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Microchip Electrophoresis (microchip + electrophoresis)
Selected AbstractsMicrochip electrophoresis in low-temperature co-fired ceramics technology with contactless conductivity measurementELECTROPHORESIS, Issue 14 2009Georg Fercher Abstract In this paper a novel micromachined contactless conductivity CE device produced in low temperature co-fired ceramics (LTCC) is introduced. The application of LTCC multilayer technology provides a promising method for the contactless detection of conductive compounds because of its increased dielectric constant compared with glass or plastics. The capacitive coupling of the excitation signal into the microchannel across the LTCC substrate is improved, resulting in better detection sensitivity. Two silver electrodes located externally at opposite sides at the end of the separation channel act as detector. Impedance variations in the channel are measured without galvanic contact between electrodes and fluid. Inorganic ions are separated in less than 1,min with this novel ceramic device. The limit of detection is 10,,M for potassium. [source] Fast quantitative determination of diuretic drugs in tablets and human urine by microchip electrophoresis with native fluorescence detectionELECTROPHORESIS, Issue 16 2007Kamal Tolba Abstract Microchip electrophoresis (MCE) with native fluorescence detection has been applied for the fast quantitative analysis of pharmaceutical formulations. For this purpose, methods for fast separation and sensitive detection of the unlabeled diuretic drugs, amiloride, triamterene, bendroflumethiazide (BFMTZ), and bumetanide were developed. An epifluorescence setup was used enabling the coupling of different lasers into a commercial fluorescence microscope. The detection sensitivity of different excitation light sources was compared utilizing either a HeCd laser (,exc,=,325,nm), a frequency quadrupled Nd:YAG laser (,exc,=,266,nm), or a mercury lamp (,exc,=,330,380,nm). At optimal conditions using the HeCd laser, the drugs were separated within 15,s with LODs less than 1,,g/mL for the four compounds. A linear relationship between concentration and peak area was obtained in the concentration range of 0.05,20,,g/mL with a mean correlation coefficient of around 0.996 for all analytes. The method was successfully applied to the analysis of the respective drugs in commercial formulations and in human urine without interference from other constituents. These data show that MCE has a great potential for reliable drug analysis. [source] Microchip electrophoresis with wall-jet electrochemical detector: Influence of detection potential upon resolution of solutesELECTROPHORESIS, Issue 24 2006Martin Pumera Dr. Abstract This report studies the electrochemical response of wall-jet detector for microchip electrophoresis (µCE). It shows that in wall-jet configuration, the electrochemical detector operates in coulometric mode and that there is an influence of detection potential upon peak width and therefore upon the resolution of solutes. Upon raising the detection potential from +0.3 to +0.9,V, the resolution between model analytes, dopamine and catechol, increases from 0.63 to 2.90. The reasons for this behavior originate in wall-jet detector design and in its typically significant higher detector volume than the volume of injected sample. The conversion efficiency of the wall-jet electrochemical detection cell was found to be 97.4% for dopamine and 98.0% for catechol. The paper brings deeper understanding of operations of wall-jet electrochemical detectors for microchip devices, and it explains previously reported significantly sharper peaks when electrocatalytic electrodes (i.e., palladium and carbon nanotube) were used in µCE-electrochemistry wall-jet detector. [source] Cover Picture: Electrophoresis 8'2010ELECTROPHORESIS, Issue 8 2010Article first published online: 20 APR 2010 Issue no. 8 is a regular issue comprising19 manuscripts distributed over four distinct parts. Part I is on proteins and proteomics and has 5 articles; Part II is on nucleic acids with 5 articles on DNA purification, sequencing, genotyping and differential gene expression; Part III has 4 articles on droplet dispensing and particle separation; Part IV is on various methodologies and applications assembling 5 