A novel sol-gel calix[4]arene-modified capillary column for open-tubular capillary electrochromatography

A novel open-tubular CEC (OT-CEC) column was prepared by immobilized 5,11,17,23-tetra-tert-butyl-25,27-diethoxy-26,28-dihydroxy-calix[4]arene (Calix[4]) on fused-silica capillary column with sol-gel technology. Calix[4] was initially reacted with gamma-glycidoxypropyltrimethoxysilane (KH-560) to form a new sol-gel precursor (calix[4]-KH-560), and then mixed with another precursor, namely tetraethoxysilane (TEOS). After hydrolysis and condensation, a sol-gel matrix was obtained, in which calix[4] was incorporated. Successful calix[4]-modified sol-gel coating was suggested by infrared (IR) spectra and greatly decreased EOF. In comparison with OT columns prepared by the sol-gel method with TEOS and KH-560 only, the calix[4]-modified sol-gel column showed greatly improved separation of isomeric toluidines, nitrophenols, picolines and neurotransmitters with structural similarity. Migration time and theoretical plate number reproducibility were satisfactory with RSDs less than 1 and 6% each for within column runs and not more than 3 and 7% each for column-to-column measurements, using toluidine and nitrophenol isomers as test solutes on this column. High separation efficiencies (96,000-300,000 plates/m) for basic toluidine isomers were obtained. This column was also successfully employed to combine extraction procedure for the determination of dopamine (DA) and norepinephrine (NE) in Portulaca oleracea L. The recoveries of DA and NE were 93.3 and 94.2%, respectively.     

Open-tubular capillary electrochromatography using capillary columns chemically bonded with the new host molecules calix[6]crown,calix[6]arene

Two host molecules, p-tert-butylcalix[6]-1,4-crown-4 and p-tert-butylcalix[6]arene were prepared and attached onto the inner surfaces of capillaries for open-tubular electrochromatography with the aid of gamma-glycidoxypropyl-trimethoxysilane (KH-560) as bridging agent. The successful bonding was confirmed by infrared (IR) results and greatly decreased electroosmotic flow (EOF). Parameters affecting separation, such as buffer pH and organic modifier were studied. The two novel stationary phases were evaluated by the separation of isomeric toluidines, a mixture of pyridine and isomeric picolines, and isomeric dihydroxybenzenes; comparisons between capillaries coated with the two stationary phases and bare capillary were investigated. The special selectivity of these two novel stationary phases showed a certain extent of supramolecular interactions between stationary phases and solutes.     

A new type of capillary column for open-tubular electrochromatography

A new type of open-tubular C(18) ester-bonded electrochromatographic column was prepared with sol-gel technology, followed by an on-column octadecyl silylation reaction. Glycidoxypropyltrimethoxysilane, a widely used and important silane agent, was used as the sol-gel precursor to form a thin coating layer on the wall of the fused-silica capillary. The C(18) groups were introduced into the coating layer by on-column esterification reaction with stearic acid. The electrochromatography behavior of the column was evaluated in terms of the separation of peptides. A high efficiency of 4.8x10(5) plates/m was achieved for a basic pentapeptide using the C(18 )ester-bonded column. In comparison with bare capillaries and glycidoxypropyltrimethoxysilane sol-gel-coated capillaries, the C(18) ester-bonded column showed the best separation of a mixture of seven pentapeptides under identical conditions of buffer, pH, and applied voltage.     

Preparation and characterization of p-tert-butylcalix[8]arene bonded capillaries for open-tubular capillary electrochromatography

p-tert-Butylcalix[8]arene bonded capillaries for open-tubular capillary electrochromatography were prepared with γ-glycidoxypropyltrimethoxysilane as a bridge. The bonded capillary displayed low and steady electroosmotic flow (EOF) values over the pH range from 4 to 9. Detection limits for direct spectrophotometric detection at 277 nm for benzenediols (at a signal to noise ratio of 2) were 0.96 mg l⁻¹ for the unbonded capillary and 1.48 mg l⁻¹ for the bonded capillary, showing that the bonded layer did not show significant absorbance and hence decreased sensitivity. The bonded capillaries showed good separation selectivity for o-, m- and p-benzenediols, α- and β-naphthols, and α- and β-naphthylamines. This selectivity was attributed to significant interactions between the analytes and the bonded p-tert-butylcalix[8]arene, which contributed to the electrochromatographic separation mechanism. The bonded capillaries gave high stability and reproducibility.     

