Preparation of Monolithic Silica and Polymer Capillary Columns with Ultrahigh Column Efficiencies and Comparisons between van Deemter Plots of Alkylbenzenes on These Two Kinds of Columns.

Monolithic silica and polymer capillary columns with ultrahigh column efficiencies were prepared. Permeability and electrochromatography performances of these two kinds of columns were compared. Monolithic silica columns bear higher permeability than polymer counterparts by two orders of magnitude. Furthermore, the van Deemter plots of alkylbenzenes on these two kinds of columns demonstrate that monolithic silica columns produce much lower plate heights of alkylbenzes than polymer columns do. Within the range of electroosmotic flow (EOF) velocity investigated, no uptrend of plate height with the increase of EOF was observed suggesting the great capacity of fast separation and high efficiency. The plate height of thiourea on monolithic silica columns is as low as 2.67 µm, representing its corresponding column efficiency is over 430,000 plates/m. As far as we know, it is the highest ever column efficiency reported in the literature. Moreover, the separation of butylbenzene isomers was obtained on the monolithic silica column.

Selective Enrichment of Low-abundance Compounds in a Mixture by Capillary Electrophoresis.

Recently, we developed a new approach for the selective enrichment of low-abundance compounds in biological samples by capillary electrophoresis. As a model test, the low-abundance compound lysozyme was successfully fractionated from a mixture containing high-abundance compound BSA (1:4500) using a custom-made apparatus. The feasibility of this approach for real complex biological samples was verified by rat serum, wherein three low-abundance proteins with high charge/mass ratios were detected.

Composite Nanostructure of Manganese Cluster and CHA-Type Silicoaluminaphosphates: Enhanced Catalytic Performance in Dimethylether to Light Olefins Conversion.

Light olefins, especially ethylene and propylene, are important chemicals in petrochemical industries with an increasing demand and play an essential role in the global consumption. In this regard, there have been extensive studies to design efficient catalysts for the light olefins productions. In this study, we report a new protocol to induce Mn nanoclusters (MnNC) into the mesopore of a CHA-type silicoaluminaphosphates via a one-pot synthesis of MnNC@SAPO-34 catalysts. The catalysts are characterized by a series of technology, such as TEM, XRD, NH3-TPD, 27Al MAS NMR, ICP-MS, XPS, and as well as N2-physical adsorption methods. The Mn nanoclusters of Mn2O3 and MnO2 species are well dispersed in the framework of the SAPO-34 silicoaluminaphosphates, modifying the porosity and acidic property of the SAPO-34: Giving rise to more mesoporous and improving the acid density. The MnNC@SAPO-34 catalysts exhibit decent 100% conversion and 92.2% olefins selectivity in the dimethyl ether to olefins (DTO) reactions, which is considerably higher than that for SAPO-34 silicoaluminaphosphates (79.6% olefins selectivity). The higher olefins selectivity over the MnNC@SAPO-34 is deemed to associate with the strong acid density and intensity of the silicoaluminaphosphates. Further, the Mn particles largely improve silicoaluminaphosphates's durability.

CuO Nanoparticle-Decorated TiO2-Nanotube Heterojunctions for Direct Synthesis of Methyl Formate via Photo-Oxidation of Methanol.

Methyl formate is widely employed as one of the important organic intermediates in C1 chemistry and as an environmentally benign chemical in the emerging area of "green chemistry". We here study the catalytic effectiveness of CuO/TiO2-nanotube nanocomposites for the direct synthesis of methyl formate via methanol photo-oxidation in this paper. The CuO/TiO2-nanotube nanocomposites with 7% CuO weight loading give a promising photocatalytic performance (over 93.3% methanol conversion and 89.4% methyl formate selectivity) at 25 °C under 365 nm UV irradiation. The turnover frequency and the apparent quantum efficiency reach up to 22.9 molmethanol gcat -1 h-1 and 57.6% over 7% CuO/TiO2-nanotube, respectively, which is mainly because of the effective minimization of the recombination of photogenerated electrons and holes by the surface ultrasmall-sized CuO particles. Furthermore, these CuO/TiO2-nanotube nanocomposites exhibit excellent durability and robust nature during the catalytic processes of the methanol photo-oxidation to methyl formate. The experimental results demonstrate the promise of the robust CuO/TiO2 nanocomposites for efficient and green production of methyl formate, which may offer guidelines for the design of efficient catalysts for selective alcohol conversion to other valuable chemicals.

