Open-tubular Capillary Electrochromatography Research Articles

Separation of antipyretic analgesics by open tubular capillary electrochromatography with homopolymer coatings.

This study developed an open-tubular capillary electrochromatography protocol for the analysis of antipyretic analgesic drugs, which used a multifunctional homopolymer as coating. A controlled/living radical polymerization strategy was adopted to obtain poly(N-acryloxysuccinimide) with a tunable chain-length. The homopolymer coating enhanced the separation performance by contributing to the hydrophobic and hydrogen-bonding interactions between the analytes and the homopolymer. The effect of polymer chain-length and buffer pH and concentration on the separation efficiency was evaluated. In this approach, baseline separation of the three test drugs was achieved within 15 min. The repeatability of the prepared homopolymer coating was investigated, with the relative standard deviations< 2.88% observed in intra- and interday runs. Good linearity in the 5-800 µM range (R2 ≥ 0.998) demonstrates that accurate quantitative analysis of real samples was achieved. Moreover, the proposed assay was used to quantify the three drugs (aminopyrine, 4-aminoantipyrine, and phenacetin) in urine samples, achieving recovery rates between 92.1 and 108.7%. This promising methodology may be used for the analysis of drugs in real bio-samples and for the development of unique homopolymer coatings for open-tubular capillary electrochromatography systems.

Modulation of Electroosmotic Flow in Open Tubular Capillary Electrochromatography by Chitosan-Assisted Titanium Oxide Nanoparticles Liquid Phase Deposition

Electroosmotic flow is one of the most important driving forces in capillary electrophoresis, which can control the migration velocity and direction of the target substance, thus affecting the separation efficiency and reproducibility of capillary electrophoresis. In this study, a facile method was developed for the preparation of TiO2/chitosan open tubular capillary electrochromatography for modulating electroosmotic flow in capillary electrophoresis. The magnitude and direction of electroosmotic flow were modulated by changing a series of coating preparation parameters, such as deposition time of (NH4)2TiF6 and H3BO3, the concentration of chitosan, and the concentration ratio of (NH4)2TiF6 and chitosan. The results showed that when the concentration of (NH4)2TiF6 was below 2 mg/mL, the surface of the capillary was not modified completely by phosphate ion by Lewis acid–base interaction, resulting in an anodic electroosmotic flow. Furthermore, the feasibility and stability of the column were assessed by separation of nucleotides and alkaloid compounds. With the TiO2/chitosan liquid phase deposition columns with the concentration ratio of 3:7 [(NH4)2TiF6:chitosan], alkaloids were baseline separated within 11 min. The baseline separation of the four nucleotides was achieved using the TiO2/chitosan-coated columns with the concentration ratio of 1.8:7 [(NH4)2TiF6:chitosan]. Furthermore, the relative standard deviations of migration times for inter-day, intra-day, and column-to-column repeatability of the TiO2/chitosan-coated columns with the concentration ratio of 3:7 [(NH4)2TiF6:chitosan] were all less than 5.8%. The strategy of liquid phase deposition of TiO2 nanoparticles provides a promising approach for modulation electroosmotic flow.

Polydopamine-assisted immobilization of a zinc(II)-derived metal-organic cage as a stationary phase for open-tubular capillary electrochromatography.

A capillary column was modified with a soluble zinc(II)-derived metal-organic cage (MOC) [Zn2L] as the stationary phase to obtain a new coating layer for use in open-tubular capillary electrochromatography. The inner surface of the capillary was first coated with a layer of polydopamine. Then, a solution of the MOC in dichloromethane was introduced into the capillary upon which it is bonded both covalently and non-covalently. The resulting coating layer on the capillary was characterized by scanning electron microscopy and Fourier transform infrared spectroscopy. The results demonstrated the successful formation of the [Zn2L] modified open-tubular column. The column showed good separation performance towards neutral compounds (such as methylbenzene, ethylbenzene, n-propylbenzene and n-butylbenzene), acidic drugs (such as ibuprofen, ketoprofen, flurbiprofen and diclofenac sodium), food additives (such as parabens, vanillin and related phenolic compounds) and small biomolecules (such as nucleosides and nucleotide bases) by π-interaction and hydrophobic interaction. It also exhibited good precision, the relative standard deviations of the retention time for intra-day, inter-day runs and column-to-column being <1.6%, 2.8%, and 4.0%, respectively. Graphical abstract Schematic presentation of the open-tubular column modified with zinc(II)-derived metal-organic cage by polydopamine-assisted strategy onto the inner wall of capillary for electrochromatographic separations.

