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At least 109 records · Page 6Linked to original sources
PubMed2000

Separation and identification of etodolac and its urinary phase I metabolites using capillary electrochromatography and on-line capillary electrochromatography-electrospray ionisation mass spectrometry coupling.

Capillary high-performance liquid chromatography (capillary HPLC), pressure-assisted capillary electrochromatography (pCEC) and capillary electrochromatography (CEC) were performed in the same capillary packed with 5 microm octadecylsilica (C18) as stationary phase. These three separation modes were compared from the viewpoint of peak efficiency and separation selectivity in order to critically evaluate the advantages which CEC may offer compared to capillary HPLC for the solution of practical biomedical problems. The separation of the non-steroidal anti-inflammatory drug etodolac (ET, 1) and its phase I metabolites, 6-hydroxy etodolac (6-OH-ET, 2), 7-hydroxy etodolac (7-OH-ET, 3) and 8-(1'-hydroxyethyl) etodolac (8-OH-ET, 4) was selected as an example. Baseline separation of all compounds was achieved in different modes and conditions. The effect of pure electrophoretic separation mechanism on the overall separation selectivity observed in CEC has been shown. A high electroosmotic flow (EOF) was observed in C18 packed capillary even at pH 2.5 in various buffers. Furthermore, these separations were coupled on-line with electrospray ionisation mass spectrometry (ESI-MS) and the parent drug and its metabolites were identified in urine. For the coupling of CEC with ESI-MS a laboratory-made electrophoretic device was used in order to overcome some technical disadvantages of commercial instrumentation.

Anti-Inflammatory Agents, Non-Steroidal↗
PubMed2003

Control of electroosmotic flow and wall interactions in capillary electrophoresis capillaries by photografted zwitterionic polymer surface layers.

A novel capillary with covalently bonded zwitterionic surface modification was prepared by photograft polymerization of the zwitterionic monomer N,N-dimethyl-N-methacryloxyethyl-N-(3-sulfopropyl)ammonium betaine, onto the inner surface of a UV-transparent fused-silica capillary. Although the zwitterionic moieties in the resulting polymeric "tentacles" comprise both a positive quaternary ammonium group and a negative sulfonate group, the coating has a net zero charge. The electroosmotic flow (EOF) was therefore extensively suppressed on the grafted capillary compared to the native silica capillary and to the silica capillary that had been activated for graft polymerization by reaction with 3-(methacryloyl)oxypropyltrimethoxysilane. It was also found that the EOF can be varied by adding chaotropic anions or divalent cations such as perchlorate ion and magnesium ion to the running buffer, due to the interaction between these ions and zwitterionic functional group. This provides a new way of altering the EOF and the wall interaction without changing the pH or the overall ionic strength of the separation buffer. The influence of pH and ionic strength of separation buffer on the EOF were also investigated to optimize the separation conditions. Good separations of a mixture containing eight inorganic anions were achieved within 5 min under optimal conditions by capillary zone electrophoresis. The newly prepared capillary was also well suited for the separation of peptides or proteins.

Anions↗
PubMed1983

An estimate of capillary anisotropy and determination of surface and volume densities of capillaries in skeletal muscles of the conger eel (Conger conger L.).

Fast and slow muscle fibres were isolated from the axial muscles of conger eel (Conger conger L.) and examined by electron microscopy. Volume densities of mitochondria, VV (mit, f); intracellular lipid stores, VV (lip, f); and myofibrils, VV (mf, f) were respectively 0.23, 0.18, 0.47 for slow and 0.03, 0.002, 0.79 for fast fibres. Factors affecting the representative sampling of capillary supply were investigated using semi-thin (0.5 micron) sections. An estimate of the degree of capillary anisotropy was made using the assumption of a Fisher axial distribution (Mathieu, Cruz-Orive, Hoppeler & Weibel, 1983). The capillary bed is highly anisotropic. Mean capillary cross-sectional area, determined from electron micrographs, was 25.7 and 30.5 microms2 for slow and fast muscle, respectively. The number of capillaries per unit cross-sectional fibre area, NA (c,f); volume VV (c,f); and surface density, SV (c,f) of capillaries was 615 mm-1 0.016, 135 cm-1 for slow and 21.3 mm-1 0.0007, 4.7 cm-1 for fast muscle, respectively. The relationship between the extent of the capillary bed and mitochondrial volume density is discussed in relation to the function of different muscle types.

Animals↗
PubMed1984

The development of glomerular capillary tufts of the bullfrog kidney from a straight interstitial vessel to an anastomosed capillary network. A scanning electron microscopic study of vascular casts.

