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At least 217 records · Page 12Linked to original sources
PubMed1990

Capillary electrophoresis in 2 and 5 microns diameter capillaries: application to cytoplasmic analysis.

Capillary electrophoresis with electrochemical detection in 2- and 5-micron capillaries has been developed to study ultrasmall biological environments. Sample volumes as low as 270 fL have been injected into the electrophoresis capillary with subattomole detection limits for easily oxidized species. We have applied this method to the analysis of single cell cytoplasm. Sampling of the cytoplasm is accomplished by inserting one end of the electrophoresis capillary directly into a single nerve cell. The high-voltage end of the electrophoresis capillary has been etched with hydrofluoric acid to form a microinjector. This injection scheme represents an improvement over those previously used for similar applications. The excellent selectivity of this method is demonstrated for catecholamine and indolamine neurotransmitters and their metabolites found in the invertebrate system, the pond snail Planorbis corneus.

Animals↗
PubMed2003

Comparison of frits used in the preparation of packed capillaries for capillary electrochromatography.

The performance of several types of frits, including sintered frits, photopolymerized frits, and frits made by sol-gel technologies, is evaluated. The frits are formed in open capillaries, and the electroosmotic mobilities are calculated and compared to those obtained in an open capillary. Run-to-run, day-to-day, and column-to-column reproducibilities are evaluated. In all cases, the run-to-run reproducibility varied by < 4%. The greatest mobility was through the capillaries with the sol-gel frits (1.29 x 10(-3) cm2/V x s) which also exhibited the best day-to-day reproducibility. The capillaries with the photopolymerized frits had the best column-to-column reproducibility, yet the slowest mobility (1.00 x 10(-3) cm2/V x s). The second central moment of benzene as a solute is calculated and plotted as a function of time to estimate the differences in peak dispersion among the various frits studied. Although the mobilities through the capillaries with the sintered frits were the least reproducible, these frits are associated with the least amount of band-broadening. An estimation of the dispersion caused by each of the different types of frits is reported.

Journal Article↗
PubMed2004

A low-makeup beveled tip capillary electrophoresis /electrospray ionization mass spectrometry interface for micellar electrokinetic chromatography and nonvolatile buffer capillary electrophoresis.

A robust interface has been developed for interfacing micellar electrokinetic chromatography (MEKC) and nonvolatile buffer capillary electrophoresis (CE) to electrospray ionization mass spectrometry (ESI-MS). The interface consists of two parallel capillaries for separation (50 microm i.d. x 155 microm o.d.) and makeup (50 microm i.d. x 155 microm o.d.) housed within a larger capillary (530 microm i.d. x 690 microm o.d.). The capillaries terminate in a single tapered tip having a beveled edge. The use of a tapered beveled edge results in a greater tip orifice diameter (75 microm) than in a previous design from our laboratory (25 microm) that used a flat tip. While maintaining a similar optimum flow rate and consequently similar sample dilution, a 75-microm beveled emitter is more rugged than a 25-microm flat tip. Furthermore, the incorporation of a sheath liquid capillary allows the compositions of the final spray solution to be controlled. The application of this novel CE/ESI-MS interface was demonstrated for MEKC using mixtures of triazines (positive ion mode) and phenols (negative ion mode). The ability to perform CE/ESI-MS using a nonvolatile buffer was demonstrated by the analysis of gangliosides with a buffer consisting of 40 mM borate and 20 mM alpha-cyclodextrin.

Journal Article↗
PubMed2006

Field amplified separation in capillary electrophoresis: a capillary electrophoresis mode.