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.

Open-tubular capillary electrochromatography with β-cyclodextrin-functionalized magnetic nanoparticles as stationary phase for enantioseparation of dansylated amino acids.

Magnetic nanoparticles (MNPs) modified with β-cyclodextrin and mono-6-deoxy-6-(1-methylimidazolium)-β-cyclodextrin tosylate (an ionic liquid), which called MNP-β-CD and MNP-β-CD-IL, were coated into the capillary inner wall. Compared to anuncoated capillary, thenew systems show good reproducibility and durability. The systems based on the use of MNP-β-CD or MNP-β-CD-IL as stationary phases were established for enantioseparation of Dns-modifiedamino acids. Improved resolutions were obtained for both CEC systems. Primary parameters such as running buffer pH value and applied voltage were systematically optimized in order to obtain optimal enantioseparations. Under the optimized conditions, the capillaries exhibited excellent chiral recognition ability for six Dns-amino acids (the DL-forms of alanine, leucine, lsoleucine, valine, methionine, glutamic acid)and provided a promising way for the preparation of chiral column. Graphical Abstract Schematic presentation of the open-tubular capillary electrochromatography systems with MNP-β-CD and MNP-β-CD-IL as stationary phases for enantioseparation of dansylatedamino acids.

In situ immobilization of sulfated-β-cyclodextrin as stationary phase for capillary electrochromatography enantioseparation

In this work, a novel sulfated-β-cyclodextrin (S-β-CD) coated stationary phase was prepared for open-tubular capillary electrochromatography (OT-CEC). The capillary was developed by attaching polydopamine/sulfated-β-cyclodextrin (PDA/S-β-CD) onto the gold nanoparticles (AuNPs) coated capillary which was pretreated with polydopamine. The results of scanning electron microscopy (SEM) and energy dispersive X-ray analysis spectroscopy (EDS) indicated that polydopamine/sulfated-β-cyclodextrin was successfully fixed on the gold nanoparticles coated capillary. To evaluate the performance of the prepared open tubular (OT) column, the enantioseparation was carried out by using ten chiral drugs as model analytes. Under the optimal conditions, salbutamol, terbutaline, trantinterol, tulobuterol, clorprenaline, pheniramine, chlorpheniramine, brompheniramine, isoprenaline and tolterodine were baseline separated with the resolution (Rs) values of 3.25, 1.76, 2.51, 1.89, 3.17, 2.17, 1.99, 1.72, 2.01 and 3.20, respectively. Repeatability of the column was studied, with the relative standard deviations for run-to-run, day-to-day and column-to-column lower than 5.7%.

Open-tubular capillary electrochromatography for the simultaneous determination of cadmium and copper in plants.

We discuss the separation and determination of cadmium and copper in plant samples such as Triticum durum (wheat) and Helianthus annuus (sunflower) using open tubular capillary electrochromatography with indirect detection. Before performing the analysis, the samples were digested by microwave-assisted methods using HNO3 . Regarding the electrophoretic system, several experimental parameters were previously evaluated such as the capillary surface, mobile phase composition, buffer, pH, and voltage applied. The baseline resolution of the studied metals was obtained within 8 min by using a capillary immobilized with carboxylic multi-walled carbon nanotubes and a background electrolyte composed of 6mM imidazole, pH 4.0. The applied voltage and the temperature were set at 20 kV and 25°C, respectively. Precision, detection, and quantification limits, along with linearity were investigated. The limits of detection and quantification were 2.20 and 7.40 μg/kg, for Cu2 ⁺ and 0.05 and 0.20 μg/kg in the case of Cd2 ⁺. A good linearity was achieved over a concentration working range of 7.5-100 and 0.2-25 μg/kg for Cu2 ⁺ and Cd2 ⁺ accordingly. Recovery data for validation studies were found in a range of 98.2-101.5% for both analytes.

A novel coating method for CE capillary using carboxymethyl-β-cyclodextrin-modified magnetic microparticles as stationary for electrochromatography enantioseparation.

