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Selected AbstractsMulti-walled carbon nanotube composites with polyacrylate prepared for open-tubular capillary electrochromatographyELECTROPHORESIS, Issue 19 2010Jian-Lian Chen Abstract A new phase containing immobilized carbon nanotubes (CNTs) was synthesized by in situ polymerization of acid-treated multi-walled CNTs using butylmethacrylate (BMA) as the monomer and ethylene dimethacrylate as the crosslinker on a silanized capillary, forming a porous-layered open-tubular column for CEC. Incorporation of CNT nanomaterials into a polymer matrix could increase the phase ratio and take advantage of the easy preparation of an OT-CEC column. The completed BMA-CNT column was characterized by SEM, ATR-IR, and EOF measurements, varying the pH and the added volume organic modifier. In the multi-walled CNTs structure, carboxylate groups were the major ionizable ligands on the phase surface exerting the EOF having electroosmotic mobility, 4.0×104,cm2,V,1,S,1, in the phosphate buffer at pH 2.8 and RSD values (n=5), 3.2, 4.1, and 4.3%, for three replicate capillaries at pH 7.6. Application of the BMA-CNT column in CEC separations of various samples, including nucleobases, nucleosides, flavonoids, and phenolic acids, proved satisfactory upon optimization of the running buffers. Their optima were found in the borate buffers at pH 9.0/50,mM, pH 9.5/10,mM/50% v/v ACN, and pH 9.5/30,mM/10% v/v methanol, respectively. The separations could also be used to assess the relative contributions of electrophoresis and chromatography to the CEC mechanism by calculating the corresponding velocity and retention factors. Discussions about interactions between the probe solutes and the bonded phase included the ,,, interactions, electrostatic repulsion, and hydrogen bonding. Furthermore, a reversed-phase mode was discovered to be involved in the chromatographic retention. [source] Dynamic coating of SU-8 microfluidic chips with phospholipid disksELECTROPHORESIS, Issue 15 2010Tiina Sikanen Abstract In this work, PEG-stabilized phosphatidylcholine lipid aggregates (disks), mimicking mammalian cell membranes, were introduced as a new biofouling resistant coating for SU-8 polymer microchannels. A rapid and simple method was developed for immobilization of PEGylated phosphatidylcholine disks in microchannels. Microfluidic chips made from SU-8, PDMS, or glass were dynamically coated with the PEGylated disks followed by characterization of their surface chemistry before and after coating. On the basis of the observed changes in EOF and nonspecific protein adsorption, the affinity of the PEGylated disks was shown to be particularly strong toward SU-8. The PEG-lipid coating enabled permanent change in EOF in SU-8 microchannels with an initial value of 4.5×10,8,m2,V,1,s,1, decreasing to 2.1×10,8,m2,V,1,s,1 (immediately after modification), and, eventually, to 1.5×10,8,m2,V,1,s,1 (7 days after modification) for 9,mM sodium borate (pH 10.5) as BGE. As determined by the Wilhelmy plate measurements and microchip-CE analysis of BSA, the PEG-lipid coating also enabled efficient biofouling shield against protein adsorption, similar to that of low amounts of SDS (3.5,mM) or Tween-20 (80,,M) as buffer additives. These results suggest that dynamically attached PEG-lipid aggregates provide stable, biomimicking surface modification that efficiently reduces biofouling on SU-8. [source] Synthesis of poly(N, N -dimethylacrylamide)- block -poly(ethylene oxide)- block -poly(N, N -dimethylacrylamide) and its application for separation of proteins by capillary zone electrophoresisELECTROPHORESIS, Issue 10 2010Jing Xu Abstract A series of well-defined triblock copolymers, poly(N, N -dimethylacrylamide)- block -poly(ethylene oxide)- block -poly(N, N -dimethylacrylamide) (PDMA- b -PEO- b -PDMA) synthesized by atom transfer radical polymerization, were used as physical coatings for protein separation. A comparative study of EOF showed that the triblock copolymer presented good capillary coating ability and EOF efficient suppression. The effects of the Mr of PDMA block in PDMA- b -PEO- b -PDMA triblock copolymer and buffer pH on the separation of basic protein for CE were investigated. Moreover, the influence of the copolymer structure on separation of basic protein was studied by comparing the performance of PDMA- b -PEO- b -PDMA triblock copolymer with PEO- b -PDMA diblock copolymer. Furthermore, the triblock copolymer coating showed higher separation efficiency and better migration time repeatability than fused-silica capillary when used in protein mixture separation and milk powder samples separation, respectively. The results demonstrated that the triblock copolymer coatings would have a wide application in the field of protein separation. [source] Capillary modified with covalently attached coating for enhanced CE separation of biopolymersELECTROPHORESIS, Issue 4 2010Yuan He Abstract ,-Gluconolactone was covalently coupled with aminopropyl-derivatized capillary, creating hydrophilic brushes on the inner wall of the capillary. The hydrophilic coating provided suppression of EOF and minimized protein adsorption, resulting in the separation of basic proteins and DNA with efficiencies up to 450,000,plates/m. The intra- and inter-day repeatabilities of the coating referring to the migration times of the four tested proteins were satisfactory with RSD of no more than 1.1 and 1.8% (n=5), respectively. Two hundred consecutive runs were performed with negligible change in migration times and efficiency. [source] Microstructure of microemulsion in