Alkynyl- and phosphine-ligated quaternary Au2Ag2 clusters featuring an Alkynyl-AuAg motif for multicomponent coupling.

The coordination motif of alkynly with a metal atom is versatile and plays a pivotal role in tailoring the kernel configuration of the atomically precise metal nanoclusters. In this study, we synthesized a new mono-valent Au(I)2Ag(I)2(C10H6NO)4(Ph3P)2 alloy cluster with a very high yield of >90%, which is well characterized by a serial of technologies, e.g. UV-vis, X-ray single crystal diffraction (SCXRD) and FT-IR. The SCXRD analysis shows the alloy cluster is composed of a quadrangular Au2Ag2 kernel protected by four alkynyl and two phosphine ligands. Intriguingly, a new divergent alkyne-metal coordination model is revealed in this cluster, the alkynyl ligands selectively bind to Au and Ag atoms via σ- and π-bond configurations and adopt a VI-shaped alkynyl-M motif. It is distinct from the convergent motif observed in big clusters featuring an IV- or V-shaped alkynyl-M motif due to the steric effect. Finally, the titanium oxide-supported Au2Ag2 cluster catalysts show good catalytic performance in the multicomponent coupling reaction of alkynes, aldehydes and amines.

Capillary electrochromatography and on-line concentration.

Capillary electrochromatography (CEC) is a micro-separation technique that combines the advantages of capillary zone electrophoresis with those of high-performance liquid chromatography. Accordingly, it has attracted extensive attention over the last decade. Among the stationary phases for CEC, monolithic stationary phase has been regarded as the most suitable stationary phase for CEC because of its simple preparation, the elimination of frits, and its excellent performance. In this chapter, procedures for preparing CEC monolithic columns with an improved configuration, in which there are stationary phases at both sides of detection window and no stationary phase at detection window, are presented. The separation of acidic and basic compounds on such monolithic columns is used as an example to demonstrate CEC separation protocol. Additionally, an on-line concentration technique in CEC is presented. As a result of the coexistence of stationary phase and electric field in a CEC column, it is possible to employ chromatographic zone sharpening and field-amplified sample stacking effects simultaneously to improve CEC detection sensitivity.

Ultrafast diagnosis of the genetic-related disorders using the combined technologies of multiplex PCR and multichannel microchip electrophoresis.

For the diagnosis of unexplained male infertility a multiplex PCR for 6 markers, which are well-known as candidate genes for studying male infertility and located on the human Y-chromosome, has been designed. The multiplex PCR products have been separated on a 12 channel microchip electrophoresis system, which can analyze different samples simultaneously. By combining the technologies of multiplex PCR with multichannel microchip electrophoresis, the number of the DNA markers that can be screened simultaneously is increased to be 72 marker (12 x 6) in a single run while the electrophoresis analysis time is reduced to be only 180 s.

Multiplex PCR with multichannel microchip electrophoresis: an ultrafast analysis for genetic diseases.

A Y chromosomal polymorphic markers screening strategy using a multiplex polymerase chain reaction (PCR) and DNA microchip electrophoresis technology has recently been developed. It is a part of the human Y chromosome haplotyping system for studying Japanese population genetics and its relationship with male spermatogenic failure. This strategy is based on optimizing and modifying the primer set concentrations while keeping all other components of the PCR mixtures and conditions similar to those of a singleplex PCR. Well-balanced PCR products are obtained without changing even the DNA oligomer melting temperatures. Here, a panel of primer sets are used to amplify two groups of Y chromosome markers. The first consists of five markers and the second consists of seven markers. Both are possibly deleted in infertile men. The microchip electrophoresis technology is fast and sensitive, enables direct molecular typing of several Y chromosomal markers, and is separated by a difference of as many as six base pairs.

Separation of acidic and basic compounds in capillary electrochromatography with polymethacrylate-based monolithic columns.

