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R Dadoo

Publications and source records attributed to R Dadoo.

7 recordsLinked to original sources

Electrophoretron: a new method for enhancing resolution in electrokinetic separations.

Two capillaries, each of which have different surface preparations on their inside walls, are joined together to form a closed loop, and electrodes are placed inside the two capillaries. When the loop is filled with liquid and a potential difference is applied between the two electrodes, a circulating flow of liquid is established inside the loop because the resistance to flow is unequal in going from one electrode to another in a clockwise versus a counterclockwise direction. Consequently, a sample injected into this device, which we call an electrophoretron, repeatedly circulates between the two electrodes and the capillary separation column becomes effectively one of unlimited length. On each cycle the separation between analytes with different mobilities increases, thus enhancing resolution of analytes having nearly the same mobilities. The operation of a prototype electrophoretron is demonstrated.

Chromatography, Micellar Electrokinetic Capillary↗

Analysis of underivatized amino acids by capillary electrophoresis using constant potential amperometric detection.

A mixture of native (underivatized) amino acids is separated by capillary electrophoresis under alkaline conditions (pH approximately 12) and amperometrically detected with a copper-disk microelectrode. A simple design facilitates capillary-electrode alignment without the need for micropositioning equipment. The limits of detection for the amino acids are in the low microM concentration range, and the signal response is linear over 2-3 orders of magnitude. This procedure is applied to analyze the amino acid hydrolysis products from cytochrome c.

Amino Acids↗

Field programming to achieve uniform sensitivity for on-line detection in electrophoresis.

Under constant applied electric field, an analyte band traverses an on-line detection zone in an electrophoretic separation at a velocity inversely proportional to the detection time. Because analyte bands migrate with different velocities, the sensitivity of the detection system to different species can vary significantly. One approach for overcoming this problem is to change the applied electric field, E, as a function of the separation time, tau, so that the velocity of an analyte band when it passes through the detection zone is independent of the analyte's electrophoretic mobility. A possible solution is to make E(tau) grow exponentially, but this procedure causes band compression. The preferred solution is to make both E(tau) and the distance between the starting position of the sample and the detection zone increase linearly with separation time. This approach can be implemented with either slab-gel or capillary electrophoresis.

Electromagnetic Fields↗

Selective determination of adenine-containing compounds by capillary electrophoresis with laser-induced fluorescence detection.

Capillary electrophoresis coupled with laser-induced fluorescence detection provides a selective analysis of mixtures of adenine-containing compounds (adenine, adenosine, cAMP, AMP, ADP, and ATP) that are derivatized using chloroacetaldehyde as a fluorogenic reagent. The components can be detected with linear response over the concentration range of 10(-4) to 10(-9) M, and the detection limit corresponds to 5 to 10 x 10(-19) mol for each nucleotide. Analysis is achieved in less than 10 min with high separation efficiencies (up to 400,000 theoretical plates) and shows good reproducibility for migration times (0.5 to 1.0%) and peak heights (2.8 to 7.7%). To demonstrate the rapid analysis of small-volume samples, the adenine-containing compounds present in an oocyte from Xenopus laevis are determined.

Adenine↗

Optimizing fluorescence detection in chemical separations for analyte bands traveling at different velocities.

In many separation techniques, such as field flow fractionation, liquid chromatography, and electrophoresis, chemical species form bands that migrate at distinct velocities. If these bands are to be quantified on-line using a shot-noise-limited detection system, then attention must be given to the data-digitization rate and to the removal rate of molecules from the analyte pool as a result of the detection process. A theory is developed for calculating the signal-to-noise ratio under such conditions, and it is specialized to the case of fluorescence detection in capillary electrophoresis. Using standard detection procedures in which the data-digitization rate and excitation intensity remain constant for the duration of a separation, detection sensitivity can vary by more than a factor of five for bands that arrive at the detection zone between migration times tau fast and 10 tau fast, where tau fast is the time after the start of the separation that the fastest migrating band arrives at the detection zone. To compensate for different band velocities, both the data-digitization rate and the excitation intensity must be decreased as separation time (tau) increases by the factor tau fast/tau. Only when these corrections are made can uniform sensitivity with the highest possible signal-to-noise ratio be achieved for each peak. These predictions are experimentally tested and compare favorably to observations.

Electrophoresis↗

The involvement of catalase in alcohol metabolism in Drosophila melanogaster larvae.

The involvement of catalase (H2O2:H2O2 oxidoreductase, EC 1.11.1.6) in the metabolism of alcohols was investigated by comparing Drosophila melanogaster larvae in which catalase was inhibited by dietary 3-amino-1,2,4-triazole (3AT) to larvae fed a diet without 3AT. 3AT inhibited up to 80% of the catalase activity with concordant small increases in the in vitro activities of sn-glycerol-3-phosphate dehydrogenase, fumarase, and malic enzyme, but with a 16% reduction in the in vivo incorporation of label from [14C]glucose into lipid. When the catalase activity was inhibited to different degrees in ADH-null larvae, there was a simple linear correlation between the catalase activity and flux from [14C]ethanol into lipid. By feeding alcohols simultaneously with 3AT, ethanol and methanol were shown to react efficiently with catalase in wild-type larvae at moderately low dietary concentrations. Drosophila catalase did not react with other longer chain alcohols. Catalase apparently represents a minor pathway for ethanol degradation in D. melanogaster larvae, but it may be an important route for methanol elimination from D. melanogaster larvae.

Alcohols↗

Electron spin resonance studies under dynamic mobile phase conditions on chemically modified silica.

The electron-spin resonance (ESR) techniques of spin labeling and probing are shown to be powerful methods for the study of bonded silicas used in chromatographic separations under dynamic solvent flow conditions. Microscopic information gained from these studies includes solvation composition of the bonded phase at various locales in the bonded phase, configuration of the bonded phase and solvent-flow effects. In the spin-label study, 2-(2,2-dimethyloxazolidine-N-oxy)hexylsilane (DOXYL hexyl siloxane) was used as the label. ESR spectra show changes in the motional freedom of the bonded surface as a function of mobile phase composition and elution time and demonstrate chain stiffening with the introduction of water, as inferred by other studies. In the spin-probe work, various nitroxide compounds of the 2,2,6,6-tetramethyl-1-piperidinyloxyl (TEMPO) family were introduced under flow conditions as a 0.1 mM solution in various water-methanol mobile phases. C18 sample columns were equilibrated with this mixture. Interpretation of the spectra yielded information about the solvated environment on the surface that was sampled by the probes as a function of both the mobile phase and the polarity of the probe.

Electron Spin Resonance Spectroscopy↗