MEEKCELECTROPHORESIS, Issue 4 2010Yuhua Cao Abstract The influences of the composition of microemulsion on the microstructure including dimensions and , potentials of microdroplets were measured in details. The average dynamic dimension of microdroplets was measured by dynamic laser light scattering, and , potential was determined to characterize average surface charge density of microdroplets. The experiment results showed that increase of the amount of surfactant resulted in decrease of microdroplet size but almost invariant , potential, which would enlarge migration time of the microdroplet in MEEKC. With increment of cosurfactant concentration, the microdroplet size had an increasing trend, whereas the , potential decreased. Thus, observed migration velocity of microdroplets increased, which made the separation window in MEEKC shortened. Neither dimension nor , potential of microdroplets changed by varying both the type and the amount of the oil phase. Adding organic solvent as modifier to microemulsion did not change the microdroplet size, but lowered , potential. The migration time of microdroplet still became larger, since EOF slowed down owing to organic solvent in capillary. So, besides increment of surfactant concentration, organic additive could also enlarge the separation window. Increase of cosurfactant concentration was beneficial for separation efficiency thanks to the looser structure of swollen microdroplet, and the peak sharpening might compensate for the resolution and peak capacity owing to a narrow separation window. Except the oil phase, tuning the composition of microemulsion would change the microstructure, eventually could be exploited to optimize the resolution and save analysis time in MEEKC. [source] Continuous intact cell detection and viability determination by CE with dual-wavelength detectionELECTROPHORESIS, Issue 2 2010Xiaomin Ren Abstract We introduce here a method for continuous intact cell detection and viability determination of individual trypan blue stained cells by CE with ultraviolet,visible dual-wavelength detection. To avoid cell aggregation or damage during electrophoresis, cells after staining were fixed with 4% formaldehyde and were continuously introduced into the capillary by EOF. The absorbance of a cell at 590,nm was used to determine its viability. An absorbance of two milli-absorbance unit at 590,nm was the clear cut-off point for living and dead Hela cells in our experiments. Good viability correlation between the conventional trypan blue staining assay and our established CE method (correlation coefficient, R2=0.9623) was demonstrated by analysis of cell mixtures with varying proportions of living and dead cells. The CE method was also used to analyze the cytotoxicity of methylmercury, and the results were in good agreement with the trypan blue staining assay and 3-(4,5-dimethyl-2-thiazyl)-2,5-diphenyl-2H-tetrazolium bromide methods. Compared with the 3-(4,5-dimethyl-2-thiazyl)-2,5-diphenyl-2H-tetrazolium bromide method, our established CE method can be easily automated to report cell viability based on the state of individual cells. Tedious manual cell counting and human error due to investigator bias can be avoided by using this method. [source] Online CIEF-ESI-MS in glycerol,water media with a view to hydrophobic protein applicationsELECTROPHORESIS, Issue 23 2009Meriem Mokaddem Abstract A new online coupling of CIEF with ESI-MS has been developed in glycerol,water media. This improved protocol provides: (i) the electric continuity during the whole analysis by a discontinuous filling of the capillary with 60:40 (cm/cm) catholyte/proteins,ampholyte mixture; (ii) the use of an anticonvective medium, i.e. 30:70 glycerol/water, v/v, compatible with MS detection and as an aid to hydrophobic protein solubilization and (iii) the use of unmodified bare fused-silica capillaries, as the glycerol/water medium strongly reduces EOF. Focusing was performed in positive polarity and cathodic mobilization was achieved by both voltage and pressure application. The setup was optimized with respect to analysis time, sensitivity and precision on pI determination. The optimized anolyte and catholyte were composed of 50,mM formic acid/1,mM glutamic acid (pH 2.35) and 100,mM NH3/1,mM lysine (pH 10.6), respectively. The effects of ampholyte concentration, focusing time and ESI parameters were presented for model proteins and discussed. This new integrated protocol should be an easy and effective additional tool in the field of proteome analysis, providing a means for the characterization of a large number of hydrophilic and hydrophobic proteins. [source] Etched succinate-functionalized silica hydride stationary phase for open-tubular CECELECTROPHORESIS, Issue 22 2009Jian-Lian Chen Abstract An open-tubular (OT) CEC column was designed to anchor ionizable succinate-functionalized ligands onto a silica hydride-based stationary phase through surface etching, silanization, and hydrosilation reactions beginning from a bare fused-silica tube. The modified columns that were produced in each step were monitored by analysis of the effect of performance of EOF on the changes of pH values, concentrations, and the amount of ACN added in the running buffers. By tracking the EOF patterns between columns, the author determined that the surface composition of the final product column was a combination of silanols, silica hydrides, and succinate ligands. Furthermore, lower loading volumes of the succinate ligands prepared for the hydrosilation reaction served to complete the mixed-mode OT-CEC columns, and subsequently