Petr Tůma

SOFSEM 2009: Theory and Practice of Computer Science

SOFSEM 2009: Theory and Practice of Computer Science: 35th Conference on Current Trends in Theory and Practice of Computer Science : Spindlerův Mlýn, Czech Republic, January 2009, Proceedings of the Conference

Determination of the spectrum of low molecular mass organic acids in urine by capillary electrophoresis with contactless conductivity and ultraviolet photometric detection--an efficient tool for monitoring of inborn metabolic disorders.

A mixture of 29 organic acids (OAs) occurring in urine was analyzed by capillary electrophoresis (CE) with capacitively coupled contactless conductivity detection (C(4)D) and UV photometric detection. The optimized analytical system involved a 100 cm long polyacrylamide-coated capillary (50 μm i.d.) and the background electrolyte of 20mM 2-morpholinoethanesulfonic acid (MES)/NaOH+10% (v/v) methanol, pH 6.0 (pH is related to the 20mM MES/NaOH buffer in water). The LOD values obtained by C(4)D for the OAs which do not absorb UV radiation range from 0.6 μM (oxalic acid) to 6.8 μM (pyruvic acid); those obtained by UV photometry for the remaining OAs range from 2.9 μM (5-hydroxy-3-indoleacetic acid) to 10.2 μM (uric acid). The repeatability of the procedure developed is characterized by the coefficients of variation, which vary between 0.3% (tartaric acid) and 0.6% (5-hydroxy-3-indoleacetic acid) for the migration time and between 1.3% (tartaric acid) and 3.5% (lactic acid) for the peak area. The procedure permitted quantitation of 20 OAs in a real urine sample and was applied to monitoring of the occurrence of the inborn metabolic fault of methylmalonic aciduria.

A Contactless Conductometric Detector with Easily Exchangeable Capillary for Capillary Electrophoresis

An alternative construction of a contactless conductivity detector in which semitubular instead of tubular electrodes were used is described. The electrodes are made of a strip of aluminium foil formed into a semitubular shape. They are positioned between two Plexiglass plates in a groove matching the outer diameter of a separation capillary. One Plexiglass plate is immovable, the other is free and can be detached from the immovable plate after the fixing screws are released. The separation capillary is inserted between the plates into the groove and flattened to the electrodes by the tightening of the fixing screws. This construction permits a simple exchange of the separation capillary and makes the connection of individual electrodes with the detector electronics easier and more reliable. The detector performance was tested under the electrophoretic conditions using model sample solutions of potassium chloride in the concentration range from 10−3 to 10−8 mol/L. It was shown (for K+ ion) that the operational characteristics of the detector with semitubular electrodes, namely dynamic range, 0.5–1000 μM, linear dynamic range, 5–1000 μM, limit of detection, 0.5 μM (LOD here represents the concentration from that the detector signal ceases to change with further concentration decrease) and sensitivity (1.2 mV/pg), are comparable with those for the detectors with common tubular electrodes.

A contactless conductivity detector for capillary electrophoresis: effects of the detection cell geometry on the detector performance.

The effect of the gap between the electrodes and of their width on the behavior of a capacitively wired contactless conductivity detector was studied. The results obtained have indicated that the detector response can be qualitatively described by a model based on the concept of the effective electrode width which is a complex parameter determined by the gap between the electrodes, the frequency of the input signal and the conductivity of the test solution. The detector sensitivity and the effect on the separation efficiency depend on the difference between the effective and geometric electrode widths. Higher detection sensitivities have been attained for detectors with wide electrodes operating at lower frequencies, however, better separation efficiencies have been achieved using detectors with narrow electrodes and higher operational frequencies. The noise increases with decreasing gap between the electrodes and increasing frequency, especially with detectors employing narrow electrodes.

Rapid monitoring of arrays of amino acids in clinical samples using capillary electrophoresis with contactless conductivity detection

CE with contactless conductivity detection has been used to separate 28 biogenic amino acids in a short capillary with an effective length of 18 cm. All the tested amino acids can be mutually separated in 0.5-10 mol/L acetic acid electrolytes. The time of analysis does not exceed 6 min and the LODs vary from 0.1 to 1.7 micromol/L. The CVs lie within the intervals 0.01-0.4% and 0.9-4% for the migration times and the analyte peak areas, respectively. The procedure has been successfully applied to the determinations of the whole amino acid spectra in blood plasma, urine, saliva and cerebrospinal fluid samples.

Rapid monitoring of mono- and disaccharides in drinks, foodstuffs and foodstuff additives by capillary electrophoresis with contactless conductivity detection.

A capillary electrophoresis (CE) procedure with contactless conductivity detection (C(4)D) has been developed for monitoring of neutral mono- and disaccharides in drinks and foodstuffs. The separation of a mixture of seven neutral saccharides (glucose, fructose, galactose, mannose, ribose, sucrose and lactose) employed a quartz capillary, 5 μm i.d., with an effective length of 18.3 cm, and 75 mM NaOH (pH 12.8) as the background electrolyte (BGE). The limit of detection (LOD) values obtained lied within a range from 0.4 μmol L(-1) for lactose to 0.9 μmol L(-1) for ribose, with a separation time shorter than 140 s. The procedure was successfully applied to determinations of saccharides in fruit juices, Coca-Cola, milk, red and white wines, yoghurts, honey and a foodstuff additive.

