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PubMed1997

Evaluation of capillary zone electrophoresis and micellar electrokinetic capillary chromatography with direct injection of plasma for the determination of cefotaxime and its metabolite.

Quantitative aspects of capillary zone electrophoresis (CZE) and micellar electrokinetic capillary chromatography (MECC) were investigated for the determination of cefotaxime (C) and its deacetyl metabolite (DA) in human plasma in a concentration range of therapeutic interest. For CZE, plasma samples spiked with C and DA were injected after deproteinization with acetonitrile, and analytes were separated in a fused silica capillary using a borate buffer at pH 9.2 as electrolyte; no suitable internal standard was found. For MECC, plasma samples spiked with C, DA, and theobromine as internal standard were directly injected after dilution with water and analyzed using a phosphate buffer, pH 8.00, containing 165 mM SDS as separation electrolyte and a fused silica capillary. Both methods gave satisfactory interday precision with respect to migration times (RSD < 1%) and gave linear responses over the concentration ranges investigated (5-100 mg L-1 C and 5-20 mg L-1 DA). For CZE, intraday RSD (n = 4 graphs) between the slopes of the calibration graphs was acceptable (5.7%) for C. The corresponding figures for interday precision (n = 4 days) were fair (16.1%) in comparison to those obtained with MECC, for which the RSD was 1.49% when theobromine was used as internal standard. A satisfactory interday precision between slopes was also obtained with MECC even without the use of an internal standard (RSD = 4.38%), which demonstrated the ruggedness of this method. Detection limits (S/N = 3) were about 2 mg L-1 (CZE) and 1 mg L-1 in plasma (MECC) for C and DA. MECC was shown to be superior with regard to simplicity, rapidity, precision, and sensitivity.

Cefotaxime↗

PubMed1997

Use of a polybrene capillary coating in capillary electrophoresis for rapid analysis of hemoglobin variants with on-line detection via an ion trap storage/reflectron time-of-flight mass spectrometer.

A polybrene capillary coating in capillary electrophoresis (CE) has been used for rapid analysis of hemoglobin variant digests. The use of the polybrene capillary coating has allowed sufficient separation to resolve the large number of digest products formed upon tryptic digestion of the whole protein, so that prior separation of the hemoglobin alpha and beta chains is not required. The resolution of the digest peaks obtained by CE is sufficient so that even single amino acid substitutions can easily be detected using UV absorption detection. The digest is further analyzed by capillary electrophoresis separation with on-line detection using electrospray ionization interfaced to the ion trap storage/reflectron time of flight device (CE/ESI-IT/reTOF), where a comparison of the total ion electropherograms and mass spectra of the mutant and normal hemoglobins can detect the presence of a mutation site. The CE separation and mass analysis can be accomplished in typically 10-15 min. The unique capability of the CE/ESI-IT/reTOF system for detection of fast separations with narrow peaks that may be under 1 s fwhm is demonstrated. The speed of this system is essential for resolution of the large number of peaks that are separated in a short time duration using CE separations.

Adult↗

PubMed2006

Capillary electrophoresis-mass spectrometry of proteins at medium pH using bilayer-coated capillaries.

The feasibility of using noncovalently bilayer-coated capillaries for capillary electrophoresis-mass spectrometry (CE-MS) of acidic proteins was investigated using background electrolytes (BGEs) of medium pH. The capillary was coated by successively rinsing the capillary with solutions of the oppositely charged polymers polybrene (PB) and poly(vinyl sulfonic acid) (PVS). Volatile BGEs containing ammonium formate and/or N-methyl morpholine were tested at pH 7.5 and 8.5. Overall, these BGEs provided relatively fast protein separations (analysis times of ca. 12 min) and showed high efficiencies (70,000-300,000 plates) when the ionic strength was sufficiently high. Migration-time reproducibilities were very favorable with RSDs of less than 1.0%. Infusion experiments showed satisfactory MS responses for studied proteins dissolved in ammonium formate (pH 8.5), however, high concentrations of N-methyl morpholine appeared to seriously suppress the MS protein signals. Evaluation of the CE-MS system was performed by analyzing a mixture of intact proteins yielding efficient separations and good-quality mass spectra. CE-MS analysis of a reconstituted formulation of the biopharmaceutical recombinant human growth hormone (rhGH) which was stored for a prolonged time, revealed one degradation product which was provisionally identified as desamido rhGH. Based on the MS responses the amount of degradation was estimated to be ca. 25%.