to carry out the separation of six phenyl alcohols. Studies on the elution order of these alcohols identified the presence of chromatographic interactions in addition to electrophoresis. Based on the employment of a solvation parameter model, these interactions likely included dispersion interactions, dipole-type interactions, and interactions arising through the polarizable electrons in the solute. The optimum buffer conditions for CEC separations of phenyl alcohols, carbonyl-substituted phenols, and a mixture of nucleosides and thymine were a phosphate buffer (50,mM, pH 10.51), a borate buffer (50,mM, pH 8.62), and a borate buffer (50,mM, pH 9.50), respectively. Overall, the hydride-based stationary phases with ionizable ligands were successfully applied to the OT-CEC separations, and these results confidently propose an ideal route to the synthesis of novel OT-CEC columns. [source] Use of coated capillaries for the electrophoretic separation of stereoisomers of a growth hormone secretagogueELECTROPHORESIS, Issue 21 2009Reine Nehmé Abstract The diastereoisomeric separation of peptidomimetics of hexarelin, a strong growth hormone secretagogue, in CE has been studied. Highly sulfated-,-CD was found to be an appropriate selector for the separation of the stereoisomers. However, non-repeatable analyses were obtained on bare fused silica capillary due to the progressive adsorption of the analytes on the capillary wall. Two types of polyelectrolyte coating agents were tested to prevent this phenomenon. Coating with neutral polyethylene oxide was found to be efficient but resulted in a very long analysis time (about 40,min). Coating with cationic poly(diallyldimethylammonium) chloride was found both to prevent analyte adsorption, reduce analysis time and alter separation selectivity. EOF measurement revealed that the highly sulfated-,-CDs were strongly adsorbed on the poly(diallyldimethylammonium) chloride coating surface yielding a stable strong cathodic EOF, which considerably reduced analysis time (about 12,min). Very good repeatability of analysis was obtained (RSDmigration time<1%). [source] Cover Picture: Electrophoresis 20'2009ELECTROPHORESIS, Issue 20 2009Article first published online: 27 OCT 200 Issue no. 20 is a regular issue with an Emphasis on "Fundamentals and Methodologies". The bulk of this issue (13 articles) is on fundamentals and methodologies covering various topics, e.g. EOF, affinity CE, structural analysis of glycosphingolipids by CE-ESI-MS, on-line concentration, monolithic columns, etc. The other 6 articles are on protein separation and proteomics. Selected articles are: Micropump based on electroosmosis of the second kind ((10.1002/elps.200900271)) A splicing model-based DNA computing approach on microfluidic chip ((10.1002/elps.200900323)) Proteomic Characterization of Plasma-derived Clotting Factor VIII , von Willebrand Factor Concentrates ((10.1002/elps.200900270)) [source] Preparation and evaluation of the highly cross-linked poly(1-hexadecane-co-trimethylolpropane trimethacrylate) monolithic column for capillary electrochromatographyELECTROPHORESIS, Issue 20 2009Minghua Lu Abstract In this paper, a novel highly cross-linked porous monolithic stationary phase having a long alkyl chain ligand (C16) was introduced and evaluated in CEC. The monolithic stationary phase was prepared by in situ copolymerization of 1-hexadecene, trimethylolpropane trimethacrylate, and 2-acrylamido-2-methyl-1-propanesulfonic acid (AMPS) in the presence of ternary porogenic solvent (cyclohexanol/1,4-butanediol/water). In preparing monoliths, the ternary cross-linker trimethylolpropane trimethacrylate was usually applied to preparing molecularly imprinted polymers or molecularly imprinted solid-phase extraction, instead of binary cross-linker ethylene dimethacrylate. 1-Hexadecene was introduced to provide the non-polar sites (C16) for chromatographic retention, while AMPS was used to generate the EOF for transporting the mobile phase through the monolithic capillary. Monolithic columns were prepared by optimizing proportion of porogenic solvent and AMPS content in the polymerization solution as well as the cross-linkers. The monolithic stationary phases could generate a strong and stable EOF in various pH values and exhibit an RP-chromatographic behavior for neutral compounds. For charged compounds, the separation was mainly based on the association of hydrophobic, electrostatic and electrophoretic interaction. [source] Multiple-point electrochemical detection for a dual-channel hybrid PDMS-glass microchip electrophoresis deviceELECTROPHORESIS, Issue 19 2009Mario Castaño-Álvarez Abstract A new PDMS-based dual-channel MCE with multiple-point amperometric detection has been evaluated. Electrophoresis has been optimised in a single-channel device. Pretreatment with 0.1,M NaOH is very important for increasing and stabilising the EOF. The precision is adequate for a day's work in terms of both peak current and migration time. The RSD of the peak current for five successive signals was 1.9, 2.4 and 3.1% for dopamine, p- aminophenol and hydroquinone. RSD for the migration time was always less than 1.3%, which demonstrates the stability of the EOF and the possibility of running multiple experiments in the same microchip. The adequate inter-microchip precision as well as the rapid and simple manufacturing procedure indicates the disposable nature of the PDMS microchips. A dual-channel device with very simple multiple-point amperometric detection is proposed here. Elasticity of the PDMS allows removing the polymer slightly and aligning gold