Use of the sol-gel technique to prepare capillary columns coated with a macrocyclic dioxopolyamine for open-tubular capillary electrochromatography

Summary Columns for open-tubular (OT) capillary electrochromatography (CEC), coated with 1,4,7,10-tetraazacyclotridecane-11,13-dione (dioxo[13]aneN₄) by use of the sol-gel technique, have been investigated for the first time. Dioxol[13]aneN₄ was reacted with 3-(2-cyclooxypropoxy)propyltrimethoxysilane and the product was then mixed with tetraethoxysilane (TEOS) and water to form a glass matrix as a network in which the stationary phase (dioxo[13]aneN₄) was incorporated. In comparison with OTCEC columns prepared by the sol-gel process with TEOS only, the sol-gel-derived macrocyclic dioxopolyamine columns enabled better separations of a mixture of isomeric nitrophenols and benzenediols, a mixture of isomeric aminophenols and diaminobenzenes, and a mixture of four biogenic monoamine neurotransmitters. High efficiencies (60 000—340 000 plates m⁻¹) were achieved for isomeric benzenediols, aminophenols, nitrophenols, and diaminobenzenes. Migration time and theoretical plate number reproducibility was satisfactory;RSD was <2% for one column and <8.5% from column to column.     

Preparation of stationary phases for open-tubular capillary electrochromatography using the sol-gel method

Capillary electrochromatography requires the deposition of a stationary phase inside the capillary. In this paper the sol-gel method is proposed for this purpose. The gels were prepared externally and injected into a fused-silica capillary, where anchorage to the capillary wall was possible through condensation reactions between the silanol groups of the capillary wall and the residual silanol groups the gel. Contrary to a commonly used practice, alkaline pretreatment of the inner capillary wall prior to the introduction and anchoring of the gel was found to be only marginally effective in improving the mechanical stability of the column. The influence of various parameters, such as the pH, the water content, the presence of alcohol (ethanol) on the formation of tetraethoxysilane (TEOS)-n-octyltriethoxysilane (C8-TEOS) hybrid gels of varied composition is discussed. The pH and the amount of water present were found to be the determining factors in the preparation of a stable gel with the desired mechanical and chromatographic properties. By carrying out the gel formation at 80 degrees C, capillary columns could be produced in 2.5 h. While an acidic pH was required during (external) gel formation, subsequent treatment of the gel inside the capillary with an alkaline solution ('aging') was found to improve separation and stationary phase capacity significantly. The capillary columns were subsequently used to separate a mixture of polycyclic aromatic hydrocarbons in less than 3 min.     

原子转移自由基聚合法制备新型亲水开管毛细管柱及在毛细管电色谱上的应用

为了增加开管毛细管柱(OTCC)的相比,提高分离效率,发展了表面引发原子转移自由基聚合法(SI-ATRP)制备葡萄糖聚合物修饰的开管毛细管柱。通过扫描电镜观察,该开管柱内壁上修饰了三维波浪状聚合物,明显增加了内壁比表面积和相比。在pH 3~11范围内,对含糖聚合物修饰的开管柱和空柱的电渗流进行了比较。修饰后开管柱的电渗流仅为空柱的1/2~1/3,且在pH 6~11范围内保持平稳。稳定的电渗流保证了分离的重复性和稳定性。用该开管毛细管柱成功实现了小分子混合物(苯丙氨酸、胸腺嘧啶、腺苷、鸟苷、5-溴尿嘧啶、水杨酸)以及蛋白质大分子(核糖核酸酶B、转铁蛋白和牛血清白蛋白)的有效分离,结果表明葡萄糖聚合物修饰的开管毛细管柱具有良好的重复性和稳定性。     

p-tert-Butylcalix[4]arene-1,3-bis(allyloxyethoxy)ether coated capillaries for open-tubular electrochromatography.