In field-amplified injection in capillary electrophoresis (CE), the capillary is filled with two buffering zones of different ionic strength; this induces an amplified electrical field in the low ionic strength zone and a lower field in the high ionic strength zone, making sample stacking feasible. The electroosmotic flow (eof) usually observed in CE, however, displaces the low field zone and induces an extra band broadening preventing any CE separation in the field-amplified zone. These limitations have originated the restricted use of field amplification in CE only for stacking purposes. For the first time, in this work it is theoretically shown and experimentally corroborated that CE separation speed and efficiency can simultaneously be increased if the whole separation is performed in the field-amplified zone, using what we have called field amplified separation in capillary electrophoresis (FAsCE). The possibilities of this new CE mode are investigated using a new and simple coating able to provide near-zero eof at the selected separation pH. Using FAsCE, improvements of 20% for separation speed and 40% for efficiency are achieved. Moreover, a modified FAsCE approach is investigated filling the capillary with the high ionic strength buffer up to the interior of the detection window. Under these conditions, an additional 3-fold increase in sensitivity is also observed. The most interesting results were obtained combining the short-end injection mode and this modified FAsCE approach. Under these conditions, a part of a 3-fold improvement in efficiency and sensitivity, the total analysis time was drastically reduced to 40 s, giving rise to a time reduction of more than 7-fold compared to normal CE. This speed enhancement brings about one of the fastest CE separations achieved using capillaries, demonstrating the great possibilities of FAsCE as a new, sensitive, efficient, and fast CE separation mode.

Journal Article↗
PubMed2004

Retardation of ice growth in glass capillaries: measurement of the critical capillary radius.

An experiment was designed to compare the freezing of an aqueous solution in glass microcapillaries and in thin films. The velocity dependence of the ice front propagation in glass capillaries with radii of 87.5 microm-1.5 microm was observed. A critical capillary radius r(0), corresponding to certain thermal conditions, was obtained, below which the ice growth inside the capillaries was retarded. This critical capillary radius is further related to lambda(0), the smallest wavelength used in the Mullins-Sekerka criterion for the instability analysis of bulk solidifications [Mullins and Sekerka, J. Appl. Phys. 35, 444 (1964)]. It was found that for the present hypothesis, r(0)=lambda(0)/4 gives good predictions. The relation between the propagation velocity (or cooling rate) and the critical radius (or pore size) is summarized in a chart for applications in capillary-porous media, such as in the freezing of biological tissues.

Journal Article↗
PubMed2004

Three-phase threshold capillary pressures in noncircular capillary tubes with different wettabilities including contact angle hysteresis.

We use the Helmholtz free energy balance and the Mayer-Stowe-Princen method to derive general expressions governing multiphase pistonlike displacements in noncircular capillary elements with arbitrary wettability. We take into account hysteresis in oil/water, gas/water, and gas/oil contact angles. We study both two- and three-phase systems. We find threshold capillary pressures for gas invasion into oil, oil invasion into gas, and water invasion into oil for capillaries with an irregular triangular cross section. Finally we study the effects of shape factor, oil/water capillary pressure, and oil/water and gas/oil contact angles on the threshold capillary pressure for gas invasion into oil for spreading and nonspreading systems. In many cases the threshold pressures of the three-phase displacements are not the same as those of the equivalent pseudo-two-phase displacements. It is possible that gas invasion can occur resulting in a configuration without oil layers, even if oil layers are geometrically possible. This emphasizes the distinction between geometric and thermodynamic stability-it is the latter that controls threshold pressures.

Journal Article↗
PubMedDate not supplied

Evaluation of capillary gas chromatography for pesticide and industrial chemical residue analysis. II. Comparison of quantitative results obtained on capillary and packed columns.

Results of pesticide and industrial chemical residue determinations, using both capillary and packed column gas chromatography (GC), in 3 Food and Drug Administration (FDA) laboratories have been compiled and compared. Samples consisted of food products collected for routine residue screening by the respective laboratories. Extracts were prepared by conventional multiresidue methodology. Capillary column systems and operating conditions were selected at the discretion of each laboratory and were therefore variable, although split/splitless injectors in the split mode were used with prescribed precautions in all cases. Packed column systems were operated as specified in the FDA Pesticide Analytical Manual (PAM). Overall correlation between the 2 systems, expressed as the average ratio of packed column result to capillary column result, was 0.99 for 120 determinations in 41 samples. The higher resolving power of the capillary systems allowed quantitation of several residues that were incompletely separated and therefore unquantifiable using the packed columns. Capillary column GC with the split injection technique, used with appropriate precautions, was found to be both reliable and advantageous for regulatory determination of pesticide and industrial chemical residues in foods and feeds.

Chromatography, Gas↗
PubMed2006

Recent developments in capillary electrophoresis and capillary electrochromatography of peptides.