Magnetic microparticles (MMPs) have been extensively studied and aroused considerable interest in separation science owing to their superior characteristics. In this paper, a novel coated capillary with carboxymethyl-β-cyclodextrin-functionalized magnetic microparticles (CD-MMPs) as stationary phase was constructed and then applied to establish an open-tubular capillary electrochromatography enantioseparation system. The preparation of the CD-MMP-coated open-tubular column was very convenient because the coating of the magnetic microparticles onto the capillary column could be easily manipulated by an external magnetic field. The preparation conditions of the coated capillary such as magnetic field intensity and coating time are discussed in detail. The new constructed CD-MMP capillary system was applied to separate enantiomers of several racemic drugs. Compared to the uncoated capillary system, obviously preferable separations of tested enantiomers were obtained. Several important parameters affecting the enantioseparation, such as CM-β-CD concentration, running buffer pH, organic solvent, and applied voltage, were systematically optimized. Furthermore, satisfactory repeatability and chemical stability of this new CD-MMP capillary system were achieved in the experiment. Graphical abstract ᅟ.

In-situ growth of a metal organic framework composed of zinc(II), adeninate and biphenyldicarboxylate as a stationary phase for open-tubular capillary electrochromatography.

This work reportson the in-situ growth of a metal organic framework (MOF) composed of zinc(II), adeninate and biphenyldicarboxylate on the inner wall of a capillary, and theuse of this MOF as a stationary phases in open-tubular capillary electrochromatography. The inner wall of a fused-silica capillary was first modified with 3-aminopropyltriethoxysilane to create surface amino sites, and then the MOF was synthesized by in-situ growth. The modified capillary was characterized by scanning electron microscopy, Fourier transform infrared spectroscopy and X-ray diffraction. The results proved the successful growth of the MOF. The resultant open-tubular column showed good separation selectivity towards neutral compounds, acidic and basic compounds including nonsteroidal anti-inflammatory drugs, sulfa drugs and small biomolecules. In addition, the modified column is stable and repeatable. The precisions (expressed as RSDs) of the retention time for intra-day (n= 5) and inter-day (n = 5) separations and between columns (n= 3) are less than 0.5, 1.6 and 4.7%, respectively. Conceivably, this new kind of MOF represents a most useful novel stationary phase in electrochromatographic separations. Graphical abstract Schematic presentation of the open-tubular column modified with bio-MOF-1 by in situ hydrothermal reaction with adeninate, Zn(II) and biphenyldicarboxylate for capillary electrochromatographic separations.

Using open-tubular capillary electrochromatography with part-coating column for binding constants determination of β2 -adrenergic receptor with seven drugs.

An open-tubular capillary electrochromatography method has been developed for the determination of binding constants between β2 -adrenergic receptor (β2 -AR) and seven drugs. β2 -AR was oriented immobilized onto one part of inner surface of capillary via microwave-assisted technical synthesis. According to the linear relationship between coating length and the apparent mobility of analyte, the binding constant (Kb ) can be obtained by related theories and equations. The order of Kb values between drugs such as adrenaline hydrochloride, norepinephrine bitartrate, and propranolol hydrochloride with β2 -AR is well consistent with that reported in the literature. By the method, Kb values between four extracts of Radix Paeoniae Rubra and β2 -AR were also successfully obtained. Subsequently, computer models were applied to interpret the CEC experiments. And the results proved to be in good agreement with the method. The work, herein, demonstrates the potential of the method in drug-receptor affinity interactions evaluation and screening of lead compounds from natural sources.

The preparation of poly-levodopa coated capillary column for capillary electrochromatography enantioseparation

Levodopa (L-DOPA) is promising as chiral stationary phase for open tubular capillary electrochromatography (OT-CEC) enantioseparation owing to its self-polymerization adhesive property and chiral recognition potential. In this work, CuSO4/H2O2 was used as a trigger agent to accelerate the polymerization process of L-DOPA and the poly-levodopa (poly-(L-DOPA)) coated column was successfully prepared for the first time by depositing it on the inner wall of fused silica capillary via the rapid and in-situ approach at room temperature. The performance of the poly-(L-DOPA) coated capillary was validated by the separation of different chiral analytes, including chiral amine drugs, neurotransmitters and amino acids, and the good enantioseparation efficiencies were achieved. For five consecutive runs, the relative standard deviations (RSDs) for the migration time of the analytes for intra-day, inter-day and column-to-column were in the range of 0.19–1.33%, 0.96–4.47%, and 2.21–7.79%, respectively. Additionally, the poly-(L-DOPA) coated capillary column could be successively used over 250 runs without observable change in the separation efficiency.