Animals↗

PubMed1967

Capillary-tube scanner for mechanized microbiology. A photoelectric scanner measures growth in agar-filled capillaries and gives a new approach to microbiology.

Growing microcolonies of bacteria can be detected by their light-scattering property. We have described a system in which growing bacteria in glass capillaries filled with nutrient agar are counted as the capillaries move through a narrow beam of light. Increased counts after incubation indicate the presence of viable organisms in the original sample; the equipment can detect colonies when they grow to a diameter of 8 microm. Aerobic, anaerobic, and facultative organisms can grow in the capillary environment. The sensitivities of several organisms to antibiotics were determined by adding antibiotics to the agar before the capillaries were filled.

Agar↗

PubMed2004

Glomerular capillary tenascin staining in renal allografts and its correlation with capillary loop 'double contours', proteinuria and graft outcome.

BACKGROUND: Glomerular capillary injuries result in proteinuria, previously reported to be a risk factor for renal allograft dysfunction. According to the Banff 97 Working Classification of Renal Allograft Pathology, 'double contours' in glomerular capillary loops (GC-DC) were suggested to be most specific for chronic transplant glomerulopathy. Tenascin-C (TN), a component of extracellular matrix, is known to be overexpressed in various conditions. METHODS: TN immunostaining was performed by the labeled streptavidin biotin method. The correlations between glomerular capillary TN (GC-TN) and proteinuria, and between GC-DC and GC-TN, were studied in 27 biopsies showing chronic allograft nephropathy (CAN) lesions. In 14 patients (serum creatinine <2.0 mg/dl at biopsy), graft outcome was studied. Graft function 3 years after biopsy was classified as stable or deteriorated; the term 'deteriorated' was used if serum creatinine had increased 20% over baseline, or if the patient required dialysis. RESULTS: In CAN, GC-TN correlated with GC-DC. Proteinuria in patients with intense GC-TN tended to increase during 1 year after biopsy. In graft outcome analysis, GC-TN was significantly lower in stable grafts than in deteriorated ones. Furthermore, all 3 patients demonstrating weak GC-TN without GC-DC returned to dialysis. CONCLUSION: TN immunostaining is a sensitive method to detect glomerular capillary injury with a predictive value for renal allograft dysfunction.

Adult↗

PubMed1991

Pulmonary "capillary" wedge pressure not the pressure in the pulmonary capillaries.

In 1949, Hellems, Haynes, and Dexter proposed that the pressure in a catheter wedged so as to occlude a pulmonary artery was an "estimate" of the pressure in the pulmonary capillaries. Their report led to the designation of this pulmonary artery wedge pressure as the pulmonary "capillary" wedge pressure. In fact, the pulmonary artery wedge pressure is a distorted measure of the pressure in the pulmonary veins. Usually this pressure differs only slightly from the capillary pressure, and the misconception fostered by the inaccurate name is inconsequential; however, sometimes this misconception leads to errors in diagnosis. This report briefly reviews the early history of pulmonary vascular catheterization, offers an explanation of pulmonary artery wedge pressure, and discusses a disease in which pulmonary artery wedge pressure is normal, even though capillary pressure is elevated.

Capillaries↗

PubMed1975

Venous serum, capillary serum, and capillary plasma compared for use in determination of lactate dehydrogenase and aspartate aminotransferase activities.

Lactate dehydrogenase and aspartate aminotransferase activities in capillary serum or plasma were significantly greater than in simultaneously assayed venous serum, the greatest differences being between capillary and venous serum. Although some difference is attributable to tissue fluid contributions, platelets seem to account for most of it, with possible small contributions from leukocytes. Capillary tube size and type appear to be important factors. We recommend that when capillary blood is to be used for enzyme assay, it should be processed as plasma.

Aspartate Aminotransferases↗

PubMed2004

Simultaneous determination of nitrate and nitrite in biological fluids by capillary electrophoresis and preliminary study on their determination by microchip capillary electrophoresis.