wires working electrodes. Injection can be performed from each of the sample reservoirs or from both simultaneously. The distance between the separation channels is critical for obtaining adequate signals as well as the introduction of a high-voltage electrode in the buffer reservoir. Simultaneous measurement of the same analytes in both channels is possible by applying the same potential. Moreover, since no cross-separation is produced, different analytes or samples can be simultaneously measured. [source] Enhancement of electrokinetically driven microfluidic T-mixer using frequency modulated electric field and channel geometry effectsELECTROPHORESIS, Issue 18 2009Deguang Yan Abstract This study reports improved electrokinetically driven microfluidic T-mixers to enhance their mixing efficiency. Enhancement of electrokinetic microfluidic T-mixers is achieved using (i) an active approach of utilizing a pulsating EOF, and (ii) a passive approach of using the channel geometry effect with patterned blocks. PDMS-based electrokinetic T-mixers of different designs were fabricated. Experimental measurements were carried out using Rhodamine B to examine the mixing performance and the micro-particle image velocimetry technique to characterize the electrokinetic flow velocity field. Scaling analysis provides an effective frequency range of applied AC electric field. Results show that for a T-mixer of 10,mm mixing length, utilizing frequency modulated electric field and channel geometry effects can increase the mixing efficiency from 50 to 90%. In addition, numerical simulations were performed to analyze the mixing process in the electrokinetic T-mixers with various designs. The simulation results were compared with the experimental data, and reasonable agreement was found. [source] Co-electroosmotic capillary electrophoresis of basic proteins with 1-alkyl-3-methylimidazolium tetrafluoroborate ionic liquids as non-covalent coating agents of the fused-silica capillary and additives of the electrolyte solutionELECTROPHORESIS, Issue 11 2009Danilo Corradini Abstract The paper reports the results of a study carried out to evaluate the use of three 1-alkyl-3-methylimidazolium-based ionic liquids as non-covalent coating agents for bare fused-silica capillaries and additives of the electrolyte solutions (BGE) for CE of basic proteins in the co-EOF separation mode. The three ionic liquids are differentiated from each other by the length of the alkyl group on the imidazolium cation, consisting of either an ethyl, butyl or octyl substituent, whereas tetrafluoroborate is the common anionic component of the ionic liquids. Coating the capillary with the ionic liquid resulted in improved peak shape and protein separation, while the EOF was maintained cathodic. This indicates that each ionic liquid is effective at masking the protein interaction sites on the inner surface of the capillary, also when its adsorption onto the capillary wall has not completely neutralized all the negative charges arising from the ionization of the silanol groups and the ionic liquid is not incorporated into the BGE employed for separation. Using the coated capillaries with BGE containing the ionic liquid employed for the coating, at concentration low enough to maintaining the EOF cathodic, both peak shape and protein separation varied to different extents, based on the particular ionic liquid used and its concentration. Fast and efficient separation of the model basic protein mixture in co-electroosmotic CE is obtained with the 1-butyl-3-methylimidazolium tetrafluoroborate coated capillary and 100,mM acetate buffer (pH 4.0) containing 4.4,mM 1-butyl-3-methylimidazolium tetrafluoroborate as the BGE. [source] Improved simultaneous enantioseparation of ,-agonists in CE using ,-CD and ionic liquidsELECTROPHORESIS, Issue 6 2009Lu Huang Abstract In this study, approaches to improve chiral resolutions in simultaneous enantioseparation of ,-agonists by CE via a CD inclusion complexation modified with ionic liquids (ILs) are described. Different types of ILs, including tetraalkylammonium-based ILs, alkylimidazolium-based ILs and alkylpyridinium-based ILs, were examined and compared for controlling the EOF in order to improve resolutions of ,-agonists enantiomers. In this regard, tetraalkylammonium-based ILs were more effective because they could be used at much higher concentrations than other types of ILs. N -octylpyridinium hexafluorophosphate gave poor resolutions of ,-agonists enantiomers. In addition, when different ILs were mixed to use, they would present particular properties of their own. Moreover, the presence of ILs was essential in the chiral separations of (±) salbutamol, (±) cimaterol and (±) formoterol, which were reportedly not enantioseparated by using the buffer electrolytes containing only ,-CD as a chiral selector. [source] Novel negatively charged tentacle-type polymer coating for on-line preconcentration of proteins in CEELECTROPHORESIS, Issue 4 2009Liang Xu Abstract A novel negatively charged tentacle-type polymer-coated capillary column was fabricated and applied for on-line extraction and preconcentration of proteins. The polymer coating was prepared by glycidyl-methacrylate graft polymerization in a silanized capillary column and the following sulfonic acid group functionalization. It had high surface area and offered high phase ratio for protein adsorption. In addition, the polymer-coated capillary column provided more stable EOF than a bare uncoated capillary. These features of the polymer coating facilitated the extraction of proteins through electrostatic