The preparation and characterization of p-tert-butylcalix[4]arene-1,3-bis(allyloxyethoxy)ether (CA[4]-BAE) chemically coated capillaries via a free-radical reaction with vinyltriethoxysilane (VTES) that was attached onto the inner wall previously were carried out. IR spectra and decreased electroosmotic flow (EOF) suggested that the capillary was successfully coated with CA[4]-BAE. A slight slope of EOF versus pH at 5 < pH < 9 would help to make the separation reproducible. The CA[4]-BAE-coated capillary showed improved separations of toluidine isomers, naphthol isomers and polycyclic aromatic hydrocarbons (PAHs) compared with an uncoated capillary. The special selectivity indicates that there is a certain extent of host-guest interactions between the solutes and the CA[4]-BAE coating.     

Comparison of separation behavior of benzodiazepines in packed capillary electrochromatography and open-tubular capillary electrochromatography

Packed column capillary electrochromatography (CEC), open-tubular CEC and microcolum liquid chromatography (LC) using a cholesteryl silica bonded phase have been studied to compare the retention behavior for benzodiazepines. It has been found that packed column CEC gives better resolution, faster analysis time than microcolumn LC for benzodiazepines maintaining similar selectivity except for some solutes which are charged species under the separation conditions. However, open-tubular CEC gave different selectivities to a larger extent for charged benzodiazepines from that which should be produced by the chromatographic properties of the cholesteryl silica phase. Charged species migration times are mainly influenced by electrophoretic mobility rather than the chromatographic interactions.     

Triazole-modified calix[4]crown as a novel fluorescent on-off switchable chemosensor

A novel fluorescent on-off switchable chemosensor 1 with two different types of cationic binding sites is synthesized, which is composed of a triazole-modified calix[4]crown in the 1,3-alternate conformation. Among 15 metal ions examined, the fluorescence of 1 was strongly quenched by Hg2+, Cu2+, Cr3+, and Pb2+; however, the revival of emission from the strongly quenched 1.Pb2+ complex was achievable by the addition of K+, Ba2+, or Zn2+ ions. Thus, metal ion exchange can trigger an on-off switchable fluorescent chemosensor.     

Calix[4] open-chain crown ether-modified, vinyl-functionalized hybrid silica monolith for capillary electrochromatography

A novel calix[4] open-chain crown ether (p-tert-butylcalix[4]arene-1,3-bis(allyloxyethy) ether)-modified, organic-inorganic hybrid silica-based monolithic column possessing vinyl ligands for CEC is described. The monolithic silica matrix containing a vinyl functionality was synthesized by in situ cocondensation of tetramethoxysilane (TMOS) and vinyltrimethoxysilane (VTMS) via sol-gel process and chemically modified with calix[4] open-chain crown ether by free radical polymerization procedure using alpha, alpha'-azobisisobutyronitrile (AIBN) as an initiator. Morphology of the monolithic column was examined by SEM and the successful incorporation of calix[4] open-chain crown ether to the vinyl-hybrid monolith was characterized by infrared (IR) spectra. Compared with an unmodified vinyl-hybrid monolithic column, slightly stronger EOF at pH >7.5 was observed for this monolithic column due to the ionization of phenolic hydroxyls on the lower rim of calix[4]arene. VTMS/TMOS ratios in the reaction mixture were varied and 1:4 was found to be optimum to obtain homogeneous monolith with good permeability. The performance of the column was evaluated by nucleotides, beta-blockers, neurotransmitters, and PAHs as test solutes and compared with that of unmodified vinyl-hybrid monolithic column. Greatly improved column performance was obtained due to the host-guest interaction and intermolecular hydrogen bonding provided by the calix[4] open-chain crown ether moiety. The column efficiencies for neurotransmitters and nucleotides are up to 120 000 and 110 000 plates/m, respectively. Migration time and theoretical plate number reproducibilities were reasonable with RSDs less than 1.0 and 1.8% each for within column runs and not more than 7.2 and 8.6% each for column-to-column measurements, using four nucleotides as test solutes.     