In order to develop a highly sensitive and high-throughput screening method for nitrogen monoxide metabolites in biological fluids, we have investigated the simultaneous determination of nitrite and nitrate, using capillary electrophoresis and microchip capillary electrophoresis. In capillary zone electrophoresis, a running buffer based on human serum components with high ionic strength has been developed for the determination of nitrite and nitrate in human serum and human saliva. We obtained successful separation of nitrite and nitrate in the serum and the saliva within 7 min under optimum analytical conditions. Linear calibration curves for nitrite and nitrate for both peak height and area were obtained by a standard addition method. The limits of detection obtained at a signal-to-noise ratio (S/N) of 3 for nitrite and nitrate in the serum were 2.6 and 1.5 microM, respectively. The values of the relative standard deviation of peak height for the serum with 9.2 microM nitrite and 20.9 microM nitrate were 5.7 and 4.1%, respectively. For on-site analysis with high-throughput screening, a microchip capillary electrophoresis method using a microchip made of quartz with a UV detector was developed. In this high-throughput format, using a running buffer with an electroosmotic flow modifier, the peaks of nitrite and nitrate in an artificial serum sample were obtained within 8 s. In high-resolution mode, using the buffer without electroosmotic flow modifier, the separation of nitrite and nitrate was obtained within 15 s. In high-resolution mode, using an artificial serum sample with 50 microM NO2- and 50 microM NO3-, the limits of detection (S/N = 3) of 41 microM for NO2- and 26 microM for NO3- were obtained. The method was applied to human serum and saliva. We obtained peaks due to nitrite and nitrate in 10-fold diluted saliva.

Electrochemistry↗

PubMed2005

Capillary density and capillary-to-fibre ratio in vastus lateralis muscle of untrained and trained men.

Muscle fibre profile area (Af), volume density (Vv), capillary-to-fibre ratio (CF) and number of capillaries per fibre square millimetre (CD) were determined from needle biopsies of vastus lateralis of twenty-four male volunteers (mean +/- SD: age 25.4+/-5.8 years, height 178.6+/-5.5 cm, body mass 72.1+/-7.7 kg) of different training background. Seven subjects were untrained students (group A), nine were national and sub-national level endurance athletes (group B) with the background of 7.8+/-2.9 years of specialised training, and eight subjects were sprint-power athletes (group C) with 12.8+/-8.7 years of specialised training. Muscle biopsies of vastus lateralis were analysed histochemically for mATPase. Capillaries were visualized and counted using CD31 antibodies against endothelial cells. There were significant differences in the Vv of type I and type II muscle fibres in both trained groups, B (51.8%; 25.6%) and C (50.5%; 26.4%). However, in untrained group A that was treated as a reference group, the difference between Vv of type I and type II fibres was less prominent, nevertheless statistically significant (42.1%; 35.1%). There was also a significant difference in CF: 1.9 in group A and 2.1 in groups B and C. The number of capillaries per mm2 (CD) was 245 (group A), 308 (group B) and 325 (group C). Significant differences (P<0.05) in CF and CD, were found only between group A (1.9; 245) and both groups of trained men, B and C (2.1; 308 and 325). However, endurance athletes (group B), such as long-distance runners, cyclists and cross country skiers, did not differ from the athletes representing short term, high power output sports (group C) such as ice hockey, karate, ski-jumping, volleyball, soccer and modern dance.

Adult↗

PubMed1992

[Capillary blood as diagnostic sample in pediatric laboratory diagnosis. Arterio-capillary-venous difference and blood collection technique].

It is preferable to use capillary blood as diagnostic material in paediatric laboratory tests rather than venous blood. The collection of capillary blood makes less demands on the child and is easy in principle. The objection to capillary blood primarily concerns differences in the analyte concentration in arterial, capillary and venous blood. Comparative studies in the same child under standardised conditions are not available for a number of parameters. However, according to the results available at present, for almost all routine clinical chemical and haematological examinations clinically usable results can be obtained. Using the example of total protein analysis for the same child we did not find any differences between arterial and venous blood. It is highly probably that the same would apply to many other components. Suspected differences are probably a result of short-comings in the collection technique.