Blood vascular tufts of immature glomeruli of the post-metamorphous bullfrog kidney were reproduced by a resin casting medium and observed by scanning electron microscopy. The simplest glomerular tufts consisted of a single straight vessel continuous with the afferent and efferent vessels. In the next stage, the single vessel dilated and bent to form a U-shaped loop; this loop sprouted free-ending protrusions. Subsequently a conglomerated network of anastomosing capillaries developed. The network became larger, and increased in number of capillaries. Even during these later stages, the capillaries possessed luminal protrusions with free endings. Small holes and narrow grooves representing endothelial intrusions into the capillary lumens were noted only rarely. The findings supported the in-growth concept, in that an interstitial vessel invaded the cleft of the S-shaped renal vesicle to form a glomerular capillary network. Secondly, the glomerular capillaries multiplied by sprouting daughter capillaries rather than by luminal splitting.

Animals↗
PubMed1992

1-Deoxymannojirimycin inhibits capillary tube formation in vitro. Analysis of N-linked oligosaccharides in bovine capillary endothelial cells.

Capillary endothelial cells can be induced to form capillary-like structures in vitro by plating on fibronectin-coated dishes (Ingber, D. E., and Folkman, J. (1989) J. Cell Biol. 109, 317-330), thereby mimicking angiogenesis. To assess the role of glycoproteins bearing asparagine-linked oligosaccharides in this process, we tested the effect of oligosaccharide processing inhibitors on the formation of capillary tubes. Deoxymannojirimycin, a compound that prevents synthesis of hybrid and complex-type oligosaccharides, inhibited the formation of capillary tubes. In contrast, swainsonine, an inhibitor that blocks synthesis of complex- but not hybrid-type oligosaccharides, did not inhibit tube formation. Lectin affinity chromatography of 2-[3H] mannose-labeled glycopeptides from endothelial cells induced to form tubes did not reveal a striking difference in the spectrum of oligosaccharides compared to uninduced cells. Since endothelial cells formed tubes normally in the presence of swainsonine, we analyzed glycopeptides from swainsonine-treated induced and uninduced cells. Cells induced to form tubes were enriched in monosialylated hybrid-type oligosaccharides sensitive to alpha-fucosidase, beta-galactosidase, and beta-N-acetylhexosaminidase, suggestive of sialyl Lewis-X determinants. We used an enzyme-linked immunoassay to measure sialyl Lewis-X epitopes on capillary endothelial cells and found that both induced and uninduced cells expressed sialyl Lewis-X epitopes. Deoxymannojirimycin and, to a lesser extent, swainsonine reduced the level of sialyl Lewis-X epitopes in cells induced to form capillary tubes, but neither compound affected the level of epitopes in cell monolayers. We conclude that synthesis of at least hybrid-type oligosaccharides is required for capillary tube formation in vitro and that an increase in monosialylated, fucosylated glycans on asparagine-linked oligosaccharides occurs during this process.

1-Deoxynojirimycin↗
PubMedDate not supplied

Capillary and pre-capillary coronary vascular growth during left ventricular hypertrophy.

An inadequate vascular growth during pressure-overload cardiac hypertrophy involves both pre-capillary and capillary vessels. With the development of hypertrophy in spontaneously hypertensive rats (SHR) coronary reserve declines as indicated by increases in minimal coronary vascular resistance (MCVR). Capillary numerical density, surface density, and volume density also decline. However, after hypertrophy becomes stabilized late in life these parameters return to normal. That elevated blood pressure plays a major role in the decrement in coronary reserve was demonstrated by treating prehypertensive SHR with hydralazine which prevented hypertension but not cardiac hypertrophy. In these animals capillary density remained depressed but coronary reserve was normal. Capillary growth in SHR was, however, stimulated by exercise training even though blood pressure and cardiac hypertrophy were unaffected. Studies on thyroxine-induced cardiac hypertrophy provided evidence of a substantial angiogenesis. In this model coronary reserve is normal or slightly elevated, while capillary density is increased. These studies suggest that angiogenesis is effected by the stimulus evoking cardiac enlargement and that a variety of factors may influence the growth of precapillary and capillary coronary vessels.

Animals↗
PubMed1998

Is the capillary microscopic determination of the visible capillary length a diagnostic criterion in thromboangiitis obliterans?