Methacrylate-based monolithic columns with electroosmotic flow (EOF) or very weak EOF are prepared by in situ copolymerization in the presence of a porogen in fused-silica capillaries pretreated with a bifunctional reagent. Satisfactory separations of acidic and basic compounds on the column with EOF at either low or high pH are achieved, respectively. With sulfonic groups as dissociation functionalities, sufficient EOF mobility still remains as high as 1.74 x 10(-4) cm2 s(-1) V(-1) at low pH. Under this condition, seven acidic compounds are readily separated within 5.7 min. Moreover, at high pH, the peak shape of basic compounds is satisfactory without addition of any masking amines into running mobile phase since the secondary interaction between the basic compounds and the monolithic stationary phase are minimized at high pH. Reversed-phase mechanism for both acidic and basic compounds is observed under investigated separation conditions. In addition, possibilities of acidic and basic compound separations on a monolithic column with extremely low EOF are discussed.

Study on the multiple sites binding of human serum albumin and porphyrin by affinity capillary electrophoresis.

Equations to describe the two sites binding between proteins and ligands were deduced. According to these equations, not only the binding constants, but also the mole fraction of proteins in different forms could be obtained. Using the published data on the interaction between human serum albumin (HSA) and three kinds of porphyrin (coproporphyrin (CP), uroporphyrin I (UP) and protoporphyrin (PP)), a further study on their binding was carried out. It was concluded that there may exist two binding sites with the binding constants at the first site, proved to be the preferential one, being 6.50 x l0(5), 1.94 x 10(6) and 8.94 x 10(5), respectively. In addition, it was also demonstrated that the two binding sites of HSA with CP and UP might be of different kinds, though those of HSA and PP were of the same kind but at different positions.

Dynamic modification of poly(methyl methacrylate) chips using poly(vinyl alcohol) for glycosaminoglycan disaccharide isomer separation.

We describe a microchip electrophoresis (MCE) method for the assay of unsaturated disaccharides of chondroitin sulfates, dermatan sulfates, and hyaluronic acid (HA). Poly(vinyl alcohol) (PVA) could be irreversibly adsorbed onto poly(methyl methacrylate) (PMMA) substrates and this approach was applicable for dynamic coating. The characteristics of the PMMA surface with PVA coating were evaluated in terms of the wettability, EOF, and adsorption of 2-aminoacridone (AMAC)-labeled disaccharide. The water contact angle decreased from 73 degrees on a pristine PMMA surface to 37.5 degrees on a PVA-coated surface, indicating that the PVA coating increased hydrophilicity. EOF was reduced approximately twofold and was relatively stable. Scanning electron microscopy and fluorescence microscopy images showed that adsorption of AMAC-labeled disaccharides was dramatically suppressed. Using the PVA coating, baseline separation of two pairs of glycosaminoglycan (GAG) disaccharide isomers, DeltaDi-diS(B)/DeltaDi-diS(D) and DeltaDi-0S/DeltaDi-HA, was achieved in Tris-borate buffer within 130 s by MCE.

Enhanced electrophoretic resolution of monosulfate glycosaminoglycan disaccharide isomers on poly(methyl methacrylate) chips.

To improve the separation of monosulfate glycosaminoglycan disaccharide isomers by microchip electrophoresis, we found that addition of 1,4-dioxane (DO) dramatically improved analyte resolution, probably due to solvation effects. Methylcellulose (MC) was tested for the ability to suppress EOF and analyte adsorption to the chip. To improve analyte resolution, buffer pH, beta-CD, and DO were systematically investigated. Fast separation was achieved by increasing the electric field strength, and field-amplified sample stacking occurred with increasing buffer concentrations. Therefore, based on our findings, we describe an efficient method for the separation of monosulfate and trisulfate unsaturated disaccharides (DeltaDi-UA2S, DeltaDi-4S, DeltaDi-6S, and DeltaDi-triS) derivatized with 2-aminoacridone hydrochloride. A mixture of monosulfate disaccharide isomers (DeltaDi-UA2S, DeltaDi-4S, and DeltaDi-6S) was baseline-separated within 75 s on a poly(methyl methacrylate) chip using a mixed buffer (DO/running buffer 57:43 v:v), 0.5% MC, pH 6.81, with an E(sep) of 558 V/cm. The theoretical plate was in the range of 5 x 10(5) to 1 x 10(6) m-1.

