Sample dispersion in isotachophoresis with Poiseuille counterflow

A particular mode of isotachophoresis (ITP) employs a pressure-driven flow opposite to the sample electromigration direction in order to anchor a sample zone at a specific position along a channel or capillary. We investigate this situation using a two-dimensional finite-volume model based on the Nernst-Planck equation. The imposed Poiseuille flow profile leads to a significant dispersion of the sample zone. This effect is detrimental for the resolution in analytical applications of ITP. We investigate the impact of convective dispersion, characterized by the area-averaged width of a sample zone, for various values of the sample P\'{e}clet-number, as well as the relative mobilities of the sample and the adjacent electrolytes. A one-dimensional model for the area-averaged concentrations based on a Taylor-Aris-type effective axial diffusivity is shown to yield good agreement with the finite-volume calculations. This justifies the use of such simple models and opens the door for the rapid simulation of ITP protocols with Poiseuille counterflow

Numerical simulation of electrophoresis separation processes

A new Petrov-Galerkin finite element formulation has been proposed for transient convection-diffusion problems. Most Petrov-Galerkin formulations take into account the spatial discretization, and the weighting functions so developed give satisfactory solutions for steady state problems. Though these schemes can be used for transient problems, there is scope for improvement. The schemes proposed here, which consider temporal as well as spatial discretization, provide improved solutions. Electrophoresis, which involves the motion of charged entities under the influence of an applied electric field, is governed by equations similiar to those encountered in fluid flow problems, i.e., transient convection-diffusion equations. Test problems are solved in electrophoresis and fluid flow. The results obtained are satisfactory. It is also expected that these schemes, suitably adapted, will improve the numerical solutions of the compressible Euler and the Navier-Stokes equations

Measurement of adenosine triphosphate and some other metabolites in blood cells by isotachophoresis. I. Preparative technique and enzymatic confirmation.

This study investigated the optimal conditions for detection of nucleotides in blood using an IP-1B capillary isotachophoretic apparatus. The system used 10 mM HCl-beta-alanine (pH 4.2) as the leading electrolyte and n-caproic acid as the terminal electrolyte. Direct application of lysed red blood cells was shown to be inaccurate, and a method of deproteinization based on heat in a microwave oven was developed. The zones for 2,3-diphosphoglycerate, ATP, inorganic phosphate, and lactate were identified enzymatically by withdrawal of pure samples of each zone via a special withdrawal cell. The quantitative values obtained by isotachophoresis were also confirmed enzymatically. The technique is now available for convenient and accurate identification of these metabolites simultaneously.</p

Bioprocessing: Prospects for space electrophoresis

The basic principles of electrophoresis are reviewed in light of its past contributions to biology and medicine. The near-zero gravity environment of orbiting spacecraft may present some unique advantages for a variety of processes, by abolishing the major source of convection in fluids. As the ground-based development of electrophoresis was heavily influenced by the need to circumvent the effects of gravity, this process should be a prime candidate for space operation. Nevertheless, while a space facility for electrophoresis may overcome the limitations imposed by gravity, it will not necessarily overcome all problems inherent in electrophoresis. These are, mainly, electroosmosis and the dissipation of the heat generated by the electric field. The NASA program has already led to excellent coatings to prevent electroosmosis, while the need for heat dissipation will continue to impose limits on the actual size of equipment. It is also not excluded that, once the dominant force of gravity is eliminated, disturbances in fluid stability may originate from weaker forces, such as surface tension

Review on the development of truly portable and in-situ capillary electrophoresis systems

Capillary electrophoresis (CE) is a technique which uses an electric field to separate a mixed sample into its constituents. Portable CE systems enable this powerful analysis technique to be used in the field. Many of the challenges for portable systems are similar to those of autonomous in-situ analysis and therefore portable systems may be considered a stepping stone towards autonomous in-situ analysis. CE is widely used for biological and chemical analysis and example applications include: water quality analysis; drug development and quality control; proteomics and DNA analysis; counter-terrorism (explosive material identification) and corrosion monitoring. The technique is often limited to laboratory use, since it requires large electric fields, sensitive detection systems and fluidic control systems. All of these place restrictions in terms of: size, weight, cost, choice of operating solutions, choice of fabrication materials, electrical power and lifetime. In this review we bring together and critique the work by researchers addressing these issues. We emphasize the importance of a holistic approach for portable and in-situ CE systems and discuss all the aspects of the design. We identify gaps in the literature which require attention for the realization of both truly portable and in-situ CE systems

