Biomedical subjects

Vladislav Kahle

Publications and source records attributed to Vladislav Kahle.

6 recordsLinked to original sources

PubMed2006

Separation of microcystins by capillary electrochromatography in monolithic columns.

Contribution on microcystin variant analysis by capillary electrochromatography (CEC) with easily affordable spectrophotometric detection is presented. Two types of reversed-phase capillary columns formed by inorganic or organic polymer monoliths were prepared for this purpose. The analyses were performed isocratically by means of tris(hydroxymethyl) aminomethane (TRIS) buffers of mildly alkaline pH containing 30% (v/v) acetonitrile as the mobile phases. The samples were injected electrokinetically and the analyses were done at the same separation field strength of 500 V/cm. Microcystins were detected at 238 nm. Although both column types differ not only in monolith quality (inorganic versus organic) but also in the length of the aliphatic moiety (C8 versus C12) similar results were achieved. The on-column preconcentration as the encouraging prospect of electrochromatographic technique was also tested. Consequently 5% of column volume was injected in contrast with 0.5% at standard injection scheme resulting in the six times enrichment of the low concentrated cyanobacterial extract at the top of the separation column. From these preliminary results can be seen that the CEC method is fully applicable for rapid microcystin screening.

Chromatography, High Pressure Liquid↗

PubMed2006

Miniaturized liquid core waveguide-based fluorimetric detection cell for capillary separation methods: application in CE of amino acids.

A miniaturized post-column fluorimetric detection cell for capillary separation methods based on optical fibers and liquid core waveguides (LCWs) is described. The main part of the detection cell is a fused-silica capillary coated with Teflon AF serving as an LCW. The optical fibers are used both for coupling the excitation source with the detection domain in the LCW and for the axial fluorescence collection from the LCW end. The latter fiber is connected with a compact CCD spectrometer that serves for the rejection of the scattered excitation light and for the fluorescence signal detection. The proposed design offers a compact fluorescence detector for various microcolumn separation techniques without optical elements such as filters or objectives. Moreover, its construction and optical adjustment are very simple and the whole system is highly miniaturized. The function of the detection cell is demonstrated by CE of amino acids labelled by fluorescein-based tags. Separations of different standard amino acid mixtures and plasma samples are presented. The comparison of plasma amino acid levels of individuals being in good health with those of patients with inherited metabolic disorders is also shown.

Amino Acids↗

PubMed2005

Fluorescence detection system for capillary separations utilizing a liquid core waveguide with an optical fibre-coupled compact spectrometer.

A fluorescence detection system for capillary liquid separation methods is described. The system is based on a silica capillary coated with a low refractive index fluoropolymer Teflon AF that serves both as a separation channel and as a liquid core waveguide (LCW). A fibre-coupled laser excites separated analytes in a detection point and arising fluorescence is collected at one end of the LCW capillary into the other optical fibre which brings it to a compact charge-coupled device (CCD) array spectrometer installed in a desktop computer. No additional components such as focusing optics or filters are necessary. This system was used for detecting isoelectrically focused fluorescent low-molecular-mass pI (isoelectric point) markers and fluorescein isothiocyanate (FITC) labelled proteins. The ability of the system to acquire fluorescent spectra is also demonstrated.

Calibration↗

PubMed2005

Preliminary study on the monitoring of glutathione S-tranferase activity toward styrene oxide by electromigration methods.

A new method has been developed for the monitoring of glutathione S-tranferase (GST) detoxification activity toward styrene oxide (SO). The enzymatic reaction was carried out directly in a thermostatted autosampler vial and the formation of conjugates between glutathione (GSH) and SO was monitored by sequential MEKC runs. The determinations were performed in a 50-microm fused silica capillary using 50 mM SDS in 20 mM phosphate 20 mM tetraborate buffer (pH 8.3) as a background electrolyte; separation voltage 28 kV (positive polarity), temperature of capillary 25 degrees C, and detection at 200 nm. The method is rapid, amenable to automation, and requires only small amounts of samples, which is especially important in the case of GST isoenzyme analyses.

Animals↗

PubMed2004

Sample injection in capillary electrochromatography by heart-cut technique.

The splitter working in heart-cut regime was used for sample injection in capillary electrochromatography. The principle was implemented in an automated microgradient system allowing to inject from microlitre down to nanolitre volumes with high repeatability and minimal extra-column band broadening. The apparatus is able to deliver discrete volumes of liquids at a preset volumetric flow rate and to stop and restore the flow at any moment. This brings a high degree of liquid manipulation flexibility. An extremely low split ratio is sufficient during the analysis, which saves mobile phase consumption substantially. The key parameters influencing the function of the heart-cut splitter were characterised. The function of the apparatus was demonstrated under isocratic, preconcentration and gradient capillary electrochromatography separation conditions. In all cases the statistic evaluation of the main parameters was performed, showing that high repeatability of retention times, peak heights and areas was achieved.

Automation↗

PubMed2003

Automated microgradient system for capillary electrochromatography.

A microprocessor controlled gradient elution system suitable for capillary electrochromatography has been developed and tested. It is based on a liquid handling device described previously which is capable of liquid transport with both low and high fluid dispersion. The low dispersion region formed by stainless steel needle 250 microm I.D. serves for sample injection, while the high dispersion region, created by steep extension of tube diameter, is used for continuous mobile phase gradient generation. A homologous series of seven alkylphenones was electrochromatographically separated on a monolithic polyacrylic column under gradient conditions. An S-shaped acetonitrile gradient (30-70%) was applied. A high reproducibility of retention times (RSD about 0.1%) was obtained, indicating accuracy of automated gradient operations.

Automation↗