Rapid multiplexing and simultaneous detection of human spermatogenetic failure with a 12 lane microchip electrophoresis system.

For the amplification and ultrafast separation of the genetic markers and DNA sequences that are related to human male infertility, a multiplex PCR for amplifying three DNA sequence-tagged sites (STS) located on the human Y chromosome with possible roles in the spermatogenesis process has been designed and applied followed by separation on a microchip. First, the optimum T(m) degree for the three DNA markers was optimized and determined experimentally, and the three DNA STS were amplified. These three DNA markers were then separated on a 12-lane microchip electrophoresis system, which can analyze the DNA markers on 12 channels simultaneously. The combination of these two technologies, multiplex PCR and microchip electrophoresis, allows the analysis of 36 DNA markers (12x3) within only 180 s.

Development of a capillary electrophoretic method for the analysis of low-molecular-weight amines from metal working fluid aerosols and ambient air.

A method for the determination of low-molecular-weight amines from indoor and ambient air was developed using a concentration device followed by CE coupled with indirect spectrophotometric and mass spectrometric detection that enables a reliable, rapid-response and easy-to-operate method. In indirect detection method, the selected amines were separated from interfering metal ions and amino alcohols present in the samples with an imidazole-based buffer with ethanol and EDTA as modifier. By replacing imidazole with ammonium, the final buffer was applicable for MS detection for the analytes with m/z higher than 50. A novel monolithic polymer material based on poly(methacrylate-acrylate) copolymer was developed for sampling short-chain amines from the gaseous phase. The selected analysis conditions were applied to quantify the selected short-chain amines with detection limits for the whole procedure determined between 1 and 2 microg/filter when 40 L air was sampled with 1 L/min velocity. Improved linearity and precision were obtained when the raw, time-scaled electropherogram data were transformed into mobility-scale applied for the determination of the performance characteristics of the methods. The applicability of the process of data transformation into the mobility scale was demonstrated by studying the matrix effect of water-miscible metal working fluid (stable water-oil emulsion) and of ambient air as real samples. CE-indirect UV and CE-MS, combined with the possibility of rapid air sampling, can be useful for the estimation of short-term exposure of the selected biogenic amines.

Analysis of lipoproteins by microchip electrophoresis with high speed and high reproducibility.

A method for the fast analysis of lipoproteins by microchip electrophoresis with light-emitting diode confocal fluorescence detection has been developed. Lipoproteins labeled with BODIPY FL C(5)-ceramide are found to strongly adsorb on the bare surface of a poly(methyl methacrylate) (PMMA) microchip. Sodium dodecyl sulfate and cetyltrimethylammonium bromide were therefore utilized to alter lipoproteins and channel surface to make them bear the same type of charge. After modification, the peak shape of lipoproteins was greatly improved, demonstrating lipoprotein adsorption on a PMMA chip dramatically reduced due to electrostatic repulsion. In addition, polymers were added into the running buffer to suppress electroosmotic flow and to serve as a sieving matrix. As a result, lipoprotein separation was manipulated by both electrophoretic mobilities and particle sizes. Various separation parameters including surfactant concentration, buffer pH, and polymer concentration as well as on-line concentration were investigated systematically. Under optimal conditions, two baseline separations of standard lipoproteins including high-density lipoprotein, low-density lipoprotein, and very low-density lipoprotein were achieved with different selectivity. This method affords high separation speed (within 100 s) and high reproducibility. The intraassay and interassay RSDs of lipoprotein migration times were in the range of 0.90-1.9%, indicating this method is highly reliable.

[Advances in packing capillary electrochromatographic columns].

The columns in capillary electrochromatography can be classified into three classes: open tubular, packed and monolithic columns. The monolithic columns can be divided into three categories: organic polymer-based monolithic columns made from the polymerization of acrylamide, styrene, acrylate or methacrylate monomers, silica-based monolithic columns generally prepared by using sol-gel technology, and packed particulate-based monolithic columns. Monolithic columns are receiving quite remarkable attention and developing rapidly with a focus on monolithic stationary phases prepared from synthetic polymers. The preparation methods for various types of capillary electrochromatographic columns and their advantages and disadvantages are reviewed according to 100 research articles. In particular, recent advances in the preparation methods of monolithic columns and their applications are discussed in details.