一种二维吖嗪共价有机骨架材料涂层毛细管的制备及其用于开管-毛细管电色谱分离硝基苯酚类环境内分泌干扰物

作为一种新型多孔晶体材料,共价有机骨架材料（COFs）由于具有比表面积大、密度小、稳定性高及孔径可调等特点而在诸多领域中得到了广泛的应用。但将其用作固定相以提高开管-毛细管电色谱（OT-CEC）分离效率的研究报道较少。鉴于此,该文参考文献方法合成了一种二维吖嗪COF（ACOF-1）,然后以ACOF-1作为固定相制备了ACOF-1涂层毛细管并以其为分离通道建立了一种分离硝基苯酚类环境内分泌干扰物（EEDs）的OT-CEC新方法。通过X射线粉末衍射、傅里叶变换红外光谱和扫描电子显微镜等表征手段证明成功合成并制备了ACOF-1和ACOF-1涂层毛细管。实验结果表明,在最佳分离条件下,所建立的OT-CEC方法可在20 min内实现2-硝基苯酚、4-硝基苯酚、2,4-二硝基苯酚和2,4,6-三硝基苯酚4种硝基苯酚分析物的基线分离。4种分析物的线性范围分别为10~500 mg/L和20~1000 mg/L,决定系数均大于0.99,检出限和定量限分别为0.13~0.23 mg/L和0.45~0.60 mg/L。迁移时间和峰面积的日内、日间及柱间相对标准偏差均不超过9.4%,表明所建立的方法重现性好,稳定性高,可用于硝基苯酚类EEDs的分离检测。分离机理研究表明ACOF-1孔结构对各分析物的尺寸选择作用是影响分离行为的主要因素。该工作证明了以COFs作为固定相的OT-CEC方法用于分离检测EEDs的可行性,后续将继续围绕COFs涂层毛细管的制备及其用于OT-CEC分离测定EEDs开展研究。     

Titanium dioxide coated surfaces for capillary electrophoresis and capillary electrochromatography

The potential of titanium dioxide coatings to control the electroosmotic flow and to affect the migration behavior of analytes in capillary electrophoresis and open-tubular capillary electrochromatography was evaluated. The inner wall of a fused-silica capillary was applied with a solution of a titanium peroxo complex, followed by heating at an elevated temperature. The resultant product was ascertained to be titanium dioxide in a crystalline form of anatase by the results of Fourier transform-infrared spectrometry (FT-IR), X-ray photoelectron spectroscopy, and Raman spectroscopy. The capillary thus made had anodic or cathodic electroosmotic flow, depending on the pH and composition of the background electrolyte used. The titanium dioxide surface of the capillary was readily modified by a silanizing reagent. The performance of the titanium dioxide surfaces with or without chemical modifications was examined with inorganic anions, neutral compounds and peptides.     

Synthesis and structure of a novel molybdenum imido calix[6]arene

Treatment of p-tert-butylcalix[6]areneH₆ (H₆L) with [Mo(OBu^t)₂{[2,2′-(N)-C₆H₄]₂(CH₂CH₂)}] in refluxing toluene affords, after work-up, the complex [Mo(2-NC₆H₄CH₂CH₂C₆H₄NHC(Me)NH-2^/)LH₂]·4MeCN (1), which contains an 11-membered metallocyclic ring as characterised by Synchrotron X-radiation.     

Synthesis of novel rigid triazine-based calix[6]arenes

In this report, the stepwise synthesis of a novel rigid and functionalised macrocycle 2 based on triazine and phenylenediamine linkers, is presented. Poor recognition of the macrocycle 2 for its substrates is observed, which shows experimentally that for the meltriazine based-calix[6]arene system, the binding ability of the melamine moiety gets more benefit from the ring flexibility derived from a xylenediamine linker 1 than from a phenylenediamine linker 2.     