Covalent organic framework TpPa-1 as stationary phase for capillary electrochromatographic separation of drugs and food additives.

Because of unique properties like permanent porosity, low density and large surface area, covalent organic frameworks are attractive microporous materials as stationary phase for CEC. Among them, COF-TpPa-1 possesses extraordinary chemical stability and common features of covalent organic frameworks. So, COF-TpPa-1 could be a promising material as stationary phase to enhance separation selectivity and tolerate the acid or alkaline electrochromatographic separation conditions. Here, we report the preparation of COF-TpPa-1 modified capillary column by in situ growth for electrochromatographic separation. The immobilization of COF-TpPa-1 on the inner wall of capillary column was confirmed by Fourier transform infrared spectroscopy, scanning electron microscopy and X-ray diffraction. The fabricated COF-TpPa-1 modified capillary column indicated good resolution for the separation of neutral analytes, nonsteroidal anti-inflammatory drugs and food additives in open-tubular CEC mode. Compared with bare capillary column, the COF-TpPa-1 modified capillary column improved separation selectivity remarkably for tested analytes, including hydrophobic, π-π and hydrogen bond interactions. Besides, the prepared capillary column showed excellent repeatability and stability. The intraday, interday and column-to-column relative standard deviations of migration time for neutral analytes were less than 3.54%. This work shows enamine-linked covalent organic frameworks have great potential as stationary phases in CEC.

In situ growth of Zr-based metal-organic framework UiO-66-NH2 for open-tubular capillary electrochromatography.

The high stability and other properties of Zr(IV)-based metal organic frameworks(MOFs) make it a promising choice for chromatographic separation, while the application in open-tubular capillary electrochromatography (OT-CEC) separation has not been explored yet. Herein, we report the first example of the in-situ growth of UiO-66-NH2 onto the capillary for open-tubular capillary electrochromatography. UiO-66-NH2 consists of ZrCl4 and 2-amino-1,4-benzenedicarboxylic acid, which is highly porous and stable in a variety of solvents. The prepared UiO-66-NH2 modified capillary was characterized by scanning electron microscopy (SEM) and Fourier transform infrared spectra (FT-IR), and the results confirmed the successful growth of the UiO-66-NH2 . The baseline separation of chlorobenzenes, phenoxyacids and two groups of phenols was achieved owing to the combined interaction of π-π interaction, hydrophobic interaction, molecular sieve effect, electrophoretic migration and hydrogen-bonding interaction etc. Besides, the prepared capillaries showed good reproducibility, with relative standard deviations (RSDs) for intra-day, inter-day and column-to-column runs in the range of 1.38-2.60%, 3.39-4.05%, and 3.47-5.03%, respectively. Our work indicates Zr(IV)-based MOFs are promising materials as stationary phase in CEC separation.

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.

Open tubular capillary electrochromatography with block co-polymer coating for separation of β-lactam antibiotics

A new open-tubular capillary electrochromatography (OT-CEC) method for analysis of β-lactam antibiotics has been developed with unique block co-polymer coating. To obtain the highly ordered block polymer chains, reversible addition fragmentation chain transfer radical polymerization method was used to synthesize poly(maleic anhydride-styrene-N-isopropylacrylamide). The prepared block co-polymer coating was characterized with NMR, fourier transform infrared spectroscopy and scanning electron microscope. Several key separation factors of OT-CEC, which including polymer amount, stability of the coating, temperature, species of organic additives, buffer pH and concentration, were investigated in detail. Our results indicated that the separation efficiency was improved greatly with the coating capillary and the three test analytes could be baseline separated. Then, the separation mechanism was briefly explored. Moreover, the proposed OT-CEC method displayed promising quantitative analysis property of the three test analytes with good linearity (R2>0.99), repeatability (relative standard deviations <0.9%) and high recovery (95.4%–106.2%). Further, the assay was applied in monitoring the three test β-lactam antibiotics (cephradine, cephalexin and amoxicillin) in serum samples, providing a useful platform for construction of novel polymer coatings in OT-CEC system and for analysis of drugs in real bio-samples.

Nano-amylose-2,3-bis(3,5-dimethylphenylcarbamate)-silica hybrid sol immobilized on open tubular capillary column for capillary electrochromatography enantioseparation.