interactions. This was used to extract proteins. The extracted analytes were then desorbed and focused by EOF in the direction opposite to the sample injection flow for subsequent CE. With this procedure, over 1500-fold sensitivity enhancement was realized for myoglobin (MB) as compared with a normal capillary zone electrophoresis. By comparison of the peak areas of the enriched protein, it was found that the polymer-coated column could capture proteins about 30 times more than the uncoated column. In addition, the separation of a protein mixture containing 0.4,,g/mL of MB and 0.4,,g/mL of insulin was demonstrated by the on-line preconcentration and electrophoretic separation with the polymer-coated column. [source] Realistic simulations of combined DNA electrophoretic flow and EOF in nano-fluidic devicesELECTROPHORESIS, Issue 24 2008Duc Duong-Hong Abstract We present a three-dimensional dissipative particle dynamics model of DNA electrophoretic flow that captures both DNA stochastic motion and hydrodynamics without requiring expensive molecular dynamics calculations. This model enables us to efficiently and simultaneously simulate DNA electrophoretic flow and local EOF (generated by counterions near the DNA backbone), in mesoscale (,,m) fluidic devices. Our model is used to study the electrophoretic separation of long DNA chains under entropic trapping conditions [Han and Craighead, Science 2000, 288, 1026,1029]. Our simulation results are in good agreement with experimental data for realistic geometries (tapered walls) and reveal that wall tapering in entropic traps has a profound impact in the DNA trapping behavior, an effect which was largely ignored in previous modeling. [source] Underivatized cyclic olefin copolymer as substrate material and stationary phase for capillary and microchip electrochromatographyELECTROPHORESIS, Issue 15 2008Omar Gustafsson Abstract We report, for the first time, the use of underivatized cyclic olefin copolymer (COC, more specifically: Topas) as the substrate material and the stationary phase for capillary and microchip electrochromatography (CEC), and demonstrate chromatographic separations without the need of coating procedures. Electroosmotic mobility measurements in a 25,,m id Topas capillary showed a significant cathodic EOF that is pH-dependent. The magnitude of the electroosmotic mobility is comparable to that found in glass substrates and other polymeric materials. Open-tubular CEC was employed to baseline-separate three neutral compounds in an underivatized Topas capillary with plate heights ranging from 5.3 to 12.7,,m. The analytes were detected using UV absorbance at 254,nm, thus taking advantage of the optical transparency of Topas at short wavelengths. The fabrication of a Topas-based electrochromatography microchip by nanoimprint lithography is also presented. The microchip has an array of pillars in the separation column to increase the surface area. The smallest features that were successfully imprinted were around 2,,m wide and 5,,m high. No plasma treatment was used during the bonding, thus keeping the surface properties of the native material. An RP microchip electrochromatography separation of three fluorescently labeled amines is demonstrated on the underivatized microchip with plate heights ranging from 3.4 to 22,,m. [source] Cover Picture: Electrophoresis 14/2008ELECTROPHORESIS, Issue 14 2008Article first published online: 23 JUL 200 Issue 14 is a regular issue including an Emphasis Section offering a series of 9 papers on ,Microfluidics and Miniaturization". These 9 research papers report on various topics including studying single DNA molecules, selective release of intracellular molecules on the single cell level, isoelectric focusing of proteins in an ordered micropillar array, sample stream focusing in a microchip, integrated microfluidic system for sensing infectious viral disease, EOF in annulus and rectangular channels, confinement effects on monolith morphology, accumulation and filtering of nanoparticles in microchannels, and carbon nanotubes disposable detectors. [source] Capillary electrophoresis-time of flight-mass spectrometry using noncovalently bilayer-coated capillaries for the analysis of amino acids in human urineELECTROPHORESIS, Issue 12 2008Rawi Ramautar Abstract A capillary electrophoresis-time of flight-mass spectrometry (CE-TOF-MS) method for the analysis of amino acids in human urine was developed. Capillaries noncovalently coated with a bilayer of Polybrene (PB) and poly(vinyl sulfonate) (PVS) provided a considerable EOF at low pH, thus facilitating the fast separation of amino acids using a BGE of 1,M formic acid (pH,1.8). The PB,PVS coating proved to be very consistent yielding stable CE-MS patterns of amino acids in urine with favorable migration time repeatability (RSDs <2%). The relatively low sample loading capacity of CE was circumvented by an in-capillary preconcentration step based on pH-mediated stacking allowing 100-nL sample injection (i.e. ca. 4% of capillary volume). As a result, LODs for amino acids were down to 20,nM while achieving satisfactory separation efficiencies. Preliminary validation of the method with urine samples showed good linear responses for the amino acids (R2 >0.99), and RSDs for peak areas were <10%. Special attention was paid to the influence of matrix effects on the quantification of amino acids. The magnitude of ion suppression by the matrix was similar for different urine samples. The CE-TOF-MS method was used for the analysis of urine samples of patients with urinary tract infection (UTI). Concentrations of a subset