BACKGROUND: The diagnosis of thromboangiitis obliterans may be difficult to establish in the everyday clinical setting. PATIENTS AND METHODS: In 16 patients with thromboangiitis obliterans vital capillary microscopy was carried out on the nailfold on all ten digits. The longest visible capillary length was noted. As controls we employed 86 healthy subjects. RESULTS: The measurement of the respective longest visible capillary on the nailfold revealed a visible capillary length of more than 0.6 mm in 38% of all patients with thromboangiitis obliterans (n = 16), such capillary lengths were not to be found among healthy subjects (n = 86). CONCLUSIONS: Vital capillary microscopic determination of the visible capillary length is a simple-to-perform, non-invasive examination, which can help to establish the diagnosis.

Adult↗
PubMedDate not supplied

Use of small-diameter capillaries for increasing peptide and protein detection sensitivity in capillary electrophoresis-mass spectrometry.

The use of small ID capillaries is shown to provide a substantial increase in sensitivity for capillary electrophoresis-electrospray ionization/mass spectrometry (CE-ESI/MS). In a comparison using capillaries ranging from either 100 to 10 microns or 50 to 5 microns ID and chemically modified with aminopropylsilane, a 25- to 50-fold increase in sensitivity was observed for both peptide and protein mixtures. This enhanced solute sensitivity allowed the detection of approximately 150 attomoles of melittin (2845 Da) with selected ion monitoring and 600 attomoles of carbonic anhydrase (29,157 Da) while scanning for CE-MS with a quadrupole mass spectrometer. For the protein mixture, mass spectra of sufficient quality for precise molecular weight determination (< or = 0.05%) were obtained for 600 attomole injections using a 5 microns ID capillary. The increase in sensitivity with small capillary diameters can be primarily attributed to a reduced mass flow rate of buffer and other background constituents into the electrospray source, which allows for greater sample ionization efficiency. A model that qualitatively accounts for the results is presented, but quantitative agreement is precluded due to difficulties in accounting for contributions due to a liquid sheath flow used with the electrospray source. The model accounts for the observation that the ESI/MS appears to function as a concentration-sensitive detector under many conditions using large-diameter capillaries. A transition occurs, however, to a regime where the ESI/MS functions as a mass flow-sensitive detector for small-diameter capillaries, where the ESI current is limited by the rate of delivery to the ESI source of charge carrying species in solution. These results suggest peptide and protein analysis at low attomole and subattomole levels should be obtainable with alternative types of mass spectrometers.

Electrophoresis↗
PubMed2003

Guidelines for capillary extraction-capillary gas chromatography: preparation of extractors and analysis of aromatic compounds in water.

The benzene, toluene, ethylbenzene and xylenes system (BTEX) in clean water is studied to verify the performance of capillary extraction as an extraction-preconcentration technique well hyphenated with GC. The approach uses pieces of coated capillaries usually 5-30 cm long, trimmed from customary high-resolution GC columns but carrying glass press-fits at their ends. The preparation of these 'capillary extractors' is explained, and their performance is discussed providing guidelines for use. Injection by capillary extraction is such that (i) band broadening in time is null, and (ii) band broadening in space cannot be higher than the extractor length. Speed, cleanliness and operative simplicity of the capillary extraction approach are remarkable, pros and cons are complementary to those of solid phase microextraction (SPME) or stir bar sorptive extraction (SBSE). Capillary extraction-capillary GC analysis of aqueous BTEX samples, in a clean water matrix, allows low part-per-billion detection limits, and does not require heated injectors or cryofocusing devices.

Benzene↗
PubMed2004

Free energy balance for three fluid phases in a capillary of arbitrarily shaped cross-section: capillary entry pressures and layers of the intermediate-wetting phase.

In this work we derive rigorously the free energy balance for three fluid phases in a straight capillary of arbitrarily shaped cross-section. This balance is then used to derive the general equation for the capillary entry pressures of all possible two-phase and three-phase displacements. Moreover, the equation provides the criterion determining the existence of layers of the intermediate-wetting phase separating the wetting and non-wetting phases in the corners or cavities of a capillary, by also treating the spreading of such layers as a capillary displacement. For a number of combinations of interfacial tensions and contact angles, illustrating all the different relevant situations, we calculate the criteria for spreading of such a layer in the corner of a capillary with polygonal cross-section. In a capillary with a cross-section in the shape of an isosceles triangle of varying corner size, these criteria are used to determine the unique capillary entry pressures for piston-like displacement from alternative solutions of the general equation. These solutions relate to displacements in the presence or absence of layers in the various differently sized corners.