Metal ion sensing novel calix[4]crown fluoroionophore with a two-photon absorption property

1,3-Alternate calix[4]arene-based fluorescent chemosensors bearing two-photon absorbing chromophores have been synthesized, and their sensing behaviors toward metal ions were investigated via absorption band shifts as well as one- and two-photon fluorescence changes. Free ligands absorb the light at 461 nm and weakly emit their fluorescence at 600 nm when excited by UV-vis radiation at 461 nm, but no two-photon excited fluorescence is emitted by excitation at 780 nm. Addition of an Al(3+) or Pb(2+) ion to a solution of the ligand causes a blue-shifted absorption and enhanced fluorescence due to a declined resonance energy transfer (RET) upon excitation by one- and two-photon processes. Addition of a Pb(2+) ion to a solution of 1.K(+) results in a higher fluorescence intensity than the original 1.Pb(2+) complex regardless of one- or two-photon excitation, due to the allosteric effect induced by the complexation of K(+) with a crown loop.     

Gold nanoparticle-coated capillaries for protein and peptide analysis on open-tubular capillary electrochromatography

We report a new method of immobilization of gold nanoparticles (AuNPs) on a fused-silica capillary through covalent binding. The resulting modified capillary was applied to electrophoretic systems to improve the efficiency of separation and the selectivity of selected solutes. The immobilization of AuNPs on the capillary wall was performed in a very simple and fast way without requiring heating. The surface features of an AuNP-coated capillary column were determined using the scanning electron microscopy. The chromatographic properties of AuNP-coated capillaries were investigated through variation of the buffer pH and separation voltage. Effective separations of synthetic peptides mixture were obtained on the AuNP-coated capillaries. The method shows a remarkable stability since it was reused about 900 times. The capacity factor was duplicated. Therefore, this modification is stable and can be applied to different separation purposes. A complex mixture of tryptic peptide fragments of HSA was analyzed in both the bare- and the AuNP-coated capillaries. Better electrophoretic peptide profile was observed when using the AuNP-coated capillary.     

Synthesis and characterization of novel calix[6]phyrin derivatives

Three novel calix[6]phyrin derivatives have been synthesized by reaction of corresponding tripyrrane with aromatic aldehyde. The reaction condition was optimized and the structures of these compounds have been characterized by NMR and MS.     

Preparation of capillary columns coated with linear polymer containing hydrophobic and charged groups for capillary electrochromatography

A linear polymer-coated capillary was prepared by in-capillary copolymerization of N-tert-butylacrylamide (TBAAm) with a charged monomer, 2-acrylamido-2-methyl-1-propanesulfonic acid (AMPS), after the capillary pretreatment with a bifunctional reagent. The coated capillaries were applied in capillary electrochromatographic (CEC) separation of small neutral compounds. Hydrophobic groups in the linear polymer, which were immobilized onto the capillary surface, functioned as the stationary phase in reversed-phase CEC separation, and charged groups in the linear polymer generated electroosmotic flow (EOF) along the column. The coated capillaries were prepared by a simple procedure. Moreover, the reproducibility with respect to EOF rate and migration times of the solutes was excellent. The results for CEC separation of small molecules using the linear polymer-coated capillaries are presented.     

Characterization and applications of etched chemically modified capillaries for open-tubular capillary electrochromatography

In this article, the effects of the stationary phase, buffer pH, organic modifier type, organic modifier composition, applied voltage, and temperature on the migration of several synthetic peptides in etched chemically modified open-tubular capillaries are discussed. With these solutes, migration is due to two effects: electrophoretic mobility and solute/bonded phase interactions. In addition, relative migration rates are evaluated for the peptide samples as a function of these experimental variables in order to determine which parameters might be useful for optimizing separations in open-tubular capillary electrochromatography (OTCEC). Some examples of synthetic peptide separations are presented where the sample contains a major component and several minor species, demonstrating how the resolution of these mixtures can be affected by the appropriate choice of experimental variables.