Frequent intravenous pulses of growth hormone together with glutamine supplementation in prolonged critical illness after multiple trauma: Effects on nitrogen balance, insulin resistance, and substrate oxidation*

Objectives:To estimate the efficacy and metabolic effects of growth hormone substitution as intravenous pulses together with alanyl-glutamine supplementation and tight blood glucose control in prolonged critical illness. Design:Prospective double-blind, randomized trial with open-label control arm. Setting:Intensive care unit of tertiary level hospital. Patients:Thirty multiple trauma patients (median Injury Severity Score 34). Interventions:Patients were randomized, at day 4 after trauma, to receive intravenous alanyl-glutamine supplementation (0.3 g/kg·day−1 from day 4 until day 17) and intravenous growth hormone (administered days 7–17, full dose 50 &mgr;g/kg·day−1 from day 10 onward) (group 1, n = 10) or alanyl-glutamine and placebo (group 2, n = 10). Group 3 (n = 10) received isocaloric isonitrogenous nutrition (proteins 1.5 g/kg·day−1) without alanyl-glutamine. Measurements and Main Results:Cumulative nitrogen balance for the whole study period was −97 ± 38 g of nitrogen for group 1, −193 ± 50 g of nitrogen for group 2, and −198 ± 77 g of nitrogen for group 3 (p < .001). This represents a daily saving of 300 g of lean body mass in group 1. Insulin-mediated glucose disposal, during euglycemic clamp, as a measure of insulin sensitivity, significantly worsened between days 4 and 17 in group 1 but improved in groups 2 and 3. Group 1 required significantly more insulin to control blood glucose, resulting in higher insulinemia (∼70 mIU in group 1 vs. ∼25 mIU in groups 2 and 3). Despite this, growth hormone treatment caused an increase in plasma nonesterified fatty acid (∼0.5–0.6 mM in group 1 in comparison with ∼0.2–0.3 mM in groups 2 and 3) but did not influence lipid oxidation. There were no differences in morbidity, mortality, or 6-month outcome among the groups. Conclusions:Treatment with frequent intravenous pulses of low-dose growth hormone together with alanyl-glutamine supplementation improves nitrogen economy in patients with prolonged critical illness after multiple trauma but worsens insulin sensitivity. Tight blood glucose control is possible but requires higher doses of insulin.

Contactless Impedance Sensors and Their Application to Flow Measurements

The paper provides a critical discussion of the present state of the theory of high-frequency impedance sensors (now mostly called contactless impedance or conductivity sensors), the principal approaches employed in designing impedance flow-through cells and their operational parameters. In addition to characterization of traditional types of impedance sensors, the article is concerned with the use of less common sensors, such as cells with wire electrodes or planar cells. There is a detailed discussion of the effect of the individual operational parameters (width and shape of the electrodes, detection gap, frequency and amplitude of the input signal) on the response of the detector. The most important problems to be resolved in coupling these devices with flow-through measurements in the liquid phase are also discussed. Examples are given of cell designs for continuous flow and flow-injection analyses and of detection systems for miniaturized liquid chromatography and capillary electrophoresis. New directions for the use of these sensors in molecular biology and chemical reactors and some directions for future development are outlined.

Large-volume sample stacking for in vivo monitoring of trace levels of γ-aminobutyric acid, glycine and glutamate in microdialysates of periaqueductal gray matter by capillary electrophoresis with contactless conductivity detection

A new variant of large-volume sample stacking injection (LVSS) was used in the capillary electrophoresis with capacitively coupled contactless conductivity detection (CE/C(4)D) determination of the neurotransmitters γ-aminobutyric acid (GABA), glycine (Gly) and glutamate (Glu) in microdialysates of periaqueductal gray matter (PAG). The separation capillary was filled to 98% from the injection side with a sample of microdialysate in acetonitrile. Simultaneously with turning on the separation voltage, the sample zone was forced out by the background electrolyte by increasing the pressure in the terminal capillary outlet vessel. As a consequence of the stacking effect, the analyte was concentrated from the large sample volume into a narrow zone at the sample/background electrolyte boundary close to the injection end of the capillary. Under these conditions, LOD values of 9, 10 and 15nM were determined in the model samples for GABA, Gly and Glu, respectively; RSD equalled 0.5% for the migration times and 1.0-1.9% for the peak areas, respectively. In analysis of microdialysates of PAG, LOD values of 29, 29 and 37nM were determined for GABA, Gly and Glu, respectively; RSD equalled 0.5-0.7% for the migration times and 2.6-8.2% for the peak areas, respectively. The determined basal levels of the neurotransmitters in PAG microdialysates are 0.08, 4.7 and 0.8μM for GABA, Gly and Glu, respectively. Carrageenan-induced hyperalgesia increases the Gly and Glu levels and reduces GABA in PAG microdialysate. Peroral administration of paracetamol in hyperalgesia effectively reduces the Gly value and has no effect on Glu and GABA.

Determination of ammonia, creatinine and inorganic cations in urine using CE with contactless conductivity detection

CE with capacitively coupled contactless conductivity detection (C(4)D) was used to determine waste products of the nitrogen metabolism (ammonia and creatinine) and of biogenic inorganic cations in samples of human urine. The CE separation was performed in two BGEs, consisting of 2 M acetic acid + 1.5 mM crown ether 18-crown-6 (BGE I) and 2 M acetic acid + 2% w/v PEG (BGE II). Only BGE II permitted complete separation of all the analytes in a model sample and in real urine samples. The LOD values for the optimized procedure ranged from 0.8 microM for Ca(2+) and Mg(2+) to 2.9 microM for NH(4)(+) (in terms of mass concentration units, from 7 microg/L for Li(+) to 102 microg/L for creatinine). These values are adequate for determination of NH(4)(+), creatinine, Na(+), K(+), Ca(2+) and Mg(2+) in real urine samples.