The article gives a comprehensive review on the recent developments in the applications of high-performance capillary electromigration methods, zone electrophoresis, isotachophoresis, isoelectric focusing, affinity electrophoresis, electrokinetic chromatography, and electrochromatography, to analysis, preparation, and physicochemical characterization of peptides. The article presents new approaches to the theoretical description and experimental verification of electromigration behavior of peptides, covers the methodological aspects of capillary electroseparations of peptides, such as rational selection of separation conditions, sample preparation, suppression of peptide adsorption, new developments in individual separation modes, and new designs of detection systems. Several types of applications of capillary electromigration methods to peptide analysis are presented: conventional qualitative and quantitative analysis, purity control, determination in biomatrices, monitoring of chemical and enzymatical reactions and physical changes, amino acid and sequence analysis, and peptide mapping of proteins. Some examples of micropreparative peptide separations are given and capabilities of capillary electromigration techniques to provide important physicochemical characteristics of peptides are demonstrated.

Animals↗
PubMed1962

The physics of blood flood in capillaries. II. The capillary resistance to flow.

A previous communication described the peculiar motion of the plasma trapped between erythrocytes in a capillary (bolus flow). In this paper the effect of this motion on capillary resistance to flow, as well as on dissipative effects associated directly with the cells, are described. The resistance that would be associated with plasma in bolus flow at high Reynolds numbers (relative to a capillary value of 0.01) was studied in a model, in which air bubbles, separated by short segments of water, passed along a glass tube. The resistance to flow, especially with short boluses, was at least ten times greater than that associated with Poiseuille flow. In a second series of experiments at lower Reynolds numbers, a single bolus of liquid was forced by air pressure along a glass tube. In these latter experiments, which more closely simulate biological conditions, the mean resistance to flow was only 30 per cent greater than that associated with Poiseuille flow. In the final series of experiments human blood and plasma, diluted in acid-citrate dextrose (A.C.D.) in varying degrees, were forced through glass micropipettes of capillary dimensions. The mean apparent viscosity of whole blood was found to exceed that of plasma by only about 5 per cent, thus verifying a conjecture to this effect made by Fahraeus and Lindqvist in 1931.

Biophysical Phenomena↗
PubMed2000

Comparative study of capillary zone electrophoresis and micellar electrokinetic capillary chromatography for the separation of naphthalenedisulfonate isomers.

We investigated the separation of a test mixture of nine substituted and unsubstituted naphthalenedisulfonate isomers by capillary electrophoresis with a UV diode array detector. In particular, we focused on how the composition of the running buffer affected the separation selectivity. When capillary zone electrophoresis was carried out, the best results were obtained when organic solvents such as ethanol or propan-2-ol were added. Eight peaks were baseline separated but in no case were all the unsubstituted isomers separated. Therefore, capillary electrophoretic separation of the compounds was examined in the presence of micellar agents, such as sodium dodecyl sulfate, polyethylene glycol dodecyl ether (Brij 35) and cetyltrimethylammonium bromide. All the substituted isomers and two of the unsubstituted isomers were well resolved within 20 min by micellar electrokinetic capillary chromatography when Brij 35 was used as micellar agent. Separations were reproducible, in terms of peak area and migration time, under these conditions.

Chromatography, Micellar Electrokinetic Capillary↗
PubMed1998

The capillary filtration coefficient for evaluation of capillary fluid permeability in cat calf muscles.

Measuring the capillary filtration coefficient (CFC) from the transvascular fluid filtration following a fixed increase in transcapillary hydrostatic pressure is a common method to estimate capillary hydraulic permeability (conductivity) in an organ. Constant flow pump perfusion with an artificial perfusate and a maximally dilated vascular bed are often used in CFC studies to avoid influence on CFC of variations in vascular tone, blood flow and perfusion pressure. The present study evaluates if capillary hydraulic conductivity can be estimated by the CFC method, when analyzed on a denervated cat skeletal muscle with quite well-preserved local vascular control and perfused with autologous blood. CFC was estimated by increasing venous pressure and by decreasing tissue pressure, and both during autoperfusion and pressure controlled pump perfusion. A constant filtration rate was achieved 3-4 min after the transcapillary pressure elevation, giving a CFC around 0.0090 mL min-1 mmHg-1 100 g. The CFC did not change with arterial pressure or with reduced vascular tone using the tissue pressure method, but decreased slightly with increased arterial pressure and reduced vascular tone using the venous pressure method. CFC variations with arterial pressure were larger during pump perfusion in which myogenic reactivity is depressed, indicating that influence of myogenic tone on CFC is small. We conclude that CFC can be used to evaluate capillary hydraulic conductivity, and also when arterial pressure, vascular tone and blood flow are altered within reasonable physiological limits during the experiment, and the tissue pressure method and autoperfusion is to be preferred.