articles on improved sample preparation method for glycan analysis by CE, measurement of intracellular accumulation chemotherapeutic drugs in cancerous cells, metabolic monitoring in microfluidic cell arrays, microchip electrophoresis for continuous monitoring of microdialysis samples, and determination of glyphosate and its metabolites in plant materials by CE. Featured articles include: Delta2D and Proteomweaver: Performance evaluation of two different approaches for two-dimensional electrophoresis analysis. ((10.1002/elps.200900766)) A Multidimensional Electrophoretic System of Separation for the Analysis of Gene Expression (MESSAGE). ((10.1002/elps.200900624)) Particle trapping using dielectrophoretically patterned carbon nanotubes. ((10.1002/elps.200900717)) [source] Cover Picture: Electrophoresis 6'2010ELECTROPHORESIS, Issue 6 2010Article first published online: 22 MAR 2010 Issue no. 6 is a regular issue consisting of 18 full research papers. The first paper is a "Fast Track" contribution involving microchip electrophoresis of Alu elements for gender determination and inference of human ethnic origin. The remaining 17 papers are distributed over 5 different parts: Part I is on some trends in CEC; Part II deals with enantioseparations; Part III describes some novel detection approaches; Part IV has investigations on proteomics and proteins; Part V tackles the subjects related to pathogens, bacteria and the binding of aptamers to small solutes. [source] Capillary and microchip electrophoresis in microdialysis: Recent applicationsELECTROPHORESIS, Issue 1 2010Elizabeth Guihen Abstract The theme of this review is to highlight the importance of microscale electrophoretic-based separation systems in microdialysis (,D). The ability of CE and MCE to yield very rapid and highly efficient separations using just nanolitre volumes of microdialysate samples will also be discussed. Recent advances in this area will be highlighted, by illustration of some exciting new applications while the need for further innovation will be covered. The first section briefly introduces the concept of ,D sampling coupled with electrophoresis-based separation and the inherent advantages of this approach. The following section highlights some specific applications of CE separations in the detection of important biomarkers such as low-molecular-weight neurotransmitters, amino acids, and other molecules that are frequently encountered in ,D. Various detection modes in CE are outlined and some of the advantages and drawbacks thereof are discussed. The last section introduces the concepts of micro-total analysis systems and the coupling of MCE and ,D. Some of the latest innovations will be illustrated. The concluding section reflects on the future of this important chemical alliance between ,D and CE/MCE. [source] Amperometric detection in microchip electrophoresis devices: Effect of electrode material and alignment on analytical performanceELECTROPHORESIS, Issue 19 2009David J. Fischer Abstract The fabrication and evaluation of different electrode materials and electrode alignments for microchip electrophoresis with electrochemical detection is described. The influences of electrode material, both metal and carbon-based, on sensitivity and LOD were examined. In addition, the effects of working electrode alignment on analytical performance (in terms of peak shape, resolution, sensitivity, and LOD) were directly compared. Using dopamine (DA), norepinephrine, and catechol (CAT) as test analytes, it was found that pyrolyzed photoresist electrodes with end-channel alignment yielded the lowest LOD (35,nM for DA). In addition to being easier to implement, end-channel alignment also offered better analytical performance than off-channel alignment for the detection of all three analytes. In-channel electrode alignment resulted in a 3.6-fold reduction in peak skew and reduced peak tailing by a factor of 2.1 for CAT in comparison to end-channel alignment. [source] Nanostructured copolymer gels for dsDNA separation by CEELECTROPHORESIS, Issue 23 2008Fen Wan Abstract Pluronics are triblock copolymers of poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) that are able to form many different ordered nanostructures at appropriate polymer concentrations and temperatures in selective solvents. These nanostructured "gels" showed desirable criteria when used as DNA separation media, especially in microchip electrophoresis, including dynamic coating and viscosity switching. A ternary system of F127 (E99P69E99)/TBE buffer/1-butanol was selected as a model system to test the sieving performance of different nanostructures in separating dsDNA by CE. The nanostructures and their lattice constants were determined by small-angle X-ray scattering. Viscosity measurements showed the sol,gel transition phenomena. In addition to the cubic structure, successful electrophoretic separation of dsDNA in 2-D hexagonally packed cylinders was achieved. Results showed that without further optimization, ,X174 DNA,Hae III digest was well separated within 15,min in a 7-cm separation channel, by using F127/TBE/1-butanol gel with a 2-D hexagonal structure. A mechanism for DNA separations by those gels with both hydrophilic and hydrophobic domains is discussed. [source] Cover Picture: Electrophoresis 19'2008ELECTROPHORESIS, Issue 19 2008Article first published online: 28 OCT 200 This issue has a dual emphasis: "APCE 2007" and "Fundamentals and Methodologies" with the aim of providing the readers of the Journal with the latest developments in the field. APCE 2007, which was held in Singapore, December 17 , 19, 2007, is the premier forum in the Asia-Pacific countries for communicating advances in capillary- and chip-based electroseparation techniques and their applications to genomics, proteomics, and chemical and biochemical analysis. The emphasis part on APCE 2007 is a "mini proceeding" that groups 7 representative research articles, which deal with microchip electrophoresis, restriction fragment length polymorphism analysis by CE, gradient MEKC, stacking and sweeping in CE, in-line pre-concentration in CE, and food and drug analysis by CE. In addition, issue 19 has a "Fast Track" article on the principles for different modes of multiple-injection CZE and provides equations that facilitate the transfer from single-injection CZE to one or more suitable modes of multiple injection CZE. [source] Confinement effects on the morphology of photopatterned porous polymer monoliths for capillary and microchip electrophoresis of proteinsELECTROPHORESIS, Issue 14 2008Mei He Abstract We find that the morphology of porous polymer monoliths photopatterned within capillaries and microchannels is substantially influenced by the dimensions of confinement. Porous polymer monoliths were prepared by UV-initiated free-radical polymerization using either the hydrophilic or hydrophobic monomers 2-hydroxyethyl methacrylate or butyl methacrylate, cross-linker ethylene dimethacrylate and different porogenic solvents to produce bulk pore diameters between 3.2 and 0.4,µm. The extent of deformation from the bulk porous structure under confinement strongly depends on the ratio of characteristic length of the confined space to the monolith pore size. The effects are similar in cylindrical capillaries and D-shaped microfluidic channels. Bulk-like porosity is observed for a confinement dimension to pore size ratio >10, and significant deviation is observed for a ratio <5. At the extreme limit of deformation a smooth polymer layer ,300 nm thick is formed on the surface of the capillary or microchannel. Surface tension or wetting also plays a role, with greater wetting enhancing deformation of the bulk structure. The films created by extreme deformation provide a rapid and effective strategy to create robust wall coatings, with the ability to photograft various surface chemistries onto the coating. This approach is demonstrated through cationic films used for electroosmotic flow control and neutral hydrophilic coatings for electrophoresis of proteins. [source] Sensitive, label-free protein assay using 1-ethyl-3-methylimidazolium tetrafluoroborate-supported microchip electrophoresis with laser-induced fluorescence detectionELECTROPHORESIS, Issue 9 2008Yuanhong Xu Abstract Based on the dimer,monomer equilibrium movement of the fluorescent dye Pyronin Y (PY), a rapid, simple, highly sensitive, label-free method for protein detection was developed by microchip electrophoresis with LIF detection. PY formed a nonfluorescent dimer induced by the premicellar aggregation of an anionic surfactant, SDS, however, the fluorescence intensity of the system increased