The development of a method to determine felinine in body fluids by capillary electrophoresis : a thesis presented in partial fulfilment of the requirements for the degree of Master of Philosophy in Chemistry at Massey University

Ion-exchange, paper-chromatography and high performance liquid chromatography were used in earlier studies for the determination of felinine in biological fluids. These methods were either inadequate and/or need laborious sample pre-treatments. A new method for the determination of felinine by capillary zone electrophoresis has been developed. Preliminary investigations were carried out to address the conditions required for the separation of felinine. The separation of felinine can be performed on a fused-silica capillary with a 20 mM phosphate buffer (pH 2.0) and detection wavelength 200 nm. The separation principle was based on the different migration times due to the different molecular weights, molecular sizes and charges under an applied potential field. The quantitative determination of felinine levels in cat urine has been achieved. The cat urine analysis was performed directly on the capillary electrophoresis without making any felinine derivative(s). The levels of felinine in different cat genders are reported. The results were compared with the results of an HPLC felinine derivatization method. Felinine levels in entire male cat urine were much higher than those in female and castrated male cat urine. A synthetic felinine was employed as standard felinine. Linear relationships between peak area and concentration of synthetic felinine calibrations are reported. Mean felinine recovery in cat urine was 95.9%. Taurine, urea, creatine and creatinine, which exist in large amounts in cat urine, showed no interference with the analysis of felinine by this method. The new capillary zone electrophoresis method was then applied to the study of felinine stability. Conditions reported to influence the stability of felinine were investigated. These conditions included oxidation, storage temperatures and times, heating, acidic and alkaline solutions. Both synthetic felinine and felinine in cat urine were investigated. Storage temperature (-20°C to 20°C) had no significant influence on the stability of felinine while higher temperatures increased the decomposition of felinine. Felinine degraded at strong acid and base conditions but was relatively stable under mild acid and base conditions. A similar stability of felinine in human urine is also reported. The capillary zone electrophoresis method was also employed to study felinine in plasma and serum. Plasma and serum as well as urine can be analysed directly on the capillary electrophoresis after sufficient dilution. Conditions (eg. protein clean up, changing of injection time, 37°C heating) that might influence of felinine behaviour in plasma and serum are discussed. This study indicated that no traces felinine be found in cat plasma, within the detection limits of this new capillary electrophoresis method

Determination of creatinine and creatine by capillary electrophoresis : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Chemistry at Massey University

The assessment of creatinine and creatine in biological fluids is important in the evaluation of renal and muscular functions. For routine creatinine determinations in the clinical laboratory, the most frequently used method is the spectrophotometric one based on the Jaffé reaction. However, this reaction is not specific for creatinine. For this reason, several methods have been proposed, but the elimination of interferences in the determination of creatinine has still not been achieved in some of these methods; others solved this problem either with expensive equipment that does not suit routine analysis or necessitates time-waste procedures. In this thesis capillary electrophoresis was the new tool investigated. It was applied in an attempt to achieve both the separation of creatinine from the non-creatinine 'Jaffé- reacting' chromogens and the determination of creatine in serum. Capillary zone electrophoresis was performed with detection at wavelength 480 nm to separate creatinine from the non-creatinine 'Jaffé-reacting' chromogens in urine. The principle was based upon the different migration times due to the different molecule weights, molecular sizes and charges under the applied high voltage. The picric acid was employed as part of the running buffer to allow reaction of creatinine and picrate to take place after the sample injection. This procedure eliminated the negative influence of the reaction time that is controlled manually in the common Jaffé reaction method. Therefore, compared to the Jaffé reaction method, the new method achieved more accuracy and precision in the determination of creatinine. Determination of creatinine in serum and urine were studied at a new wavelength 417 nm, which gave a higher sensitivity of detection than at 480 nm. This wavelength shift made the determination of creatinine in serum possible by capillary zone electrophoresis without the non-creatinine 'Jaffé-reacting' chromogens interfering. In this method, serum only needed a simple filtration before the analysis. Creatine was discovered to have absorption at 417 nm in alkaline medium. Moreover, specific sample stacking was introduced in this method. The sample was dissolved in a mixture of two-volumes acetonitrile and one-volume 3 % ammonium chloride to give a 10-fold enhancement of detection sensitivity