Animals↗
PubMed1978

Simultaneous measurements of capillary diffusion and filtration exchange during shifts in filtration-absorption and at graded alterations in the capillary permeability surface area products (PS).

The diffusion exchange of Cr-EDTA, using the single injection indicator diffusion method, was followed simultaneously with estimations of the capillary filtration capacity (CFC) in an "isogravimetric" rat hindquarter preparation during artificial perfusion and maximal dilatation. Measurements were performed at constant flow and during 1) shifts in filtration-absorbtion, 2) alterations of perfused capillary wall area (graded rarification of capillary network by microsphere injection) and 3) during alterations of permeability (i.a. infusion of histamine). At maximal vasodilatation CFC was 0.037 +/- 0.001 ml/min X mmHg X 100 g and PS for Cr-EDTA 5.67 +/- 0.13 ml/min X 100 g. During filtration or absorbtion, Cr-EDTA transfer from vessels to interstitium changed only slightly but the situation may well be different for solute transfer from interstitium to vessels. Alterations in capillary wall area resulted in proportional changes in PS for Cr-EDTA while the CFC changes were always relatively smaller. Histamine increased CFC some threefold with a marked increase in protein transfer, while PS for Cr-EDTA increased only marginally. This histamine effect could be ascribed mainly to an increase in the number of large pores which, because of their relative paucity, are of little importance for small molecular diffusion exchange but highly important for convective and macromolecular exchange.

Absorption↗
PubMed1999

Determination of adenosine deaminase activity in human erythrocytes by on-column capillary isotachophoresis-capillary zone electrophoresis in the presence of electroosmotic flow.

Transient capillary isotachophoresis (CITP)-capillary zone electrophoresis (CZE) in presence of electroosmotic flow (EOF) was utilized for the measurement of adenosine deaminase activity in human erythrocytes. Phosphates, dominant anions of the sample matrix, were used as leading ions for transient isotachophoresis, and borates (0.3 M, pH 10) were used as terminating ions and background electrolyte for CZE. Final experimental conditions made it possible to inject 70% of the total capillary volume (1.45 microL) with the sample. Enzymatic conversion products (inosine and hypoxanthine), present in the sample in the low-micromolar range, were determined using optimized conditions. The limit of detection was 28 nM using UV detection at 202 nm. The presented data shows that CITP-CZE can be performed in uncoated capillaries in the presence of strong EOF.

Adenosine Deaminase↗
PubMed1999

Chiral separations in capillary high-performance liquid chromatography and nonaqueous capillary electrochromatography using helically chiral poly(diphenyl-2-pyridylmethyl methacrylate) as chiral stationary phase.

Enantioseparations in nonaqueous capillary electrochromatography (CEC) are reported in this study for the first time, using wide-pore aminopropyl silica gel coated with helically chiral poly(diphenyl-2-pyridylmethyl methacrylate) (PDPM) as chiral stationary phase (CSP). The anodic electroosmotic flow (EOF) in a methanolic solution of ammonium acetate was used for the migration of neutral analytes through the packed bed in the capillaries. Four different techniques, high-performance liquid chromatography (HPLC) in common-size columns, capillary HPLC, pressure-assisted CEC and CEC were compared from the viewpoint of separation parameters. The latter three were performed with the same experimental setup, varying the relative contribution of the pressure-driven and the electrically driven flow to the overall mobility of the analyte. Capillary HPLC offers clear advantages compared to enantioseparations in common-size columns. However, for a given particle size of the packing material, CEC was not obviously advantageous compared to pressure-driven separations.

Chromatography, High Pressure Liquid↗
PubMed2000

Fraction collection in micropreparative capillary zone electrophoresis and capillary isoelectric focusing.