Journal Article↗
PubMed2000

Capillary electrophoresis with an integrated on-capillary tubular detector based on a carbon sol-gel-derived platform

An integrated on-capillary tubular electrochemical detector for capillary electrophoresis systems has been fabricated based on sol-gel technique. It consists of a sol-gel carbon composite tubular electrode attached permanently onto the outlet of the separation capillary. The device greatly eases the setting up of capillary electrophoresis with electrochemical detection (CEEC) as it makes possible electrode/capillary alignment without the aid of a micromanipulator since this integrated unit can be simply immersed in the CE separation buffer in an ordinary three-electrode stationary cell. To improve analytical performance of the integrated unit, the external wall of the exit capillary was etched with HF after the polyimide coating of the capillary had been removed. Influences of the working electrode length and the wall thickness at the outlet of capillary on the separation efficiency and amperometric sensitivity were assessed and optimized. The practical applicability of this configuration is demonstrated with the detection of both catecholamines and carbohydrates. The advantages, namely, versatility, convenience, ease of operation, and low-cost, of the new design combined with an excellent performance lead to high stability and low detection limits.

Journal Article↗
PubMed2005

Latex-coated polymeric monolithic ion-exchange stationary phases. 1. Anion-exchange capillary electrochromatography and in-line sample preconcentration in capillary electrophoresis.

A sulfonated methacrylate monolithic polymer has been synthesized inside fused-silica capillaries of diameters 50-533-microm i.d. and coated with 65-nm-diameter fully functionalized quaternary ammonium latex particles (AS18, Dionex Corp.) to form an anion-exchange stationary phase. This stationary phase was used for ion-exchange capillary electrochromatography of inorganic anions in a 75-microm-i.d. capillary with Tris/perchlorate electrolyte and direct UV detection at 195 nm. Seven inorganic anions (bromide, nitrate, iodide, iodate, bromate, thiocyanate, chromate) could be separated over a period of 90 s, and the elution order indicated that both ion exchange and electrophoresis contributed to the separation mechanism. Separation efficiencies of up to 1.66 x 10(5) plates m(-1) were achieved, and the monoliths were stable under pressures of up to 62 MPa. Another latex-coated monolith in a 250-microm-i.d. capillary was used for in-line preconcentration by coupling it to a separation capillary in which the EOF had been reversed using a coating of either a cationic polymer or cationic latex particles. Several capillary volumes of sample were loaded onto the preconcentration monolith, and the analytes (inorganic anions) were then eluted from the monolith with a transient isotachophoretic gradient before being separated by electrophoresis in the separation capillary. Linear calibration curves were obtained for aqueous mixtures of bromide, nitrite, nitrate, and iodide. Recoveries of all analytes except iodide were reduced significantly when the sample matrix contained high levels of chloride. The preconcentration method was applied to the determination of iodide in open ocean water and provided a limit of detection of 75 pM (9.5 ng/L) calculated at a signal-to-noise ratio of 3. The relative standard deviation for migration time and peak area for iodide were 1.1 and 2.7%, respectively (n = 6). Iodide was eluted as an efficient peak, yielding a separation efficiency of 5.13 x 10(7) plates m(-1). This focusing was reproducible for repeated analyses of seawater.

Journal Article↗
PubMed2000

A decade of capillary electrophoresis.

Since the introduction of the first commercial capillary electrophoresis (CE) instrument a decade ago, CE applications have become widespread. Today, CE is a versatile analytical technique which is successfully used for the separation of small ions, neutral molecules, and large biomolecules and for the study of physicochemical parameters. It is being utilized in widely different fields, such as analytical chemistry, forensic chemistry, clinical chemistry, organic chemistry, natural products, pharmaceutical industry, chiral separations, molecular biology, and others. It is not only used as a separation technique but to answer physicochemical questions. In this review, we will discuss different modes of CE such as capillary zone electrophoresis, micellar electrokinetic chromatography, capillary gel electrophoresis, capillary isoelectric focusing, and capillary electrochromatography, and will comment on the future direction of CE, including array capillary electrophoresis and array microchip separations.

Amino Acids↗
PubMed2000

[Advances of capillary electrophoresis and capillary electrochromatography using ionic coating column].

Capillary electrochromatography (CEC) is a promising micro-separation technique, which combines the high selectivity of HPLC and the high efficiency of HPCE. In comparing to CE with fused silica capillary and CEC with ODS stationary phase, capillary electrophoresis and capillary electrochromatography with ionic coating columns can cause EOF more controllable, and provide an alternative separation mechanism. Therefore, the new CEC and CE modes are in favor of optimizing the separation and broadening the range of samples for analysis. The development of capillary electrophoresis and capillary electrochromatography using ionic coating column is reviewed with 48 references.

Amino Acids↗
PubMed2001

Discrimination between peak spreading in capillary zone electrophoresis of proteins due to interaction with the capillary wall and due to protein microheterogeneity.