of amino acids were determined and compared with concentrations in urine of healthy controls. Furthermore, partial least squares,discriminant analysis (PLS,DA) of the CE-TOF-MS dataset in the 50,450,m/z region showed a distinctive grouping of the UTI samples and the control samples. Examination of score and loadings plot revealed a number of compounds, including phenylalanine, to be responsible for grouping of the samples. Thus, the CE-TOF-MS method shows good potential for the screening of body fluids based on the analysis of endogenous low-molecular weight metabolites such as amino acids and related compounds. [source] Macrocyclic polyamine-modified poly(glycidyl methacrylate- co -ethylene dimethacrylate) monolith for capillary electrochromatographyELECTROPHORESIS, Issue 11 2008Yun Tian Abstract 1,4,10,13,16-Pentaazatricycloheneicosane-9,17-dione (macrocyclic polyamine)-modified polymer-based monolithic column for CEC was prepared by ring opening reaction of epoxide groups from poly(glycidyl methacrylate- co -ethylene dimethacrylate) (GMA- co -EDMA) monolith with macrocyclic polyamine. Conditions such as reaction time and concentration of macrocyclic polyamine for the modification reaction were optimized to generate substantial EOF and enough chromatographic interactions. Anodic EOF was observed in the pH range of 2.0,8.0 studied due to the protonation of macrcyclic polyamine at the surface of the monolith. Morphology of the monolithic column was examined by SEM and the incorporation of macrocyclic polyamine to the poly(GMA- co -EDMA) monolith was characterized by infrared (IR) spectra. Successful separation of inorganic anions, isomeric benzenediols, and benzoic acid derivatives on the monolithic column was achieved for CEC. In addition to hydrophobic interaction, hydrogen bonding and electrostatic interaction played a significant role in the separation process. [source] Analyses of gibberellins in coconut (Cocos nucifera L.) water by partial filling-micellar electrokinetic chromatography-mass spectrometry with reversal of electroosmotic flowELECTROPHORESIS, Issue 10 2008Liya Ge Abstract In this paper, we present the results of simultaneous screening of eight gibberellins (GAs) in coconut (Cocos nucifera L.) water by MEKC directly coupled to ESI-MS detection. During the development of MEKC-MS, partial filling (PF) was used to prevent the micelles from reaching the mass spectrometer as this is detrimental to the MS signal, and a cationic surfactant, cetyltrimethylammonium hydroxide, was added to the electrolyte to reverse the EOF. On the basis of the resolution of the neighboring peaks, different parameters (i.e., the pH and concentration of buffer, surfactant concentrations, length of the injected micellar plug, organic modifier, and applied separation voltage) were optimized to achieve a satisfactory PF-MEKC separation of eight GA standards. Under optimum conditions, a baseline separation of GA standards, including GA1, GA3, GA5, GA6, GA7, GA9, GA12, and GA13, was accomplished within 25,min. Satisfactory results were obtained in terms of precision (RSD of migration time below 0.9%), sensitivity (LODs in the range of 0.8,1.9,,M) and linearity (R2 between 0.981 and 0.997). MS/MS with multiple reaction monitoring (MRM) detection was carried out to obtain sufficient selectivity. PF-MEKC-MS/MS allowed the direct identification and confirmation of the GAs presented in coconut water (CW) sample after SPE, while, the quantitative analysis of GAs was performed by PF-MEKC-MS approach. GA1 and GA3 were successfully detected and quantified in CW. It is anticipated that the current PF-MEKC-MS method can be applicable to analyze GAs in a wide range of biological samples. [source] Capillary electrophoretic separation of biologically active amines and acids using nanoparticle-coated capillariesELECTROPHORESIS, Issue 9 2008Yu-Fen Huang Abstract This manuscript describes dynamic coating of capillaries with poly(L -lysine) (PLL) and silica nanoparticles (SiO2 NPs) and use of the as-prepared capillaries for the separation of biogenic amines and acids by CE in conjunction with LIF detection. The directions of EOF are controlled by varying the outmost layer of the capillaries with PLL and SiO2 NPs, respectively. Over the pH range 3.0,5.0, the (PLL,SiO2NP)n,PLL capillaries have an EOF toward the anodic end and are more suitable for the separation of acids with respect to speed, while the (PLL,SiO2NP)n capillaries have an EOF toward the cathodic end and are more suitable for the separation of biogenic amines regarding speed and sensitivity. The separations of standard solutions containing five amines and two acids by CE with LIF detection using (PLL,SiO2NP)2,PLL and (PLL,SiO2NP)3 capillaries were accomplished within 10 and 7,min, providing plate numbers of 3.8 and 5.0×104,plates/m for 5-hydroxytryptamine (5-HT), respectively. The LODs for 5-HT and 5-hydroxyindole-3-acetic acid (5-HIAA) are 32 and 2,nM and 0.2 and 1.5,nM when using the (PLL,SiO2NP)2,PLL and (PLL,SiO2NP)3 capillaries, respectively. Identification and quantification of 5-HIAA, homovanillic acid, and DL -vanillomandelic acid in urine samples from a male before and after drinking green tea were tested to validate practicality of the present approach. The results show that the (PLL,SiO2NP)2,PLL capillary provides greater resolving power, while the (PLL,SiO2NP)3 capillary provides better sensitivity, higher efficiency, and longer durability for the separation of the amines and acids. [source] Rapid capillary electrophoresis time-of-flight mass spectrometry separations of peptides and proteins