was then introduced and can covalently react with polydopamine, realizing immobilization of graphene as a result. To enhance the modification efficiency of polydopamine, a conductive polymer, polyaniline (PANI) was introduced to be a sub-layer; polydopamine was then introduced following with GO, to generate a multilayer GO-PDA-PANI@capillary. Interestingly, separation efficiency of the graphene-based capillary was enhanced significantly by using conductive PANI as a sub-layer. The morphology of different layers modified on the capillary column was characterized by scanning electron microscopy (SEM). The electroosmotic flow (EOF) characteristics of capillaries modified with different layers were also investigated by varying the pH value of mobile phase. GO-PDA-PANI@capillary showed good separation efficiency towards alkylbenzenes by OT-CEC mode, with theoretic plate numbers up to 133,918 for benzene. The separation was found to follow a reversed-phase chromatographic retention mechanism. Repeatability of the GO-PDA-PANI@capillary was studied, with relative standard deviations for intra-day and inter-day runs less than 2.89%, and column-to-column runs less than 6.17%. The separation performance of GO-PDA-PANI@capillary was also compared with that of the reported graphene modified capillary.

Facile preparation of protein stationary phase based on polydopamine/graphene oxide platform for chip-based open tubular capillary electrochromatography enantioseparation

A novel chip-based enantioselective open-tubular capillary electrochromatography (OT-CEC) was developed employing bovine serum albumin (BSA) conjugated polydopamine–graphene oxide (PDA/GO) nanocomposites (PDA/GO/BSA) as stationary phase. After the poly(dimethylsiloxane) (PDMS) microfluidic chip was filled with a freshly prepared solution containing dopamine and graphene oxide, PDA/GO nanocomposites were formed and deposited on the inner wall of microchannel as permanent coating via the oxidation of dopamine by the oxygen dissolved in the solution. The PDA/GO-coated PDMS microchips not only have the adhesion of PDA that make them easily immobilized in the microchannel, but also have the larger surface and excellent biocompatibility of graphene which can incorporate much more biomolecules and well maintain their biological activity. In addition, incorporation of GO in PDA film can make surface morphology more rough, which is beneficial for enhancing the loading capacity of proteins in the microchannels and increasing sample capacity of OT-CEC columns. BSA was stably immobilized in the PDMS microchannel to fabricate a protein-stationary phase. Compared with the native PDMS microchannels, the modified surfaces exhibited much better wettability, more stable electroosmotic mobility, and less nonspecific adsorption. The efficient separation of chiral amino acids (tryptophan and threonine) and chiral dipeptide demonstrate that the constructed OT-CEC columns own ideal enantioselectivity. The presented strategy using PDA/GO coating as a versatile platform for facile conjugation of proteins may offer new processing strategies to prepare a functional surface designed on microfluidic chips.

An open tubular capillary electrochromatography column with porous inner surface for protein separation

An open tubular capillary electrochromatography (OTCEC) column using sole porogen to form porous inner surface has been developed. The porous layer was coated on the capillary inner wall by in situ polymerization in the presence of porogen. The results show that the columns using 1-propanol as sole porogen are appropriate for protein separation. It has higher separation efficiency than the column with the usual coporogen due to much more micropores and mesopores on the porous surface and a higher specific surface area. In addition, the sensitivity of the prepared OTCEC column was improved greatly compared with the dynamically coated capillary with polyvinylpyrrolidone.