dramatically when proteins such as BSA, bovine hemoglobin, cytochrome c, and trypsin were added to the solution due to the transition of dimer to fluorescent monomer. Furthermore, 1-ethyl-3-methylimidazolium tetrafluoroborate (EMImBF4) instead of PBS was applied as running buffers in microchip electrophoresis. Due to the excellent properties of EMImBF4, not only nonspecific protein adsorption was more efficiently suppressed, but also approximately ten-fold higher fluorescence intensity enhancement was obtained than that using PBS. Under the optimal conditions, detection limits for BSA, bovine hemoglobin, cytochrome c, and trypsin were 1.00×10,6, 2×10,6, 7×10,7, and 5×10,7 mg/mL, respectively. Thus, without covalent modification of the protein, a protein assay method with high sensitivity was achieved on microchips. [source] Fast quantitative determination of diuretic drugs in tablets and human urine by microchip electrophoresis with native fluorescence detectionELECTROPHORESIS, Issue 16 2007Kamal Tolba Abstract Microchip electrophoresis (MCE) with native fluorescence detection has been applied for the fast quantitative analysis of pharmaceutical formulations. For this purpose, methods for fast separation and sensitive detection of the unlabeled diuretic drugs, amiloride, triamterene, bendroflumethiazide (BFMTZ), and bumetanide were developed. An epifluorescence setup was used enabling the coupling of different lasers into a commercial fluorescence microscope. The detection sensitivity of different excitation light sources was compared utilizing either a HeCd laser (,exc,=,325,nm), a frequency quadrupled Nd:YAG laser (,exc,=,266,nm), or a mercury lamp (,exc,=,330,380,nm). At optimal conditions using the HeCd laser, the drugs were separated within 15,s with LODs less than 1,,g/mL for the four compounds. A linear relationship between concentration and peak area was obtained in the concentration range of 0.05,20,,g/mL with a mean correlation coefficient of around 0.996 for all analytes. The method was successfully applied to the analysis of the respective drugs in commercial formulations and in human urine without interference from other constituents. These data show that MCE has a great potential for reliable drug analysis. [source] Integration of continuous-flow sampling with microchip electrophoresis using poly(dimethylsiloxane)-based valves in a reversibly sealed deviceELECTROPHORESIS, Issue 14 2007Michelle W. Li Abstract Here we describe a reversibly sealed microchip device that incorporates poly(dimethylsiloxane) (PDMS)-based valves for the rapid injection of analytes from a continuously flowing stream into a channel network for analysis with microchip electrophoresis. The microchip was reversibly sealed to a PDMS-coated glass substrate and microbore tubing was used for the introduction of gas and fluids to the microchip device. Two pneumatic valves were incorporated into the design and actuated on the order of hundreds of milliseconds, allowing analyte from a continuously flowing sampling stream to be injected into an electrophoresis separation channel. The device was characterized in terms of the valve actuation time and pushback voltage. It was also found that the addition of sodium dodecyl sulfate (SDS) to the buffer system greatly increased the reproducibility of the injection scheme and enabled the analysis of amino acids derivatized with naphthalene-2,3-dicarboxaldehyde/cyanide. Results from continuous injections of a 0.39,nL fluorescein plug into the optimized system showed that the injection process was reproducible (RSD of 0.7%, n,=,10). Studies also showed that the device was capable of monitoring off-chip changes in concentration with a device lag time of 90,s. Finally, the ability of the device to rapidly monitor on-chip concentration changes was demonstrated by continually sampling from an analyte plug that was derivatized upstream from the electrophoresis/continuous flow interface. A reversibly sealed device of this type will be useful for the continuous monitoring and analysis of processes that occur either off-chip (such as microdialysis sampling) or on-chip from other integrated functions. [source] Enhanced electrophoretic resolution of monosulfate glycosaminoglycan disaccharide isomers on poly(methyl methacrylate) chipsELECTROPHORESIS, Issue 3 2007Yong Zhang