The chiral organic-inorganic hybrid materials can exhibit a high loading, and the chiral selector nanoparticles can create efficient stationary phases for open-tubular capillary electrochromatography (OT-CEC). Hence, a novel protocol for the preparation of an OT column coated with nano-amylose-2,3-bis(3,5-dimethylphenylcarbamate) (nano-ABDMPC)-silica hybrid sol through in situ layer-by-layer self-assembly method was developed for CEC enantioseparation. By controlling the assembly cycle number of nano-ABDMPC-silica hybrid sol, a homogeneous, dense and stable coating was successfully prepared, which was confirmed by SEM and elemental analysis. As the main parameter influencing the chiral separating effect, the nano-ABDMPC bearing 3-(triethoxysilyl)propyl residues concentration was investigated. The experimental results showed that 10.0 mg/mL nano-ABDMPC bearing 3-(triethoxysilyl)propyl residues coated OT capillary column possessed chiral recognition ability toward the six enantiomers (phenylalanine, tyrosine, tryptophan, phenethyl alcohol, 1-phenyl-2-propanol, and Tröger's base) at some of the different conditions tested. Additionally, the coated OT column revealed adequate repeatability concerning run-to-run, day-to-day and column-to-column. These results demonstrated the promising applicability of nano-ABDMPC-silica hybrid sol coated OT column in CEC enantioseparations.

Homochiral zeolite-like metal-organic framework with DNA like double-helicity structure as stationary phase for capillary electrochromatography enantioseparation

In recent years, artificial materials with double helix structure have attracted widespread attention due to their unique properties such as the DNA like double-helicity, intrinsic chirality and diverse functional groups. Developing novel chiral stationary phases (CSPs) for capillary electrochromatography enantioseparation is of intriguing interest. Herein, a novel homochiral zeolite-like metal-organic framework (ZMOF) JLU-Liu23 with unique DNA like double-helicity structure was firstly utilized as the CSP in open tubular capillary electrochromatography (OT-CEC) for enantioseparation of chiral monoamine neurotransmitters and analogues. Owing to the unique DNA like double-helicity structure of the homochiral ZMOF JLU-Liu23, the good enantioseparation of four monoamine neurotransmitters and analogues was achieved on the prepared homochiral ZMOF JLU-Liu23 coated capillary column. The relative standard deviations (RSDs) of the analytes migration time for intra-day, inter-day and column-to-column were in the range of 0.3–0.6%, 0.8–2.2% and 3.5–6.5%, respectively.

Highly uniform porous silica layer open-tubular capillary columns produced via in-situ biphasic sol–Gel processing for open-tubular capillary electrochromatography

We report a highly uniform porous layer open tubular (PLOT) column for capillary electrochromatography (CEC) analysis. The PLOT column is easily fabricated using a single-step in-situ biphasic reaction, producing homogeneous porous-layer modified surface with ∼240 nm thickness in a 50 μm-id capillary. CEC performance of the PLOT column has been investigated and optimized under various experimental parameters. Using a mixture of naphthalene and biphenyl as the test sample, we show that the PLOT column exhibits good separation efficiency with resolution >3.0 and theoretical plate numbers over 6 × 104, as well as good intra-/inter-day repeatability and column-to-column repeatability. The column has been successfully applied for CEC analysis of three different types of samples without any further modification of the columns, including complicated peptide products from tryptic-digestion of proteins (lysozyme and BSA), β-blockers (basic samples) and polycyclic aromatic hydrocarbons (neutral samples). Efficient separation has been achieved, which could be attributed to the enhanced surface-to-volume ratio of the PLOT column that will increase the interaction between solid phase and mobile phase in CEC. In addition, base-line separation of neutral samples indicates the reversed phase chromatographic property of the PLOT column, which could be induced by the residue of hexadecyltrimethylammonium bromide used in the fabrication process. Our study show that the present PLOT column is a promising approach that can significantly enhance CEC separation efficiency and could be of potential value in analysis of various different samples.

Enantioselective open-tubular capillary electrochromatography using a β-cyclodextrin–gold nanoparticles–polydopamine coating as a stationary phase

A novel β-cyclodextrin–gold nanoparticles–polydopamine modified open tubular column was prepared for the chiral separation of enantiomers in capillary electrochromatography.

Application of avidin-phospholipid vesicle complex as the stationary phase for chiral separation in open-tubular capillary electrochromatography

Application of avidin-phospholipid vesicle complex as the stationary phase for chiral separation in open-tubular capillary electrochromatography