using a monoquaternarized piperazine compound (M7C4I) for capillary coatingsELECTROPHORESIS, Issue 8 2008Anisa Elhamili Abstract A monoquaternarized piperazine, 1-(4-iodobutyl) 4-aza-1-azoniabicyclo[2,2,2] octane iodide (M7C4I), has been evaluated as a surface derivatization reagent for CE in combination with TOF MS for the analysis of proteins, peptides, and protein digests. The M7C4I piperazine, at alkaline pH, forms a covalent bond via alkylation of the ionized silanols producing a cationic surface with a highly stable and reversed EOF. The obtained surface yields rapid separations (less than 5,min) of peptides and proteins at acidic pH with high separation efficiencies (up to 1.1×106 plates/m for peptides and up to 1.8×106 plates/m for proteins) and no observed bleeding of the coating reagent into the mass spectrometer. The simplicity of the coating procedure also enables fast (2,min) regeneration of the surface, if necessary. This is useful in the analysis of complex samples in order to prevent possible memory effects. The potential of using M7C4I-coated capillaries for MS analysis of complex samples is demonstrated by the separation of peptides, proteins, and protein digests. Even more, the spectacular thing in which large intact proteins with molecular masses over 0.5,MDa could be separated. The coating showed good ability to handle these large proteins with high efficiency and retained peak shape as demonstrated by separation of IgG1 (150,kDa) and thyroglobulin (669,kDa). [source] Use of quasi-isoelectric buffers as anolyte and catholyte to improve capillary isoelectric focusing performancesELECTROPHORESIS, Issue 8 2008Martine Poitevin Abstract The use of quasi-isoelectric anolytes and catholytes has been investigated to improve CIEF performances. Narrow pH cuts of carrier ampholytes (NC) have been compared to more conventional couples of anolytes/catholytes (phosphoric acid/sodium hydroxide and glutamic acid/lysine). First, a CIEF setup that consists in a bare silica capillary and 70:30 water/glycerol separation medium has been used. The experiments have shown that when using NC instead of more classical anolytes and catholytes, an increase in the protein detection time was observed and the resolutions obtained for neutral and acidic proteins were doubled. Moreover, according to the NC fraction used, the resolution was modified. In order to investigate further the mechanisms involved, a second setup using a capillary coated with hydroxypropylcellulose was used. With this setup no difference has been observed when changing anolyte and catholyte nature. A simple methodology has then been developed to evaluate EOF during focusing and mobilization steps of CIEF experiments. It highlighted the crucial role played by EOF when using a bare silica capillary. EOF indeed decreased by 33% during mobilization step when using NC instead of classical anolytes and catholytes. [source] Photobleaching-based flow measurement in a commercial capillary electrophoresis chip instrumentELECTROPHORESIS, Issue 6 2008Guiren R. Wang Professor Abstract For microfluidic analytical instruments, a facile, fast, and accurate instrument test is highly demanded. The test includes the quantitative verification of the relationship between pressure drop and flow velocity for the hydrodynamic pump, between the electric voltage and electroosmotic flow (EOF) for the high-voltage supply, and the chip quality. The key point for the test is the measurement of the flow velocity. However, most currently available velocimetries cannot be directly used without any instrumental modification or adding extra instruments. We applied a recently developed Laser Induced Fluorescence Photobleaching Anemometer (LIFPA) for the instrument test through measuring fluid flow velocity in a microfluidic instrument with optical measurement without any modification and extra instrument. We have successfully used the method to test Caliper HTS 250 System from Caliper Life Sciences (Hopkinton, MA) with its own light source and detector. The experimental result demonstrates that this single-point method of measuring flow velocity can be easily used for accurate test of a microfluidic instrument in less than 10,min at extremely low cost without any modification and extra instrument. [source] Compositional effects on electrophoretic and chromatographic figures of merit in electrokinetic chromatography with cetyltrimethylammonium bromide/sodium octyl sulfate vesicles as the pseudostationary phase.ELECTROPHORESIS, Issue 5 2008Part 1: Effect of the phase ratio Abstract The effect of the phase ratio on the electrophoretic and chromatographic properties of unilamellar vesicles comprised of cetyltrimethylammonium bromide (CTAB) and sodium octyl sulfate (SOS) was investigated in EKC. The surfactant concentration of the vesicles was 0.9, 1.2, 1.5, and 1.8% w/v, with a mole ratio of 1:3.66 (CTAB/SOS). Results were compared to those obtained using SDS micelles at concentrations of 1.0% (w/v, 35,mM) and 1.5% (52,mM). The CTAB/SOS vesicles (0.9,1.8% w/v) provided a significantly larger elution range (5.7,,,tves/t0,,,8.7) and greater hydrophobic (methylene) selectivity (2.8,,,,CH2,,,3.1) than SDS micelles (3.1,,,tmc/t0,,,3.3; ,CH2,=,2.2). Whereas the larger elution range can be attributed to the 25% reduction in EOF due to the interaction of unaggregated CTAB cations and the negatively charged capillary wall, the higher methylene selectivity is likely due to the lower concentration of water expected in the CTAB/SOS vesicle bilayer compared to the Palisades layer of SDS micelles. For a given phase