Abstract 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, ,-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 (,Di-UA2S, ,Di-4S, ,Di-6S, and ,Di-triS) derivatized with 2-aminoacridone hydrochloride. A mixture of monosulfate disaccharide isomers (,Di-UA2S, ,Di-4S, and ,Di-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 Esep of 558,V/cm. The theoretical plate was in the range of 5×105 to 1×106,m,1. [source] Microchip electrophoresis with wall-jet electrochemical detector: Influence of detection potential upon resolution of solutesELECTROPHORESIS, Issue 24 2006Martin Pumera Dr. Abstract This report studies the electrochemical response of wall-jet detector for microchip electrophoresis (µCE). It shows that in wall-jet configuration, the electrochemical detector operates in coulometric mode and that there is an influence of detection potential upon peak width and therefore upon the resolution of solutes. Upon raising the detection potential from +0.3 to +0.9,V, the resolution between model analytes, dopamine and catechol, increases from 0.63 to 2.90. The reasons for this behavior originate in wall-jet detector design and in its typically significant higher detector volume than the volume of injected sample. The conversion efficiency of the wall-jet electrochemical detection cell was found to be 97.4% for dopamine and 98.0% for catechol. The paper brings deeper understanding of operations of wall-jet electrochemical detectors for microchip devices, and it explains previously reported significantly sharper peaks when electrocatalytic electrodes (i.e., palladium and carbon nanotube) were used in µCE-electrochemistry wall-jet detector. [source] A robust cross-linked polyacrylamide coating for microchip electrophoresis of dsDNA fragmentsELECTROPHORESIS, Issue 19 2006Joann J. Lu Abstract Surface derivatization plays an important role in microchip electrophoresis. It not only enhances the resolution, but also improves the reproducibility. So far, the most popularly used derivatization method for glass microchannels is to covalently attach a layer of linear polyacrylamide,(LPA) to the channel surfaces. However, LPA coating has two problems: incomplete coverage and limited lifetime. To address these issues, we have recently developed a cross-linked polyacrylamide,(CPA) derivatization protocol and demonstrated it for high-resolution protein separations by CIEF, CGE, and CZE. In this report, we used this protocol to coat microchip channels and exhibited the reliability and robustness of CPA coating for microchip electrophoresis of DNA molecules. dsDNA fragments were used as our test samples. High resolutions were obtained for fragments ranging from 100,bp to 10,kpb. After more than 800,runs, the CPA-coated microchannels still performed well and comparable resolutions were maintained throughout these runs. [source] Modified Hadamard transform microchip electrophoresisELECTROPHORESIS, Issue 16 2005Renato Guchardi Abstract Sensitivity is a crucial point in the development applications for medicine or environmental samples in which the analytes are present in the nanomolar range. Besides further technical development of detection systems, the multiplex sample injection technique can be applied for enhancing the signal-to-noise ratio. Hadamard transform is easily applied to microchip electrophoresis due to the fact that sample injection is generally achieved through cross, double-tee, or tee injector structures. This paper reports the first demonstration of a modified Hadamard transform electrophoresis on a microchip by using an amperometric detector. Contrary to the previous Hadamard applications, the resolution (number of points per unit of time) of electropherograms obtained is independent of the number of injections. [source] Analysis of chicken and turkey ovalbumins by microchip electrophoresis combined with exoglycosidase digestionELECTROPHORESIS, Issue 18 2003Xiuli Mao Abstract The polypeptide and carbohydrate patterns of two glycoproteins, chicken ovalbumin (CO) and turkey ovalbumin (TO), were analyzed by microchip electrophoresis (ME), following digestion with proteases and exoglycosidases. Glycopeptides derived from ovalbumin were obtained by digestion with Pronase, followed by dialysis, and then separated by ME. Using CO as model, the method was developed to deduce the