A new fraction collection system for capillary zone electrophoresis (CZE) and capillary isolelectric focusing (CIEF) is described. Exact timing of the collector steps was based on determining the velocity of each individual zone measured between two detection points close to the end of the capillary. Determination of the zone velocity shortly before collection overcame the need for constant analyte velocity throughout the column. Consequently, sample stacking in CZE with large injection volumes as well as zone focusing in CIEF could be utilized with high collection accuracy. Capillaries of 200 microm inner diameter (ID) were employed in CZE and 100 microm ID in CIEF for the micropreparative mode. A sheath flow fraction collector was used to maintain permanent electric current during the collection. The bulk liquid flow due to siphoning, as well as the backflow arising from the sheath flow droplet pressure, were suppressed by closing the separation system at the inlet with a semipermeable membrane. In the CZE mode, the performance of the fraction collector is demonstrated by isolation of individual peaks from a fluorescently derivatized oligosaccharide ladder. In the CIEF mode, collection of several proteins from a mixture of standards is shown, followed by subsequent analysis of each protein fraction by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS).

Animals↗
PubMed2000

Miniaturization of capillary electrochromatography using an ultrashort capillary column.

The instrumentation for miniaturization of capillary electrochromatography was devised and an injection method for this apparatus was proposed. By using an ultra short capillary column (15 mm packed length, 36 mm total length, 75 microm inner diameter, packed with cation exchange supports), the separation of five biochemical compounds was performed within 1 min. The high separation efficiency of 9780 plates was achieved by using an ultrashort capillary column. The miniaturized instrumentation for capillary electrochromatography might be utilized as one of the possible methods in microfabricated analysis or in an alternative approach to it.

Biochemistry↗
PubMed2001

Effects of electrolyte modification and capillary coating on separation of glycoprotein isoforms by capillary electrophoresis.

The capillary electrophoresis (CE) running electrolyte composition was optimized for the separation of selected glycoproteins. A good separation of the ovalbumin (OV) and alpha-acid glycoprotein (AAG) isoforms was achieved in 20 mmol x L(-1) N-(2-hydroxyethyl)piperazine-2'-(2-ethanesulfonic acid) (HEPES) at pH 7.0, in 20 mmol x L(-1) phosphate, pH 7.0, or in 25 mmol x L(-1) borate, pH 7.9. Various ways of suppression of the glycoprotein adsorption onto the capillary wall were compared. Alpha, omega-diamine alkanes and bis(aminoalkyl) amines were added to the CE buffers, the optimized concentration being 1 mmol x L(-1) in 20 mmol x L(-1) phosphate buffer. The OV and AAG isoforms could be separated using all the alpha,omega-diamine alkanes or bis(2-aminoethyl)amine. The length of the alkyl chain in the diaminoalkane did not influence the separation. The separation of the isoforms of pollen allergens was also tested. The effects of modification of the capillary wall by succinyl-poly-L-lysine and hydrophilic CElect-P1 capillary were compared. A decrease in the glycoprotein and protein adsorption resulted in an improved separation of the isoforms.

Buffers↗
PubMed2001

Michaelis-Menten analysis of bovine plasma amine oxidase by capillary electrophoresis using electrophoretically mediated microanalysis in a partially filled capillary.

A method for determining bovine plasma amine oxidase (PAO; EC 1.4.3.6) activity with benzylamine (Bz) as substrate is described. Electrophoretically mediated microanalysis (EMMA) combined with micellar electrokinetic capillary chromatography (MEKC) was used to perform an on-capillary enzymatic reaction and to separate the generated benzaldehyde from the other reaction products. The capillary was only partially filled with the separation solution, since the enzyme was unstable in the presence of the applied surfactant. The initial reaction velocity of the enzyme-catalyzed reaction was estimated from the peak area of the enzyme product, benzaldehyde. An amplification step was introduced by means of an on-capillary incubation of 15 min, in order to accumulate enough reaction product to detect spectrophotometrically at 254 nm. This set-up resulted in a fully automated assay, which can be carried out in less then 35 min. Using the Lineweaver-Burk equation, an average Michaelis constant (K(M)) for PAO was calculated to be 0.74 mM +/- 0.05 mM, which is consistent with previously reported values.

Amine Oxidase (Copper-Containing)↗