ratio, CTAB/SOS vesicles are somewhat less retentive than SDS micelles, although retention factors comparable to those observed in 1.0,1.5% SDS can be obtained with 1.5,1.8% CTAB/SOS. A linear relationship was observed between phase ratio and retention factor, confirming the validity of the phase ratio model for these vesicles. Unique polar group selectivities and positional isomer shape selectivities were obtained with CTAB/SOS vesicles, with both types of selectivities being nearly independent of the phase ratio. For four sets of positional isomers, the elution order was always para < ortho < meta. Finally, the thermodynamics of solute retention was qualitatively similar to that reported for other surfactant aggregates (micelles and microemulsions); the enthalpic contribution to retention was consistently favorable for all compounds, whereas the entropic contribution was favorable only to hydrophobic solutes. [source] Protein separations using polyelectrolyte multilayer coatings with molecular micelles in open tubular capillary electrochromatographyELECTROPHORESIS, Issue 4 2008Candace A. Luces Abstract Novel polyelectrolyte multilayer (PEM) coatings for enhanced protein separations in open tubular CEC (OT-CEC) are reported. Use of four cationic polymers (poly- L -lysine, poly- L -ornithine, poly- L -lysine-serine, and poly- L -glutamic acid-lysine), and three anionic molecular micelles, sodium poly(N -undecanoyl- L -leucyl-alaninate) (poly- L -SULA), sodium poly(N -undecanoyl- L -leucyl-valinate) (poly- L -SULV), and sodium poly(undecylenic sulfate) (poly-SUS) were investigated in PEM coatings for protein separations. The simultaneous effects of cationic polymer concentration, number of bilayers, temperature, applied voltage, and pH of the BGE on the separation of four basic proteins (,-chymotrypsinogen A, lysozyme, ribonuclease A, and cytochrome c) were analyzed using a Box Behnken experimental design. The influence of NaCl on the run-to-run reproducibility was investigated for PEM coatings containing each cationic polymer. All coatings exhibited excellent reproducibilities with a %RSD of the EOF less than 1% in the presence of NaCl. Optimal conditions were dependent on both the cationic and anionic polymers used in the PEM coatings. Poly- L -glutamic acid-lysine produced the highest resolution and longest migration time. The use of molecular micelles to form PEM coatings resulted in better separations than single cationic coatings. Chiral poly- L -SULA and poly- L -SULV resulted in higher protein resolutions as compared to the achiral, poly-SUS. Furthermore, the use of poly- L -SULV reversed the elution order of lysozyme and cytochrome c when compared to poly- L -SULA and poly-SUS. [source] On-line concentration of proteins by SDS-CGE with LIF detectionELECTROPHORESIS, Issue 2 2008Cheng-Ju Yu Abstract We present a simple approach for on-line concentration of SDS-protein complexes by using poly(vinyl alcohol) (PVA) solution in CGE. In comparison to the coated capillary, the presence of EOF in CGE omitted the need to fill the capillaries with polymer solutions prior to the analysis. More importantly, we found that highly reproducible separation of eight proteins by 3.5% PVA was achieved between runs and without the regeneration of high bulk EOF; the RSD of migration times was less than 0.7%. To further improve the concentration sensitivity, neutral PVA was introduced into the capillary with the help of EOF to act as sieving matrix. The occurrence of stacking at the boundary between the PVA and the sample zone is mainly due to the retardation of proteins by PVA. As a result, the LODs at an S/N of 3 for SDS,protein complexes are of the order of sub-nM to several nM. For example, the LOD for BSA is 0.78 nM, which is a 91-fold sensitivity enhancement over the normal injection. In addition, our stacking method has been applied to the analyses of proteins in Escherichia coli cells. The peak for ,-galactosidase (E. coli) was observed after 0.1 ,M ,-galactosidase was spiked into the E. coli samples. [source] Application of CE with novel dynamic coatings and field-amplified sample injection to the sensitive determination of isomeric benzoic acids in atmospheric aerosols and vehicular emissionELECTROPHORESIS, Issue 19 2007Ewa Dabek-Zlotorzynska Dr. Abstract A simple and reliable CE method with direct UV detection has been developed to separate eight isomeric benzoic acids in atmospheric aerosols and vehicular emission without complex sample pretreatment. Optimal electrophoretic conditions, with migration times under 5,min, were obtained by using a 50,mM acetate buffer (pH,4.7) containing a dynamic surface coating EOTrolÔ LN (0.005% w/v). The separations were carried out in a cathode to anode direction (,30,kV) allowing the low cathodal EOF (,1×10,9,m2V,1s,1) to extend the effective separation by slowing the movement of the studied aromatic acids. Moreover, the sensitivity of the method at 200,nm was enhanced by using a field-amplified sample injection (FASI) with electrokinetic (EK) sample injection (,2,kV, 60,s). Prior to sample injection, a short water plug (3,s at 0.5,psi) was introduced. Under these conditions, the method was capable of detecting the analytes in deionized water with LODs (S/N,=,3) as low as 0.1,,g/L for most of the studied acids. In the presence of 10,mg/L of sulphate (added to simulate a sample matrix), LODs ranged from 0.26 to 0.62,,g/L. The validation of the method has proven an excellent separation performance and accuracy for the determination of isomeric benzoic acids in the studied matrices. [source] | ||||||||||||||||