Our study attempts to find an approach to distinguishing between the contribution to peak spreading in capillary zone electrophoresis (CZE) due to protein microheterogeneity and that due to interaction with the capillary wall, by analyzing correlations between observed peak spreading and peak asymmetry. The peak asymmetry was measured as ln[(tm-t1)/(t2-tm)] where tm, t1, and t2 are migration times at the mode of the peak and at the intersection of the peak width at half-height with the ascending and descending limbs, respectively. Two isoforms of recombinant green fluorescent protein (GFP-1 and GFP-2, 27 kDa molecular mass), glucose-6-phosphate dehydrogenase (GPD, 104 kDa), and the naturally fluorescent protein R-phycoerythrin (PHYCO, 240 kDa) were subjected to CZE in polyacrylamide-coated fused-silica capillaries of 50 and 100 microns diameters under varying conditions of protein concentration, field strength, and the initial zone length. Under conditions such that contributions to peak spreading from axial diffusion, thermal effects, and electrophoretic dispersion are negligible, the analysis of the interrelations between peak width and peak asymmetry was found to allow a conclusion as to the cause of peak spreading in CZE of protein. It appears that the peak width of GFP-2 originates mostly in protein microheterogeneity while that of GFP-1 is due to protein-capillary wall interactions. For PHYCO, both microheterogeneity and protein-capillary wall interactions contribute to peak spreading. GPD exhibits relatively little microheterogeneity or interaction with capillary walls. Thus, its peak width appears to be mostly affected by an extracolumn source of spreading such as the initial zone length.

Electrophoresis, Capillary↗
PubMed2001

Assessment of the capillary zone electrophoretic behavior of proteins in the presence of electroosmotic modifiers: protein-polyamine interaction studied using a polyacrylamide-coated capillary.

The use of polyamines as electroosmotic modifiers has been shown to be effective in enhancing resolution of protein glycoforms in capillary zone electrophoresis (CZE) using a bare capillary tube. In this study, effectiveness was evaluated by using a polyacrylamide-coated capillary tube instead of a bare capillary tube. Electropherograms obtained in the presence of polyamines were inferior to those obtained in their absence with respect to resolution. Electrophoretic mobility of the proteins decreased and their peaks were broadened by polyamines bound to them. This unfavorable effect was dependent on both the species of polyamines and the pH values of the electrolyte buffer. The reduction of resolution caused by polyamines was in the following order: spermidine (SPD) approximately spermidine-tri-hydrochloride (SPD-HCI) > putrescine (PUT) > hexamethonium chloride (HMC). The observed effect can be ascribed to the formation of complexes between the proteins and the polyamines. In addition, for the bare capillary tube the complexes showed interaction with the inner surface, resulting in local suppression of electroosmosis and poor resolution. The high resolution obtained in the coated capillary tube was reduced in the presence of the polyamines. Thus, the use of the polyamines has a negative effect on the analysis of protein microheterogeneity as a result of protein-polyamine interaction.

Acrylic Resins↗
PubMed2001

Metal speciation using capillary electrophoresis--inductively coupled plasma atomic emission spectrometry and polytetrafluoroethylene capillaries.

A capillary electrophoresis--inductively coupled plasma atomic-emission spectrometry (CE-ICP-AES) system using a polytetrafluoroethylene (PTFE) capillary has been developed. The CE-ICP interface was a modified concentric nebulizer. The PTFE capillary (50 microm internal diameter) was used as the central capillary of the nebulizer. Using the PTFE capillaries, the solution flow rate induced by the carrier gas flow was smaller than that of glass capillary. Solution flow was mainly induced by the CE electric field. Baseline separation of Ba2+/Mg2+ ion pair using simple buffer solution of 0.014 M sodium acetate was reported. Separation and correlation of metal species in metallothioneins (MT-1 and MT-2 in MT) of rabbit liver using the CE-ICP system were also discussed.

Electrophoresis, Capillary↗
PubMed1996

Use of wide-bore capillaries in constant denaturant capillary electrophoresis.

A serious limitation of capillary electrophoresis in separating mutant from wild-type DNA sequences is the amount of DNA which may be loaded on narrow-bore columns. Because we sought to examine mutant sequences derived from more than 10(9) human cells, we were obliged to overcome this limitation. Thus we examined the feasibility of using wide-bore capillaries of 250, 320 and 540 microns in constant denaturant capillary electrophoresis (CDCE) using conditions which successfully limited the effect of Joule heating. Excellent separations were observed using the wide-bore capillaries comparable to those obtained using a 75 microns capillary. DNA loading capacities were increased almost 100-fold as the bore of capillary increased from 75 to 540 microns.

Adenomatous Polyposis Coli↗