Blood Chemical Analysis↗

PubMed1979

Comparison of capillary ear blood and arterial blood to validate capillary sampling as an accurate assay of blood gas in swine.

Capillary sampling in swine can be performed as an accurate assay of arterial blood gases. Studies with swine provided results similar to, or slightly more favorable than, those reported for human beings, depending upon which cutaneous technique was used on human beings. On the basis of free flow or arterilization of the cutaneous sample and of the correlation between capillary and arterial pH, CO2 partial pressure (PCO2), and O2 partial pressure (PO2) values, the capillary sampling technique of complete incisement of a 2-mm section from the tip of the warmed porcine ear could be a substitution technique for arterial blood sampling. Free flow with this technique was maximized and high correlation coefficients (r) for pH (r = 0.96), PCO2 (r = 0.82), and PO2 (r = 0.90) capillary-arterial values (n = 37) were obtained.

Animals↗

PubMed1997

Synergism of capillary isotachophoresis and capillary zone electrophoresis.

The combination of capillary isotachophoresis and capillary zone electrophoresis may enhance greatly the performance of analytical capillary electrophoresis with respect to both separation power and the concentration sensitivity. The concentrating effects and the separation power of isotachophoresis allow the analysis of diluted samples and the elimination of interferences due to bulk components. The separation process of zone electrophoresis enables one to resolve the stack of trace analytes and detect the resulting individual zones with high sensitivity. The transition of isotachophoresis into zone electrophoresis plays the key role in the overall performance of this hyphenated technique. This article describes the dynamics of the conversion of isotachophoresis into zone electrophoretic mode and shows that the key role is played by the segments of the leading and terminating zones from the isotachophoretic stage. The magnitude of these segments directly effects the detection time as well as the separation width of the peaks of analytes. It is shown that these effects are also important in the analyses by capillary zone electrophoresis where isotachophoresis is induced by the sample itself. Finally, the paper presents a list of recommended, user-friendly, electrolyte systems which enable one to simply predict the performance of the combination isotachophoresis-zone electrophoresis.

Electrolytes↗

PubMed1999

Capillary electrochromatography with novel stationary phases: II. Studies of the retention behavior of nucleosides and bases on capillaries packed with octadecyl-sulfonated-silica microparticles.

An octadecyl-sulfonated silica (ODSS) stationary phase specially designed for performing capillary electrochromatography (CEC) at relatively strong electroosmotic flow (EOF) proved useful for the separations of some nucleosides and bases. The ODSS stationary phase is composed of a hydrophilic, negatively charged sublayer to which a nonpolar top layer containing octadecyl ligands is covalently attached. The charged sublayer contains sulfonic acid groups which ensure a relatively strong EOF. Due to the presence of permanently charged sulfonic acid groups in the sublayer, the hydrophilic nature of the sublayer and the hydrophobic character of the top octadecyl layer, retention and selectivity of charged and relatively polar nucleosides and bases on the ODSS stationary phase are based on electrostatic interaction, hydrophilic interaction, and reversed-phase mechanisms. This yielded for the ODSS stationary phase a unique selectivity towards the nucleosides and bases, thus allowing their rapid separation. To gain insight into the chromatographic behavior of nucleosides and bases on the ODSS stationary phase, the results were compared to those obtained on an octadecyl-silica (ODS) capillary under otherwise the same elution conditions. Due to the difference in the nature of the organic layers on the surface of the ODSS and ODS stationary phases, the elution order on both stationary phases differed significantly, and the ODSS capillary proved more suitable for the separation of the nucleosides and bases than the ODS capillary.

Base Pairing↗

PubMed2005

Dynamically coated capillaries for enantioselective separations by capillary electrophoresis.

The level of variability in migration times and peak areas often encountered in capillary electrophoresis (CE) has restricted its use for routine analysis. Cationic analytes also tend to interact with the capillary wall, resulting in skewed peak shapes and reduced efficiency. We investigated a dynamic coating procedure that involves a two-step process to modify the capillary and that results in a higher but stable electro-osmotic flow (EOF) at low pH. We compared enantioselective separations of several basic compounds on unmodified and dynamically coated capillaries. A derivatized cyclodextrin was used as the chiral additive. When the dynamic coating was used, peak symmetry improved, and migration times were significantly shorter. The dynamic coating procedure also resulted in improved migration time and peak area precision.