structure of glycans from glycoproteins by comparing the electropherograms of glycopeptides with and without digestion of exolycosidases. Applying the same approach, the structure of oligosaccharides linked to TO was determined. TO was found to contain high-mannose type oligosaccharides and oligosaccharides with terminal N -acetylglucosamine residues. The complete primary analysis of CO and TO by ME described in this paper provides a basis for an analysis of glycoproteins with an integrated microfluidic chip. [source] Quantification of D -Asp and D -Glu in rat brain and human cerebrospinal fluid by microchip electrophoresisJOURNAL OF SEPARATION SCIENCE, JSS, Issue 17 2009Yong Huang Abstract A microchip electrophoresis (MCE) method with LIF detection was presented for quantification of D -aspartic acid (D -Asp) and D -glutamate (D -Glu) in biological samples. D -Asp and D -Glu were determined after precolumn derivatization with FITC. The chiral separation was performed on a glass/PDMS hybrid microfluidic chip using ,-CD as chiral selector in the running buffer. High sensitive detection was obtained by the LIF detection. The LODs (S/N = 3) for D -Asp and D -Glu were 6.0×10,8 and 4.0×10,8 M, respectively. Using this method, the levels of D -Asp and D -Glu in rat brain and human cerebrospinal fluid (CSF) were determined. [source] Quantitative evaluation of the interaction between netropsin and double stranded oligodeoxynucleotides by microfabricated capillary array electrophoresisJOURNAL OF SEPARATION SCIENCE, JSS, Issue 10 2007Zheng Shen Abstract Microfabricated capillary array electrophoresis (,-CAE) was applied to study the interaction between minor groove binder netropsin and a non-selfcomplementary 12 mer double stranded oligodeoxynucleotide: d(CCCCTATACCGC)·d(GCGGTATAGGGG). ESI-MS was used to provide an independent verification of the microchip electrophoresis derived data. Simultaneous parallel quantitative assay of multiple samples was performed in a single run (<50 s) on the self-developed ,-CAE device. The binding constant and stoichiometry calculated from Scatchard plot were (2.88 ± 0.23)×105 M,1 and 1:1, respectively. The values showed a good quantitative agreement with the results determined by ESI-MS and those using other methods reported in the literature. [source] Triple light chain antibodies: Factors that influence its formation in cell cultureBIOTECHNOLOGY & BIOENGINEERING, Issue 4 2010Natalia Gomez Abstract THIOMABs are recombinant antibodies engineered with reactive cysteines, which can be covalently conjugated to drugs of interest to generate targeted therapeutics. During the analysis of THIOMABs secreted by stably transfected Chinese Hamster Ovary (CHO) cells, we discovered the existence of a new species,Triple Light Chain Antibody (3LC). This 3LC species is the product of a disulfide bond formed between an extra light chain and one of the engineered cysteines on the THIOMAB. We characterized the 3LC by size exclusion chromatography, mass spectrometry, and microchip electrophoresis. We also investigated the potential causes of 3LC formation during cell culture, focusing on the effects of free light chain (LC) polypeptide concentration, THIOMAB amino acid sequence, and glutathione (GSH) production. In studies covering 12 THIOMABs produced by 66 stable cell lines, increased free LC polypeptide expression,evaluated as the ratio of mRNA encoding for LC to the mRNA encoding for heavy chain (HC),correlated with increased 3LC levels. The amino acid sequence of the THIOMAB molecule also impacted its susceptibility to 3LC formation: hydrophilic LC polypeptides showed elevated 3LC levels. Finally, increased GSH production,evaluated as the ratio of the cell-specific production rate of GSH (qGSH) to the cell-specific production rate of THIOMAB (qp),corresponded to decreased 3LC levels. In time-lapse studies, changes in extracellular 3LC levels during cell culture corresponded to changes in mRNA LC/HC ratio and qGSH/qp ratio. In summary, we found that cell lines with low mRNA LC/HC ratio and high qGSH/qp ratio yielded the lowest levels of 3LC. These findings provide us with factors to consider in selecting a cell line to produce THIOMABs with minimal levels of the 3LC impurity. Biotechnol. Bioeng. 2010. 105: 748,760. © 2009 Wiley Periodicals, Inc. [source] | ||||||||||