Comparison of gene expression measurements from cDNA and 60-mer oligonucleotide microarrays.

As the data generated by microarray technology continue to amass, it is necessary to compare and combine gene expression data from different platforms. To evaluate the performance of cDNA and long oligonucleotide (60-mer) arrays, we generated gene expression profiles for two cancer cell lines and compared the data between the two platforms. All 6182 unique genes represented on both platforms were included in the analysis. A limited correlation (r = 0.4708) was obtained and the difference in measurement of low-expression genes was considered to contribute to the limited correlation. Further restriction of the data set to differentially expressed genes detected in cDNA microarrays (1205 genes) and oligonucleotide arrays (1325 genes) showed modest correlations of 0.7076 and 0.6441 between the two platforms. Quantitative real-time PCR measurements of a set of 10 genes showed better correlation with oligonucleotide arrays. Our results demonstrate that there is substantial variation in the data generated from cDNA and 60-mer oligonucleotide arrays. Although general agreement was observed in measurements of differentially expressed genes, we suggest that data from different platforms could not be directly amassed.

Separation of selected humic degradation compounds by capillary electrochromatography with monolithic and packed columns.

The separation of selected lignin/humic substance (HS) degradation compounds by capillary electrochromatography (CEC) with a methacrylate-based monolithic column and a conventional column packed with 5 microm octadecyl silica (ODS) particles is presented. The effects of organic modifier concentration, pH of the mobile phase, ionic strength, applied voltage, and temperature on the separation were investigated to determine the optimal separation conditions. With the increase of pH in the mobile phase, some of analytes start to ionize and both chromatographic partition and electrophoresis can play roles in separation simultaneously. Accordingly, different selectivity from high-performance liquid chromatography (HPLC) and capillary electrophoresis (CE) could be achieved. The performances of both kinds of columns were compared. The results showed that the peaks of compounds obtained on the former column were much wider than those on the latter one, although good separation efficiency of alkylbenzenes could be readily achieved; the most probable reasons for this behavior and method to solve this problem were briefly discussed. The CEC of a soil fulvic acid with a monolithic column produced partly resolved broad bands; by means of nuclear magnetic resonance (NMR) analysis a wide range of oxygen derived aromatic substitution patterns was found with prominent contributions from phenolic and carboxylic groups.

[Micellar electrokinetic capillary chromatographic separation of amino acids derivatized with 2-(9-carbazole)-ethyl chloroformate as a pre-column labeling reagent].

2-(9-Carbazole)-ethyl chloroformate (CEOC) has been used for the pre-column derivatization of amino acids in micellar electrokinetic capillary chromatography (MEKC). Nine amino acid standards (Ser, Thr, Gly, Glu, Asp, Val, Ile, Leu, Phe) were labeled with CEOC and separated in a buffer system containing 20 mmol/L borate-30 mmol/L sodium lauryl sulfate (SDS) and 3% acetonitrile at pH 9.0 with an applied voltage of 18 kV and column temperature of 25 degrees C. The derivatives were detected at UV 214 nm. Under optimal conditions, this method offers a baseline separation for nine CEOC-derivatized amino acids within 14 min. The linearity range was from 0.025 to 0.25 mmol/L and the detection limits were from 2.15 to 2.46 micromol/L.

Characteristics of electroosmotic flow and migration of neutral solutes under stepwise gradient elution of capillary electrochromatography.

A theoretical study on the velocity of electroosmotic flow (EOF) and the retention times of neutral solutes under multiple-step gradient of capillary electrochromatography (CEC) was carried out, focusing on that with three kinds of mobile phases. Through the model computations, the detaining time of the second kind of mobile phase in the column was proved to play an important role in affecting EOF. The variation speed of EOF was shown to be determined by the differences among dead times in different steps. In addition, the prediction of the retention times of 13 aromatic compounds under gradient mode was performed with the deduced equations. A relative error below 3.3% between the calculated and experimental values was obtained, which demonstrated the rationality of the theoretical deduction. Our study could not only improve the comprehension of stepwise gradient elution, but also be of significance for the further optimization of separation conditions in the analysis of complex samples.