Journal Article↗

PubMed1995

Quantitative analysis of major whey proteins by capillary electrophoresis using uncoated capillaries.

A method that allows separation and quantitation of the main whey proteins by capillary electrophoresis using uncoated capillaries is proposed. Separations are performed using 100 mM borate buffer, pH 8.2, containing 30 mM sodium sulfate. The use of high pH and high ionic strength buffer reduces adsorption of proteins on the capillary wall, making their separation possible. Reproducibility of migration times and areas of peaks are improved by optimizing the capillary equilibration protocol and by using an internal standard. Relative standard deviations ranging between 0.74 and 1.03% for migration times and 2.14 to 5.23% for areas of major peaks are obtained. Detection limits equal to or lower than 0.5 mg/100 mL are achieved. Linear relationships of peak area versus concentration have been used to quantitate bovine serum albumin (BSA), alpha-LA (alpha-lactalbumin), beta-LG A (beta-lactoglobulin A) and beta-LG B (beta-lactoglobulin B) in cow's milk subjected to different thermal treatments. Electrophoretic profiles of these milk samples show peaks from other peptides besides those from main proteins. Characteristic patterns for whey from different species are obtained.

Animals↗

PubMed1995

Fused-silica capillaries with surface-bound dextran layer crosslinked with diepoxypolyethylene glycol for capillary electrophoresis of biological substances at reduced electroosmotic flow.

This report is concerned with the introduction of novel surface modification involving the covalent attachment of branched, high molecular weight dextrans and subsequent crosslinking with polyether chains on the inner surface of fused silica capillaries with the aim of producing hydrophilic capillaries with reduced electroosmotic flow (EOF). Three different molecular mass dextrans, namely 45, 71 or 150 kDa were covalently attached to the capillary surface, and subsequently crosslinked with diepoxypolyethylene glycol (PEG). This chemistry produced stable coating over a wide range of conditions including high and low pH aqueous solutions. Moreover, the various dextran-PEG-coated capillaries exhibited reduced electroosmotic flow. These features of the dextran-PEG coatings were useful for the separations of basic and acidic proteins, and provided high resolution separation for closely related acidic monosaccharides and sialooligosaccharides.

Carbohydrate Sequence↗

PubMed2003

Direct determination of bromide ions in seawater by capillary zone electrophoresis using polyethyleneimine-coated capillaries.

A rapid and simple capillary electrophoretic method was developed for the direct determination of bromide ion in seawater. We have found an effective method, based on the use of polyethyleneimine-coated capillaries and the addition of sodium chloride to the background electrolyte. The use of coated capillaries with a cationic polymer changes the direction of the electroosmotic flow in the capillary, which favors the migration speed of the bromide ion and enables the use of low salt concentrations in the separation electrolyte. Bromide ion in seawater can be determined within 2 min using this system and 20 mmol L(-1)NaCl-containing separation electrolyte. The detection limit for the bromide ion was 0.45 microg ml(-1). The method was applied to the determination of bromide ion in seawater samples collected from the Bosphorus and the Black Sea. Bromide contents in samples from 0 to 72 m depths varied between 33.2 and 72.8 mg L(-1) with a mean 3.0% RSD.

Journal Article↗

PubMed1996

Capillary electrophoresis and capillary flow injection analysis with electrochemical detection.

Measurements by capillary flow injection analysis (CFIA) and capillary electrophoresis (CE) in conjunction with electrochemical detection are described. The detection is based on an end-column electrode arrangement. Several novel electrodes, such as a spherical gold electrode and a dual-microdisk electrode, are presented and characterized regarding their analytical utility. In order to improve the selectivity of CFIA, dual-electrode and multiple-pulse detection are studied using couples of cyanometallates or metallocenes. Capillary electrophoretic experiments with amperometric detection are performed using 50 microm i.d. capillaries without any electrical-field decoupler. The practicality and analytical characteristics of this detection strategy are illustrated for the separation of serotonin and some biological precursors and metabolites of neurotransmitter substances.

Journal Article↗